acid-phosphatase and Disease-Models--Animal

Osteoprotegerin (OPG) acts as a decoy receptor for the receptor activator of the nuclear factor κ B (RANK) ligand (RANKL) , preventing its association with RANK and inhibiting osteoclastogenesis. Therefore, mice homozygous for targeted disruption of the OPG gene reveal stimulated bone resorption and bone formation, resulting in enhanced bone remodeling. The OPG deficient (OPG( - / - )) mouse showed the diturbed distribution of collagen fibers and complex meshwork of cement lines, which implies weakened strength of OPG( - / - ) bone against mechanical stress. In addition, the abnormally promoted remodeling of the OPG( - / - ) bone caused the disorganized distribution of osteocyte lacunar canalicular system (OLCS) . Histochemical assessment revealed the markedly reduced synthesis of sclerostin in the OPG( - / - ) OLCS while the synthesis of dentin matrix protein-1 was not extremely affected by the OPG deficiency. Taken together, OPG deficient mouse appears to be a valid model for extremely-stimulated bone remodeling, and would provided important clues for better understanding for activities of bone cells in a pathological state in bone.

Topics: Acid Phosphatase; Adaptor Proteins, Signal Transducing; Alkaline Phosphatase; Animals; Bone and Bones; Bone Remodeling; Bone Resorption; Disease Models, Animal; Extracellular Matrix Proteins; Fibroblast Growth Factor-23; Fibroblast Growth Factors; Glycoproteins; Intercellular Signaling Peptides and Proteins; Mice; Osteoblasts; Osteoclasts; Osteogenesis; Osteopontin; Osteoprotegerin; RANK Ligand

We targeted the MVNP gene to the OCL lineage in transgenic mice. These mice developed abnormal OCLs and bone lesions similar to those found in Paget's patients. These results show that persistent expression of MVNP in OCLs can induce pagetic-like bone lesions in vivo.. Paget's disease (PD) is one of the most exaggerated examples of abnormal bone remodeling, with increased bone resorption and excessive new bone formation. However, its etiology is unclear. A viral etiology for PD has been suggested based on the presence of paramyxoviral-like nuclear inclusions, detection of measles virus nucleocapsid (MVNP) mRNA or protein in osteoclasts (OCLs) from PD lesions, and in vitro studies showing that transfection of normal OCL precursors with the MVNP gene results in formation of OCLs that express a pagetic phenotype (increased numbers of OCLs; increased responsivity to 1,25(OH)(2)D(3), RANKL, and TNF-alpha; increased expression of the TAF(II)-17 gene, and increased bone resorption capacity).. We targeted MVNP to cells in the OCL lineage in transgenic mice using the TRACP promoter.. Histomorphometric analysis showed that there was a 64% increase in OCL perimeter (p = 6.0002) and 37% increase in osteoblast (OBL) perimeter in MVNP mice. In a mouse that was 14 months of age, there was a 225% increase in OBL perimeter and 149% in OBL perimeter. This was accompanied by increased cancellous bone volume (83%) and trabecular width (47%) and number (25%), with a marked increase in the amount of woven bone. In contrast, cancellous bone volume decreased between 3 and 12 months in wildtype (WT) mice, whereas cancellous bone volume in MVNP mice increased over the same time period. Ex vivo studies showed that the numbers of OCLs formed in marrow cultures from MVNP mice were increased, and the OCLs were hyper-responsive to 1,25(OH)(2)D(3) and had an increased bone resorbing capacity compared with WT cultures.. These results show that expression of MVNP in OCL in vivo results in a bone phenotype that is characteristic of PD.

Topics: Acid Phosphatase; Animals; Calcitriol; Cells, Cultured; Disease Models, Animal; Humans; Isoenzymes; Mice; Mice, Transgenic; Osteitis Deformans; Osteoclasts; Promoter Regions, Genetic; Tartrate-Resistant Acid Phosphatase

Prostate cancer is a significant health problem and one of the leading causes of cancer-related death among men. Given the typically long natural history of the disease, there is considerable interest in developing new therapies to treat or prevent metastatic disease, and cancer vaccines are a particularly attractive immune-based approach. Early clinical studies using non-specific immunomodulatory treatments have met with limited success, but also suggest that improved immunologic approaches might be useful in treating human prostate cancer. Over the last decade, the identification of immune cells responsible for actual destruction of prostate tissue and advances in immunologic and molecular techniques have led to a variety of vaccination approaches that are currently being evaluated in human clinical trials. The present article discusses the rationale in animal models for particular immunization strategies and describes the vaccines currently being used in patients with prostate cancer. The ongoing identification of tumor antigens and proteins involved in prostate cancer progression and the development of better immunologic animal models suggest a hopeful future for the design of effective prostate cancer vaccines.

Topics: Acid Phosphatase; Animals; Antigens, Surface; Antigens, Tumor-Associated, Carbohydrate; Cancer Vaccines; Carboxypeptidases; Dendritic Cells; Disease Models, Animal; Glutamate Carboxypeptidase II; Humans; Male; Mice; Neoplasms, Experimental; Prostate-Specific Antigen; Prostatic Neoplasms; Rats

Chediak-Higashi syndrome (CHS) is an inherited immunodeficiency disorder characterized by giant lysosomal granules in all granule-containing cells. Prior examination of lysosomal enzyme activities in granulocytes and other cells derived from patients with CHS have revealed multiple abnormalities, with the predominant finding being diminished activity of many of the enzymes tested. Abnormalities in lysosomal enzyme activity are also found in animal models of CHS (cattle, aleutian mink, and beige mice). In this study, we have examined lymphoblastoid cell lines derived from a patient with CHS and from an individual heterozygous for the CHS gene for acid phosphatase, beta-glucuronidase, and alpha-mannosidase activity. These cell lines have recently been shown to be satisfactory in vitro models for the disease. Acid phosphatase activity was increased in the heterozygous-derived cell line when compared to control while other enzyme activities were normal both in the CHS- and heterozygous-derived cell lines. We have reviewed the literature and summarized published abnormalities of lysosomal enzyme activities in humans and animals with CHS.

Topics: Acid Phosphatase; alpha-Mannosidase; Animals; Cell Line, Transformed; Cells, Cultured; Chediak-Higashi Syndrome; Disease Models, Animal; Glucuronidase; Humans; Lysosomes; Mannosidases

The W9 peptide has been shown to act as a receptor activator for nuclear factor-κB ligand (RANKL) antagonist and tumor necrosis factor (TNF)-α antagonist, which can promote bone formation and inhibit bone resorption. Studies on the W9 peptide at the cellular level have mainly focused on osteoblasts, and little research on the mechanism by which the W9 peptide regulates osteoclasts has been reported, which was the aim of this work. In this study, a rat mandibular defect model was established in vivo and implanted with hydrogel containing the W9 peptide for 2 weeks and 4 weeks, and histochemical staining was used to evaluate the formation of new bone and the changes in osteoclasts. RAW264.7 cells were cultured in vitro for osteoclast induction, and different concentrations of W9 peptide were added. Tartrate resistant acid phosphatase staining, monodansylcadaverine staining, TdT-mediated dUTP Nick-End Labeling assay, real-time PCR and Western blot were used to detect osteoclast differentiation, autophagy and apoptosis. Our results showed that the W9 peptide could reduce osteoclastogenesis and osteoclast activity induced by RANKL, and these effects were partly due to the inhibition of osteoclast autophagy. On the other hand, the W9 peptide could promote mature osteoclast apoptosis, in which autophagy might play an antagonistic role. Taken together, these results suggest that the W9 peptide inhibits osteoclastogenesis and osteoclast activity by downregulating osteoclast autophagy and promoting osteoclast apoptosis. Our results will benefit the development and application of new small molecule peptides for the treatment of bone resorption diseases.

Topics: Acid Phosphatase; Animals; Apoptosis; Autophagy; Blotting, Western; Cells, Cultured; Disease Models, Animal; Down-Regulation; Immunohistochemistry; In Situ Nick-End Labeling; Male; Mandibular Diseases; Osteoclasts; Osteogenesis; Peptides, Cyclic; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction

Lysosomal acid phosphatase 2 (

Topics: Acid Phosphatase; Animals; Cell Differentiation; Cell Proliferation; Cerebellar Ataxia; Cerebellar Cortex; Cytoplasmic Granules; Disease Models, Animal; Gene Expression Regulation, Developmental; Hedgehog Proteins; Humans; Lysosomes; Mice; Mutation; N-Myc Proto-Oncogene Protein; Neurons; Purkinje Cells; Signal Transduction

Prostate cancer is a candidate for immunotherapy because cancer cells express tissue-specific proteins that can be therapeutic targets. However, immune checkpoint inhibitors and active immunization have performed poorly in clinical trials. We developed a novel virus-like particle (VLP) vaccine composed of bovine papillomavirus L1 protein engineered to display surface docking sites. We decorated VLPs with peptides encoding T cell epitopes from two prostate cancer-associated tumor antigens, prostate stem cell antigen (PSCA), and prostatic acid phosphatase (PAP-1 and PAP-2), and a neo-antigen, stimulator of prostatic adenocarcinoma-specific T cells (SPAS-1). The VLP vaccines induced a mean frequency of antigen-specific IFN-γ secreting CD8 + T cells of 2.9% to PSCA, 9.5% to SPAS-1, 0.03% to PAP-1, and 0.03% to PAP-2 in tumor-bearing TRAMP mice. We treated TRAMP mice at 19-20 weeks of age, when mice have advanced stages of carcinogenesis, with either VLP vaccine, anti-PD1 antibody, or combination immunotherapy. The VLP vaccine alone or in combination with anti-PD1 antibody significantly reduced tumor burden, while anti-PD1 antibody had a modest non-significant therapeutic effect. All treatments significantly increased CD3 + and CD8 + T cell infiltration into tumor tissue compared to control mice, and combination therapy resulted in significantly greater CD3 + and CD8 + T cell infiltration than monotherapy. Reduction in tumor burden in vaccine-treated mice was inversely correlated with CD8 + T cell numbers in tumor tissue. No other immunotherapy has shown efficacy in this animal model of advanced prostate cancer, making bovine papillomavirus VLPs an attractive vaccine technology to test in patients with metastatic prostate cancer.

Topics: Acid Phosphatase; Animals; Antigens, Neoplasm; Cancer Vaccines; Capsid Proteins; CD8-Positive T-Lymphocytes; Disease Models, Animal; Epitopes, T-Lymphocyte; GPI-Linked Proteins; Humans; Interferon-gamma; Male; Mice, Transgenic; Neoplasm Proteins; Prostate-Specific Antigen; Prostatic Neoplasms; Treatment Outcome; Vaccination; Vaccines, Virus-Like Particle

This study aimed to radiolabel finasteride, a novel 5α-reductase inhibitor, to evaluate its cancer targeting potential in experimental model of prostate carcinogenesis. Finasteride was effectively radiolabeled with

Topics: Acid Phosphatase; Animals; Carcinogenesis; Carcinogens; Disease Models, Animal; Electrophoresis; Finasteride; Hydrogen-Ion Concentration; Kinetics; Male; Methylnitrosourea; Prostate; Prostatic Neoplasms; Radiopharmaceuticals; Rats; Rats, Sprague-Dawley; Technetium; Time Factors; Tissue Distribution

Alisma canaliculatum is a herb commonly used in traditional Korean medicine, and has been shown in scientific studies to have antitumor, diuretic hepatoprotective, and antibacterial effects. Recently, the anti-osteoclastogenesis of alisol A 24-acetate from Alisma canaliculatum was investigated in vitro. However, the influence of alisol A 24-acetate on osteoporosis in animals has not been investigated. The present study was undertaken to investigate the anti-osteoporotic effect of alisol A 24-acetate on bone mass in ovariectomized (OVX) mice and to identify the mechanism responsible for its effects. OVX mice were treated daily with 0.5 or 2 μg/g of alisol A 24-acetate for a period of six weeks. It was found that these administrations significantly suppressed osteoporosis in OVX mice and improved bone morphometric parameters. The serum estradiol, bone alkaline phosphatase levels, regulatory T/Th17 cell numbers were significantly increased by alisol A 24-acetate as compared with untreated OVX mice. In addition, TRAP activity was inhibited by alisol A 24-acetate in OVX mice. These results suggest alisol A 24-acetate effectively prevents bone loss in OVX mice, and that it can be considered a potential therapeutic for the treatment of postmenopausal osteoporosis.

Topics: Acid Phosphatase; Alisma; Alkaline Phosphatase; Animals; Bone Density; Bone Density Conservation Agents; Bone Resorption; Cholestenones; Disease Models, Animal; Estradiol; Female; Femur; Humans; Isoenzymes; Lymphocyte Count; Mice; Mice, Inbred C3H; Osteoporosis, Postmenopausal; Ovariectomy; Phytotherapy; Plant Extracts; T-Lymphocytes, Regulatory; Tartrate-Resistant Acid Phosphatase; Th17 Cells

The aim of this study was to evaluate skeletal pain associated with osteoporosis and to examine the inhibitory effect of bisphosphonate (BP) on pain in an ovariectomized (OVX) mouse model. We evaluated skeletal pain in OVX mice through an examination of pain-like behavior as well as immunohistochemical findings. In addition, we assessed the effects of alendronate (ALN), a potent osteoclast inhibitor, on those parameters. The OVX mice showed a decrease in the pain threshold value, and an increase in the number of c-Fos immunoreactive neurons in laminae I-II of the dorsal horn of the spinal cord. Alendronate caused an increase in the pain threshold value and inhibited c-Fos expression. The serum level of tartrate-resistant acid phosphatase 5b, a marker of osteoclast activity, was significantly negatively correlated with the pain threshold value. Furthermore, we found that an antagonist of the transient receptor potential channel vanilloid subfamily member 1, which is an acid-sensing nociceptor, improved pain-like behavior in OVX mice. These results indicated that the inhibitory effect of BP on osteoclast function might contribute to an improvement in skeletal pain in osteoporosis patients.

Topics: Acid Phosphatase; Alendronate; Animals; Diphosphonates; Disease Models, Animal; Female; Isoenzymes; Mice; Mice, Inbred C57BL; Osteoclasts; Osteoporosis; Ovariectomy; Pain; Pain Threshold; Posterior Horn Cells; Proto-Oncogene Proteins c-fos; Tartrate-Resistant Acid Phosphatase; Transient Receptor Potential Channels

Juvenile idiopathic arthritis (JIA) of the temporomandibular joint (TMJ) can cause severe growth disturbances of the craniomandibular system. Antigen-induced arthritis (AIA) of the rabbit TMJ is simulating the inflammatory process of the TMJ in JIA. The aim of this study was to investigate the effect of a systemic administration of the tumor necrosis factor-alpha (TNF-α) antagonist etanercept on AIA in rabbits by means of three different histological staining methods.. After sensitization, a bilateral arthritis of the TMJ was induced and maintained by repeated intra-articular administrations of ovalbumin in 12 New Zealand white rabbits aged 10 weeks. From the 13th week of age, 6 of the 12 rabbits received weekly subcutaneous injections of etanercept, and the other 6 animals remained without therapy. Another 6 animals served as controls, receiving no treatment or intra-articular injections at all. After euthanasia at the age of 22 weeks, all TMJs were retrieved en bloc. Sagittal sections were cut and stained with hematoxylin-eosin (H-E), Safranin-O for the evaluation of the Mankin score, and tartrate-resistant acid phosphatase (TRAP).. In the arthritis group, a chronic inflammation with degeneration of the articular cartilage was visible. In the etanercept group, the signs of cartilage degeneration were significantly reduced but present. In contrast, the joints in the control group were inconspicuous. A strong correlation between the Mankin score and TRAP-positive cells could be found.. Antigen-induced arthritis causes severe damage in the TMJ of young rabbits. An improvement seems to be achievable by a systemic administration of etanercept.

Topics: Acid Phosphatase; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Arthritis, Juvenile; Biomarkers; Cartilage, Articular; Coloring Agents; Disease Models, Animal; Etanercept; Female; Freund's Adjuvant; Injections, Intra-Articular; Injections, Subcutaneous; Isoenzymes; Mandibular Condyle; Osteoclasts; Ovalbumin; Phenazines; Rabbits; Random Allocation; Tartrate-Resistant Acid Phosphatase; Temporomandibular Joint Disorders; Time Factors

Receptor activator of nuclear factor kappa-B (RANK) and RANK-ligand are relevant targets for the treatment of polyethylene particle-induced osteolysis. This study assessed the local administration of siRNA, targeting both human RANK and mouse Rank transcripts in a mouse model. Four groups of mice were implanted with polyethylene (PE) particles in the calvaria and treated locally with 2.5, 5 and 10 μg of RANK siRNA or a control siRNA delivered by the cationic liposome DMAPAP/DOPE. The tissues were harvested at day 9 after surgery and evaluated by micro-computed tomography, tartrate-resistant acid phosphatase (TRAP) immunohistochemistry for macrophages and osteoblasts, and gene relative expression of inflammatory and osteolytic markers. 10 μg of RANK siRNA exerted a protective effect against PE particle-induced osteolysis, decreasing the bone loss and the osteoclastogenesis, demonstrated by the significant increase in the bone volume (P<0.001) and by the reduction in both the number of TRAP(+) cells and osteoclast activity (P<0.01). A bone anabolic effect demonstrated by the formation of new trabecular bone was confirmed by the increased immunopositive staining for osteoblast-specific proteins. In addition, 5 and 10 μg of RANK siRNA downregulated the expression of pro-inflammatory cytokines (P<0.01) without depletion of macrophages. Our findings show that RANK siRNA delivered locally by a synthetic vector may be an effective approach for reducing osteolysis and may even stimulate bone formation in aseptic loosening of prosthetic implants.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Gene Expression Regulation; Genetic Vectors; HEK293 Cells; Humans; Isoenzymes; Liposomes; Mice; Osteoblasts; Osteolysis; Polyethylene; Receptor Activator of Nuclear Factor-kappa B; RNA, Small Interfering; Tartrate-Resistant Acid Phosphatase

Bone protective effects of withaferin A (WFA) from leaves of Withania somnifera (L.) were evaluated in preventive model of Balb/c mice with 17 β-estradiol (E2) and alendronate (ALD).. Adult female Balb/c mice, 7 to 9 wk, were bilaterally ovariectomized (OVx) to mimic the state of E2 deficiency. Immediately after surgery mice were administrated WFA at doses of 1, 5, 10 mg/kg/d while other two OVx groups received ALD or E2 for 2 mo. Sham and OVx groups with vehicle and no treatment served as controls.. WFA administration increased new bone formation, as well as improving microarchitecture and biomechanical strength of the bones. It prevented bone loss by reducing expression of osteoclastic genes tartrate resistant acid phosphatase (TRAP) and receptor activator of nuclear factor κ B (RANK). Increase in bone turnover marker, osteocalcin (OCN) and inflammatory cytokine tumor necrosis factor-alpha (TNF-α) because of ovariectomy were reduced with WFA treatment, with effects comparable to E2 administration. Histomorphometric analysis of uterus shows that WFA was not fraught with estrogenic or antiestrogenic effects. At cellular level, WFA promoted differentiation of bone marrow cells (BMCs) and increased mineralization by inducing expression of osteogenic genes. WFA has bone protective potential as its treatment prevents bone loss that is comparable to ALD and E2.. It is surmised that WFA in preclinical setting is effective in preserving bone loss by both inhibition of resorption and stimulation of new bone formation before onset of osteoporosis with no uterine hyperplasia.

Topics: Acid Phosphatase; Alendronate; Animals; Biomarkers; Bone and Bones; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Estradiol; Female; Isoenzymes; Mice; Mice, Inbred BALB C; Osteocalcin; Osteoclasts; Osteoporosis; Ovariectomy; Plant Leaves; Plants, Medicinal; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha; Withania; Withanolides

The purpose of this study is to evaluate whether β-tricalcium phosphate (β-TCP) could be a promising modality to help augment alveolar bone in periodontal tissue regeneration by bone marrow mesenchymal stem cells (BMMSCs).. Expanded BMMSCs and atelocollagen (Col) were mixed together (MSC/Col). A combination of β-TCP with MSC/Col was also prepared (MSC/Col/TCP). MSC/Col/TCP or MSC/Col was transplanted into experimental periodontal Class III furcation defects that had been exposed to inflammation in beagle dogs. Periodontal tissue regeneration was evaluated by histologic and morphometric analyses at 4 and 8 weeks after transplantation.. MSC/Col and MSC/Col/TCP enhanced periodontal tissue regeneration compared to Col and TCP/Col according to hematoxylin and eosin staining. The percentage of new cementum length in the MSC/Col/TCP group was not significantly different from that in the MSC/Col group at 4 and 8 weeks. On the other hand, the percentage of new bone area in the MSC/Col/TCP group was much higher than that in the MSC/TCP group at 4 weeks. However, at 8 weeks, no significant difference in new bone area was found between the two groups. In the MSC/Col/TCP group, β-TCP was surrounded by newly formed bone. Multinucleated cells, which were positive for osteopontin and tartrate-resistant acid phosphatase, were present in the interconnected macropores of β-TCP.. These findings suggest that β-TCP is applicable as a scaffold for BMMSCs transplantation and helps augment alveolar bone without impairing regeneration of cementum.

Topics: Acid Phosphatase; Alveolar Process; Animals; Bone Regeneration; Calcium Phosphates; Cell Culture Techniques; Cementogenesis; Collagen; Collagen Type I; Dental Cementum; Disease Models, Animal; Dogs; Female; Furcation Defects; Giant Cells; Guided Tissue Regeneration, Periodontal; Isoenzymes; Mesenchymal Stem Cell Transplantation; Organ Size; Osteogenesis; Osteopontin; Tartrate-Resistant Acid Phosphatase; Tissue Scaffolds; Tooth Ankylosis; Tooth Root

The synergistic effects of vitamin D3 and vitamin K2 on bone loss prevention have been reported. This study evaluates the effects of vitamin D3 and vitamin K2 supplementation in conjunction with conventional periodontal therapy (scaling and root planing [SRP]) on gingival interleukin (IL)-1β and IL-10, serum bone alkaline phosphatase (B-ALP) and tartrate-resistant acid phosphatase 5b (TRAP-5b), and calcium and alveolar bone levels in rats with experimentally induced periodontitis.. Seventy-two rats were divided into the following groups: 1) healthy; 2) periodontitis; 3) SRP; 4) SRP + vitamin D3; 5) SRP + vitamin K2; and 6) SRP + vitamins K2 and D3. Periodontitis was induced by ligature placement for 7 days, and vitamin K2 (30 mg/kg) and/or vitamin D3 (2 μg/kg) were administered for 10 days in the SRP + vitamin D3, SRP + vitamin K2, and SRP + vitamins K2 and D3 groups by oral gavage. On day 18, the animals were sacrificed, serum B-ALP, TRAP-5b, and calcium levels were measured, gingiva specimens were extracted for IL-1β and IL-10 analysis, and distances between the cemento-enamel junction and alveolar bone crest were evaluated.. Alveolar bone levels in the periodontitis group were significantly greater than those in the other five groups. No significant differences were found in gingival IL-1β and IL-10, serum B-ALP and TRAP-5b, and calcium and alveolar bone levels between the groups receiving SRP and vitamins and the group receiving SRP alone.. Within the limitations of this study, vitamin D3 and K2 alone or in combination did not affect gingival IL-1β and IL-10, serum B-ALP and TRAP-5b levels, or alveolar bone compared with conventional periodontal therapy alone.

Topics: Acid Phosphatase; Alkaline Phosphatase; Alveolar Process; Animals; Bone Density Conservation Agents; Calcium; Cholecalciferol; Combined Modality Therapy; Dental Scaling; Disease Models, Animal; Gingiva; Interleukin-10; Interleukin-1beta; Isoenzymes; Male; Periodontitis; Random Allocation; Rats; Rats, Wistar; Root Planing; Tartrate-Resistant Acid Phosphatase; Tooth Cervix; Vitamin K 2; Vitamins

Long-term effects of glucocorticoid treatment in humans induce bone loss and increase the risk of fracture in the skeleton. The pathogenic mechanisms of glucocorticoid-induced osteoporosis (GIOP) are still unclear. The GIOP and its effects have been reproduced in several animal models including Danio rerio (zebrafish) embryo. The treatment of adult fish with prednisolone (PN) has shown a dose-dependent decrease of mineralized matrix in the scales. Large resorption lacunae are characterized by single TRAP-positive cells which migrate to the margin of the scale merging into a multinucleated structures. The treatment with PN of cultured scales did not increase TRAP activity suggesting that the massive presence of osteoclasts in the resorption sites could be likely the result of a systemic recruitment of monocyte-macrophage precursors. We observed that treatment with PN induced a significant decrease of the alkaline phosphatase (ALP) activity in scale scleroblasts if compared with untreated controls. Then, we investigated the total mineral balance under prednisolone treatment using a time-dependent double live staining. The untreated fish fully repaired the resorption lacuna induced by prednisolone, whereas treated fish failed. The presence of osteoclast resorption fingerprints on new matrix suggested that the osteoclast activity counterbalances the osteodepositive activity exerted by scleroblasts. The treatment with PN in association with alendronate (AL) has surprisingly resulted in a significant decrease of TRAP activity and increase of ALP compared to PN-treated fish in biochemical and histological assays confirming the action of alendronate against GIOP in fish as well in humans.

Topics: Acid Phosphatase; Alendronate; Alkaline Phosphatase; Animals; Biomarkers; Bone Density Conservation Agents; Bone Matrix; Bone Remodeling; Disease Models, Animal; Glucocorticoids; Isoenzymes; Male; Osteoclasts; Osteoporosis; Phenotype; Prednisolone; Tartrate-Resistant Acid Phosphatase; Time Factors; Tissue Culture Techniques; Zebrafish; Zebrafish Proteins

The aim of this study was to evaluate the influence of previous exposure to Cyclosporine A (CsA) on experimental periodontitis in rats.. Forty rats were divided into 4 groups: Control (CON); Cyclosporine A (CsA), which received daily doses of 10mg/kg CsA; Ligature (LIG), which received an insertion of a cotton ligature around the mandibular 1st molar at day 30; and Ligature and CsA (CsAL), which were treated with CsA and the cotton ligature. At day 60 of the experiment, animals were sacrificed, and groups were compared with regards to Alkaline Phosphatase (AP) activity, gingival overgrowth, periodontal bone support (PBS), bone resorption at furcation ligament area (LA) and TRAP+ cells. Data were analyzed by ANOVA/Tukey and Kruskal-Wallis and were considered to be statistically significant at 5% level.. CsA and LIG groups showed similar gingival area, which was higher than that in the CON and lower than in the CsAL group (p=0.001). The ratio between epithelial area and connective area for the CON group was similar to the CsA group and higher than that for the CsAL and LIG groups (p=0.0334). Mean percentage of PBS for the CON group was similar to that for the CsAL group and higher than that of the CsA and LIG groups (p=0.0007). No difference was observed regarding AP (p=0.2806) and TRAP+ cells (p=0.3995) among experimental groups. Mean values for LA of CON were similar to CsA, and both were statistically lower than the CsAL and LIG groups (p=0.0172).. Based on these results, we posit that previous exposure to CsA may influence gingival overgrowth, but not bone loss, in rats with experimental periodontitis.

Topics: Acid Phosphatase; Alkaline Phosphatase; Alveolar Bone Loss; Animals; Bone Resorption; Cyclosporine; Disease Models, Animal; Gingival Overgrowth; Isoenzymes; Ligation; Male; Mandible; Molar; Periodontitis; Radiography; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Our study aimed to investigate the antiosteoporotic properties of the ethanol extract of Podocarpium podocarpum (DC.) Yang et Huang (PE) in ovariectomized (OVX) rats and to characterize the active constituents. As a result, PE significantly inhibited the increased urinary Ca excretion and activity of bone resorption markers including tartrate-resistant acid phosphatase (TRAP), deoxypyridinoline crosslinks and cathepsin K in OVX rats, whereas exhibited little effects on the body, uterus and vagina weight. Detailed micro-CT analysis showed that PE notably enhanced bone quality, with increased bone mineral content (BMC), bone volume fraction (BVF), connectivity density (CD), tissue mineral content (TMC), tissue mineral density (TMD) and trabecular number (Tb. N), and decreased trabecular separation (Tb. Sp), in OVX animal. Those findings implied that PE had notable antiosteoporotic effect, especially effective in preventing bone resorption, with little side-effects on reproductive tissue. Further chemical investigation led to the isolation of 17 flavonoids, most of which showed significantly stimulatory effect on osteoblastic proliferation, ALP activity and mineralized nodes formation as well as inhibitory effect on osteoclastic TRAP activity in osteoblastic and osteoclastic cells. Our results indicated that PE, with abundant flavonoids, had remarkable antiosteoporotic activity and therefore can be a promising candidate for the treatment of postmenopausal osteoporosis induced by estrogen deficiency through herbal remedy.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Density; Cells, Cultured; Disease Models, Animal; Fabaceae; Female; Femur; Flavonoids; Isoenzymes; Molecular Structure; Osteoblasts; Osteoclasts; Osteoporosis; Ovariectomy; Plant Extracts; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

A major challenge to understanding osteoarthritis (OA) pathology is identifying the cellular events that precede the onset of cartilage damage. The objective of this study is to determine the effect of joint destabilization on early changes to fibrocartilage in the joint.. The anterior cruciate ligament was transected in collagen reporter mice (Col1CFP and ColXRFP). Mineralization labels were given every 2 weeks to measure new mineralized cartilage apposition. Novel fluorescent histology of mineralized tissue was used to characterize the changes in fibrocartilage at 2 and 4 weeks post-injury.. Changes in fibrocartilaginous structures of the joint occur as early as 2 weeks after injury and are well developed by 4 weeks. The alterations are seen in multiple entheses and in the medial surface of the femoral and tibial condyles. In the responding entheses, mineral apposition towards the ligament midsubstance results in thickening of the mineralize fibrocartilage. These changes are associated with increases in ColX-RFP, Col1-CFP reporter activity and alkaline phosphatase enzyme activity. Mineral apposition also occurs in the fibrocartilage of the non-articular regions of the medial condyles by 2 weeks and develops into osteophytes by 4 weeks post-injury. An unexpected observation is punctate expression of tartrate resistant acid phosphatase activity in unmineralized fibrochondrocytes adjacent to active appositional mineralization.. These observations suggest that fibrocartilage activates prior to degradation of the articular cartilage. Thus clinical and histological imaging of fibrocartilage may be an earlier indicator of disease initiation and may indicate a more appropriate time to start preventative treatment.

Topics: Acid Phosphatase; Animals; Anterior Cruciate Ligament Injuries; Calcification, Physiologic; Cartilage, Articular; Chondrocytes; Disease Models, Animal; Female; Femur; Fibrocartilage; Genes, Reporter; Green Fluorescent Proteins; Isoenzymes; Joint Instability; Mice, Transgenic; Tartrate-Resistant Acid Phosphatase; Tibia

Osteonecrosis of the jaws (ONJ) is a significant complication of antiresorptive medications, such as bisphosphonates and denosumab. Antiresorptive discontinuation to promote healing of ONJ lesions remains highly controversial and understudied. Here, we investigated whether antiresorptive discontinuation alters ONJ features in mice, employing the potent bisphosphonate zoledronic acid (ZA) or the receptor activator of NF-κB ligand (RANKL) inhibitor OPG-Fc, utilizing previously published ONJ animal models. Mice were treated with vehicle (veh), ZA, or OPG-Fc for 11 weeks to induce ONJ, and antiresorptives were discontinued for 6 or 10 weeks. Maxillae and mandibles were examined by μCT imaging and histologically. ONJ features in ZA and OPG-Fc groups included periosteal bone deposition, empty osteocyte lacunae, osteonecrotic areas, and bone exposure, each of which substantially resolved 10 weeks after discontinuing OPG-Fc but not ZA. Full recovery of tartrate-resistant acid phosphatase-positive (TRAP+) osteoclast numbers occurred after discontinuing OPG-Fc but not ZA. Our data provide the first experimental evidence demonstrating that discontinuation of a RANKL inhibitor, but not a bisphosphonate, reverses features of osteonecrosis in mice. It remains unclear whether antiresorptive discontinuation increases the risk of skeletal-related events in patients with bone metastases or fracture risk in osteoporosis patients, but these preclinical data may nonetheless help to inform discussions on the rationale for a "drug holiday" in managing the ONJ patient.

Topics: Abscess; Acid Phosphatase; Animals; Bisphosphonate-Associated Osteonecrosis of the Jaw; Bone Resorption; Denosumab; Diphosphonates; Disease Models, Animal; Imidazoles; Immunoglobulin Fc Fragments; Isoenzymes; Male; Mandible; Maxilla; Mice; Mice, Inbred C57BL; Osteoclasts; Osteoprotegerin; RANK Ligand; Rats; Recombinant Fusion Proteins; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography; Zoledronic Acid

Aseptic loosening associated with wear particle-induced inflammation is a major cause of joint implant failure. Recent studies have demonstrated the therapeutic effects of p38 mitogen-activated protein kinase (MAPK)-based therapies on chronic inflammatory diseases. The purpose of this study was to investigate whether SB203580, a p38 MAPK inhibitor, inhibits wear debris-induced inflammatory osteolysis in mice through downregulation of receptor activator of nuclear factor kβ (RANK)/RANK ligand (RANKL). We used a murine osteolysis model to study the effect of SB203580 on RANKL/RANK signaling and titanium particle-induced osteolysis in vivo. Pouch membranes with intact bone implants were analyzed using histological analysis and transmission electron microscopy, and the levels of RANK and RANKL protein and mRNA were evaluated by immunohistological staining and real-time reverse transcriptase-polymerase chain reaction. SB203580 had less of an effect on RANK and RANKL expression under wear debris-induced conditions. The number of TRAP-positive cells was remarkably reduced in Ti-particle-induced pouch tissues. These effects were confirmed through the transmission electron microscopy results. These results suggest that p38 MAPK-based therapies are beneficial in preventing aseptic loosening associated with total joint replacement by modulating RANK-RANKL signaling.

Topics: Acid Phosphatase; Animals; Bone and Bones; Colonic Pouches; Disease Models, Animal; Down-Regulation; Female; Immunohistochemistry; Inflammation; Isoenzymes; Mice, Inbred BALB C; Osteolysis; p38 Mitogen-Activated Protein Kinases; Prostheses and Implants; Protective Agents; Protein Kinase Inhibitors; RANK Ligand; Real-Time Polymerase Chain Reaction; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; Titanium

Subchondral bone cyst (SBC) growth, caused by osteoclast activity during early knee osteoarthritis (OA) pathogenesis, should be treated to prevent further progressions of OA. In the present study, we evaluated the effects of gentle treadmill walking on subchondral bone and cartilage changes in an experimental rat model of destabilized medial meniscus (DMM).. Twelve-week-old Wistar rats underwent DMM surgery in their right knee and sham surgery in their left knee and were assigned to either the sedentary group or walking group (n = 42/group). Animals in the walking group were subjected to treadmill exercise 2 days after surgery, which included walking for 12 m/min, 30 min/day, 5 days/week for 1, 2, and 4 week(s). Subchondral bone and cartilage changes were evaluated by micro-CT analysis, histological analysis, and biomechanical analysis.. Treadmill walking had a tendency to suppress SBC growth, which was confirmed by micro-CT (P = 0.06) and positive staining for tartrate-resistant acid phosphatase (TRAP) activity for the osteoclast number per bone surface (P = 0.09) 4 weeks after surgery. These changes coincide with the prevention of cartilage degeneration as evaluated by the Osteoarthritis Research Society International (OARSI) score (P < 0.05) and biomechanically softening (P < 0.05). Furthermore, treadmill walking could suppressed increasing osteocyte deaths (P < 0.01), which was positively correlated with the OARSI score (r = 0.77; P < 0.01).. These results indicate biomechanical and biological links exist between cartilage and subchondral bone; preventive effects of treadmill walking on subchondral bone deterioration might be partly explained by the chondroprotective effects.

Topics: Acid Phosphatase; Animals; Apoptosis; Cartilage, Articular; Cell Death; Disease Models, Animal; Exercise Test; Immunohistochemistry; In Situ Nick-End Labeling; Male; Menisci, Tibial; Osteoarthritis; Osteoarthritis, Knee; Osteocytes; Osteophyte; Rats; Rats, Wistar; Tibia; Walking; X-Ray Microtomography

Integrin-based (β3 ) attachments to the extracellular matrix (ECM) on osteocyte cell processes have recently been proposed to play an important role in facilitating osteocyte mechanosensation. However, it is not yet known whether integrin expression is altered in the mechanoregulatory osteocytes during osteoporosis. The objective of this study was to test the hypothesis that the expression of integrin-based mechanosensory complexes (β1 and β3 integrins) is altered as a direct response to estrogen deficiency, in an estrogen deficient animal model of osteoporosis. Four weeks post-operatively, immunohistochemistry was used to detect for β1 and β3 integrin subunits in bone tissue and marrow of ovariectomized (OVX; N = 4) and SHAM (N = 4) operated animals. A tartrate resistant acid phosphatase (TRAP) control stain was performed to quantify the presence of osteoclasts in the bone marrow and bone surfaces. Image analysis was performed to quantify expression patterns in different biological compartments, that is, bone marrow, endosteum, and cortical bone. Our results showed that β1 integrins were ubiquitously expressed throughout the bone and marrow, for both OVX and SHAM groups. β3 integrin subunit expression was lower in bone cells from osteoporotic animals compared to controls, whereas β3 expression in marrow cells did not differ significantly between groups. At the endosteum no difference was observed in β3 integrin subunit expression. As expected, the number of osteoclasts was higher in the OVX group validating an imbalance in bone remodeling. We propose that a reduction in β3 integrin expression in osteocytes might impair mechanosensation by bone cells during estrogen deficiency.

Topics: Acid Phosphatase; Animals; Bone Marrow; Bone Remodeling; Disease Models, Animal; Estrogens; Female; Femur; Humans; Immunohistochemistry; Integrin beta1; Integrin beta3; Isoenzymes; Mechanotransduction, Cellular; Osteoclasts; Osteoporosis, Postmenopausal; Ovariectomy; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Tibia; Time Factors

The potential of increasing bone mass and preventing fractures in osteoporosis using stem cell therapy is currently an area of intense focus. However, there are very little data available regarding the postfracture bony defect healing efficacy under osteoporotic conditions. This study aims to investigate whether critical-sized segmental bone defects in a rabbit model of osteoporosis could be repaired using an allogenic stem cell-based tissue engineering (TE) approach and to investigate the potential influence of osteoporosis on the treatment efficacy. Rabbit fetal bone marrow mesenchymal stem cells (BMSCs) were harvested and expanded in vitro. Decalcified bone matrix (DBM) scaffolds were then seeded with allogenic fetal BMSCs and cultivated in osteogenic media to engineer BMSC/DBM constructs. Critical-sized radial defects were created in ovariectomized (OVX) rabbits and the defects were repaired either by insertion of BMSC/DBM constructs or by DBM scaffolds alone. Also, nonovariectomized age-matched (non-OVX) rabbits were served as control. At 3 months post-treatment under the osteoporotic condition (OVX rabbits), the BMSC/DBM constructs inserted within the defect generated significantly more bone tissue when compared to the DBM scaffold as demonstrated by the X-ray, microcomputed tomography, and histological analyses. In addition, when compared to a normal nonosteoporotic condition (age-matched non-OVX rabbits), the defect treatment efficacy was adversely affected by the osteoporotic condition with significantly less bone regeneration. This study demonstrated the potential of allogenic fetal BMSC-based TE strategy for repairing bone defects in an osteoporotic condition. However, the treatment efficacy could be considerably compromised in the OVX animals. Therefore, a more sophisticated strategy that addresses the complicated pathogenic conditions associated with osteoporosis is needed.

Topics: Acid Phosphatase; Animals; Bone Matrix; Cell Shape; Disease Models, Animal; Female; Isoenzymes; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Osteoporosis; Ovariectomy; Rabbits; Radiography; Radius; Sus scrofa; Tartrate-Resistant Acid Phosphatase; Tissue Engineering; Treatment Outcome; Wound Healing

This study aims to develop a reproducible rat model for post-traumatic bisphosphonate-related osteonecrosis of the jaw (BRONJ). In our previous studies using dental extraction as an inducing factor, only 30%-60% of zoledronate-treated animals fulfilled the definition of clinical BRONJ. We modified the zoledronate regimen and introduced repeated surgical extraction to illicit quantifiable BRONJ in all animals. Eighty retired-breeder female Sprague-Dawley rats were divided between the treatment (i.v. zoledronate; 80 μg/kg/week for 13 weeks) and control (saline) groups. On week 13, the left mandibular first molar was surgically extracted, followed by the second molar a week later. Animals were euthanized at 1-week, 2-weeks, and 8-weeks following extraction. The occurrence and severity of BRONJ were scored in each animal based on gross and MicroCT analysis. Parameters of bone formation and osteoclast functions at the extraction site were compared between groups. All zoledronate-treated animals developed a severe case of BRONJ that fulfilled the clinical definition of the condition in humans. Osteoclast attachment continued to be defective eight weeks after stopping the treatment. There were no signs of kidney or liver toxicity. Our data confirmed that repeated surgical extraction (major trauma) by itself consistently precipitated massive bone necrosis in ZA-treated animals, eliminating the need to induce pre-existing infection or comorbidity. These results will be the basis for further studies examining the in-vivo pathogenesis and prevention of BRONJ.

Topics: Acid Phosphatase; Animals; Bisphosphonate-Associated Osteonecrosis of the Jaw; Diphosphonates; Disease Models, Animal; Female; Imidazoles; Isoenzymes; Kidney; Liver; Mandible; Osteoclasts; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Tooth Extraction; Wound Healing; Wounds and Injuries; X-Ray Microtomography; Zoledronic Acid

In the current study, we investigated the improvement of phosphorylated peptides from Antarctic krill Euphausia superba (PP-AKP) on osteoporosis in ovariectomized rats. PP-AKP was supplemented to ovariectomized Sprague-Dawley rats for 90 days. The results showed that PP-AKP treatment remarkably prevented the reduction of bone mass and improved cancellous bone structure and biochemical properties. PP-AKP also significantly decreased serum contents of tartrate-resistant acid phosphatase (TRACP), cathepsin K (Cath-k), matrix metalloproteinases-9 (MMP-9), deoxypyridinoline (DPD), C-terminal telopeptide of collagen I (CTX-1), Ca, and P. Mechanism investigation revealed that PP-AKP significantly increased the osteoprotegerin (OPG)/receptor activator of nuclear factor κB ligand (RANKL) ratio in mRNA expression, protein expression, and serum content. Further research suggested that NF-κB signaling pathways were inhibited by suppressing the mRNA and protein expressions of nuclear factor of activated T-cells (NFATc1) and tumor necrosis factor receptor-associated factor 6 (TRAF6), diminishing the mRNA expression and phosphorylation of nuclear factor κB p65 (NF-κB p65), three key transcription factors in NF-κB pathways. These results suggest that PP-AKP can improve osteoporosis by inhibiting bone resorption via suppressing the activation of osteoclastogenesis related NF-κB pathways.

Topics: Acid Phosphatase; Animals; Antarctic Regions; Bone Density; Bone Density Conservation Agents; Disease Models, Animal; Estrogens; Euphausiacea; Female; Humans; Isoenzymes; Matrix Metalloproteinase 9; Osteoporosis, Postmenopausal; Ovariectomy; Peptides; Rats; Rats, Sprague-Dawley; Signal Transduction; Tartrate-Resistant Acid Phosphatase

We evaluated the side effects of bisphosphonate (BP) on tooth extraction socket healing in spontaneously diabetic Torii (SDT) rats, an established model of non-obese type 2 diabetes mellitus, to develop an animal model of BP-related osteonecrosis of the jaws (BRONJ).. Male Sprague-Dawley (SD) rats and SDT rats were randomly assigned to the zoledronic acid (ZOL)-treated groups (SD/ZOL or SDT/ZOL) or to the control groups (SD/control or SDT/control). Rats in the SD/ZOL or SDT/ZOL groups received an intravenous bolus injection of ZOL (35 μg/kg) every 2 weeks. Each group consisted of 6 rats each. Twenty-one weeks after ZOL treatment began, the left maxillary molars were extracted. The rats were euthanized at 2, 4, or 8 weeks after tooth extraction, and the total maxillae were harvested for histological and histochemical studies.. In the oral cavity, bone exposure persisted at the tooth extraction site in all rats of the SDT/ZOL group until 8 weeks after tooth extraction. In contrast, there was no bone exposure in SD/control or SDT/control groups, and only 1 of 6 rats in the SD/ZOL group showed bone exposure. Histologically, necrotic bone areas with empty lacunae, microbial colonies, and less invasion by inflammatory cells were observed. The number of tartrate-resistant acid phosphatase-positive osteoclasts was lower in the SDT/ZOL group than in the SD/control group. The mineral apposition rate was significantly lower in the SDT/ZOL group compared with the SD/control group.. This study demonstrated the development of BRONJ-like lesions in rats and suggested that low bone turnover with less inflammatory cell infiltration plays an important role in the development of BRONJ.

Topics: Acid Phosphatase; Animals; Bisphosphonate-Associated Osteonecrosis of the Jaw; Bone Density; Bone Remodeling; Diabetes Mellitus, Type 2; Diphosphonates; Disease Models, Animal; Gene Expression; Humans; Imidazoles; Injections, Intravenous; Isoenzymes; Male; Maxilla; Molar; Osteoclasts; Rats; Rats, Sprague-Dawley; Rats, Transgenic; Tartrate-Resistant Acid Phosphatase; Tooth Extraction; Zoledronic Acid

Occlusal trauma (OT) and smoking are both factors that alter alveolar bone metabolism and therefore could synergistically act on alveolar bone loss. The aim of this experimental study was to evaluate the influence of short-term cigarette smoke inhalation (CSI) on inter-radicular alveolar bone loss promoted by primary OT in a rat model.. Forty-eight animals were randomly assigned to one of three groups based on treatment type: OT + CSI (n = 16), animals were exposed to CSI three times per day, for 8 min per exposure, and they concomitantly received unilateral vertical augmentation creating an occlusal interference inducing experimental OT; OT (n = 16), animals received only unilateral vertical augmentation; negative control (NC; n = 16), animals maintained for equal periods to achieve periodontal baseline values of periodontal ligament dimension. Each group was divided into two subgroups (n = 8) based on treatment length: 7 or 14 d.. After 7 d, the OT + CSI group exhibited significantly higher bone loss compared to the NC group (p = 0.0022). After 14 d, the OT (p < 0.0001) and OT + CSI (p < 0.0001) groups presented significantly higher bone loss compared to the NC group, and OT + CSI resulted in significantly higher bone loss than OT alone (p = 0.0241). The number of tartrate-resistant acid phosphatase-positive cells on the linear surface of the bone crest after 7 d was significantly higher in the OT + CSI group as compared to the NC and OT groups (p < 0.0001 and p = 0.0045, respectively) and remained significantly higher in the OT + CSI group after 14 d, compared to the OT group (p < 0.0001).. Short-term CSI increases early bone loss in association with OT after 7 d, and this worsens in severity after 14 d of exposure.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Dental Occlusion, Traumatic; Disease Models, Animal; Disease Progression; Furcation Defects; Isoenzymes; Male; Random Allocation; Rats; Rats, Wistar; Smoking; Tartrate-Resistant Acid Phosphatase; Time Factors

Studies have demonstrated that a moderate intake of amino acids is associated with development of bone health. Methionine, a sulphur-containing essential amino acid, has been largely implicated for improving cartilage formation, however its physiological significance on bone integrity and functionality have not been elucidated. We investigated whether methionine can prevent osteoporotic bone loss.. The anti-resorptive effect of methionine, (250 mg kg(-1) body wt administered in drinking water for 10 weeks), was evaluated in ovariectomized (OVX) rats by monitoring changes in bone turnover, formation of osteoclasts from blood-derived mononuclear cells and changes in the synthesis of pro-osteoclastogenic cytokines.. Methionine improved bone density and significantly decreased the degree of osteoclast development from blood mononuclear cells in OVX rats, as indicated by decreased production of osteoclast markers tartarate resistant acid phosphatase b (TRAP5b) and MIP-1α. siRNA-mediated knockdown of myeloid differentiation primary response 88 [MyD88], a signalling molecule in the toll-like receptor (TLR) signalling cascade, abolished the synthesis of both TRAP5b and MIP-1α in developing osteoclasts. Methionine supplementation disrupted osteoclast development by inhibiting TLR-4/MyD88/NF-κB pathway.. TLR-4/MyD88/NF-κB signalling pathway is integral for osteoclast development and this is down-regulated in osteoporotic system on methionine treatment. Methionine treatment could be beneficial for the treatment of postmenopausal osteoporosis.

Topics: Acid Phosphatase; Administration, Oral; Alendronate; Animals; Bone Density; Bone Density Conservation Agents; Bone Remodeling; Cells, Cultured; Chemokine CCL3; Disease Models, Animal; Dose-Response Relationship, Drug; Down-Regulation; Drug Therapy, Combination; Female; Inflammation Mediators; Isoenzymes; Methionine; Myeloid Differentiation Factor 88; NF-kappa B; Osteoclasts; Osteoporosis; Ovariectomy; Rats; Rats, Sprague-Dawley; RNA Interference; Signal Transduction; Tartrate-Resistant Acid Phosphatase; Time Factors; Toll-Like Receptor 4

Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase and have anti-inflammatory effects independent of cholesterol lowering. Recent clinical studies have indicated that statin intake has a beneficial effect on periodontal disease. However, the underlying mechanisms have not been well understood. In the current study, we employed a rat model with lipopolysaccharide (LPS)-induced periodontal disease and determined the effect of simvastatin, a commonly prescribed statin, on osteoclastogenesis, gingival inflammation and alveolar bone loss.. Sprague-Dawley rats were injected with Aggregatibacter actinomycetemcomitans LPS in periodontal tissue three times per week for 8 wk and part of the rats with LPS injection were also given simvastatin via gavage. After the treatments, the rat maxillae were scanned by microcomputed tomography and the images were analyzed to determine alveolar bone loss. To explore the underlying mechanisms, the effect of simvastatin on osteoclastogenesis and gingival expression of proinflammatory cytokines were also determined by tartrate-resistant acid phosphatase staining and real-time polymerase chain reaction assays, respectively.. Results showed that LPS treatment markedly increased bone loss, but administration of simvastatin significantly alleviated the bone loss. Results also showed that LPS treatment stimulated osteoclastogenesis and the expression of inflammatory cytokines, but simvastatin significantly modulates the stimulatory effect of LPS on osteoclastogenesis and cytokine expression.. This study demonstrated that simvastatin treatment inhibits LPS-induced osteoclastogenesis and gingival inflammation and reduces alveolar bone loss, indicating that the intake of simvastatin may hinder the progression of periodontal disease.

Topics: Acid Phosphatase; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Anti-Inflammatory Agents; Cytokines; Disease Models, Animal; Gingiva; Gingivitis; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Inflammation Mediators; Isoenzymes; Lipopolysaccharides; Matrix Metalloproteinase 9; Maxillary Diseases; Osteoclasts; Periodontal Diseases; Rats; Rats, Sprague-Dawley; Simvastatin; Tartrate-Resistant Acid Phosphatase; Toll-Like Receptors; X-Ray Microtomography

The objectives of this study were to establish a bisphosphonate-related osteonecrosis of the jaw (BRONJ) rat model and to analyse the effects of teriparatide (TP) on this model.. Sprague-Dawley rats were divided into three groups: I-zoledronic acid (ZA, n = 10); II-ZA and teriparatide (ZA + TP, n = 10); III-control (n = 10). Osteonecrosis was induced by administering zoledronic acid to groups ZA and ZA + TP. A week after the injections, rats underwent extraction of the first left mandibular molar. Following a four week period, TP was administered to the ZA + TP group for 28 days. Upon killing, extraction sockets were examined clinically, radiologically and histopathologically.. Clinical examination revealed necrotic bone exposure in none of the animals. MicroCT (µCT) examination showed that bone mineral density of the newly formed bone in the extraction socket was lower in the ZA group than in the ZA + TP group (p < 0.05). Histopathological examination revealed that only the ZA and ZA + TP groups developed osteonecrosis, and the osteonecrotic bone area in the ZA group was larger than that in the ZA + TP group (p < 0.05). Tartrate-resistant acid phosphatase (TRAcP) enzyme histochemistry revealed that the number of detached and large osteoclasts were higher in the ZA group than in other groups, whereas the number of apoptotic osteoclasts in both ZA and ZA + TP groups were higher than in the control group (p < 0.05).. Our data indicate that bisphosphonate-related osteonecrosis of the jaw model used in the present study is an attractive model to investigate treatment modalities and that TP might be an effective treatment in BRONJ.

Topics: Acid Phosphatase; Alveolar Process; Animals; Apoptosis; Biomarkers; Bisphosphonate-Associated Osteonecrosis of the Jaw; Bone Density; Bone Density Conservation Agents; Bone Marrow; Cell Adhesion; Cell Count; Diphosphonates; Disease Models, Animal; Female; Imidazoles; Injections, Intraperitoneal; Injections, Subcutaneous; Isoenzymes; Molar; Osteoclasts; Osteogenesis; Random Allocation; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Teriparatide; Tooth Extraction; Tooth Socket; X-Ray Microtomography; Zoledronic Acid

There is evidence that histamine released during inflammation plays a role in bone metabolism via the H2 receptor, stimulating bone resorption. The purpose of this study is to evaluate whether cimetidine, a histamine H2-receptor antagonist, interferes with the initiation and progression of induced periodontal disease in rat molars.. Forty male rats received 100 mg/kg body weight of cimetidine (cimetidine group [CimG]) or saline solution (sham group [SG]). Periodontal disease was induced in the maxillary left first molars (PDSG and PDCimG); maxillary right molars were used as non-ligature controls. After 7, 15, 30, and 50 days, maxillary fragments were embedded in paraffin. The sections were stained with Masson trichrome and hematoxylin and eosin and subjected to the tartrate-resistant acid phosphatase (TRAP) method. The distances between the cemento-enamel junction (CEJ) and alveolar process (AP) crest, as well as between the CEJ and junctional epithelium (JE) level, were measured; the number of inflammatory cells was computed. Receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) immunohistochemistry was carried out, and the RANKL/OPG ratio was calculated.. In PDSG and PDCimG, a significant increase (P ≤0.05) was observed in CEJ-AP and CEJ-JE distances. However, the increases in both distances were significantly less in PDCimG compared with PDSG at 15, 30, and 50 days. Numerous TRAP-positive osteoclasts were found in the PDSG and PDCimG. In PDCimG, the volume density of inflammatory cells and the RANKL/OPG ratio were significantly lower (P ≤0.05) than in PDSG.. Cimetidine exerts a beneficial effect on periodontal disease in rats, decreasing the RANKL/OPG ratio in gingival connective tissue and reducing alveolar bone resorption.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Cimetidine; Coloring Agents; Connective Tissue; Disease Models, Animal; Disease Progression; Epithelial Attachment; Fibroblasts; Gingiva; Histamine H2 Antagonists; Isoenzymes; Male; Maxilla; Molar; Osteoprotegerin; Periodontitis; RANK Ligand; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Cervix

Occlusal alignment is known clinically to have a widespread influence on the stomatognathic system, including the temporomandibular joint and masticatory muscles. However, while occlusion is still an important determinant of most dental treatments, the exact effect of occlusal alignment is unclear because of a lack of conclusive scientific evidence. In this study, a malocclusion model system is used to examine the cellular and histologic alterations in the contralateral condyle of mice after a malocclusion was induced by a build-up of resin on the left maxillary molars. A significant decrease in the thickness of the condylar cartilage was found in the 1-week experimental group, together with increased apoptosis and decreased proliferation in the condylar head, which included cartilage and subchondral bone. Additionally, the number of TRAP-positive osteoclasts and MPO- and F4/80-positive inflammatory cells in the subchondral bone were significantly higher in the 1-week experimental group. Unbalanced malocclusion caused increased bone remodeling, as evidenced by increased osteoclastic activity and inflammatory responses (macrophages and neutrophils, respectively). However, these alterations in the 1-week experimental group were subsequently attenuated and restored almost to the baseline at 3 weeks after the induction of the malocclusion.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Imaging, Three-Dimensional; In Situ Nick-End Labeling; Isoenzymes; Ki-67 Antigen; Male; Malocclusion; Mandibular Condyle; Mice; Mice, Inbred C57BL; Peroxidase; Tartrate-Resistant Acid Phosphatase

Tartrate-resistant acid phosphatase (TRAP) is known as an osteoclast marker, but osteoblasts and osteocytes in the vicinity of bone remodeling sites also express TRAP. Cell culture studies suggest that osteoblasts endocytose osteoclastic TRAP for inactivation. To evaluate whether changes in osteoclast activity could alter TRAP expression in osteoblasts and/or osteocytes in vivo, we studied the ovariectomized and vitamin D-deficient rat (Ovx-D) and rats healing from rickets. Bone sections were analyzed for TRAP gene expression by in situ hybridization, TRAP protein by immunogold labeling, and TRAP enzyme activity using the fluorescent substrate ELF97. Osteoblasts and osteocytes close to intracortical remodeling sites and bone surfaces demonstrated TRAP, most prominently in cancellous bone and osteocytes. Intracellular TRAP was located to electron-dense vesicles with similar morphology in both cell types. Ovx-D increased osteoclast activity (p < 0.001) and ELF97⁺ osteocytes (p < 0.05) in cancellous bone, but no corresponding increase was observed in the osteocyte lacunar area. The level of TRAP⁺ vesicles in cortical osteoblasts (p < 0.01) in Ovx-D rats was also increased. Enhanced osteoclast activity was noted in healing rickets after 72 h (p < 0.05), but no differences in TRAP expression were detected in osteoblasts or osteocytes. Thus, increased osteoclast activity does not affect TRAP expression in osteoblasts and osteocytes, favoring the notion that increased TRAP in these cells is rather due to increased synthesis. Although the role of TRAP in osteoblasts and osteocytes remains elusive, we speculate that the function is related to the capability of the enzyme to regulate the phosphorylation of proteins known to be expressed by these cells.

Topics: Acid Phosphatase; Animals; Bone Remodeling; Disease Models, Animal; Female; Fluorescent Antibody Technique; Humans; Immunohistochemistry; In Situ Hybridization; Isoenzymes; Microscopy, Electron, Transmission; Osteoblasts; Osteoclasts; Osteocytes; Osteoporosis, Postmenopausal; Rats; Rickets; Tartrate-Resistant Acid Phosphatase

Femoral head osteonecrosis is frequently observed in patients treated with excessive corticosteroids. The objective of the current study was to establish a rat model to investigate the disruption of immune response in steroid-induced femoral head osteonecrosis via Toll-like receptor 4 (TLR4) signaling pathway.. Male SD rats were divided into the treatment group (group A) and the model group (group B) consisting of 24 rats each, and were injected intramuscularly with 20 mg/kg methylprednisolone (MP) for 8 weeks, once a week. The rats in group A were injected intravenously with 7.5 mg/kg TAK242 before each MP administration. A control group (group N) consisted of 12 rats were received saline injection. All animals were sacrificed 8, 10 and 12 weeks from the first MP injection, respectively. Histopathological analysis was performed and the concentration of tartrate-resistant acid phosphatase (TRAP) in serum was tested. The signaling molecules including TLR4, MyD88, NF-κB p65 and MCP-1 were detected by immunohistochemistry, quantitative real-time PCR and Western blot.. Femoral head osteonecrosis was observed in the model rats, and the concentration of TRAP and positive staining of all signaling molecules increased significantly in group B compared with that in group A and group N. Compare with the control group, the mRNA expressions and protein levels of all signaling molecules were enhanced significantly in group B, but no significant in group A.. Corticosteroids can induce femoral head osteonecrosis by disturbing the immune response via TLR4 signaling pathway. These findings suggest that the disruption of immune response play a role in the pathogenesis of osteonecrosis.

Topics: Acid Phosphatase; Adrenal Cortex Hormones; Animals; Bone and Bones; Chemokine CCL2; Disease Models, Animal; Femur Head Necrosis; Isoenzymes; Male; Methylprednisolone; Myeloid Differentiation Factor 88; Rats; Rats, Sprague-Dawley; RNA, Messenger; Signal Transduction; Sulfonamides; Tartrate-Resistant Acid Phosphatase; Time Factors; Toll-Like Receptor 4; Transcription Factor RelA

To investigate the changes in condylar cartilage and subchondral bone of the temporomandibular joint (TMJ) in a mouse model of incisor malocclusion.. By bonding a single (single group) or a pair (pair group) of metal tube(s) to the left incisor(s), a crossbite-like relationship was created between left-side incisors in mice. The morphological changes in the TMJ condyles were examined by hematoxylin and eosin and toluidine blue staining. Indices of osteoclastic activity, including tartrate-resistant acid phosphatase (TRAP) staining and macrophage colony stimulating factor (M-CSF) were investigated by histochemistry or real-time polymerase chain reaction (PCR). The osteoblastic activity was indexed by osteocalcin expression. Expressions of semaphorin 4D and its receptor, Plexin-B1, were detected by real-time PCR. Two-way analysis of variance was used to assess the differences between groups.. One week and 3 weeks after bonding the metal tube(s), cartilage degradation and subchondral bone loss were evident histologically. Both indices of osteoclastic activity (TRAP and M-CSF) were significantly increased in cartilage and subchondral bone after bonding the metal tube(s). Osteocalcin expression in cartilage was significantly increased at week 3, while its expression in subchondral bone was significantly increased at week 1 but decreased at week 3. The semaphorin 4D expression in cartilage and subchondral bone was significantly decreased at week 1 but significantly increased at week 3. For Plexin-B1 expression, a significant increase was detected in subchondral bone at week 3.. Bonding a single or a pair of metal tube(s) to left incisor(s) is capable of inducing remodeling in the TMJ, which involved cartilage degradation and alteration of osteoclastic and osteoblastic activity.

Topics: Acid Phosphatase; Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Bone Remodeling; Bone Resorption; Cartilage, Articular; Chondrocytes; Coloring Agents; Disease Models, Animal; Fluorescent Dyes; Incisor; Isoenzymes; Macrophage Colony-Stimulating Factor; Malocclusion; Mandibular Condyle; Mice; Mice, Inbred BALB C; Nerve Tissue Proteins; Osteoblasts; Osteocalcin; Osteoclasts; Receptors, Cell Surface; Semaphorins; Tartrate-Resistant Acid Phosphatase; Temporomandibular Joint

It appears there are no studies evaluating the influence of the bisphosphonate tiludronic acid (TIL) on periodontitis. The purpose of this study is to evaluate via microtomographic, histopathologic, histometric, and immunohistochemical analyses the effects of local administration of TIL on ligature-induced periodontitis in rats.. Forty-eight rats were divided into six groups: C (control), EP (experimental periodontitis), EP-Saline, EP-TIL0.1, EP-TIL0.3, and EP-TIL1. In EP, a ligature was placed around maxillary second molars. In EP-TIL0.1, EP-TIL0.3, and EP-TIL1, TIL solutions of 0.1, 0.3, and 1 mg/kg body weight, respectively, were injected into the subperiosteal palatal area adjacent to maxillary second molars every other day. EP-Saline received 0.9% NaCl solution instead. Animals were euthanized at day 11. Bone changes were evaluated by microtomographic and histometric analyses. Histopathologic analysis and immunohistochemical detection of tartrate-resistant acid phosphatase (TRAP) were also performed. Data were statistically analyzed (analysis of variance or Kruskal-Wallis, P <0.05).. Histometric and microtomographic analyses (at buccal, interproximal, and furcation sites) demonstrated that EP-TIL1 presented less alveolar bone loss (ABL) than EP (P <0.05), whereas EP-TIL0.1 and EP-TIL0.3 did not demonstrate significant differences in alveolar bone level compared to EP (P >0.05). Also, EP-TIL1 showed significantly fewer TRAP-positive multinucleated osteoclasts than EP and EP-Saline (P <0.05).. It can be concluded that locally administered TIL solution (1 mg/kg body weight) reduced alveolar bone loss in experimental periodontitis and the dosage of TIL may influence its anti-inflammatory and antiresorptive properties.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Anti-Inflammatory Agents; Biomarkers; Bone Density Conservation Agents; Connective Tissue; Diphosphonates; Disease Models, Animal; Epithelial Attachment; Gingiva; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Immunohistochemistry; Injections; Isoenzymes; Male; Molar; Osteoclasts; Periodontitis; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Time Factors; X-Ray Microtomography

This study investigated the effects of seahorse (Hippocampus spp.) extracts in a rat model of benign prostatic hyperplasia (BPH) and mouse model of oligospermatism. Compared to the sham operated group, castration and testosterone induced BPH, indicated by increased penile erection latency; decreased penis nitric oxide synthase (NOS) activity; reduced serum acid phosphatase (ACP) activity; increased prostate index; and epithelial thickening, increased glandular perimeter, increased proliferating cell nuclear antigen (PCNA) index and upregulation of basic fibroblast growth factor (bFGF) in the prostate. Seahorse extracts significantly ameliorated the histopathological changes associated with BPH, reduced the latency of penile erection and increased penile NOS activity. Administration of seahorse extracts also reversed epididymal sperm viability and motility in mice treated with cyclophosphamide (CP). Seahorse extracts have potential as a candidate marine drug for treating BPH without inducing the side effects of erectile dysfunction (ED) or oligospermatism associated with the BPH drug finasteride.

Topics: Acid Phosphatase; Animals; Biological Products; Castration; Cyclophosphamide; Disease Models, Animal; Female; Fibroblast Growth Factor 2; Male; Mice; Nitric Oxide Synthase; Oligospermia; Penis; Proliferating Cell Nuclear Antigen; Prostate; Prostatic Hyperplasia; Rats, Sprague-Dawley; Smegmamorpha; Sperm Count; Sperm Motility; Testosterone

Craniometaphyseal dysplasia (CMD) is a rare genetic disorder encompassing hyperostosis of craniofacial bones and metaphyseal widening of tubular bones. Dental abnormalities are features of CMD that have been little discussed in the literature. We performed dentofacial examination of patients with CMD and evaluated consequences of orthodontic movement in a mouse model carrying a CMD knock-in (KI) mutation (Phe377del) in the Ank gene. All patients have a history of delayed eruption of permanent teeth. Analysis of data obtained by cone-beam computed tomography showed significant bucco-lingual expansion of jawbones, more pronounced in mandibles than in maxillae. There was no measurable increase in bone density compared with that in unaffected individuals. Orthodontic cephalometric analysis showed that patients with CMD tend to have a short anterior cranial base, short upper facial height, and short maxillary length. Microcomputed tomography (micro-CT) analysis in homozygous Ank (KI/KI) mice, a model for CMD, showed that molars can be moved by orthodontic force without ankylosis, however, at a slower rate compared with those in wild-type Ank (+/+) mice (p < .05). Histological analysis of molars in Ank (KI/KI) mice revealed decreased numbers of TRAP(+) osteoclasts on the bone surface of pressure sides. Based on these findings, recommendations for the dental treatment of patients with CMD are provided.

Topics: Acid Phosphatase; Animals; Bone Density; Bone Diseases, Developmental; Cephalometry; Cone-Beam Computed Tomography; Craniofacial Abnormalities; Disease Models, Animal; Gene Knock-In Techniques; Humans; Hyperostosis; Hypertelorism; Isoenzymes; Mandible; Maxilla; Mice; Mutation; Osteoclasts; Phenylalanine; Phosphate Transport Proteins; Sequence Deletion; Skull Base; Tartrate-Resistant Acid Phosphatase; Tooth Abnormalities; Tooth Movement Techniques; Vertical Dimension; X-Ray Microtomography

To investigate the effect of Lithothamnium sp (LTT) supplement, a calcium-rich alga widely used for mineral reposition, on strain-induced (orthodontic tooth movement [OTM]) and infection-induced bone resorption (periodontal disease [PD]) in mice.. Mice were divided into two bone resorption models: one with an orthodontic appliance and the other with PD induced by the oral inoculation of Aggregatibacter actinomycetencomitans (Aa). Both groups were fed a regular diet (vehicle), LTT-rich diet (LTT), or calcium-rich diet (CaCO3). Alveolar bone resorption (ABR), the number of osteoclasts, and the levels of tumor necrosis factor α (TNF-α), calcium, and vitamin D3 were evaluated.. The number of osteoclasts was reduced in LTT and CaCO3 mice, which led to diminished OTM and infection-induced alveolar bone loss. In addition, LTT- and calcium-treated groups also presented decreased levels of TNF-α in periodontal tissues and increased levels of calcium in serum.. These results indicate that the LTT supplement influences ABR, probably due to its calcium content, by affecting osteoclast function and local inflammatory response, thus modulating OTM and PD.

Topics: Acid Phosphatase; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Alveolar Process; Animals; Bone Density Conservation Agents; Calcitriol; Calcium; Calcium Carbonate; Calcium, Dietary; Cell Count; Dietary Supplements; Disease Models, Animal; Isoenzymes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Osteoclasts; Pasteurellaceae Infections; Periodontal Diseases; Rhodophyta; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques; Tumor Necrosis Factor-alpha

Treatments to reduce fracture rates in adults with osteogenesis imperfecta are limited. Sclerostin antibody, developed for treating osteoporosis, has not been explored in adults with OI. This study demonstrates that treatment of adult OI mice respond favorably to sclerostin antibody therapy despite retention of the OI-causing defect.. Osteogenesis imperfecta (OI) is a heritable collagen-related bone dysplasia, characterized by brittle bones with increased fracture risk. Although OI fracture risk is greatest before puberty, adults with OI remain at risk of fracture. Antiresorptive bisphosphonates are commonly used to treat adult OI, but have shown mixed efficacy. New treatments which consistently improve bone mass throughout the skeleton may improve patient outcomes. Neutralizing antibodies to sclerostin (Scl-Ab) are a novel anabolic therapy that have shown efficacy in preclinical studies by stimulating bone formation via the canonical wnt signaling pathway. The purpose of this study was to evaluate Scl-Ab in an adult 6 month old Brtl/+ model of OI that harbors a typical heterozygous OI-causing Gly > Cys substitution on Col1a1.. Six-month-old WT and Brtl/+ mice were treated with Scl-Ab (25 mg/kg, 2×/week) or Veh for 5 weeks. OCN and TRACP5b serum assays, dynamic histomorphometry, microCT and mechanical testing were performed.. Adult Brtl/+ mice demonstrated a strong anabolic response to Scl-Ab with increased serum osteocalcin and bone formation rate. This anabolic response led to improved trabecular and cortical bone mass in the femur. Mechanical testing revealed Scl-Ab increased Brtl/+ femoral stiffness and strength.. Scl-Ab was successfully anabolic in an adult Brtl/+ model of OI.

Topics: Acid Phosphatase; Adaptor Proteins, Signal Transducing; Anabolic Agents; Animals; Antibodies, Neutralizing; Body Mass Index; Bone Density; Bone Morphogenetic Proteins; Disease Models, Animal; Drug Evaluation, Preclinical; Femur; Genetic Markers; Isoenzymes; Male; Mice, Mutant Strains; Osteocalcin; Osteogenesis; Osteogenesis Imperfecta; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography

Whole body vibration (WBV) stimulation has a beneficial effect on the recovery of osteoporotic bone. We aimed to investigate the immediate effect of WBV on lipopolysaccharide (LPS)-mediated inflammatory bone loss by varying the exposure timing. Balb/C mice were divided into the following groups: control, LPS (L), and LPS with vibration (LV). The L and LV groups received LPS (5 mg/kg) by 2 intraperitoneal injections on days 0 and 4. The LV group was exposed to WBV (0.4 g, 45 Hz) either during LPS treatment (LV1) or after cessation of LPS injection (LV2) and then continued WBV treatment for 10 min/d for 3 d. Evaluation based on micro-computed tomography was performed 7 d after the first injection, when the L group showed a significant decrease in bone volume (-25.8%) and bone mineral density (-33.5%) compared with the control group. The LV2 group recovered bone volume (35%) and bone mineral density (19.9%) compared with the L group, whereas the LV1 group showed no improvement. This vibratory signal showed a suppressive effect on the LPS-mediated induction of inflammatory cytokines such as IL-1β or TNF-α in human mesenchymal stem cells in vitro. These findings suggest that immediate exposure to WBV after the conclusion of LPS treatment efficiently reduces trabecular bone loss, but WBV might be less effective during the course of treatment with inflammatory factor.

Topics: Acid Phosphatase; Adult; Animals; Bone Density; Bone Regeneration; Cells, Cultured; Disease Models, Animal; Femur; Humans; Interleukin-1beta; Isoenzymes; Lipopolysaccharides; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Organ Size; Osteoporosis; Tartrate-Resistant Acid Phosphatase; Tibia; Time Factors; Tumor Necrosis Factor-alpha; Vibration; X-Ray Microtomography

Despite the worldwide increased prevalence of osteoporosis, no data are available evaluating the effect of an enamel matrix derivative (EMD) on the healing of periodontal defects in patients with osteoporosis. This study aims to evaluate whether the regenerative potential of EMD may be suitable for osteoporosis-related periodontal defects.. Forty female Wistar rats (mean body weight: 200 g) were used for this study. An osteoporosis animal model was carried out by bilateral ovariectomy (OVX) in 20 animals. Ten weeks after OVX, bilateral fenestration defects were created at the buccal aspect of the first mandibular molar. Animals were randomly assigned to four groups of 10 animals per group: 1) control animals with unfilled periodontal defects; 2) control animals with EMD-treated defects; 3) OVX animals with unfilled defects; and 4) OVX animals with EMD-treated defects. The animals were euthanized 28 days later, and the percentage of defect fill and thickness of newly formed bone and cementum were assessed by histomorphometry and microcomputed tomography (micro-CT) analysis. The number of osteoclasts was determined by tartrate-resistant acid phosphatase (TRAP), and angiogenesis was assessed by analyzing formation of blood vessels.. OVX animals demonstrated significantly reduced bone volume in unfilled defects compared with control defects (18.9% for OVX animals versus 27.2% for control animals) as assessed by micro-CT. The addition of EMD in both OVX and control animals resulted in significantly higher bone density (52.4% and 69.2%, respectively) and bone width (134 versus 165μm) compared with untreated defects; however, the healing in OVX animals treated with EMD was significantly lower than that in control animals treated with EMD. Animals treated with EMD also demonstrated significantly higher cementum formation in both control and OVX animals. The number of TRAP-positive osteoclasts did not vary between untreated and EMD-treated animals; however, a significant increase was observed in all OVX animals. The number of blood vessels and percentage of new vessel formation was significantly higher in EMD-treated samples.. The results from the present study suggest that: 1) an osteoporotic phenotype may decrease periodontal regeneration; and 2) EMD may support greater periodontal regeneration in patients suffering from the disease. Additional clinical studies are necessary to fully elucidate the possible beneficial effect of EMD for periodontal regeneration in patients suffering from osteoporosis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Bone Density; Bone Regeneration; Cell Count; Cementogenesis; Dental Enamel Proteins; Disease Models, Animal; Female; Humans; Imaging, Three-Dimensional; Isoenzymes; Mandible; Neovascularization, Physiologic; Osteoclasts; Osteogenesis; Osteoporosis, Postmenopausal; Ovariectomy; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography

Although osteonecrosis of the jaws (ONJ), a serious complication of antiresorptive medications, was reported a decade ago, the exact mechanisms of disease pathophysiology remain elusive. ONJ-like lesions can be induced in animals after antiresorptive treatment and experimental interventions such as tooth extraction or periapical or periodontal disease. However, experimental induction and manipulation of disease progression does not always reflect clinical reality. Interestingly, naturally occurring maxillofacial abscesses, inducing aggressive inflammation of the peri-radicular mucosa with significant osteolysis and alveolar bone expansion, have been reported in mice. Here, we aimed to explore whether osteonecrotic lesions would develop in areas of maxillary peri-radicular infections, in mice on antiresorptive medications with distinct pharmacologic action, thus establishing a novel ONJ animal model. Mice were treated with RANK-Fc or OPG-Fc that bind to RANKL or with the potent bisphosphonate zoledronic acid (ZA). Maxillae were assessed radiographically and histologically. μCT imaging of vehicle mice revealed several maxillae with altered alveolar bone morphology, significant ridge expansion and large lytic areas. However, in RANK-Fc, OPG-Fc and ZA treated animals the extent of bone loss was significantly less, but exuberant bone deposition was noted at the ridge periphery. BV and BV/TV were increased in the diseased site of antiresorptive vs. veh animals. Histologically, extensive inflammation, bone resorption and marginal gingival epithelium migration were seen in the diseased site of vehicle animals. Rank-Fc, OPG-Fc and ZA reduced alveolar bone loss, increased periosteal bone formation, and induced areas of osteonecrosis, and bone exposure that in many animals covered significant part of the alveolar bone. Collectively, our data demonstrate ONJ-like lesions at sites of maxillary peri-radicular infection, indistinguishable in mice treated with RAKL inhibitors vs. zoledronate. This novel mouse model of spontaneous ONJ supports a central role of osteoclast inhibition and infection/inflammation in ONJ pathogenesis and validates and complements existing animal models employing experimental interventions.

Topics: Acid Phosphatase; Alveolar Process; Animals; Bisphosphonate-Associated Osteonecrosis of the Jaw; Bone Resorption; Diphosphonates; Disease Models, Animal; Imidazoles; Isoenzymes; Male; Maxilla; Mice, Inbred C57BL; Periodontium; Radiography; Recombinant Fusion Proteins; Tartrate-Resistant Acid Phosphatase; Zoledronic Acid

The effect of type 2 diabetes mellitus (T2DM) on bone is controversial. Therefore, the present study investigated whether T2DM causes osteoporosis and explored the underlying mechanisms involved in this process. The effects of T2DM on bone physiology were analyzed in a mouse model of T2DM; KK/Upj‑Ay/J (KK‑Ay) mice develop diabetes after 8 weeks and exhibit stable diabetes symptoms and signs after 10 weeks when fed a KK‑Ay mouse maintenance fodder. Diabetic mice exhibited hyperglycemia, hyperinsulinemia and increased body and fat pad weight in comparison with C57BL/6 non-diabetic mice. Furthermore, diabetic mice demonstrated low bone weight and bone mineral density in the femur, tibia and fifth lumbar vertebra. Using von Kossa and tartrate-resistant acid phosphatase (TRAP) staining, alkaline phosphatase and TRAP activity analyses and gene profiling it was demonstrated that osteoblastogenesis and osteoclastogenesis were impaired in diabetic mice. To evaluate the bone biomechanics, the ultimate load of the bone was analyzed. It was found that the ultimate load of the tibia in diabetic mice was lower than that in the controls. The results from the present study suggest that bone metabolism is impaired in T2DM, resulting in decreased osteoblastogenesis, osteoclastogenesis and bone mass.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone and Bones; Bone Density; Cell Differentiation; Cells, Cultured; Diabetes Mellitus, Experimental; Disease Models, Animal; Femur; Isoenzymes; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Osteogenesis; Spine; Tartrate-Resistant Acid Phosphatase; Tibia; Tomography, X-Ray Computed

Cryptococcus neoformans strains isolated from patients with AIDS secrete acid phosphatase, but the identity and role of the enzyme(s) responsible have not been elucidated. By combining a one-dimensional electrophoresis step with mass spectrometry, a canonically secreted acid phosphatase, CNAG_02944 (Aph1), was identified in the secretome of the highly virulent serotype A strain H99. We created an APH1 deletion mutant (Δaph1) and showed that Δaph1-infected Galleria mellonella and mice survived longer than those infected with the wild type (WT), demonstrating that Aph1 contributes to cryptococcal virulence. Phosphate starvation induced APH1 expression and secretion of catalytically active acid phosphatase in the WT, but not in the Δaph1 mutant, indicating that Aph1 is the major extracellular acid phosphatase in C. neoformans and that it is phosphate repressible. DsRed-tagged Aph1 was transported to the fungal cell periphery and vacuoles via endosome-like structures and was enriched in bud necks. A similar pattern of Aph1 localization was observed in cryptococci cocultured with THP-1 monocytes, suggesting that Aph1 is produced during host infection. In contrast to Aph1, but consistent with our previous biochemical data, green fluorescent protein (GFP)-tagged phospholipase B1 (Plb1) was predominantly localized at the cell periphery, with no evidence of endosome-mediated export. Despite use of different intracellular transport routes by Plb1 and Aph1, secretion of both proteins was compromised in a Δsec14-1 mutant. Secretions from the WT, but not from Δaph1, hydrolyzed a range of physiological substrates, including phosphotyrosine, glucose-1-phosphate, β-glycerol phosphate, AMP, and mannose-6-phosphate, suggesting that the role of Aph1 is to recycle phosphate from macromolecules in cryptococcal vacuoles and to scavenge phosphate from the extracellular environment.. Infections with the AIDS-related fungal pathogen Cryptococcus neoformans cause more than 600,000 deaths per year worldwide. Strains of Cryptococcus neoformans isolated from patients with AIDS secrete acid phosphatase; however, the identity and role of the enzyme(s) are unknown. We have analyzed the secretome of the highly virulent serotype A strain H99 and identified Aph1, a canonically secreted acid phosphatase. By creating an APH1 deletion mutant and an Aph1-DsRed-expressing strain, we demonstrate that Aph1 is the major extracellular and vacuolar acid phosphatase in C. neoformans and that it is phosphate repressible. Furthermore, we show that Aph1 is produced in cryptococci during coculture with THP-1 monocytes and contributes to fungal virulence in Galleria mellonella and mouse models of cryptococcosis. Our findings suggest that Aph1 is secreted to the environment to scavenge phosphate from a wide range of physiological substrates and is targeted to vacuoles to recycle phosphate from the expendable macromolecules.

Topics: Acid Phosphatase; Acquired Immunodeficiency Syndrome; Animals; Biological Transport; Cryptococcosis; Cryptococcus neoformans; Disease Models, Animal; Female; Fungal Proteins; Gene Deletion; Glucosephosphates; Glycerophosphates; Green Fluorescent Proteins; Humans; Hydrogen-Ion Concentration; Mannosephosphates; Mice; Mice, Inbred BALB C; Monocytes; Moths; Proteomics; Vacuoles

Lycopene is a member of the carotenoid family and has strong anti-oxidant properties. Lycopene occurs in tomato-based food products primarily as an all-E isomer (80-97%),but its Z-isomers accounts for 79 to 88% of total lycopene in benign or malignant prostate tissues, while the specific biological functions of Z-isomers are still not clarified at present. This study was to examine the bioactive potency of Z-isomers on benign prostatic hyperplasia (BPH) in mice and to make a comparison of effective inhibition between Z-isomers and all-E isomer.. Mice were divided into the Saline group, Vehicle control group and testosterone propionate induced BPH mice group (BPH model group, vehicle BPH model group, lycopene treated (5 mg/kg and 2.5 mg/kg), Z-isomers (57%) treated, Z-isomers (86%) treated, finasteride treated). The drugs were orally administered once a day consecutively for 30 days. The inhibitory effects on BPH of all-E lycopene and Z-isomers were evaluated by prostatic index, prostatic acid phosphatase (PAP), estradiol, testosterone and dihydrotestosterone (DHT) levels in serum and histopathology examination.. Compared with the BPH model group, E/Z isomers exhibited significant differences in prostatic index, PAP, estradiol, testosterone and DHT levels in serum and similar histological aspects observed in the mice of the control group. The present research also shows that Z-isomers may be more potent inhibitors than all-E isomers in BPH treatment.

Topics: Acid Phosphatase; Animals; Carotenoids; Disease Models, Animal; Estradiol; Finasteride; Lycopene; Male; Mice; Mice, Inbred ICR; Prostate; Prostatic Hyperplasia; Protein Tyrosine Phosphatases; Solanum lycopersicum; Testosterone

The objective of this study was to investigate whether intermittent administration of parathyroid hormone [1-34] (PTH[1-34]) promotes tendon-bone healing after anterior cruciate ligament (ACL) reconstruction in vivo. A rat model of ACL reconstruction with autograft was established at the left hind leg. Every day, injections of 60 μg PTH[1-34]/kg subcutaneously were given to the PTH group rats (n=10) for four weeks, and the controls (n=10) received saline. The tendon-bone healing process was evaluated by micro-CT, biomechanical test, histological and immunohistochemical analyses. The effects of PTH[1-34] on serum chemistry, bone microarchitecture and expression of the PTH receptor (PTH1R) and osteocalcin were determined. Administration of PTH[1-34] significantly increased serum levels of calcium, alkaline phosphatase (AP), osteocalcin and tartrate-resistant acid phosphatase (TRAP). The expression of PTH1R on both osteocytes and chondrocyte-like cells at the tendon-bone interface was increased in the PTH group. PTH[1-34] also enhanced the thickness and microarchitecture of trabecular bone according to the micro-CT analysis. The results imply that systematically intermittent administration of PTH[1-34] promotes tendon-bone healing at an early stage via up-regulated PTH1R. This method may enable a new strategy for the promotion of tendon-bone healing after ACL reconstruction.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Anterior Cruciate Ligament; Anterior Cruciate Ligament Injuries; Anterior Cruciate Ligament Reconstruction; Bone Density; Calcium; Disease Models, Animal; Isoenzymes; Male; Osteocalcin; Parathyroid Hormone; Rats; Rats, Sprague-Dawley; Receptor, Parathyroid Hormone, Type 1; Recombinant Proteins; Tartrate-Resistant Acid Phosphatase; Tibia; Tomography, X-Ray Computed; Transplantation, Autologous; Up-Regulation; Wound Healing

Therapeutic and prophylactic properties ozonated of sea buckthorn oil in the experiment on the model of generalized periodontitis in Wistar rats induced by action of extracted products of incomplete combustion of tobacco smoke was investigated. It is proved that the proposed method of ozone therapy in combination with fitooil prevents and corrects metabolic disturbances in the periodontal tissues, caused a by high therapeutic effect of the drug.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Hippophae; Humans; Male; Nicotiana; Ozone; Periodontitis; Plant Oils; Rats; Rats, Wistar; Smoke; Tobacco Use Disorder

Optimum stresses for a favorable response to orthodontics are still unknown. Here, we compared the effects of initial periodontal ligament (PDL) stresses over time in orthodontic external root resorption (OERR), necrosis, and the TRAP+ cell population. Forty-two rats (Fischer CDF) were treated with 10 cN of force for 5 different time periods. Finite element (FE) models of the first maxillary molars were constructed from μCT scans to calculate initial PDL stresses. The scans were also used for OERR measurements before histology. Time, stress, and their interaction were significant to result in an OERR increase only in the regions of medium and high stress. OERR was not significantly different between control and treated animals over time in the region of low stress. After 30 days, OERR was increased by 5- and 3-fold in the zone of high- and medium-stress regions, respectively. The TRAP+ cell population initially followed the stress gradient, but changed after bone and necrotic tissue resorption. In the 30-day modeling cycle, the correspondent 3rd principal stress range to promote direct bone resorption and insignificant OERR was between -9.92 and -7.75 KPa. These translate to approximate forces of 30 to 40 cN applied at the bracket level (tipping) of a human maxillary canine.

Topics: Acid Phosphatase; Animals; Dental Stress Analysis; Disease Models, Animal; Finite Element Analysis; Isoenzymes; Maxilla; Molar; Orthodontics; Periodontal Ligament; Rats; Rats, Inbred F344; Root Resorption; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Movement Techniques; Tooth Root

Obesity is markedly associated with abnormal bone density indicating the importance of adipocytes in bone metabolism. However, the specific function of adipocytes remains unclear, with marked discrepancies in observations of previous studies. In the present study, the effect of adipocytes on osteoblasts/osteoclasts was analyzed. A mouse model of obesity was established and an in vitro co-culture system was utilized containing adipocyte and MC3T3/RAW 264.7 cells in a Transwell plate. Compared with control mice, obese mice exhibited low body weight and bone mineral density of the tibia and fat cells were observed to accumulate in bone marrow. MC3T3/RAW 264.7 cells were co-cultured with adipocytes and the mRNA and protein expression of alkaline phosphatase and osteocalcin was found to be decreased in MC3T3-E1 cells and mRNA and protein expression of tartrate-resistant acid phosphatase and cathepsin K was significantly increased in RAW 264.7 cells. In addition, the effect of adipocytes on the osteoprotegerin (OPG)/receptor activator of nuclear factor κB ligand (RANKL)/RANK system indicated that the RANKL/OPG ratio secreted by osteoblasts increased and RANK expression by osteoclasts increased, leading to increased osteoclastogenesis. These results indicate that bone metabolism is impaired in obese mice leading to decreased osteoblastogenesis and marked increases in osteoclastogenesis and low bone mass.

Topics: Acid Phosphatase; Adipocytes; Alkaline Phosphatase; Animals; Bone Density; Bone Marrow Cells; Cathepsin K; Cells, Cultured; Coculture Techniques; Disease Models, Animal; Male; Mice; Mice, Inbred C57BL; Mice, Obese; Obesity; Osteocalcin; Osteogenesis; Osteoprotegerin; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B

Pathological alterations in the balance of bone metabolism are central to the progression of inflammatory bone diseases such as periodontal disease. We have developed and characterized a novel ex vivo murine mandible model of inflammatory bone destruction. Slices of mandible were cultured for 14 days in the presence or absence of P. gingivalis lipopolysaccharide (LPS) or pro-inflammatory cytokines. Following culture, cell viability and tissue histomorphometry were assessed with quantification of matrix proteins, resident osteoclasts, ligament cells, monocytes, macrophages, and neutrophils. In the absence of inflammatory factors, culture viability, osteoclasts, and matrix components were maintained. LPS or TNFα stimulation demonstrated an increase in cellular proliferation, monocyte cells, osteoclast differentiation, and matrix degradation. Pathophysiological bone metabolism can be induced via exposure to LPS and direct influence of TNFα within the model despite the absence of systemic circulation, providing a model for inflammatory bone destruction and investigation of the effects of novel therapeutics.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Cell Differentiation; Cell Proliferation; Cell Survival; Collagen Type I; Disease Models, Animal; Extracellular Matrix Proteins; Inflammation Mediators; Integrin-Binding Sialoprotein; Interleukin-23; Interleukin-6; Isoenzymes; Lipopolysaccharides; Macrophages; Male; Mandibular Diseases; Mice; Monocytes; Neutrophils; Organ Culture Techniques; Osteocalcin; Osteoclasts; Osteopontin; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

Ticks of Rhipicephalus sanguineus species have great medical and veterinary importance for being a vector of various diseases. In an attempt to minimize their action on the host, people have resorted to chemical control by using various acaricides, such as selamectin. Although previous studies have demonstrated its toxic action in domestic animals, no studies focused on the detection of cell death when exposed to selamectin. For this reason, the technique for detecting autophagic cell death was used in order to demonstrate the responses of rabbits' skin tissues pre-infested with R. sanguineus and exposed to different concentrations of selamectin. The obtained results when exposed to 100 and 80% concentrations of selamectin showed a strong mark of acid phosphatase on the cells of the connective tissue of the dermis and hair follicles, whereas the ones exposed to the 50% concentration had a weak mark on the cells of the connective tissue of the dermis and moderate staining in hair follicles. It became clear that, when used at high concentrations (100 and 80%), selamectin is capable to induce a large scale occurrence of the autophagic cell death process. On the other hand, the concentration of 50% causes minor morphophysiological changes in the skin of rabbit hosts when evaluated the cell death process. Therefore, the data confirms that selamectin is a powerful dose-dependent toxic agent causes increased activity of the enzyme acid phosphatase.

Topics: Acaricides; Acid Phosphatase; Animals; Autophagy; Disease Models, Animal; Ectoparasitic Infestations; Ivermectin; Rabbits; Rhipicephalus sanguineus; Skin

The molecular mechanisms underlying prostate carcinogenesis are poorly understood. Prostatic acid phosphatase (PAP), a prostatic epithelial secretion marker, has been linked to prostate cancer since the 1930's. However, the contribution of PAP to the disease remains controversial. We have previously cloned and described two isoforms of this protein, a secretory (sPAP) and a transmembrane type-I (TMPAP). The goal in this work was to understand the physiological function of TMPAP in the prostate. We conducted histological, ultra-structural and genome-wide analyses of the prostate of our PAP-deficient mouse model (PAP(-/-)) with C57BL/6J background. The PAP(-/-) mouse prostate showed the development of slow-growing non-metastatic prostate adenocarcinoma. In order to find out the mechanism behind, we identified PAP-interacting proteins byyeast two-hybrid assays and a clear result was obtained for the interaction of PAP with snapin, a SNARE-associated protein which binds Snap25 facilitating the vesicular membrane fusion process. We confirmed this interaction by co-localization studies in TMPAP-transfected LNCaP cells (TMPAP/LNCaP cells) and in vivo FRET analyses in transient transfected LNCaP cells. The differential gene expression analyses revealed the dysregulation of the same genes known to be related to synaptic vesicular traffic. Both TMPAP and snapin were detected in isolated exosomes. Our results suggest that TMPAP is involved in endo-/exocytosis and disturbed vesicular traffic is a hallmark of prostate adenocarcinoma.

Topics: Acid Phosphatase; Adenocarcinoma; Animals; Cell Line, Tumor; Cell Transformation, Neoplastic; Disease Models, Animal; Male; Mice; Mice, Knockout; Models, Biological; Prostate; Prostatic Neoplasms; Protein Binding; Protein Transport; Protein Tyrosine Phosphatases; Pseudopodia; Vesicular Transport Proteins

Fresh bone marrow cells have already exhibited its advantages as osteogenic donor cells, but the combination between fresh bone marrow cells and other donor cells utilized for bone healing has not been fully explored. To highlight the impact of fresh bone marrow cells on scaffold-based bone regeneration, single or a combination of calvarial osteoprogenitor cells (OPCs) and bone marrow cells (BMCs) were used as donor cells combined with collagen-apatite scaffold for calvarial defect healing. The host and donor contributions to bone formation were assessed using histological and GFP imaging analysis. Although the amount of new bone formed by different cell sources did not show significant differences, the origin of the bone formation in the defects mainly depended on the types of donor cells employed: when only calvarial OPCs were used as donor cells, a donor-derived bone healing instead of host-derived bone ingrowth was observed; when only fresh BMCs were loaded, the host bone could grow into the defect along the lamellar structure of the scaffolds, but the amount of new bone formed was significantly lower than the defect loaded with calvarial OPCs only. The combination of calvarial OPCs and fresh BMCs had similar amount of new bone formation as the group loaded with calvarial osteoprogenitors alone, but did not induce any host-derived bone formation. These results provide compelling evidence of the importance of fresh BMCs to induce host-implant integration in bone tissue engineering.

Topics: Acid Phosphatase; Animals; Apatites; Bone Marrow Cells; Bone Marrow Transplantation; Collagen; Disease Models, Animal; Green Fluorescent Proteins; Isoenzymes; Mice; Osteoclasts; Prosthesis Implantation; Rats; Regenerative Medicine; Skull; Staining and Labeling; Stem Cell Transplantation; Stem Cells; Tartrate-Resistant Acid Phosphatase; Tissue Scaffolds; X-Ray Microtomography

We administered recombinant interleukin (IL)-4 and recombinant IL-13 locally into the air pouch of mice to improve bone resorption induced by ultra-high-molecular-weight polyethylene (UHMWPE) particles.. Air pouches were established on the back of BALB/c mice, followed by the surgical introduction of a section of calvaria from a syngeneic mouse donor. We stimulated the bone-implanted pouches with the UHMWPE suspension. We divided UHMWPE-containing mice into four study groups to receive injections of phosphate-buffered saline (control), IL-4 alone, IL-13 alone, or IL-4 and IL-13 into the pouches. We harvested the tissues at 14 d after treatment for molecular and histological analyses.. The inhibitory effect of IL-4 was stronger than that of IL-13 toward osteoclast differentiation and osteoblast for the induction of osteoprotegerin production and down-regulation of receptor for activation of nuclear factor-κB ligand production. Furthermore, the combined treatment with both IL-4 and 1L-13 had a more important role in inhibiting bone resorption in these pouches with UHMWPE stimulation, compared with IL-4 or IL-13 treatment alone.. Local administration of recombinant IL-4 and IL-13 may be a feasible and effective therapeutic candidate to treat or prevent wear debris-associated osteolysis.

Topics: Acid Phosphatase; Animals; Cell Differentiation; Disease Models, Animal; Female; Interleukin-13; Interleukin-4; Isoenzymes; Mice; Mice, Inbred BALB C; Osteoclasts; Osteolysis; Polyethylenes; Receptor Activator of Nuclear Factor-kappa B; Recombinant Proteins; Tartrate-Resistant Acid Phosphatase

Osteogenesis imperfecta (OI) is a genetic bone dysplasia characterized by osteopenia and easy susceptibility to fracture. Symptoms are most prominent during childhood. Although antiresorptive bisphosphonates have been widely used to treat pediatric OI, controlled trials show improved vertebral parameters but equivocal effects on long-bone fracture rates. New treatments for OI are needed to increase bone mass throughout the skeleton. Sclerostin antibody (Scl-Ab) therapy is potently anabolic in the skeleton by stimulating osteoblasts via the canonical wnt signaling pathway, and may be beneficial for treating OI. In this study, Scl-Ab therapy was investigated in mice heterozygous for a typical OI-causing Gly→Cys substitution in col1a1. Two weeks of Scl-Ab successfully stimulated osteoblast bone formation in a knock-in model for moderately severe OI (Brtl/+) and in WT mice, leading to improved bone mass and reduced long-bone fragility. Image-guided nanoindentation revealed no alteration in local tissue mineralization dynamics with Scl-Ab. These results contrast with previous findings of antiresorptive efficacy in OI both in mechanism and potency of effects on fragility. In conclusion, short-term Scl-Ab was successfully anabolic in osteoblasts harboring a typical OI-causing collagen mutation and represents a potential new therapy to improve bone mass and reduce fractures in pediatric OI.

Topics: Acid Phosphatase; Adaptor Proteins, Signal Transducing; Animals; Antibodies; Biomarkers; Biomechanical Phenomena; Body Weight; Calcification, Physiologic; Disease Models, Animal; Femur; Fluoresceins; Glycoproteins; Intercellular Signaling Peptides and Proteins; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Nanotechnology; Organ Size; Osteocalcin; Osteogenesis; Osteogenesis Imperfecta; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography

Micro-CT provides three-dimensional details and has been widely used for biomedical assessments. This study aimed to determine the most appropriate threshold method for quantitatively assessing the dynamics of periodontal destruction.. Inflammation was induced by submerging a silk ligature in the sulcus of the maxillary second molars of rats, and the animals were killed prior to ligature placement and after 7 and 21 days. The maxillae were examined for the bone resorptive activities by micro-CT, histology and tartrate-resistant acid phosphatase staining. The imaging threshold was determined by CT phantom, global and local algorithms. A bone fraction measurement from each threshold-determining technique was compared with histomorphometry. The reliability and reproducibility were examined by the intraclass correlation coefficient (ICC) and the coefficient of variation.. Significant reduction of inflammatory infiltration (p < 0.01) and active osteoclastic resorption (p < 0.05) from Day 7 to Day 21 were noted. High inter- and intraexaminer agreement were demonstrated in both histomorphometric and micro-CT assessments (ICC > 0.98). The algorithm-based technique demonstrated stronger correlation to histomorphometry than phantom-based thresholds, and the highest agreement was presented by the local algorithm (ICC > 0.96). This, however, was considerably computationally expensive.. The local threshold-determining algorithm is suggested for examining inflammation-induced bone loss. Further investigation will be aimed at enhancing computational efficiency.

Topics: Acid Phosphatase; Algorithms; Alveolar Bone Loss; Animals; Biomarkers; Collagen; Coloring Agents; Connective Tissue; Disease Models, Animal; Gingiva; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Isoenzymes; Male; Maxillary Diseases; Molar; Osteoclasts; Periodontitis; Phantoms, Imaging; Pilot Projects; Rats; Rats, Sprague-Dawley; Reproducibility of Results; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Cervix; X-Ray Microtomography

Pulsed electromagnetic field (PEMF) has been shown to increase bone mineral density in osteoporosis patients and prevent bone loss in ovariectomized rats. But the mechanisms through which PEMF elicits these favorable biological responses are still not fully understood. Receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) are cytokines predominantly secreted by osteoblasts and play a central role in differentiation and functional activation of osteoclasts. The purpose of this study was to investigate the effects of PEMF on RANKL and OPG expression in ovariectomized rats. Thirty 3-month-old female Sprague-Dawley rats were randomly divided into three groups: sham-operated control (Sham), ovariectomy control (OVX), and ovariectomy with PEMF treatment (PEMF). After 12-week interventions, the results showed that PEMF increased serum 17β-estradiol level, reduced serum tartrate-resistant acid phosphatase level, increased bone mineral density, and inhibited deterioration of bone microarchitecture and strength in OVX rats. Furthermore, PEMF could suppress RANKL expression and improve OPG expression in bone marrow cells of OVX rats. In conclusion, this study suggests that PEMF can prevent ovariectomy-induced bone loss through regulating the expression of RANKL and OPG.

Topics: Absorptiometry, Photon; Acid Phosphatase; Animals; Biomarkers; Biomechanical Phenomena; Bone and Bones; Bone Density; Bone Marrow Cells; Disease Models, Animal; Down-Regulation; Electromagnetic Fields; Estradiol; Female; Isoenzymes; Osteoporosis; Osteoprotegerin; Ovariectomy; RANK Ligand; Rats; Rats, Sprague-Dawley; RNA, Messenger; Tartrate-Resistant Acid Phosphatase; Time Factors; Up-Regulation

The aim of this study was to investigate how atopic dermatitis (AD) contributes to root resorption during orthodontic tooth movement.. Atopic dermatitis model mice and wild-type mice were subjected to an excessive orthodontic force (OF) to induce movement of the upper first molars. The expression levels of the tartrate-resistant acid phosphatase (TRAP), IL-17, IL-6, and RANKL proteins were determined in the periodontal ligament (PDL) by an immunohistochemical analysis. Furthermore, the effects of the compression force on co-cultures of CD4(+) cells from AD patients or healthy individuals and human PDL cells were investigated with regard to the levels of secretion and mRNA expression of IL-17, IL-6, RANKL, and osteoprotegerin.. The immunoreactivities for TRAP, IL-17, IL-6, and RANKL in the AD group were found to be significantly increased. The double immunofluorescence analysis for IL-17/CD4 detected immunoreaction. The secretion of IL-17, IL-6, and RANKL, and the mRNA levels of IL-6 and RANKL in the AD patients were increased compared with those in healthy individuals.. Th17 cells may therefore be associated with the deterioration of root resorption of AD mice, and may explain why AD patients are more susceptible to root resorption than healthy individuals when an excessive OF is applied.

Topics: Acid Phosphatase; Adult; Animals; CD4-Positive T-Lymphocytes; Cell Culture Techniques; Coculture Techniques; Dermatitis, Atopic; Disease Models, Animal; Female; Humans; Immunoglobulin E; Interleukin-17; Interleukin-6; Isoenzymes; Male; Mice; Mice, Inbred BALB C; Osteoprotegerin; Periodontal Ligament; RANK Ligand; Root Resorption; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Th17 Cells; Tooth Movement Techniques; Young Adult

The purpose of this study was to examine histological alterations on osteoblasts from the alveolar bone of transgenic mice with targeted ablation of osteoctyes. Eighteen weeks-old transgenic mice based on the diphtheria toxin (DT) receptor-mediated cell knockout (TRECK) system were used in these experiments. Mice were injected intraperitoneally with 50 µg/kg of DT in PBS, or only PBS as control. Two weeks after injections, mice were subjected to transcardiac perfusion with 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), and the available alveolar bone was removed for histochemical analyses. Approximately 75% of osteocytes from alveolar bones became apoptotic after DT administration, and most osteocytic lacunae became empty. Osteoblastic numbers and alkaline phosphatase (ALP) activity were markedly reduced at the endosteum of alveolar bone after DT administration compared with the control. Osteoblastic ALP activity in the periodontal ligament region, on the other hand, hardly showed any differences between the two groups even though numbers were reduced in the experiment group. Silver impregnation showed a difference in the distribution of bone canaliculi between the portions near the endosteum and the periodontal ligament: the former appeared regularly arranged in contrast to the latter's irregular distribution. Under transmission electron microscopy (TEM), the osteoblasts in the periodontal ligament showed direct contact with the Sharpey's fibers. Thus, osteoblastic activity was affected by osteocyte ablation in general, but osteoblasts in contact with the periodontal ligament were less affected than endosteal osteoblasts.

Topics: Ablation Techniques; Acid Phosphatase; Alkaline Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Bone Density; Diphtheria Toxin; Disease Models, Animal; Isoenzymes; Mandibular Diseases; Mice; Mice, Transgenic; Osteoclasts; Osteocytes; Periodontal Ligament; RANK Ligand; Silver Staining; Tartrate-Resistant Acid Phosphatase

Previous studies have shown that fracture healing depends on gender and that in females, ovariectomy-induced osteoporosis impairs the healing process. There is no information, however, whether the alteration of fracture healing in osteoporosis also depends on gender.. Therefore, we herein studied fracture healing in female and male senescence-accelerated osteoporotic mice, strain P6 (SAMP6), including biomechanical, histomorphometric, and protein biochemical analysis.. Bending stiffness was reduced in male and female SAMP6 mice compared with senescence-resistant strain 1 (SAMR1) controls. This was associated with elevated serum concentrations of tartrate-resistent acid phosphatase form 5b (TRAP) in both female and male SAMP6 mice. Callus size, however, was significantly larger in female SAMP6 mice compared with male SAMP6 mice and female SAMR1 controls. This indicates a delayed remodeling process in female SAMP6 mice. The delay of callus remodeling in female SAMP6 mice was associated with a significantly higher osteoprotegerin (OPG) callus tissue expression and increased serum concentrations of osteocalcin (OC) and deoxypyridinoline (DPD), indicating elevated osteoblast and osteoclast activities.. The present study shows that remodeling during fracture healing in female, but not in male, SAMP6 mice is delayed, most probably due to an increased osteoblast and osteoclast activity.

Topics: Acid Phosphatase; Aging; Amino Acids; Animals; Biomechanical Phenomena; Bone Remodeling; Bony Callus; Disease Models, Animal; Female; Fracture Healing; Isoenzymes; Male; Mice; Mice, Mutant Strains; Osteoblasts; Osteocalcin; Osteoclasts; Osteoporosis; Osteoprotegerin; Sex Characteristics; Tartrate-Resistant Acid Phosphatase

This study examined the efficacy of hydroalcoholic extract of dried clove buds, which is rich in phenolic compounds namely eugenol and eugenol derivatives (precursors of flavones, isoflavones and flavonoids), on different primary and secondary osteoporotic marker changes in an ovariectomised (OVX) rat model of osteoporosis. Female Wistar rats were randomly divided into three groups: sham-operated control (A), OVX (B) and OVX plus 50% hydroalcoholic extract of dried clove buds for 4 weeks (C). Results indicated that, compared to control, serum alkaline phosphatase (AP; 48.25%, p < 0.01), serum tartrate-resistant acid phosphatase (TRAP; 63.48%, p < 0.01), urinary calcium (14.70%, p < 0.01), urinary phosphate (50.30%, p < 0.01) and urinary creatinine (122.44%, p < 0.01) were significantly altered in OVX rats. All these altered responses were significantly restored (AP: 27.53%, p < 0.01; TRAP: 33.51%, p < 0.01; calcium: 53.15%, p < 0.01; phosphate: 27.49%, p < 0.01; creatinine: 46.40%, p < 0.01) by supplementation with hydroalcoholic extract of dried clove buds. Results of bone density, bone mineral content, bone tensile strength and histological analysis also showed similar trend of results, which supported initial observations of this study. It is proposed that hydroalcoholic extract of dried clove buds has bone-preserving efficacy against hypogonadal osteoporosis.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Density; Calcium; Clove Oil; Creatinine; Disease Models, Animal; Drug Evaluation, Preclinical; Eugenol; Female; Isoenzymes; Osteoporosis; Ovariectomy; Phosphates; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Tensile Strength; Tibia

Long-term sequential expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegrin (OPG) in lipopolysaccharide (LPS)-induced rat periapical lesions has not been studied..   Seventy-two 4-week-old Wistar rats were divided into eight experimental groups and one control group (eight animals in each).. Lipopolysaccharide-induced periapical lesions were produced in rats by occlusal exposure of the pulp of their lower first molars in all experimental groups but not the control group. The extent of periapical destruction was measured by radiographic imaging. RANKL and OPG mRNA were measured in all tissue sections containing the periapical lesions as well as the control group every week from week 1 to week 8 by real-time quantitative reverse transcription polymerase chain reaction. RANKL and OPG protein were determined by immunohistochemistry. Osteoclasts were identified by enzyme histochemistry.. The sequential changes in the mRNA and protein expression of RANKL and OPG were largely compatible with the occurrence of osteoclasts histologically and enzymes histochemically, as well as the mean areas of the periapical lesions radiographically during long-term observation of the LPS-induced rat periapical lesions.. This study may be the first to demonstrate the long-term RANKL and OPG expression every week from week 1 to week 8 using LPS to produce periapical infection in a Wistar rat model. The long-term findings of high expressions of RANKL and OPG further extend the potential application of the Wistar rat model for future experimental trials using RANKL inhibitor to evaluate the treatment outcome for LPS-induced rat periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Cell Count; Dental Pulp Exposure; Disease Models, Animal; Escherichia coli; Giant Cells; Image Processing, Computer-Assisted; Immunohistochemistry; Isoenzymes; Lipopolysaccharides; Male; Osteoclasts; Osteoprotegerin; Periapical Diseases; Radiography, Bitewing; RANK Ligand; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Tartrate-Resistant Acid Phosphatase; Time Factors

We report a direct comparison of receptor activator of nuclear factor kappa B ligand (RANKL) inhibition (RANK-Fc) with bisphosphonate treatment (alendronate, ALN) from infancy through early adulthood in a mouse model of osteogenesis imperfecta. Both ALN and RANK-Fc decreased fracture incidence to the same degree with increases in metaphyseal bone volume via increased number of thinner trabeculae.. The potential therapeutic benefit of RANKL inhibitors in osteogenesis imperfecta (OI) is under investigation. We report a direct comparison of RANKL inhibition (RANK-Fc) with bisphosphonate treatment (ALN) from infancy through early adulthood in a model of OI, the oim/oim mouse.. Two-week-old oim/oim, oim/+, and wildtype (+/+) mice were treated with RANK-Fc 1.5 mg/kg twice per week, ALN 0.21 mg/kg/week or saline (n = 12-20 per group) for 12 weeks.. ALN and RANK-Fc both decreased fracture incidence (9.0 ± 3.0 saline 4.4 ± 2.7 ALN, 4.3 ± 3.0 RANK-Fc fractures per mouse). Serum TRACP-5b activity decreased to 65% after 1 month in all treated mice, but increased sacrifice with RANK-Fc to 130-200% at sacrifice. Metaphyseal density was significantly increased with ALN in +/+ and oim/oim mice (p < 0.05) and tended to increase with RANK-Fc in +/+ mice. No changes in oim/oim femur biomechanical parameters occurred with treatment. Both ALN and RANK-Fc significantly increased trabecular number (3.73 ± 0.77 1/mm for oim/oim saline vs 7.93 ± 0.67 ALN and 7.34 ± 1.38 RANK-Fc) and decreased trabecular thickness (0.045 mm ± 0.003 for oim/oim saline vs 0.034 ± 0.003 ALN and 0.032 ± 0.002 RANK-Fc) and separation in all genotypes (0.28 ± 0.08 mm for oim/oim saline vs 0.12 ± 0.010 ALN and 13 ± 0.03 RANK-Fc)., with significant increase in bone volume fraction (BVF) with ALN, and a trend towards increased BVF in RANK-Fc.. Treatment of oim/oim mice with either a bisphosphonate or a RANK-Fc causes similar decreases in fracture incidence with increases in metaphyseal bone volume via increased number of thinner trabeculae.

Topics: Acid Phosphatase; Alendronate; Animals; Biomechanical Phenomena; Bone Density; Bone Density Conservation Agents; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Isoenzymes; Male; Mice; Osteogenesis Imperfecta; Osteoporotic Fractures; RANK Ligand; Recombinant Fusion Proteins; Tartrate-Resistant Acid Phosphatase; Weight Gain; X-Ray Microtomography

Neurofibromatosis type 1 (NF1) is a common genetic condition caused by mutations in the NF1 gene. Patients often suffer from tissue-specific lesions associated with local double-inactivation of NF1. In this study, we generated a novel fracture model to investigate the mechanism underlying congenital pseudarthrosis of the tibia (CPT) associated with NF1. We used a Cre-expressing adenovirus (AdCre) to inactivate Nf1 in vitro in cultured osteoprogenitors and osteoblasts, and in vivo in the fracture callus of Nf1(flox/flox) and Nf1(flox/-) mice. The effects of the presence of Nf1(null) cells were extensively examined. Cultured Nf1(null)-committed osteoprogenitors from neonatal calvaria failed to differentiate and express mature osteoblastic markers, even with recombinant bone morphogenetic protein-2 (rhBMP-2) treatment. Similarly, Nf1(null)-inducible osteoprogenitors obtained from Nf1 MyoDnull mouse muscle were also unresponsive to rhBMP-2. In both closed and open fracture models in Nf1(flox/flox) and Nf1(flox/-) mice, local AdCre injection significantly impaired bone healing, with fracture union being <50% that of wild type controls. No significant difference was seen between Nf1(flox/flox) and Nf1(flox/-) mice. Histological analyses showed invasion of the Nf1(null) fractures by fibrous and highly proliferative tissue. Mean amounts of fibrous tissue were increased upward of 10-fold in Nf1(null) fractures and bromodeoxyuridine (BrdU) staining in closed fractures showed increased numbers of proliferating cells. In Nf1(null) fractures, tartrate-resistant acid phosphatase-positive (TRAP+) cells were frequently observed within the fibrous tissue, not lining a bone surface. In summary, we report that local Nf1 deletion in a fracture callus is sufficient to impair bony union and recapitulate histological features of clinical CPT. Cell culture findings support the concept that Nf1 double inactivation impairs early osteoblastic differentiation. This model provides valuable insight into the pathobiology of the disease, and will be helpful for trialing therapeutic compounds.

Topics: Acid Phosphatase; Animals; Bone Morphogenetic Protein 2; Cell Differentiation; Cell Lineage; Cell Proliferation; Disease Models, Animal; Female; Fibrosis; Fracture Healing; Gene Deletion; HEK293 Cells; Humans; Integrases; Isoenzymes; Mice; Mice, Inbred C57BL; Mice, Knockout; Muscles; Neurofibromatosis 1; Neurofibromin 1; Osteoblasts; Osteoclasts; Osteogenesis; Pseudarthrosis; Recombinant Proteins; Reproducibility of Results; Tartrate-Resistant Acid Phosphatase; Tibia; Transforming Growth Factor beta

Several studies have shown that cathepsin K (CTK) is overexpressed in osteoarthritic (OA) cartilage and subchondral bone. However, it has not been well established whether CTK expression is harmful or beneficial. We undertook this study to investigate the direct involvement of CTK in OA development using Ctsk-knockout (Ctsk(-/-)) mice in a joint instability-induced model of OA.. We analyzed the natural course of the phenotype of 25-week-old Ctsk(-/-) mice. OA development was evaluated with a modified Mankin histologic score up to 8 weeks after surgery was performed to destabilize the knee in Ctsk(-/-) and Ctsk(+/+) mice. Histologic analysis was used to evaluate expression of CTK, matrix metalloproteinase 13 (MMP-13), ADAMTS-5, and tartrate-resistant acid phosphatase (TRAP) proteins in chondrocytes, synovial cells, and osteoclasts. Bone architecture was analyzed by histomorphometry.. Bone mineral content and bone volume were higher in Ctsk(-/-) mice at 25 weeks, whereas OA did not develop spontaneously in either Ctsk(-/-) or Ctsk(+/+) mice. In a model of destabilization-induced OA, OA progression was significantly delayed in Ctsk(-/-) mice. CTK was overexpressed in chondrocytes and synovial cells of knee joints developing OA in Ctsk(+/+) mice. MMP-13 and ADAMTS-5 were less strongly expressed in chondrocytes of Ctsk(-/-) mice, and MMP-13 was less strongly expressed in synovial cells. TRAP-positive osteoclasts were overexpressed in Ctsk(-/-) mice.. These results indicate that CTK plays crucial direct roles in the early to intermediate stage of OA development. CTK-positive chondrocytes and synovial cells may be a possible target to prevent disease progression in OA.

Topics: Acid Phosphatase; ADAM Proteins; Animals; Bone and Bones; Bone Density; Cartilage, Articular; Cathepsin K; Chondrocytes; Disease Models, Animal; Disease Progression; Isoenzymes; Knee Joint; Matrix Metalloproteinase 13; Mice; Mice, Knockout; Osteoarthritis, Knee; Osteoclasts; Phenotype; Synovial Membrane; Tartrate-Resistant Acid Phosphatase

Prostate cancer (PCa) is the most commonly diagnosed type of cancer in men in western industrialized countries. As a public health burden, the need for the invention of new cost-saving PCa immunotherapies is apparent. In this study, we present a DNA vaccine encoding for the prostate-specific antigen prostatic acid phosphatase (PAP) linked to the J-domain and the SV40 enhancer sequence. The PAP DNA vaccine induced a strong PAP-specific cellular immune response after electroporation (EP)-based delivery in C57BL/6 mice. Splenocytes from mice immunized with PAP recognized the naturally processed PAP epitopes, indicating that vaccination with the PAP-J gene broke its self-tolerance against PAP. Remarkably, DNA vaccination with PAP-J inhibited tumor growth in the Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) mouse model that closely resembled human PCa. Therefore, this study highlights a novel cancer immunotherapy approach with the potential to control PCa in clinical settings.

Topics: Acid Phosphatase; Amino Acid Motifs; Amino Acid Sequence; Animals; Antibodies; Antigens, Neoplasm; Cancer Vaccines; Cell Line; Disease Models, Animal; Disease Progression; Epitopes; Genetic Vectors; H-2 Antigens; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Peptides; Prostatic Neoplasms; Protein Binding; Protein Tyrosine Phosphatases; Self Tolerance; T-Lymphocytes, Cytotoxic; Vaccines, DNA

Mi-64, a high molecular weight protein (130 kDa), obtained from the tissue homogenate of marine polychaete (Mastobranchus indicus) collected from the Indian Sunderban has antiarthritic activity in experimental animals. The FCA-induced arthritis model was developed in Wistar albino rats to evaluate the antiarthritic effects of Mi-64. After FCA induction, the rats were treated with Mi-64 (0.25 and 0.5 mg kg(-1) body weight) for 10 days. We have determined the paw/ankle swellings, urinary hydroxyproline and glucosamine, serum acid and alkaline phosphatases to assess the antiarthritic activity. The levels of interleukin-1 beta (IL-1β), IL-6, cytokine-induced neutrophil chemoattractant-1 (CINC-1), tumor necrosis factor-alpha (TNF-α), and IL-10 were measured by ELISA. Results showed that Mi-64 significantly reduced paw/ankle swellings and restored the urinary hydroxyproline/glucosamine and serum phosphatases. Mi-64 significantly inhibited the overproduction of IL-1β, IL-6, CINC-1, and TNF-α and augmented IL-10 production. The data suggest that Mi-64 produced significant antiarthritic effects that may be mediated by balancing the pro- and anti-inflammatory cytokines.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Ankle Joint; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Experimental; Chemokine CXCL1; Disease Models, Animal; Glucosamine; Hydroxyproline; Interleukin-10; Interleukin-1beta; Interleukin-6; Male; Polychaeta; Proteins; Random Allocation; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

The present study investigated whether bacteria infecting the root canal can activate any infiltrating T cells to produce receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL).. Using a mouse model of periapical lesion induced by artificial dental pulp exposure, the presence of RANKL-positive T cells and osteoclasts in the periapical lesion was examined by an immunohistochemical approach. The bacteria colonizing the exposed root canal were identified by 16S ribosomal RNA (rRNA) sequence analysis. The isolated endodontic bacteria were further immunized to normal mice, and soluble activator of NF-κB ligand (sRANKL) production by the T cells isolated from the immunized mice was evaluated by ex vivo culture system.. RANKL-positive T cells along with TRAP+ osteoclasts were identified in periapical bone resorption lesions. The gram-negative bacterium Pasteurella pnumotropica, which was most frequently detected from the root canal of exposed pulp, showed remarkably elevated serum immunoglobulin G (IgG)-antibody response in pulp-exposed mice compared with control nontreated mice. Immunization of mice with P. pneumotropica induced not only serum IgG-antibody but also primed bacteria-reactive T cells that produced sRANKL in response to ex vivo exposure to P. pneumotropica.. T cells infiltrating the periapical region express RANKL, and the endodontic bacteria colonizing the root canal appear to induce RANKL expression from bacteria-reactive T cells, suggesting the possible pathogenic engagement of the immune response to endodontic bacteria in the context of developing bone resorptive periapical lesions.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Antibodies, Bacterial; Biomarkers; CD3 Complex; Dental Pulp Cavity; Dental Pulp Exposure; Disease Models, Animal; Enterococcus; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Immunization; Immunoglobulin G; Immunologic Memory; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Microscopy, Confocal; Osteoclasts; Pasteurella Infections; Pasteurella pneumotropica; Periapical Diseases; RANK Ligand; RNA, Bacterial; RNA, Ribosomal, 16S; T-Lymphocytes; Tartrate-Resistant Acid Phosphatase

Root resorption is a ubiquitous although undesirable sequela to orthodontic treatment. Current methods to investigate the pathophysiology have certain limitations. In pursuit to understand and develop treatment modalities for orthodontically induced root resorption, the ability to manipulate cells within their natural extracellular matrix in a three dimensional organotypic model is invaluable. The study aimed to develop a laboratory-based organotypic model to investigate the effect of orthodontic forces on the periodontium.. Mandibular slices of male Wistar rats were maintained in Trowel-typed cultures at 37°C in 5% carbon dioxide in air for 7 days with test specimens subjected to compressive forces at 50 g and 100g by stainless steel springs. Tissue architecture and cell viability were maintained under culture conditions.. Osteoclast numbers increased significantly in both test groups whilst odontoclasts increased in the 50 g group. Immunohistochemistry demonstrated increased dentine sialoprotein expression in both test groups, suggesting changes in mineralization-related activity due to mechanical strain.. The study showed initial cellular and molecular changes of key markers that relate to root resorption in response to mechanical loading.. Severe root resorption may occur when forces applied are heavy or transmitted over an extended period and could lead to mobility and tooth loss. This ex vivo model can be used to investigate cellular and molecular processes during orthodontic tooth movement which may advance the clinical management of root resorption.

Topics: Acid Phosphatase; Animals; Biomarkers; Biomechanical Phenomena; Bone Marrow; Cell Count; Cell Survival; Dental Pulp; Disease Models, Animal; Extracellular Matrix Proteins; Immunohistochemistry; Isoenzymes; Male; Mandible; Organ Culture Techniques; Orthodontic Wires; Osteoclasts; Periodontal Ligament; Phosphoproteins; Rats; Rats, Wistar; Root Resorption; Sialoglycoproteins; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques

The purpose of this study is to investigate the effects of dexamethasone on repair of a critical size defect of the mandible in male Sprague-Dawley rats.. Fifty rats were divided into 2 groups: saline control and dexamethasone-treated groups. A 1 mm × 3 mm full-thickness bone defect was created at the inferior border of the mandible. Saline or dexamethasone was administered once a day for 5 days after postoperative palinesthesia. On days 1, 3, 6, 10 and 17, after cessation of drug administration, 5 samples from each group were analysed. The bone defect healing process was examined and analysed by stereology, radiology, histology and histochemical staining for total collagen, tartrate-resistant acid phosphatase staining for osteoclasts and immunohistochemical staining for the COX-2, RUNX2 and osteocalcin antigens.. The dexamethasone-treated rats exhibited significantly lower radiopacity properties compared to the control rats. Histological staining revealed that the osteogenic differentiation and maturation of a callus in the defect region was significantly delayed from day 1 to day 10 in the dexamethasone group after cessation of drug administration compared to the control group. Consistent with the histological data, the level of total collagen protein was significantly lower in the dexamethasone group than in the control group. However, there was no significant difference between the 2 groups at day 17. Immunohistochemical analysis of COX-2, RUNX2 and osteocalcin expression showed that, at day 1, COX-2 and RUNX2 expression in the dexamethasone group was significantly lower than in the control group. There was no significant difference in osteocalcin expression between the two groups at each time point. There was no significant difference in the number of osteoclasts between the two groups.. In a model of bone healing of a mandible defect, dexamethasone-treated rats exhibited impaired osteogenic differentiation and maturation due to the inhibition of COX-2, osteogenic gene, RUNX2 and collagen protein expression, which resulted in delayed bone repair. Although perioperative short-term therapy did not exhibit long-term effects on wound healing of the maxillofacial bone, the application of glucocorticoids should be cautiously considered in the clinic.

Topics: Acid Phosphatase; Animals; Cell Differentiation; Collagen; Dexamethasone; Disease Models, Animal; Glucocorticoids; Isoenzymes; Male; Mandible; Osteocalcin; Osteogenesis; Rats; Rats, Sprague-Dawley; RNA, Messenger; Tartrate-Resistant Acid Phosphatase; Wound Healing

In osteoprotegerin-deficient (OPG-/-) mice, osteoclast activity causes bone resorption to outpace bone formation, leading to the development of severe osteoporosis. Such mice are therefore useful for investigating the alveolar bone of patients with osteoporosis. Reveromycin A (RM-A) was recently identified as the unique agent acting on osteoclast activation. This study aimed to analyze the effect of RM-A on the orthodontic treatment of OPG-/- mice (a model of osteoporosis patients with high levels of bone turnover). We examined alveolar bone remodeling in OPG-/- and wild-type (WT) mice during continuous tooth movement. The orthodontic force was induced by means of a Ni-Ti closed-coil spring to move the maxillary first molar for 14 days. RM-A sodium salt (1 mg/kg) was administered intraperitoneally twice daily. In OPG-/- mice, the tooth movement distance was longer, alveolar bone resorption was enhanced, the osteoclast count was greater, and serum alkaline phosphatase and tartrate-resistant acid phosphatase levels were higher relative to those in WT mice. However, the administration of RM-A in OPG-/- mice reduced these parameters. We conclude that RM-A normalizes bone metabolism and loss of alveolar bone during continuous tooth movement in OPG-/- mice.

Topics: Acid Phosphatase; Alkaline Phosphatase; Alveolar Process; Animals; Biomarkers; Bone Density Conservation Agents; Bone Remodeling; Bone Resorption; Cell Count; Dental Alloys; Disease Models, Animal; Injections, Intraperitoneal; Isoenzymes; Male; Maxilla; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Knockout; Mutation; Nickel; Orthodontic Wires; Osteitis Deformans; Osteoclasts; Osteoporosis; Osteoprotegerin; Pyrans; Spiro Compounds; Tartrate-Resistant Acid Phosphatase; Titanium; Tooth Movement Techniques; X-Ray Microtomography

Vascular calcification is a hallmark of atherosclerosis, a major cause of morbidity and mortality in the United States. We have previously reported that the osteogenic transcription factor Runx2 is an essential and sufficient regulator of calcification of vascular smooth muscle cells (VSMC) in vitro.. To determine the contribution of osteogenic differentiation of VSMC to the pathogenesis of vascular calcification and the function of VSMC-derived Runx2 in regulating calcification in vivo.. SMC-specific Runx2-deficient mice, generated by breeding SM22α-Cre mice with the Runx2 exon 8 floxed mice, exhibited normal aortic gross anatomy and expression levels of SMC-specific marker genes. Runx2 deficiency did not affect basal SMC markers, but inhibited oxidative stress-reduced expression of SMC markers. High-fat-diet-induced vascular calcification in vivo was markedly inhibited in the Runx2-deficient mice in comparison with their control littermates. Runx2 deficiency inhibited the expression of receptor activator of nuclear factor κB ligand, which was accompanied by decreased macrophage infiltration and formation of osteoclast-like cells in the calcified lesions. Coculture of VSMC with bone marrow-derived macrophages demonstrated that the Runx2-deficient VSMC failed to promote differentiation of macrophages into osteoclast-like cells.. These data have determined the importance of osteogenic differentiation of VSMC in the pathogenesis of vascular calcification in mice and defined the functional role of SMC-derived Runx2 in regulating vascular calcification and promoting infiltration of macrophages into the calcified lesion to form osteoclast-like cells. Our studies suggest that the development of vascular calcification is coupled with the formation of osteoclast-like cells, paralleling the bone remodeling process.

Topics: Acid Phosphatase; Animals; Atherosclerosis; Bone Remodeling; Calcinosis; Cell Differentiation; Cells, Cultured; Coculture Techniques; Core Binding Factor Alpha 1 Subunit; Diet, High-Fat; Disease Models, Animal; Exons; Female; Isoenzymes; Macrophages; Male; Mice; Mice, Knockout; Muscle, Smooth, Vascular; Mutagenesis; Osteoclasts; RANK Ligand; Tartrate-Resistant Acid Phosphatase

Receptor activator of nuclear factor-κB ligand (RANKL) inhibitors are being considered for use in children with osteogenesis imperfecta (OI). We sought to assess efficacy of two doses of a RANKL inhibitor, osteoprotegerin-immunoglobulin Fc segment complex (OPG-Fc), in a growing animal model of OI, the col1α2-deficient mouse (oim/oim) and its wild-type controls (+/+).. Treated mice showed runting and radiographic evidence of osteopetrosis with either high- (20 mg/kg twice weekly) or low-dose (1 mg/kg/week) OPG-Fc. Because of this adverse event, OPG-Fc treatment was halted, and the mice were killed or monitored for recovery with monthly radiographs and assessment of serum osteoclast activity (tartrate-resistant acid phosphatase 5b, TRACP-5b) until 25 wk of age.. Twelve weeks of OPG-Fc treatment resulted in radiographic and histologic osteopetrosis with no evidence of bone modeling and negative tartrate-resistant acid phosphatase staining, root dentin abnormalities, and TRACP-5b activity suppression. Signs of recovery appeared 4-8 wk post-treatment.. Both high- and low-dose OPG-Fc treatment resulted in osteopetrotic changes in infant mice, an outcome that was not seen in studies with the RANKL inhibitor RANK-immunoglobulin Fc segment complex (RANK-Fc) or in studies with older animals. Further investigations of RANKL inhibitors are necessary before their consideration for use in children.

Topics: Acid Phosphatase; Age Factors; Animals; Biomarkers; Bone Remodeling; Collagen Type I; Dentin; Disease Models, Animal; Female; Immunoconjugates; Immunoglobulin Fc Fragments; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Osteoclasts; Osteogenesis Imperfecta; Osteopetrosis; Osteoprotegerin; Radiography; RANK Ligand; Risk Assessment; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Eruption; Weight Gain

Soy with its isoflavones has been shown to positively influence bone mineral density in female ovariectomized rats; hence, we hypothesized a similar effect in orchidectomized (ORX) male rats. Forty male Sprague-Dawley rats, aged 95 days, were divided into 4 groups and were either sham operated (Sham) or ORX. The ORX groups were fed a soy protein-based diet (SOY), an isoflavone-depleted soy protein diet (SOY-), or a casein based diet for 65 days after surgery. Orchidectomy increased the rate of bone turnover, resulting in reduced bone mineral density and bone mineral content by 3.5% and 14%, respectively, and compromised biomechanical properties. The mean femoral length of ORX animals was also significantly shorter than Sham animals, but ORX rats that were fed SOY diet did not experience this reduction in bone length, implicating a role for soy protein in bone growth (4.02 ± 0.02, 3.93 ± 0.01, 3.99 ± 0.02, 3.91 ± 0.01 for Sham, ORX, SOY, SOY-, respectively). The SOY and SOY- positively influenced the biomechanical properties of bone such as yield and ultimate force, the measures of bone elasticity, and plasticity. In terms of bone histomorphometry, the data indicate that SOY- tends to reduce ORX-induced increase in bone turnover as evidenced by suppressed bone formation rate/mineralized surface by about 9%. Overall, our results indicated that soy protein, regardless of its isoflavone content, was unable to prevent the ORX-induced femoral decrease in bone density and mineral content. However, soy may enhance the quality of bone as indicated by increased yield force.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Density; Caseins; Disease Models, Animal; Energy Intake; Glycine max; Gonadal Hormones; Isoenzymes; Isoflavones; Male; Orchiectomy; Osteoporosis; Rats; Rats, Sprague-Dawley; Soybean Proteins; Tartrate-Resistant Acid Phosphatase

Osteoprotegerin (OPG), as an osteoclast antagonist, limits mineralised tissue resorption under physiological conditions. Previous work investigating OPG in a rat periodontal ligament (PDL) ankylosis model found no inhibitory effect on osteoclasts when OPG was administered at a dosage of 2.5mg/kg.. The object of this study was to determine whether dosages higher than 2.5 mg/kg of OPG were required to limit osteoclastic activity in an aseptic inflammatory model in rats.. Dry ice was applied for 15 minutes to the upper right first molar crown of eighteen, 8-week-old, male Sprague-Dawley rats. Three groups of 3 were injected with OPG at dosages of 2.5, 5.0 and 7.5 mg/kg of body weight immediately following the thermal insult. After 7 days, the rats were sacrificed and each maxilla processed for histological examination and stained for osteoclastic activity using tartrate-resistant acid phosphatase (TRAP). Osteoclast population numbers were estimated via light microscopy and results were analysed using a comparative mixed model statistical analysis.. Results showed OPG inhibited osteoclastic activity in a dose-dependent manner. From 2.5 mg/kg to 7.5 mg/kg, osteoclast populations were linearly reduced by 39.78% (p < 0.05). OPG did not appear to affect the inflammatory process and had varied efficacy in different regions of individual teeth.. Although osteoclastic activity reduced, it was not completely eliminated, perhaps because dosages were still inadequate, or additional factors might influence OPG and osteoclast activation in the aseptic inflammatory model.

Topics: Acid Phosphatase; Animals; Biomarkers; Cell Count; Disease Models, Animal; Dose-Response Relationship, Drug; Dry Ice; Freezing; Inflammation; Isoenzymes; Male; Maxilla; Molar; Necrosis; Odontoblasts; Osteoclasts; Osteoprotegerin; Rats; Rats, Sprague-Dawley; Root Resorption; Tartrate-Resistant Acid Phosphatase; Tooth Crown

Despite the clinical adoption of distraction osteogenesis (DO), studies examining the bone healing process at the distraction gap in osteoporotic bone are limited. We examined the effect of osteoporosis in the ovariectomized rat on DO.. Mid-diaphyseal osteotomies were performed on the femurs of ovariectomized (OVX) rats. External distractors were placed on these rats and also on sham-ovariectomized rats. After a 7-day latency period, distraction was carried out at a rate of 0.5mm/day for 10 days. The bone volume (BV) of the distraction gap was measured by Micro-focused X-ray computed tomography (micro-CT) at 0, 2, and 4 weeks after completion of the distraction, and the distraction gap was examined histologically.. The BV of the distraction gap in the OVX group was significantly lower than that in the sham group at 2 and 4 weeks after completion of distraction (p<0.01). On histological examination, the distraction gap in the OVX group was filled with scattered smaller bone trabeculae than those seen in the sham group at 4 weeks after completion of distraction. Osteoclast numbers at the distraction gap in the OVX group were significantly increased when compared to the sham group (p<0.01).. Bone turnover with osteoclast predominance in ovariectomized rats is likely to be the cause of a reduction in new bone formation at the distraction gap.

Topics: Acid Phosphatase; Animals; Biomarkers; Body Weight; Bone Density; Bone Marrow; Bone Matrix; Cartilage; Connective Tissue; Diaphyses; Disease Models, Animal; External Fixators; Female; Femur; Isoenzymes; Osteoclasts; Osteogenesis; Osteogenesis, Distraction; Osteoporosis; Ovariectomy; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Time Factors; Wound Healing; X-Ray Microtomography

Orthodontic tooth movement is achieved by the remodeling of alveolar bone in response to mechanical loading. Type 1 diabetes results in bone remodeling, suggesting that this disease might affect orthodontic tooth movement. The present study investigated the effects of the diabetic state on orthodontic tooth movement. An orthodontic appliance was placed in normoglycemic (NG), streptozotocin-induced diabetes (DB), and insulin-treated DB (IT) C57BL6/J mice. Histomorphometric analysis and quantitative PCR of periodontium were performed. The DB mice exhibited greater orthodontic tooth movement and had a higher number of tartrate-resistant acid phosphate (TRAP) -positive osteoclasts than NG mice. This was associated with increased expression of factors involved in osteoclast activity and recruitment (Rankl, Csf1, Ccl2, Ccl5, and Tnfa) in DB mice. The expression of osteoblastic markers (Runx2, Ocn, Col1, and Alp) was decreased in DB mice. Reversal of the diabetic state by insulin treatment resulted in morphological findings similar to those of NG mice. These results suggest that the diabetic state up-regulates osteoclast migration and activity and down-regulates osteoblast differentiation, resulting in greater orthodontic tooth movement.

Topics: Acid Phosphatase; Alveolar Process; Animals; Bone Remodeling; Cell Differentiation; Cell Movement; Chemokines; Cytokines; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Disease Models, Animal; Insulin; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Osteoblasts; Osteoclasts; Periodontal Ligament; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques

Bone-lining tissues contain a population of resident macrophages termed osteomacs that interact with osteoblasts in vivo and control mineralization in vitro. The role of osteomacs in bone repair was investigated using a mouse tibial bone injury model that heals primarily through intramembranous ossification and progresses through all major phases of stabilized fracture repair. Immunohistochemical studies revealed that at least two macrophage populations, F4/80(+) Mac-2(-/low) TRACP(-) osteomacs and F4/80(+) Mac-2(hi) TRACP(-) inflammatory macrophages, were present within the bone injury site and persisted throughout the healing time course. In vivo depletion of osteomacs/macrophages (either using the Mafia transgenic mouse model or clodronate liposome delivery) or osteoclasts (recombinant osteoprotegerin treatment) established that osteomacs were required for deposition of collagen type 1(+) (CT1(+)) matrix and bone mineralization in the tibial injury model, as assessed by quantitative immunohistology and micro-computed tomography. Conversely, administration of the macrophage growth factor colony-stimulating factor 1 (CSF-1) increased the number of osteomacs/macrophages at the injury site significantly with a concurrent increase in new CT1(+) matrix deposition and enhanced mineralization. This study establishes osteomacs as participants in intramembranous bone healing and as targets for primary anabolic bone therapies.

Topics: Acid Phosphatase; Animals; Bone Matrix; Calcification, Physiologic; Clodronic Acid; Disease Models, Animal; Inflammation; Isoenzymes; Liposomes; Macrophage Colony-Stimulating Factor; Macrophages; Membranes; Mice; Mice, Inbred C57BL; Mice, Transgenic; Osteoblasts; Osteogenesis; Osteoprotegerin; Surface Properties; Tartrate-Resistant Acid Phosphatase; Tibia; Wound Healing

Nitric oxide (NO) and reactive oxygen species (ROS) are key molecules in resistance to pathogens. Little is known about their role in pathogenesis of periapical lesions. To address this issue, we induced periapical lesions in mice lacking nitric oxide synthase (iNOS(-/-)) or phagocyte oxidase (PHOX(-/-)). iNOS(-/-) mice expressed higher levels of IL-1β, TNF-α, RANK, RANKL, and MCP-1 than C57BL/6 and PHOX(-/-). Apical thickening of the periodontal ligament was also greater in iNOS(-/-) compared with other groups. Interestingly, ROS production did not interfere in periapical lesion progression, but seemed to be essential for the appearance of multinucleated TRAP-positive cells. Thus, periapical lesion progression in iNOS(-/-) was associated with an imbalance of pro-inflammatory cytokines (IL-1β and TNF-α), bone-resorptive modulators (RANK and RANKL), and MCP-1. We conclude that NO, but not ROS, controls progression of bone resorption in a murine experimental model of apical periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Chemokine CCL2; Cytokines; Disease Models, Animal; Disease Progression; Inflammation Mediators; Interleukin-1beta; Isoenzymes; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Knockout; NADPH Oxidases; Nitric Oxide; Nitric Oxide Synthase Type II; Periapical Periodontitis; Periodontal Ligament; Phagocytes; RANK Ligand; Reactive Oxygen Species; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

This study examined the impact of an interleukin-6 (IL-6) knockout on fracture healing in terms of histological and biomechanical responses. Following IACUC approval, tibial fractures were produced in 4- to 6-week-old IL-6 knockouts (n = 35) and wild-type mice (n = 36) and harvested along with contralateral limbs at 2 and 6 weeks postsurgery. Histology quantified stage of healing, lymphocyte infiltration, TRAP+ cells, and osteocalcin deposition. Bend testing established maximum load and stiffness. Based on normality assessments, Mann-Whitney U or independent t-tests were used for data analysis using a p-value threshold of 0.05. Stage of healing, lymphocyte infiltration, and osteocalcin deposition were similar for all time points (p ≥ 0.243). TRAP+ cell counts were reduced approximately 10-fold in the knockout at 2 weeks (p = 0.015) but were similar at 6 weeks (p = 0.689). Force-to-failure in knockouts was approximately 40% that of wild-type mice at 2 weeks (p = 0.040) but similar at 6 weeks (p = 0.735). Knockout bone was about 25% less stiff at 2 weeks but approximately 60% stiffer at 6 weeks (p ≥ 0.110). The absence of IL-6 during early fracture healing significantly reduced osteoclastogenesis and impaired callus strength. By 6 weeks, most histological and biomechanical parameters were similar to fractures in wild-type bone.

Topics: Acid Phosphatase; Animals; Bony Callus; Disease Models, Animal; Elasticity; Fracture Healing; Interleukin-6; Isoenzymes; Lymphocytes; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Osteocalcin; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tibial Fractures

Pumpkins are thought to be useful in the management of benign prostatic hyperplasia (BPH). The ability of a 15% Telfairia occidentalis seeds incorporated diet to inhibit hormonal induction of BPH in rats was studied.. Twenty male Wistar rats were divided into 4 equal groups - one test group and three control groups. The test group was placed on the test diet and was given subcutaneous injections of dihydrotestosterone (DHT) and estradiol valerate (ratio 10:1) every other day for 28 days. One control group, 'no test diet' (ND) group, received the hormones, but was placed on a normal diet. The other two control groups, 'no hormone' (NH) and 'no hormone/test diet' (NHD), received subcutaneous olive oil (vehicle) for the same duration and were placed on the test and normal diets, respectively. Markers of BPH and hormone profile were determined using standard methods.. The mean relative prostate weight (×10(3)) was reduced in the test group (3.6 ± 0.2) relative to the ND group (4.0 ± 0.4). The protein content (mg/tissue) of the rats' prostates decreased significantly (p < 0.05) from 68.3 ± 2.7 in the ND group to 43.4 ± 3.9 in the test group. Serum prostatic acid phosphatase levels (U/l) decreased significantly (p < 0.05) from 4.8 ± 0.4 in the ND group to 4.0 ± 0.9 in the test group. Histological findings corroborate these data. The testosterone:estradiol ratio (×10(3)) was significantly (p < 0.05) increased from 7.1 ± 0.1 in the ND group to 8.4 ± 0.4 in the test group.. The test diet inhibited the induction of BPH in rats and may act by increasing the testosterone:estradiol ratio.

Topics: Acid Phosphatase; Animal Feed; Animals; Cucurbita; Disease Models, Animal; Estradiol; Male; Prolactin; Prostate; Prostatic Hyperplasia; Protein Tyrosine Phosphatases; Rats; Rats, Wistar; Seeds; Testosterone; Time Factors

Clinical studies have revealed a blunting of the bone anabolic effects of parathyroid hormone treatment in osteoporotic patients in the setting of pre- or cotreatment with the antiresorptive agent alendronate (ALN). Sclerostin monoclonal antibody (Scl-Ab) is currently under clinical investigation as a new potential anabolic therapy for postmenopausal osteoporosis. The purpose of these experiments was to examine the influence of pretreatment or cotreatment with ALN on the bone anabolic actions of Scl-Ab in ovariectomized (OVX) rats. Ten-month-old osteopenic OVX rats were treated with ALN or vehicle for 6 wk, before the start of Scl-Ab treatment. ALN-pretreated OVX rats were switched to Scl-Ab alone or to a combination of ALN and Scl-Ab for another 6 wk. Vehicle-pretreated OVX rats were switched to Scl-Ab or continued on vehicle to serve as controls. Scl-Ab treatment increased areal bone mineral density, volumetric bone mineral density, trabecular and cortical bone mass, and bone strength similarly in OVX rats pretreated with ALN or vehicle. Serum osteocalcin and bone formation rate on trabecular, endocortical, and periosteal surfaces responded similarly to Scl-Ab in ALN or vehicle-pretreated OVX rats. Furthermore, cotreatment with ALN did not have significant effects on the increased bone formation, bone mass, and bone strength induced by Scl-Ab in the OVX rats that were pretreated with ALN. These results indicate that the increases in bone formation, bone mass, and bone strength with Scl-Ab treatment were not affected by pre- or cotreatment with ALN in OVX rats with established osteopenia.

Topics: Acid Phosphatase; Alendronate; Animals; Antibodies, Monoclonal; Bone and Bones; Bone Density; Bone Density Conservation Agents; Bone Diseases, Metabolic; Bone Morphogenetic Proteins; Disease Models, Animal; Female; Genetic Markers; Isoenzymes; Osteocalcin; Osteogenesis; Ovariectomy; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

Omentin-1 (also known as intelectin-1) is a recently identified visceral adipose tissue-derived cytokine that is inversely related to obesity. Our previous study showed that omentin-1 inhibits osteoblastic differentiation of calcifying vascular smooth muscle cells (CVSMCs) in vitro. This study was undertaken to investigate the effects of omentin-1 on arterial calcification and bone metabolism in vivo.. In vitro, omentin-1 stimulated production of osteoprotegerin (OPG) and inhibited production of receptor activator for nuclear factor κB ligand (RANKL) in both CVSMCs and osteoblasts. In vivo, adenovirus-mediated over-expression of omentin-1 attenuated arterial calcification and bone loss in OPG(-/-) mice. All these in vitro and in vivo actions were abrogated by blockade of the PI3K-Akt signalling pathway. Furthermore, omentin-1 reduced serum levels of RANKL, tartarate-resistant acid phosphatase-5b and osteocalcin, all of which are increased dramatically in OPG(-/-) mice.. These data suggest that omentin-1 ameliorates arterial calcification and bone loss in vivo through the regulation of the RANK signalling pathway.

Topics: Acid Phosphatase; Adiponectin; Animals; Bone Density; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cytokines; Disease Models, Animal; Down-Regulation; Female; Genetic Therapy; GPI-Linked Proteins; Humans; Isoenzymes; Lectins; Mice; Mice, Inbred C57BL; Mice, Knockout; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Osteoblasts; Osteocalcin; Osteoporosis; Osteoprotegerin; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; RANK Ligand; Recombinant Proteins; Signal Transduction; Tartrate-Resistant Acid Phosphatase; Time Factors; Vascular Calcification

The aim of our study was to see the efficacy of petroleum ether extract of Cissus quadrangularis (CQ) on development of osteopenia in ovariectomy induced Wistar rats.. The female Wistar rats were ovariectomized or Sham operated. The rats were anesthetized with pentobarbital sodium (40 mg/ kg b.w, i.p.), the ovaries were removed bilaterally. Sham-operation was performed in the same manner but only exposing the ovaries (sham operated (SHAM) group). A day later, the ovariectomized rats were randomly divided into four groups of eight animals each. The groups are 1. Sham operated (SHAM), 2. Ovariectomized (OVX), 3. Ovariectomized and treated with 25 mg/kg b.w of raloxifene (OVX+RAL), 4. Ovariectomized and treated with 500 mg/kg b.w of petroleum ether extract of CQ (OVX+CQ). The treatment continued for 30 days. At the end of the treatment, rats in all groups were sacrificed by cervical dislocation. Before sacrifice, blood was collected for the estimation of serum ALP, TRAP, Calcium and hydroxyproline; where as the left femur was used for histomorphometrical analysis.. The findings assessed on the basis of animal weight, morphology of femur, histomorphometry and biochemical analysis. As compared to SHAM group, OVX group animals showed a significant rise in serum ALP, TRAP and hydroxyproline levels at the end of 1 month following ovariectomy while no significant change was seen in the serum calcium levels. ALP and TRAP levels of OVX + RAL and OVX + CQ groups showed a further increase following administration of raloxifene and Cissus quadrangularis. The serum hydroxyproline content was found to be increased in the OVX + CQ compared to SHAM group. CQ significantly increased the thickness of both cortical (p <0.001) and trabecular bone (p <0.001).This action of CQ is comparable to action of Raloxifene. )These data suggest a strong anti-osteoporotic activity of CQ.. The results confirm, at least in part, for the use of Cissus quadrangularis in folk medicine to treat osteoporosis.

Topics: Acid Phosphatase; Alkaline Phosphatase; Alkanes; Animals; Biomarkers; Bone Diseases, Metabolic; Bone Resorption; Calcium; Cissus; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Femur; Humans; Hydroxyproline; Isoenzymes; Osteoporosis, Postmenopausal; Ovariectomy; Phytotherapy; Plant Extracts; Raloxifene Hydrochloride; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

In the present study, we evaluated the osteogenic potential of an autogenous bone marrow graft combined with beta-tricalcium phosphate (beta-TCP) in a rat calvarial bone defect model. The bone marrow harvested from the tibia of 7-week-old rats was grafted autogenously in a calvarial defect together with beta-TCP (=BTG group, n=16) or without beta-TCP (=BG group, n=16). Groups of animals were also treated with beta-TCP alone (=TG group, n=16) and control animals (n=8) received no graft implanted into the defect. We then observed the process of bone formation by histology, enzyme histochemistry and immunohistochemistry. Five days after grafting, in the BTG and BG groups, cell proliferation and osteogenic differentiation were observed. From 5 to 10 days after surgery, active Runx2, osteopontin (OPN), and TRAP- positive cells appeared in the BTG and BG groups. New bone formation started in the defect in both the BTG and BG groups. At 30 days after grafting, the BTG group showed new bone development and replacement of beta-TCP to fill the bone defect. New bone formation in the BTG group was significantly greater than in the BG group (P<0.01). The TG group showed no marked bone formation in the defect. The combination graft of bone marrow with beta-TCP showed marked bone formation in rat calvarial defects. Our results indicate that the combination grafts of bone marrow with beta-TCP may be an effective technique for repairing bone defects Beta-TCPgraft (TG) group.

Topics: Acid Phosphatase; Animals; Biocompatible Materials; Bone Marrow Cells; Bone Marrow Transplantation; Bone Regeneration; Bone Substitutes; Calcium Phosphates; Cell Differentiation; Cell Proliferation; Core Binding Factor Alpha 1 Subunit; Disease Models, Animal; Isoenzymes; Male; Osteogenesis; Osteopontin; Rats; Skull; Tartrate-Resistant Acid Phosphatase; Tissue Engineering; Wound Healing

To investigate the effects of systemically administered alendronate, one of the most potent bisphosphonates (BPs), on alveolar bone resorption and angiogenesis in rats subjected to experimental periapical lesions over two time periods.. Forty adult Sprague-Dawley (SD) rats were divided equally into control and experimental groups, and the pulp chambers of mandibular first molars of all rats were exposed to the oral environment to induce periapical lesions. The experimental group received daily subcutaneous injections of alendronate at a dose of 0.25 mg kg(-1), whereas the control group received only the saline vehicle. These injections were initiated 1 week before the periapical lesion induction and then continued daily throughout the entire experimental period. After 2 or 4 weeks following pulp exposure, the rats were killed, and the mandibles were examined histologically for periapical bone loss area, number of microvascular vessels (NMV) and tartrate-resistant acid phosphatase (TRAP) activity.. Overall, periapical bone loss area and the number of TRAP-positive cells (osteoclasts) were significantly decreased at 2 and 4 weeks, respectively, after daily subcutaneous injection of alendronate compared with the control group (P < 0.05). There was no significant decrease change in NMV (P > 0.05).. Administration of alendronate to rats might inhibit alveolar bone resorption associated with periapical disease, which might not lead to impairment of angiogenesis. However, because of the differences between rats and humans, one has to consider the possible consequences of this treatment in the clinic.

Topics: Acid Phosphatase; Alendronate; Alveolar Bone Loss; Alveolar Process; Animals; Bone Density Conservation Agents; Bone Resorption; Disease Models, Animal; Drug Administration Schedule; Isoenzymes; Male; Mandible; Microvessels; Neovascularization, Physiologic; Periapical Diseases; Rats; Rats, Sprague-Dawley; Statistics, Nonparametric; Tartrate-Resistant Acid Phosphatase

Periostin is a gamma-carboxyglutamic acid protein preferentially expressed in periosteum and bone mesenchymal stem cells. Lack of a precise assay for measuring circulating levels impairs the investigation of its biological significance. We developed a new ELISA and studied changes of periostin levels both locally at the bone site and systemically in circulating blood during growth and after bisphosphonate-induced inhibition of bone remodeling in the mouse. The ELISA we developed is based on an affinity-purified polyclonal antibody that was raised against the C-terminal sequence of mouse periostin. Reproducibility, repeatability, precision, and accuracy tests met standards of acceptance. Serum periostin and levels of the bone turnover markers osteocalcin, PINP, CTX-I, and TRAP5b were measured in (1) 4-, 6-, 8-, 10-, and 12-week-old wild-type female Balb/c mice and (2) adult ovariectomized female Balb/c mice treated with zoledronic acid or vehicle. Serum periostin decreased during growth and stabilized from 8 weeks and older, its levels correlating with bone turnover markers. Immunohistochemistry in bones from different growth stages showed that periostin localized specifically at the sites of endochondral and intramembranous ossification, especially at the periosteal envelopes. Zoledronic acid induced a marked decrease in bone remodeling markers but did not alter serum periostin levels or periostin immunostaining pattern. The novel ELISA is highly specific and allows accurate and precise measurements of serum periostin levels in mice.

Topics: Acid Phosphatase; Animals; Biomarkers; Bone Density Conservation Agents; Bone Remodeling; Cell Adhesion Molecules; Collagen Type I; Diphosphonates; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Fluorescent Antibody Technique, Direct; Imidazoles; Immunoenzyme Techniques; Isoenzymes; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovariectomy; Peptide Fragments; Peptides; Procollagen; Reproducibility of Results; Tartrate-Resistant Acid Phosphatase; Tibia; Zoledronic Acid

The purpose of this study was to investigate the underlying mechanism in favorable prognosis following pediatric condylar fractures managed by closed procedures. Seventy-five 1-month-old male Wistar rats were used in this experiment. Unilateral medially rotated condyle fracture in growing rats was adopted as the condyle fracture model to investigate the mechanism in favorable healing of pediatric condylar fractures. The entire fracture healing process was investigated. The rotated subcondylar fractures in young rats healed by means of callus formation, with simultaneous and prompt repositioning of the condyle. The positive outcome in these condyle fractures was also associated with active cell proliferation potential in the condyle, as well as the condyle's remodeling capability. The growth potential and remodeling capability of the condyle during the growing period might be the intrinsic factor for favorable healing following pediatric condylar fractures managed by closed procedures.

Topics: Acid Phosphatase; Animals; Azo Compounds; Biomarkers; Body Weight; Bone Remodeling; Bony Callus; Cartilage, Articular; Cell Proliferation; Chondrocytes; Coloring Agents; Disease Models, Animal; Eosine Yellowish-(YS); Fracture Healing; Isoenzymes; Joint Dislocations; Male; Mandibular Condyle; Mandibular Fractures; Methyl Green; Osteogenesis; Phenazines; Prognosis; Proliferating Cell Nuclear Antigen; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Treatment Outcome

Periodontal disease is characterised by alveolar bone loss. Some studies have suggested the involvement of sympathetic nervous system in the deterioration of periodontal disease. Noradrenaline, released from sympathetic nerve terminals due to various stimuli, binds to specific adrenergic receptors on immune cells. Recently, we reported that restraint stress augmented the alveolar bone loss induced by Porphyromonas gingivalis infection. In this study, we investigated the effects of the beta-blocker (propranolol) on alveolar bone loss induced by P. gingivalis infection to examine the involvement of sympathetic nerves in periodontal breakdown.. Sprague-Dawley rats were treated as follows: saline injection (Group A), propranolol injection (Group B), saline injection and oral challenge with P. gingivalis (Group C), and propranolol injection and oral challenge with P. gingivalis (Group D). Horizontal alveolar bone loss was evaluated by measuring the distance between the cemento-enamel junction and the alveolar bone crest. Specimens from periodontal tissue were evaluated by staining with hematoxylin-eosin and tartrate-resistant acid phosphatase.. Blockade of beta-receptors in periodontal tissue by propranolol inhibited osteoclast differentiation and prevented alveolar bone loss induced by P. gingivalis infection. Histological study revealed that the number of osteoclasts detected was proportional to the level of bone loss.. These results indicate that the sympathetic nervous system is involved in the development of periodontitis and suggest that sympathetic signal modulation with beta-blockers enables the control of alveolar bone mass metabolism.

Topics: Acid Phosphatase; Adrenergic beta-Antagonists; Alveolar Bone Loss; Alveolar Process; Animals; Bacteroidaceae Infections; Biomarkers; Body Weight; Cell Differentiation; Coloring Agents; Disease Models, Animal; Fluorescent Dyes; Isoenzymes; Male; Organ Size; Osteoclasts; Periodontitis; Porphyromonas gingivalis; Propranolol; Rats; Rats, Sprague-Dawley; Spleen; Sympathetic Nervous System; Tartrate-Resistant Acid Phosphatase; Thymus Gland

Osteosarcoma (OS) is the most common primary malignant bone tumour, and mainly affects adolescents and young adults. Although there has been substantial improvement in management of OS with surgery and chemotherapy, further survival increase has not been achieved over the past two decades.. We focused on the receptor activator of nuclear factor kappaB ligand (RANKL)-osteoclast (OCL) system as a biological target for OS. RANKL is a critical factor for OCL formation and bone resorption activity. The primary lesion in bone and ensuing metastasis in OS both require the induction of OCLs. RANK-Fc is a potent RANKL antagonist and inhibitor of OCL formation and activity.. In an orthotopic model in Balb/c nu/nu mice, a twice weekly dosing regimen of 350 microg of RANK-Fc per mouse subcutaneously (n= 5) reduced lung metastasis (P > 0.05), preserved bone structure and reduced tartrate-resistant acid phosphatase (TRAP)(+) OCLs (P < 0.005) in OS-bearing bone. In vitro, RANK-Fc suppressed OCL formation (P < 0.005), bone resorption activity (P < 0.005) and RANKL-induced anti-apoptosis (P < 0.5) of OCLs.

Topics: Acid Phosphatase; Animals; Apoptosis; Bone and Bones; Bone Neoplasms; Bone Resorption; Disease Models, Animal; Humans; Isoenzymes; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Knockout; Osteoclasts; Osteosarcoma; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Recombinant Fusion Proteins; Tartrate-Resistant Acid Phosphatase

Breast cancer metastases to bone are common in advanced stage disease. We have recently demonstrated that vitamin D deficiency enhances breast cancer growth in an osteolytic mouse model of breast cancer metastasis. In this study, we examined the effects of vitamin D deficiency on tumor growth in an osteosclerotic model of intra-skeletal breast cancer in mice.. The effects of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] on proliferation and apoptosis of MCF-7 breast cancer cells, and changes in the expression of genes within the vitamin D metabolic pathway (VDR, 1α- and 24-hydroxylase) were examined in vitro. MCF-7 breast cancer cells were injected intra-tibially into vitamin D deficient and vitamin D sufficient mice co-treated with and without osteoprotegerin (OPG). The development of tumor-related lesions was monitored via serial X-ray analysis. Tumor burden and indices of proliferation and apoptosis were determined by histology along with markers of bone turnover and serum intact PTH levels.. In vitro, MCF-7 cells expressed critical genes for vitamin D signalling and metabolism. Treatment with 1,25(OH)(2)D(3) inhibited cell growth and proliferation, and increased apoptosis. In vivo, osteosclerotic lesions developed faster and were larger at endpoint in the tibiae of vitamin D deficient mice compared to vitamin D sufficient mice (1.49±0.08 mm(2) versus 1.68±0.15 mm(2), P<0.05). Tumor area was increased by 55.8% in vitamin D deficient mice (0.81±0.13 mm(2) versus 0.52±0.11 mm(2) in vitamin D sufficient mice). OPG treatment inhibited bone turnover and caused an increase in PTH levels, while tumor burden was reduced by 90.4% in vitamin D sufficient mice and by 92.6% in vitamin D deficient mice. Tumor mitotic activity was increased in the tibiae of vitamin D deficient mice and apoptosis was decreased, consistent with faster growth.. Vitamin D deficiency enhances both the growth of tumors and the tumor-induced osteosclerotic changes in the tibiae of mice following intratibial implantation of MCF-7 cells. Enhancement of tumor growth appears dependent on increased bone resorption rather than increased bone formation induced by these tumors.

Topics: Acid Phosphatase; Adipose Tissue; Animals; Apoptosis; Bone and Bones; Bone Neoplasms; Bone Remodeling; Breast Neoplasms; Calcitriol; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Humans; Isoenzymes; Mice; Osteolysis; Osteosclerosis; Radiography; Tartrate-Resistant Acid Phosphatase; Tumor Burden; Vitamin D Deficiency; Xenograft Model Antitumor Assays

Parathyroid hormone-related protein (PTHrP) is an important regulator of bone formation and remodeling. Our recent findings demonstrate that PTHrP (107-111) (osteostatin) loaded onto silica-based ordered mesoporous SBA15 materials exhibit osteogenic features in osteoblastic cell cultures. We aimed here to elucidate whether these peptide-coated materials might be suitable for promoting bone repair following a cavitary defect in the rabbit femur. Histological examination revealed the absence of significant inflammation or bone resorption within the time of study (4 and 8 weeks) after implantation. At 8 weeks, the peptide-unloaded materials were still separated from the bone marrow by a fibrous cap, which was greatly diminished by the presence of the PTHrP peptide. By using μCT analysis, new bone formation was evident at different distances from the implants, mainly for the latter peptide-loaded biomaterials. This was confirmed by performing immunostaining for different osteoblast markers. Our findings demonstrate that these PTHrP (107-111)-loaded bioceramics significantly improve local bone induction, as compared to that observed with the unloaded material.

Topics: Acid Phosphatase; Animals; Bone Regeneration; Coated Materials, Biocompatible; Core Binding Factor Alpha 1 Subunit; Disease Models, Animal; Femur; Humans; Immunohistochemistry; Implants, Experimental; Isoenzymes; Osteocalcin; Osteogenesis; Osteopontin; Parathyroid Hormone-Related Protein; Peptide Fragments; Porosity; Proliferating Cell Nuclear Antigen; Rabbits; Silicates; Tartrate-Resistant Acid Phosphatase; Vascular Endothelial Growth Factor A; X-Ray Microtomography

Squamous cell carcinoma (SCC) is the most common form of oral cancer. Destruction and invasion of mandibular and maxillary bone frequently occurs and contributes to morbidity and mortality. We hypothesized that the bisphosphonate drug zoledronic acid (ZOL) would inhibit tumor-induced osteolysis and reduce tumor growth and invasion in a murine xenograft model of bone-invasive oral SCC (OSCC) derived from an osteolytic feline OSCC. Luciferase-expressing OSCC cells (SCCF2Luc) were injected into the perimaxillary subgingiva of nude mice, which were then treated with 100 μg/kg ZOL or vehicle. ZOL treatment reduced tumor growth and prevented loss of bone volume and surface area but had no effect on tumor invasion. Effects on bone were associated with reduced osteolysis and increased periosteal new bone formation. ZOL-mediated inhibition of tumor-induced osteolysis was characterized by reduced numbers of tartrate-resistant acid phosphatase-positive osteoclasts at the tumor-bone interface, where it was associated with osteoclast vacuolar degeneration. The ratio of eroded to total bone surface was not affected by treatment, arguing that ZOL-mediated inhibition of osteolysis was independent of effects on osteoclast activation or initiation of bone resorption. In summary, our results establish that ZOL can reduce OSCC-induced osteolysis and may be valuable as an adjuvant therapy in OSCC to preserve mandibular and maxillary bone volume and function.

Topics: Acid Phosphatase; Animals; Bone Density Conservation Agents; Bone Resorption; Calcium; Carcinoma, Squamous Cell; Cats; Diphosphonates; Disease Models, Animal; Imidazoles; Isoenzymes; Luciferases; Male; Mice; Mice, Nude; Mouth Neoplasms; Osteoclasts; Osteolysis; Tartrate-Resistant Acid Phosphatase; Transplantation, Heterologous; X-Ray Microtomography; Zoledronic Acid

PTH has been shown to enhance fracture repair; however, exactly when and where PTH acts in this process remains to be elucidated. Therefore, we conducted a longitudinal, region-specific analysis of bone regeneration in mature, osteopenic rats using a cortical defect model. Six-month-old rats were ovariectomized, and allowed to lose bone for 2 months, before being subjected to bilateral 2-mm circular defects in their femoral diaphyses. They were then treated for 5 wk with hPTH1-38 at doses of 0, 3, 10, or 30 microg/kg . d and scanned weekly by in vivo quantitative computed tomography. Quantitative computed tomography analyses showed temporal, dose-dependent increases in mineralization in the defects, intramedullary (IM) spaces, and whole diaphyses at the defect sites. Histomorphometry confirmed PTH stimulation of primarily woven bone in the defects and IM spaces, but not the periosteum. After necropsy, biomechanical testing identified an increase in strength at the highest PTH dose. Serum procollagen type I N-terminal propeptide concentration showed a transient increase due to drilling, but procollagen type I N-terminal propeptide also increased with PTH treatment, whereas tartrate-resistant acid phosphatase unexpectedly decreased. Analyses of lumber vertebra confirmed systemic efficacy of PTH at a nonfracture site. In summary, PTH dose dependently induced new bone formation within defects, at endocortical surfaces, and in IM spaces, resulting in faster and greater bone healing, as well as efficacy at other skeletal sites. The effects of PTH were kinetic, region specific, and most apparent at high doses that may not be entirely clinically relevant; therefore, clinical studies are necessary to clarify the therapeutic utility of PTH in bone healing.

Topics: Acid Phosphatase; Animals; Biomechanical Phenomena; Bone Density; Bone Regeneration; Collagen Type I; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Femur; Isoenzymes; Ovariectomy; Parathyroid Hormone; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Tomography Scanners, X-Ray Computed

Male hypogonadism is frequently associated with testopathy in patients with type 2 diabetes and in middle-aged males. We hypothesized that abnormal matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in testis have large roles to play in male hypogonadism. It has been found in diabetic rats that a novel compound, strontium fructose 1,6-diphosphate (FDP-Sr), with extra high energy supply, could reverse male hypogonadism by normalizing MMP-9 and TIMPs in the testis. We investigated whether FDP-Sr could be promising in treating diabetic testopathy.. Adult male Sprague-Dawley rats were administered a single dose of streptozocin (65 mg/kg, i.p.) to induce diabetes. The diabetic rats were treated with FDP-Sr in three doses or testosterone propionate in the final four weeks during the eight-week study.. Serum testosterone, activity of marker enzymes, and mRNA of MMPs and TIMPs and protein of MMP-9 in the testis were detected. After eight weeks, the activity of acid phosphatase, lactate dehydrogenase, succinate dehydrogenase and g-glutamyl transpeptidase in testis were significantly decreased (P < 0.01), accompanied by down-regulated mRNA and activity of MMP-2 and MMP-9 (P < 0.01) and upregulated mRNA of TIMP-1 and TIMP-2. Downregulated MMP-9 protein and degenerative changes in histology were predominant in diabetic testis.. FDP-Sr or testosterone propionate significantly normalized expression and activity of the MMPs-TIMPs system to attenuate changes in serum testosterone, marker enzymes and histology in testis. Effects of FDP-Sr were dose-dependent and comparable with those of testosterone propionate. By supplying extra energy, FDP-Sr could be promising in treating diabetic testopathy by normalizing abnormal MMP-9 and its endogenous inhibitors in testes.

Topics: Acid Phosphatase; Animals; Diabetes Complications; Diabetes Mellitus, Type 2; Disease Models, Animal; Dose-Response Relationship, Drug; Fructosediphosphates; gamma-Glutamyltransferase; Gene Expression; Hyperglycemia; Hypogonadism; L-Lactate Dehydrogenase; Male; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases; Rats; Rats, Sprague-Dawley; RNA, Messenger; Streptozocin; Succinate Dehydrogenase; Testis; Testosterone; Tissue Inhibitor of Metalloproteinases

It has been suggested that hormones released after nutrient absorption, such as glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide 2 (GLP-2), could be responsible for changes in bone resorption. However, information about the role of GLP-1 in this regard is scanty. Diabetes-related bone loss occurs as a consequence of poor control of glucose homeostasis, but the relationship between osteoporosis and type 2 diabetes remains unclear. Since GLP-1 is decreased in the latter condition, we evaluated some bone characteristics in streptozotocin-induced type 2 diabetic (T2D) and fructose-induced insulin-resistant (IR) rat models compared to normal (N) and the effect of GLP-1 or saline (control) treatment (3 days by osmotic pump). Blood was taken before and after treatment for plasma measurements; tibiae and femora were collected for gene expression of bone markers (RT-PCR) and structure (microCT) analysis. Compared to N, plasma glucose and insulin were, respectively, higher and lower in T2D; osteocalcin (OC) and tartrate-resistant alkaline phosphatase 5b were lower; phosphate in IR showed a tendency to be higher; PTH was not different in T2D and IR; all parameters were unchanged after GLP-1 infusion. Bone OC, osteoprotegerin (OPG) and RANKL mRNA were lower in T2D and IR; GLP-1 increased OC and OPG in all groups and RANKL in T2D. Compared to N, trabecular bone parameters showed an increased degree of anisotropy in T2D and IR, which was reduced after GLP-1. These findings show an insulin-independent anabolic effect of GLP-1 and suggest that GLP-1 could be a useful therapeutic agent for improving the deficient bone formation and bone structure associated with glucose intolerance.

Topics: Acid Phosphatase; Animals; Bone and Bones; Bone Resorption; Diabetes Mellitus, Type 2; Disease Models, Animal; Glucagon-Like Peptide 1; Glucose; Insulin; Insulin Resistance; Isoenzymes; Male; Osteocalcin; Osteoprotegerin; Parathyroid Hormone; Peptide Fragments; RANK Ligand; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Craniometaphyseal dysplasia (CMD) is a monogenic human disorder characterized by thickening of craniofacial bones and flaring metaphyses of long bones. Mutations for autosomal dominant CMD have been identified in the progressive ankylosis gene ANKH. Previous studies of Ank loss-of-function models, Ank(null/null) and Ank(ank/ank) mice, suggest that Ank plays a role in the regulation of bone mineralization. However, the mechanism for Ank mutations leading to CMD remains unknown. We generated the first knockin (KI) mouse model for CMD expressing a human mutation (Phe377 deletion) in ANK. Homozygous Ank knockin mice (Ank(KI/KI)) replicate many typical features of human CMD including hyperostosis of craniofacial bones, massive jawbones, decreased diameters of cranial foramina, obliteration of nasal sinuses, fusion of middle ear bones, and club-shaped femurs. In addition, Ank(KI/KI) mice have increased serum alkaline phosphatase and TRACP5b, as reported in CMD patients. Biochemical markers of bone formation and bone resorption, N-terminal propeptide of type I procollagen and type I collagen cross-linked C-terminal telopeptide, are significantly increased in Ank(KI/KI) mice, suggesting increased bone turnover. Interestingly, Ank(KI/KI) bone marrow-derived macrophage cultures show decreased osteoclastogenesis. Despite the hyperostotic phenotype, bone matrix in Ank(KI/KI) mice is hypomineralized and less mature, indicating that biomechanical properties of bones may be compromised by the Ank mutation. We believe this new mouse model will facilitate studies of skeletal abnormalities in CMD at cellular and molecular levels.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Diseases, Developmental; Collagen Type I; Disease Models, Animal; Humans; Isoenzymes; Macrophages; Membrane Proteins; Mice; Mice, Transgenic; Phosphate Transport Proteins; Sequence Deletion; Skull; Tartrate-Resistant Acid Phosphatase

Rett syndrome (RTT), a neurological disorder characterized by neurological impairment and a high frequency of osteopenia which often manifests early in childhood, most often is caused by inactivating mutations in the X-linked gene encoding a regulator of epigenetic gene expression, methyl CpG binding protein, MeCP2. Clinical data show that, along with neurological defects, females with RTT frequently have marked decreases in bone mineral density (BMD) beyond that expected from disuse atrophy. To investigate the relationship between loss of Mecp2 and reduced BMD, we used a Mecp2 null mouse model, Mecp2 (-/yBIRD), for our histological and biochemical studies. Mecp2 (-/yBIRD) mice have significantly shorter femurs and an overall reduced skeletal size compared to wild-type mice by post-natal day 60 (P60). Histological and histomorphometric studies identified growth plate abnormalities as well as decreased cortical and trabecular bone in P21 and especially in P60 Mecp2 (-/yBIRD) mice. Dynamic histomorphometry revealed decreased mineral apposition rates (MAR) in Mecp2 null femoral trabecular bone as well as in calvarial bone samples. While changes in MAR of cortical bone were not significant, loss of Mecp2 significantly reduced cortical, trabecular and calvarial bone volume compared with age-matched wild-type animals. These differences indicate that Mecp2 deficiency leads to osteoblast dysfunction, which translates into reduced osteoid deposition accounting for the reduced bone volume phenotype. While individual variations were observed in OPG and Rankl concentrations, molar ratios of OPG:Rankl at P21 and P60 were comparable between wild-type and Mecp2 (-/yBIRD) mice and showed a consistent excess of OPG. In tibial sections, TRAP staining demonstrated equivalent osteoclast number per bone surface measurements between wild-type and null animals. Our work with a Mecp2 null mouse model suggests epigenetic regulation of bone in the Mecp2 (-/yBIRD) mice which is associated with decreased osteoblast activity rather than increased osteoclastic bone loss.

Topics: Acid Phosphatase; Animals; Bone and Bones; Cell Count; Disease Models, Animal; Femur; Growth Plate; Isoenzymes; Male; Methyl-CpG-Binding Protein 2; Mice; Organ Size; Osteoclasts; Osteogenesis; Osteoprotegerin; RANK Ligand; Rett Syndrome; Skull; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography

To examine whether grape seed proanthocyanidin extract (GSPE) which is known to act as an antioxidant has therapeutic effect on collagen-induced arthritis (CIA) in mice, an animal model of rheumatoid arthritis. Mice were treated with an intraperitoneal injection of GSPE (10, 50, or 100 mg/kg) or saline. Clinical, histological, and biochemical parameters were assessed. The effects of GSPE on osteoclastogenesis were determined by tartrate-resistant acid phosphatase (TRAP) staining of the inflamed joints and bone-marrow cells cultured with the receptor activator of nuclear factor B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). Intracellular levels of hydrogen peroxide were determined using carboxy-dichlorodihydrofluorescein diacetate. GSPE treatment significantly attenuated the severity of CIA in a dose-dependent manner and reduced the histology scores for synovial inflammation, cartilage erosion, bone erosion, and the number of TRAP+ osteoclasts. GSPE treatment significantly reduced the numbers of tumor necrosis factor alpha (TNF-alpha)- or interleukin 17 (IL-17)-producing cells in the synovial tissue and the spontaneous production of TNF-alpha and IL-17 by splenocytes compared with those in the control mice. The serum levels of type-II-collagen-specific IgG2a and plasma levels of 8-isoprostane in the GSPE-treated mice were significantly lower than those in the control mice. GSPE dose-dependently suppressed osteoclastogenesis in vitro. GSPE significantly reduced hydrogen peroxide production by anti-CD3-monoclonal-antibody-stimulated CD4+ splenocytes. These results indicate that intraperitoneal injection of GSPE attenuated CIA in mice. GSPE may be useful in the treatment of rheumatoid arthritis.

Topics: Acid Phosphatase; Animals; Ankle Joint; Antibodies; Arthritis, Experimental; Arthritis, Rheumatoid; Cells, Cultured; Collagen Type II; Disease Models, Animal; Grape Seed Extract; Hydrogen Peroxide; Interleukin-17; Isoenzymes; Isoprostanes; Macrophage Colony-Stimulating Factor; Mice; Mice, Inbred DBA; Osteoclasts; Plant Extracts; Proanthocyanidins; RANK Ligand; Spleen; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

Intermittent administration of the parathyroid hormone (1-34) has an anabolic effect on bone and it has been shown to reduce alveolar bone loss in experimental periodontitis models. The aim of the present study was to investigate the effect of parathyroid hormone on tissue degradation-related factors in an experimental periodontitis model in rats.. Periodontitis was induced in seventy-six male Wistar rats using ligature around the lower right first molars. The animals were then treated with parathyroid hormone (1-34) (T-group) or vehicle (C-group), three times a week for 15 d (C15, T15) or 30 d (C30, T30). At each experimental time-point, the 19 rats were killed in each group and the gingival tissue around the first lower molar was removed and prepared for the following analyses: mRNA expression of interleukin-1 beta, interleukin-6, matrix metalloproteinase (MMP)-2 and MMP-9, and gelatinolytic activity of MMP-2 and MMP-9. Hemimandibles were decalcified, and serial sections were processed and analyzed for interleukin-6 immohistochemistry. Samples were also histochemically stained by tartrate-resistant acid phosphatase (TRAP) to evaluate the number of osteoclasts present.. Parathyroid hormone-treated samples showed decreased of levels of mRNA for interleukin-6 in the T30 group (p < 0.01) and of MMP-2 in the T15 and T30 groups (p < 0.05). Zymography assays demonstrated that treatment with parathyroid hormone led to a decrease in MMP-9 activity (p < 0.01). TRAP staining of alveolar bone revealed that osteoclasts were present in higher numbers (p < 0.05) in the groups not treated with parathyroid hormone.. These data suggest that intermittent administration of parathyroid hormone can down-regulate the expression of biomarkers responsible for connective tissue breakdown and bone resorption, and potentially affect alveolar bone resorption activity.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Cell Count; Connective Tissue; Disease Models, Animal; Down-Regulation; Gingiva; Injections, Subcutaneous; Interleukin-1beta; Interleukin-6; Isoenzymes; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Osteoclasts; Parathyroid Hormone; Periodontitis; Rats; Rats, Wistar; RNA, Messenger; Tartrate-Resistant Acid Phosphatase; Time Factors

Oral malodor is mainly attributed to volatile sulphur compounds (VSCs) such as hydrogen sulphide (H(2)S), methyl mercaptan and dimethyl sulphide. VSC accelerate periodontal soft tissue destruction. However, there is little information about the potential role of H(2)S in alveolar bone loss. The purpose of this animal study was to examine the effects of sodium hydrogen sulphide (NaHS), H(2)S donor drug, on osteoclast differentiation in rat periodontal tissue.. Twenty-four male Wistar rats (8 weeks old) were divided into four groups: a control group and three experimental groups, which were examined at 3h, 1 day, and 3 days after topical application of 3microl NaHS (lM in physiological saline) into the gingival sulcus of rat first molar. Expression of tumour necrosis factor (TNF)-alpha, RANKL, NF-kappaB and tartrate-resistant acid phosphatase (TRAP) was evaluated in the periodontal tissue.. Three hours after NaHS application, TNF-alpha expression increased in the periodontal ligament. The numbers of RANKL-positive osteoblasts and TRAP-positive osteoclasts significantly increased progressively with time and reached a maximum level after 1 day. Significant up-regulation of RANKL and NF-kappaB mRNA was observed at 3h after NaHS application.. H(2)S application caused a transient increase of osteoclast differentiation with up-regulation of RANKL expression in osteoblasts. H(2)S, which is primarily responsible for halitosis, may also contribute to alveolar bone resorption through RANKL expression.

Topics: Acid Phosphatase; Administration, Topical; Alveolar Process; Animals; Biomarkers; Cell Differentiation; Connective Tissue; Disease Models, Animal; Epithelial Attachment; Extracellular Space; Gingiva; Hydrogen Sulfide; Isoenzymes; Male; NF-kappa B; Osteoclasts; Periodontal Ligament; Periodontium; Random Allocation; RANK Ligand; Rats; Rats, Wistar; Sulfides; Tartrate-Resistant Acid Phosphatase; Time Factors; Tumor Necrosis Factor-alpha; Up-Regulation

Orthodontic therapy is known to have an aggravating effect on the progression of destructive periodontitis if oral hygiene is not maintained. However, it is largely unknown how active periodontitis affects the velocity of orthodontic tooth movement. In this study, we examined the effect of periodontal inflammation on orthodontic tooth movement using a mouse model. Orthodontic force was applied on the maxillary first molar of mice, with or without ligature wire to induce experimental periodontitis. The distance moved by the first molar was significantly reduced by the ligature-induced experimental periodontitis. Tartrate-resistant acid phosphatase staining revealed that the number of osteoclasts present during orthodontic treatment was lower in the pressure zone of alveolar bone in the presence of periodontal inflammation. Consistently, the expression level of receptor activator of nuclear factor-kappaB ligand (RANKL) in the pressure zone was decreased in the ligature group. By contrast, experimental periodontitis increased the expression of cyclooxygenase-2 mRNA in the periodontal tissues, while in vitro treatment with prostaglandin E(2) decreased extracellular signal-regulated kinase phosphorylation and RANKL expression induced by mechanical stress in osteoblasts. Taken together, these results suggest that the orthodontic force-induced osteoclastogenesis in alveolar bone was inhibited by the accompanying periodontal inflammation, at least partly through prostaglandin E(2), resulting in reduced orthodontic tooth movement.

Topics: 3T3 Cells; Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Cyclooxygenase 2; Dinoprostone; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Isoenzymes; Male; Maxilla; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Molar; Osteoblasts; Osteoclasts; Periodontitis; Phosphorylation; RANK Ligand; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques

Bone metastasis is a frequent and catastrophic consequence of prostate cancer for which only palliative treatment is available. Animal models of bone metastatic prostate cancer are necessary for understanding disease mechanisms but few models exist.. We have used the murine prostate carcinoma cell line RM1 to generate a bone metastatic model of prostate cancer. Repeated intracardiac injection of RM1 cells followed by isolation of cells from bone tumors has yielded a cell line with strong bone-metastatic potential, RM1 bone metastatic (BM).. This cell line metastasizes to multiple bony sites in over 95% of injected C57BL/6 mice and is far less tropic to soft tissues. Bone tumors produced by the RM1(BM) cell line show no preference for particular skeletal sites as most bones are affected. Histology, and micro-computed tomography show that RM1(BM) cells form osteolytic tumors, but with evidence of osteoblastic changes. In vitro the RM1 cells express E-cadherin but not vimentin, do not form colonies in soft agar, are non-invasive but are more motile than the parent cell line.. This model provides a novel means for identifying cellular and molecular mechanisms that contribute to bone metastasis and allow for preclinical testing of therapies to prevent and treat tumor metastasis to bone. Finally as the syngeneic tumor cells are injected into immunocompetent mice, this model will provide a means to study interactions between the immune system, tumors and bone, and therapies that target such interactions.

Topics: Acid Phosphatase; Animals; Bone Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Movement; Disease Models, Animal; Histocytochemistry; Immunocompetence; Male; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence; Osteocalcin; Prostatic Neoplasms; Tomography, X-Ray Computed

A new spontaneous mouse mutant (ntl) with autosomal-recessive osteopetrosis was characterized. These mice formed tartrate-resistant acid phosphate (TRAP)-positive osteoclasts but their osteoclasts had no ruffled border and did not resorb bone. These mice displayed no tooth eruption or tooth root formation. Adult mutant mice developed odontoma-like proliferations near the proximal ends of the incisors. Intraperitoneal injection of progenitor cells from the liver of 16.5 days postcoitum wild-type embryos into newborn mutants rescued the osteopetrosis phenotype, indicating that the defects were intrinsic to the osteoclasts. Our findings not only provide further support for a critical role of osteoclasts in tooth eruption and tooth root development, but also suggest that the perturbation of the homeostasis of the odontogenic precursors of the incisors is primarily responsible for the development of the odontoma-like proliferations in this osteopetrosis mutant. Genetic mapping has narrowed down the location of the mutant allele to a genetic interval of 3.2 cM on mouse chromosome 17.

Topics: Acid Phosphatase; Alleles; Animals; Biomarkers; Bone Resorption; Cells, Cultured; Chloride Channels; Chromosomes, Mammalian; Disease Models, Animal; Genes, Recessive; Genetic Linkage; Genotype; Homeostasis; Incisor; Isoenzymes; Liver; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Mutant Strains; Mutation; Odontogenesis; Odontoma; Osteoclasts; Osteopetrosis; Phenotype; Stem Cells; Tartrate-Resistant Acid Phosphatase; Tooth Eruption; Tooth Root

Rheumatoid arthritis (RA) is a kind of chronic inflammatory autoimmune disease. The degradation of extracellular matrix and cartilage pave way in understanding the molecular mechanisms in RA. Degradation of cartilage is a more complex event involving the local release of metallaoproteases and lysosomal enzymes that mediate inflammation in joints and in the synovial fluid in RA.. In the present study, the efficacy of a Siddha preparation named Kalpaamruthaa (KA) in ameliorating the disease process via markedly reducing the joint destruction was demonstrated in adjuvant induced arthritis rat model. KA consists of Semecarpus anacardium nut milk extract (SA), dried powder of Emblica officinalis fruit and honey.. Both SA and KA were administered at dose of 150 mg/kg b.wt. for 14 days after 14 days of adjuvant injection in rats. The activity of lysosomal enzymes, the level of collagen, glycosaminoglycans (GAGs) and its degradative products were analyzed in control and experimental animals.. The study revealed that KA exhibited a profound reduction (p<0.05) in the activities of lysosomal enzymes and thereby decreasing (p<0.05) the levels of GAGs and its fractions when compared to arthritis rats. The latter was confirmed by Safrannin O staining for GAGs in the interphalangeal joints of control and experimental animals. The effect of KA was found to be improved than SA and this might be due to the combined interactions of phytoconstituents present in KA.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Arthritis, Rheumatoid; Aspartic Acid Endopeptidases; Blood Proteins; Cathepsin D; Collagen; Disease Models, Animal; Enzyme Activation; Glycosaminoglycans; Glycoside Hydrolases; Hip Joint; Lysosomes; Male; Plant Extracts; Rats; Rats, Wistar

External apical root resorption (EARR) is an unwanted sequelae of orthodontic treatment. Genetic factors account for approximately 64% of the EARR variation in humans. Inbred mice offer a model to control the environmental factors and genetic heterogeneity that complicate human genetic studies. Genetically distinct inbred mice and their offspring (F1s) were analyzed to examine the mode of inheritance and the influence of parental sex on the susceptibility to root resorption associated with orthodontic force (RRAOF).. RRAOF was determined histologically for male and female mice of the A/J, DBA/2J, and BALB/cJ strains, and the A/JxDBA/2J and A/JxBALB/cJ crosses (10 males and 10 females/reciprocal cross). RRAOF was induced by tipping the maxillary first molar mesially for 9 days.. Sex differences were observed only among the mice of the BALB/cJ strain. Two patterns of inheritance were observed; F1s from the A/JxBALB/cJ cross were resistant, suggesting that the A/J have dominant resistance alleles. On the other hand, F1s from the A/JxDBA/2J cross showed RRAOF intermediate between their parental mice, suggesting a polygenic trait.. These results provide evidence of a traceable and polygenetic component affecting RRAOF in mice.

Topics: Acid Phosphatase; Alleles; Animals; Biomarkers; Crosses, Genetic; Disease Models, Animal; Female; Genes, Dominant; Genetic Predisposition to Disease; Isoenzymes; Male; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred DBA; Mice, Inbred Strains; Molar; Multifactorial Inheritance; Root Resorption; Sex Factors; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques

To study the effects of estrogen withdrawal on osteoclast number and osteoclast activity in the rat ovariectomy (OVX) model.. We first cultured human CD34+ osteoclast precursor cells on bovine bone slices, allowing them to differentiate into mature resorbing osteoclasts. Secreted tartrate-resistant acid phosphatase 5b (TRACP 5b) and C-terminal cross-linked telopeptides of type I collagen (CTX) were determined from the culture medium. TRACP 5b correlated strongly with osteoclast number and CTX with osteoclast activity, facilitating their subsequent use in the rat OVX model. An 8 week OVX study was then performed including sham-operated rats receiving vehicle, OVX rats receiving vehicle, and OVX rats receiving 10 microg/kg/day 17 beta-estradiol (E2). Trabecular bone parameters were determined from the tibial metaphysis using peripheral quantitative computed tomography and histomorphometry. Osteoclast number was normalized with bone perimeter (N.Oc/B.Pm) and tissue area (N.Oc/T.Ar, indicating absolute number of osteoclasts). TRACP 5b and CTX were determined from fasting serum samples.. Trabecular bone parameters indicated substantial bone loss after OVX that was prevented by E2. N.Oc/B.Pm increased after OVX, while N.Oc/T.Ar and TRACP 5b decreased, and TRACP 5b correlated strongly with N.Oc/T.Ar. However, CTX values increased after OVX, and the "resorption index" CTX/TRACP 5b showed more substantial changes than either CTX or TRACP 5b alone.. These results show that TRACP 5b is a reliable marker of osteoclast number, and the index CTX/TRACP 5b is a useful parameter in rat OVX model. The high elevation of CTX/TRACP 5b values by OVX demonstrates that estrogen withdrawal generates high activity of osteoclasts in the rat OVX model.

Topics: Acid Phosphatase; Animals; Biomarkers; Bone Resorption; Cattle; Cell Count; Cells, Cultured; Disease Models, Animal; Estradiol; Female; Humans; Isoenzymes; Osteoclasts; Ovariectomy; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

Osteoprotegerin (OPG) is a novel secreted member of the tumor necrosis factor receptor family which plays a crucial role in negative regulation of osteoclastic bone resorption. OPG-deficient (OPG-/-) mice develop severe osteoporosis caused by significant enhancement of bone resorption by osteoclasts. We investigated the effect of administering bisphosphonate on mandibular growth and development in OPG-/- mice. Eight-week-old male OPG-/- mice and wild-type (WT) mice were administered bisphosphonate (1.25 mg/kg body weight) intraperitoneally once every 3 days for 30 days. All bone formation-related parameters and bone resorption-related parameters were significantly lower in OPG-/- mice with bisphosphonate than in those without bisphosphonate. The volume of the whole condyle and the mandibular length in OPG-/- mice without bisphosphonate were significantly smaller than in WT mice without bisphosphonate. Bisphosphonate treatment of the OPG-/- mice resulted in an increase in the volume of the mandibular condyle and mandibular ramus length. In fact, the mandibular ramus length in OPG-/- mice with bisphosphonate was similar to the length in WT mice without bisphosphonate. Histologically, the surface irregularity of the mandibular condyle that was observed in the OPG-/- mice without bisphosphonate tended to be less marked in the OPG-/- mice with bisphosphonate, and the proportion of the area of the cartilage layer relative to the whole condyle was significantly larger in OPG-/- mice with bisphosphonate than in those without bisphosphonate. In conclusion, bisphosphonate treatment results in an increase in mandibular condylar dimensions and normalization of mandibular ramus growth.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Biomarkers; Bone Density Conservation Agents; Bone Resorption; Cell Count; Diphosphonates; Disease Models, Animal; Gene Silencing; Injections, Intraperitoneal; Isoenzymes; Male; Mandibular Condyle; Mice; Mice, Inbred C57BL; Mice, Knockout; Osteitis Deformans; Osteocalcin; Osteoclasts; Osteogenesis; Osteoprotegerin; Tartrate-Resistant Acid Phosphatase

The preferential proliferation of cancer cells in the bone microenvironment is poorly characterised. Expression pattern of bone marrow and other organ microenvironment in contact with osteolytic (Walker W256) and osteoblastic (MatLyLu MLL) metastases were investigated. Fisher and Copenhagen rats received, respectively, W256 and MLL cells injection. Bone and soft tissues were analysed by immunochemistry for DKK1, cathepsin K, RANKL, MCSF or IL6 expression. Tartrate-resistant acid phosphatase (TRAcP)-positive cells were detected by a histoenzymatic technique. In bone, expressions of MCSF and DKK1 were shown in stromal cells of the bone marrow, in contact with metastatic foci of both tumours. Many stromal cells were found RANKL positive in the vicinity of the tumours. Cells expressing cathepsin K and multinucleated TRAcP+ cells were found in direct contact with trabeculae but also in bone marrow spaces near metastatic cells. In extraosseous tumours, cells in contact with malignant cells did not expressed DKK1, MCSF, cathepsin K and IL6. Some RANKL+ cells were found in the periphery of subcutaneous tumours but may represent Langerhans cells. Abnormal presence of TRAcP+ cells was never observed in the vicinity of malignant cells. Interaction between stromal and cancer cells induces the expression on the formers of characteristics leading to osteoclastogenesis only in the bone microenvironment.

Topics: Acid Phosphatase; Animals; Bone Neoplasms; Cathepsin K; Cathepsins; Cell Line, Tumor; Disease Models, Animal; Female; Gene Expression Profiling; Immunoenzyme Techniques; Interleukin-6; Isoenzymes; Macrophage Colony-Stimulating Factor; Mammary Neoplasms, Experimental; Osteoblasts; Osteoclasts; RANK Ligand; Rats; Rats, Inbred F344; Stromal Cells; Tartrate-Resistant Acid Phosphatase

Because osteoporotic patients are prone to fractures, it must be considered whether or not patients undergoing drug therapies should discontinue treatment after sustaining a non-vertebral fracture. This study has tested the effect of novel active vitamin D3 analog, 1alpha,25-dihydroxy-2beta(3-hydroxypropoxy)vitamin D3 (ED-71), on the fracture healing comparing with a powerful anti-resorptive agent, alendronate, using a rat femoral fracture model.. Female SD rats (n=201) allocated into 6 groups were treated with MCT-vehicle and ED-71 at 0.025 and 0.05 microg/kg/day (EDL and EDH groups), and with saline-vehicle and alendronate at 5 and 10 microg/kg/day (ALL and ALH groups). After 4 weeks of pretreatment, osteotomy of the femur was performed. Treatment was continued until sacrifice at 6 and 16 weeks post-fracture. Fracture callus was evaluated by soft X-ray radiography, pQCT, biomechanical testing and histomorphometry.. At 16 weeks post-fracture, new cortical shell appeared in 100% of Control (MCT and saline-vehicle), EDL and EHL, and in 67% and 56% of ALL and ALH, respectively. ED-71 treatment showed insignificantly large callus area only at 6 weeks, while alendronate treatment induced bigger callus at both 6 and 16 weeks post-fracture. The lamellar/callus area was decreased only at 6 weeks by ED-71 treatment, but both at 6 and 16 weeks by alendronate treatment. Osteoclast number in callus surface was decreased in both ED-71 and alendronate treatment groups at 6 weeks and in EDH, ALL and ALH at 16 weeks, indicating that ED-71 inhibits osteoclastic bone resorption, but its effect is less prominent than alendronate. Almost complete callus remodeling was observed in ED-71-treated groups at 16 weeks without any significant change in structural and material properties of fractured bone.. ED-71 suppression of callus remodeling by inhibiting osteoclastic bone resorption was mild and dose-dependent and did not interfere with natural fracture healing process at 16 weeks post-fracture.

Topics: Acid Phosphatase; Animals; Bone Remodeling; Bony Callus; Calcitriol; Disease Models, Animal; Female; Femoral Fractures; Femur; Fracture Healing; Isoenzymes; Radiography; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Vitamin D

A severely osteopenic rat model was obtained by combining orchidectomy (ORX) and disuse (due to local paralysis induced by botulinum toxin [BTX] in the quadriceps muscle).. Forty-two aged male rats (5-6 months old) were randomized into three groups: 18 were SHAM operated; 6 were ORX; and 18 were ORX and BTX injected in the right hindlimb. One, two, and three months after surgery, bone mass (BV/TV) and microarchitectural parameters (Tb.Th, Tb.N, Tb.Sp, Tb.Pf, and structure model index [SMI]) were measured by microcomputed tomography (microCT) on the primary and secondary spongiosa of the femur. Osteoid parameters (OS/BS, O.Th), the number of osteoclasts (Nb.Oc), and the mineral apposition rate (Ct.MAR, Cn.MAR) were measured by histology. The serum tartrate-resistant acid phosphatase (TRAcP) 5b activity was measured by immunoassay.. ORX induced a decrease of BV/TV, Tb.N and an increase of Tb.Sp, Tb.Pf, and SMI on both primary and secondary spongiosa. ORX and BTX had cumulative effects on bone loss, since differences were maximized on the right femur. The decrease in BV/TV reached -65%. Osteoid parameters and mineral apposition rate increased during the time course of the study. A peak of serum TRAcP was found at 7 days post-ORX. TRAcP levels reached the highest values in the ORX-BTX groups and the effect lasted longer than in the group with ORX alone. The association of ORX-BTX induced a greater bone resorption, due to the removal of complete trabeculae, compared to ORX alone.. This model induced a severe and rapid bone loss and can be used to explore pharmacological- and biomaterial-based countermeasures.

Topics: Acid Phosphatase; Animals; Botulinum Toxins; Disease Models, Animal; Imaging, Three-Dimensional; Isoenzymes; Male; Orchiectomy; Osteoclasts; Osteoporosis; Paralysis; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Tomography, X-Ray Computed

Mongolian gerbils (Meriones unguiculatus) were grouped into two experimental groups: GEx.01 suffered orchiectomy and after 30 days received doses of testosterone cipionate (T), while GEx.02 received weekly and alternated doses of the anti-androgens cyproterone acetate and flutamide for 30 days, and the animals were then euthanized. Structural evaluation reveals a more intense reduction in epithelial height in GEx.02. Smooth muscle cells (SMC) presented a star-shaped aspect after 30 days of hormonal ablation and basal membrane was shown to be more intensely grooved in GEx.01. In both groups, after hormonal replacement, recovery in epithelial height could be noted and the SMC presented its phenotypes, but an increase in RER was seen, characterizing a modulation from its contractile to secreting phenotype. In conclusion, the prostate presented involution capacity after androgen ablation and the ability to reorganize after hormonal replacement, but events resulting from orchiectomy and subsequent T replacement were shown to be more aggressive to the prostate.

Topics: Acid Phosphatase; Androgen Antagonists; Animals; Cyproterone; Disease Models, Animal; Endoplasmic Reticulum, Rough; Endothelium; Flutamide; Gerbillinae; Immunohistochemistry; Male; Microscopy, Electron, Transmission; Orchiectomy; Organ Size; Prostate; Prostatic Hyperplasia; Protein Tyrosine Phosphatases; Receptors, Androgen; Testosterone

To find out the active principles against ethanol-induced toxicity in mice, Andrographis paniculata Nees. (Ap) was chosen and isolated andrographolide (ANDRO) and arabinogalactan proteins (AGPs). ANDRO was detected by HPTLC, FTIR and quantified by HPLC (10mg/g of Ap powder). AGPs was detected by beta-glucosyl Yariv staining of SDS-PAGE gel, FTIR and quantified by single radial gel diffusion assay with beta-glucosyl Yariv reagent (0.5mg/g Ap powder). The mice are pretreated intra-peritoneally (i.p.) with different doses (62.5, 125, 250, and 500mg/kg) of body weight of mice] of ANDRO and AGPs for 7 days and then ethanol (7.5g/kg of body weight) was injected, i.p. Besides, silymarin was used as standard hepatoprotective agent for comparative study with ANDRO and AGPs. The ameliorative activity of ANDRO and AGP against hepatic renal alcohol toxicity was measured by assessing GOT, GPT, ACP, ALP and LP levels in liver and kidney. It has been observed that pretreatment of mice with ANDRO and AGPs at 500mg/kg of body weight and 125mg/kg of body weight respectively could able to minimize the toxicity in compare to ethanol treated group as revealed by the different enzymatic assay in liver and kidney tissues and the results were comparable with silymarin. Hence, out of several ill-defined compounds present in Ap, ANDRO and AGPs are the potential bioactive compounds responsible for protection against ethanol-induced toxicity.

Topics: Acid Phosphatase; Alkaline Phosphatase; Andrographis; Animals; Aspartate Aminotransferases; Central Nervous System Depressants; Chromatography, High Pressure Liquid; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Ethanol; Galactans; Glucosides; India; Kidney Diseases; Lipid Peroxidation; Liver Diseases, Alcoholic; Male; Mice; Phloroglucinol; Protective Agents; Silymarin; Spectroscopy, Fourier Transform Infrared

Malaria leads to pathophysiological and biochemical alterations in placenta and blood of pregnant mice. A significant decrease in the sugar, protein and lipid levels in the placental homogenate of pregnant-infected mice was observed compared to the pregnant mice. However, serum protein content was not altered much in the pregnant-infected mice as compared to the levels in control mice. The serum lipid level enhanced significantly in both pregnant and non pregnant-infected mice. The enzymatic activities of alkaline phosphatase and acid phosphatase altered significantly in malaria-infected placenta. Our study clearly highlights the possible role of these enzymes in damaging the placenta which in turn may jeoparadise the fetal growth together with altered biochemistry of placenta. Therefore biochemical along with pathological alterations occurring during malaria infection in pregnancy may account for compromised maternal fetal relationship.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Female; Fetal Development; Lipids; Malaria; Mice; Placenta; Plasmodium berghei; Pregnancy; Pregnancy Complications, Parasitic

Prevention of alveolar bone destruction is a clinical challenge in periodontal disease treatment. The receptor activator of nuclear factor-kappa B ligand (RANKL) inhibitor osteoprotegerin (OPG) inhibits osteoclastogenesis and suppresses bone resorption.. To study the effects of RANKL inhibition on alveolar bone loss, an experimental ligature-induced model of periodontitis was used. A total of 32 rats were administered human OPG-Fc fusion protein (10 mg/kg) or vehicle by subcutaneous delivery twice weekly for 6 weeks. Negative or positive controls received no treatment or disease through vehicle delivery, respectively. Biopsies were harvested after 3 and 6 weeks, and mandibulae were evaluated by microcomputed tomography (microCT) and histology. Serum levels of human OPG-Fc and tartrate-resistant acid phosphatase-5b (TRAP-5b) were measured throughout the study by enzyme-linked immunosorbent assay (ELISA). Statistical analyses included analysis of variance (ANOVA) and Tukey tests.. Human OPG-Fc was detected in the sera of OPG-Fc-treated animals by 3 days and throughout the study. Serum TRAP-5b was sharply decreased by OPG-Fc treatment soon after OPG-Fc delivery and remained low for the observation period. Significant preservation of alveolar bone volume was observed among OPG-Fc-treated animals compared to the controls at weeks 3 and 6 (P <0.05). Descriptive histology revealed that OPG-Fc significantly suppressed osteoclast surface area at the alveolar crest.. Systemic delivery of OPG-Fc inhibits alveolar bone resorption in experimental periodontitis, suggesting that RANKL inhibition may represent an important therapeutic strategy for the prevention of progressive alveolar bone loss.

Topics: Acid Phosphatase; Alveolar Bone Loss; Analysis of Variance; Animals; Disease Models, Animal; Gene Expression Regulation; Humans; Isoenzymes; Male; Mandible; Osteoprotegerin; Periodontitis; RANK Ligand; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Statistics, Nonparametric; Tartrate-Resistant Acid Phosphatase

Cyclosporine A is an immunosuppressive drug that is widely used in organ transplant patients as well as to treat a number of autoimmune conditions. Bone loss is reported as a significant side-effect of cyclosporine A use because this can result in serious morbidity of the patients. As we have shown that cyclosporine A-associated bone loss can also affect the alveolar bone, the purpose of this study was to evaluate the effect of the concomitant administration of alendronate on alveolar bone loss in a rat model.. Forty Wistar rats (10 per group) were given cyclosporine A (10 mg/kg, daily), alendronate (0.3 mg/kg, weekly), or both cyclosporine A and alendronate, for 60 d. The control group received daily injections of sterile saline. The expression of proteins associated with bone turnover, including osteocalcin, alkaline phosphatase and tartrate-resistant acid phosphatase (TRAP), and also the calcium levels, were evaluated in the serum. Analysis of the bone volume, alveolar bone surface, the number of osteoblasts per bone surface and the number of osteoclasts per bone surface around the lower first molars was also performed.. The results indicate that cyclosporine A treatment was associated with bone resorption, represented by a decrease in the bone volume, alveolar bone surface and the number of osteoblasts per bone surface and by an increase in the number of osteoclasts per bone surface and TRAP-5b. These effects were effectively counteracted by concomitant alendronate administration.. It is concluded that concomitant administration of alendronate can prevent cyclosporine A-associated alveolar bone loss.

Topics: Acid Phosphatase; Alendronate; Alkaline Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Bone Density; Bone Density Conservation Agents; Calcium; Cell Count; Cyclosporine; Disease Models, Animal; Immunosuppressive Agents; Isoenzymes; Male; Osteoblasts; Osteocalcin; Osteoclasts; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

The effect of insulin-dependent diabetes mellitus (IDDM) on bone metabolism was evaluated using the streptozotocin (STZ)-induced diabetic rat 1 week after the induction of diabetes. The urinary excretion of cross-linked N-telopeptides of type I collagen (NTx) and deoxypyridinoline (Dpd) in diabetic rats increased to 3.6-fold and 1.2-fold the control level, respectively. The amount of hydroxyproline and calcium in the distal femur of diabetic rats significantly decreased to 76% and 90% of the control, respectively. The levels of serum osteocalcin and alkaline phosphatase (ALP) activity in the distal femur of the diabetic rats were significantly reduced to about 40% and 70% of the control levels, respectively. The decrease in the expression osteocalcin was observed in distal femur of the diabetic rats, although the level of ALP mRNA was unchanged. The activity and the mRNA level of tartrate-resistant acid phosphatase (TRAP) increased to 1.5- and 2.3-fold the control level, respectively, in distal femur of the diabetic rats. The activity, protein, and mRNA levels of cathepsin K of diabetic rats also elevated to about 2-, 2.3-, and 2-fold the control levels, respectively. These results suggest that IDDM contributes to bone loss through changes in gene expression of TRAP and cathepsin K in osteoclasts as well as osteocalcin in osteoblasts resulting in increased bone resorptive activity and decreased bone formation.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Biomarkers; Body Weight; Calcium; Cathepsin K; Cathepsins; Diabetes Mellitus, Experimental; Disease Models, Animal; Enzyme Activation; Female; Femur; Gene Expression Regulation, Enzymologic; Glucose; Hydroxyproline; Isoenzymes; Osteocalcin; Rats; Rats, Wistar; RNA, Messenger; Streptozocin; Tartrate-Resistant Acid Phosphatase

A study using rat spondylolisthesis models.. To clarify pathomechanism of vertebral rounding deformity in pediatric spondylolisthesis.. For high-grade slippage, rounding of sacrum surface associated with L5 spondylolisthesis is reported to be the most responsible risk factor. However, the exact pathomechanism of the rounding deformity is yet to be clarified.. Spondylolisthesis rat model (4-week-old) was used. Radiographs were taken weekly for 5 weeks after the surgery. The lumbar spines were harvested for histology. Hematoxylin and eosin, alcian blue staining, and tartrate-resistant acid phosphatase staining were used. Immunohistochemically, the growth plate cartilage was studied for type II and X collagen. A modified bone histomorphometric analysis was also performed.. Radiographs showed slippage 1 week after surgery. Rounding deformity was obvious 2 weeks after surgery. The rounding deformity progressed with time. Three weeks after surgery, the specific columns of growth plate were unclear at the anterior corner, which corresponded to the rounding surface observed on radiographs. Instead, a huge mass of cartilage was observed at that site. Tartrate-resistant acid phosphatase-positive cells were observed in the vicinity of the growth plate except in relation with the anterior corner. The growth plate and cartilage mass at the anterior corner stained positive for type II collagen. Chondrocytes in the hypertrophied layer stained positively for type X collagen; however, staining was faint at the anterior corner. The results suggested that the chondrocytes at the anterior did not form, morphologically and functionally, the normal growth plate. From histomorphometrical analysis, the normal posterior growth plate made endochondral bone growth in 510 +/- 20 microm for a week, whereas the anterior corner in 200 +/- 15 microm.. Deficient endochondral ossification of the growth plate in the anterior upper corner of the vertebra could be the pathomechanism of the rounding deformity of the sacrum.

Topics: Acid Phosphatase; Age Factors; Animals; Cartilage; Chondrocytes; Collagen Type II; Collagen Type X; Disease Models, Animal; Female; Growth Plate; Immunohistochemistry; Isoenzymes; Lumbar Vertebrae; Ossification, Heterotopic; Radiography; Rats; Rats, Wistar; Spondylolisthesis; Tartrate-Resistant Acid Phosphatase; Time Factors

Root resorption (RR) is an unwanted sequela of orthodontic treatment. Despite rigorous investigation, no single factor or group of factors that directly causes RR has been identified. The purpose of this study was to examine the effect of the genotype on susceptibility or resistance to develop RR secondary to orthodontic force. Nine-week-old male mice from eight inbred strains were used and randomly distributed into control (C) or treatment (T) groups as follows: A/J (C = 9,T = 9), C57BL/6J (C = 7,T = 8), C3H/HeJ (C = 8,T = 6), BALB/cJ (C = 8,T = 6), 129P3/J (C = 6,T = 8), DBA/2J (C = 8,T = 9), SJL/J (C = 8,T = 10), and AKR/J (C = 9,T = 8). Each of the treated mice received an orthodontic appliance to tip the maxillary left first molar mesially for 9 days. Histological sections of the tooth were used to determine RR and tartrate resistant acid phosphatase (TRAP) activity. The Wilcoxon ranked-sum non-parametric test was used to evaluate differences between the groups. The results showed that the DBA/2J, BALB/cJ, and 129P3/J inbred mouse strains are highly susceptible to RR, whereas A/J, C57BL/6J and SJL/J mice are much more resistant. The variation in the severity of RR associated with orthodontic force among different inbred strains of mice when age, gender, food, housing, and orthodontic force magnitude/duration are controlled support the hypothesis that susceptibility or resistance to RR associated with orthodontic force is a genetically influenced trait.

Topics: Acid Phosphatase; Animals; Biomarkers; Dental Stress Analysis; Disease Models, Animal; Genetic Predisposition to Disease; Genotype; Isoenzymes; Male; Mice; Mice, Inbred Strains; Random Allocation; Reproducibility of Results; Root Resorption; Tartrate-Resistant Acid Phosphatase; Tooth Movement Techniques

The purpose of this study was to examine the antiosteoporosis effects of garlic oil in an ovariectomized (Ovx) rat model of osteoporosis and to compare its efficacy with lovastatin (a synthetic hypocholesterolemic drug) and 17beta-estradiol (a potent antiosteoporotic agent). Animals were divided into five groups: sham-operated control, ovariectomized, ovariectomized supplemented with lovastatin, ovariectomized supplemented with garlic oil and ovariectomized supplemented with 17beta-estradiol. In our study, the development of a high rate of bone turnover and osteoporosis in the ovariectomized animals were confirmed by significant alterations of serum alkaline phosphatase activity, serum tartrate-resistant acid phosphatase activity, urinary excretion of calcium, phosphate, hydroxyproline and urinary calcium to creatinine ratio, when compared with the sham-operated control group. Supplementation of these animals with either garlic oil or lovastatin or 17beta-estradiol, in addition to their hypocholesterolemic effect, could counterbalance all these changes. The results revealed that all three compounds significantly protected the hypogonadal bone loss as reflected by higher bone densities and higher bone mineral contents than the ovariectomized group of animals. The results emphasize that, like 17beta-estradiol, the hypocholesterolemic compounds garlic oil and lovastatin are also effective in suppressing bone loss owing to estrogen deficiency and their efficacy in the order of lower to higher is garlic < lovastatin < 17beta-estradiol.

Topics: Acid Phosphatase; Alkaline Phosphatase; Allyl Compounds; Animals; Bone Density; Calcium; Cholesterol; Creatinine; Disease Models, Animal; Estradiol; Female; Humans; Hydroxyproline; Isoenzymes; Lovastatin; Osteoporosis, Postmenopausal; Ovariectomy; Phosphates; Rats; Recovery of Function; Sulfides; Tartrate-Resistant Acid Phosphatase

The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)-positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular matrix by mediating osteopontin/CD44 interaction, and internalize the extracellular matrices. Microsc.

Topics: Acid Phosphatase; Animals; Bone Neoplasms; Carcinoma, Small Cell; Cathepsin K; Cathepsins; Cytoplasmic Vesicles; Disease Models, Animal; Extracellular Matrix; Femur; Golgi Apparatus; Humans; Hyaluronan Receptors; Immunohistochemistry; Male; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Mice, Nude; Microscopy, Immunoelectron; Osteoclasts; Osteolysis; Osteopontin; Sialoglycoproteins; Tibia

An experiment evaluated the effect of citrus juice on enhancing serum antioxidant status and on osteoporosis prevention in orchidectomized rats.. Thirty-six 1-y-old male rats were randomized to two groups: a sham-control group (n = 9) and an orchidectomized group (n = 27). The orchidectomized group was divided into three groups of nine and assigned to one of the following treatments: orchidectomy, orchidectomy plus orange juice, and orchidectomy plus grapefruit juice. Sixty days after initiation of the study, all rats were killed, blood was collected, and serum was harvested for total antioxidant status and indices of bone formation and resorption. Femoral density and biomechanical properties were monitored.. Orchidectomy decreased (P < 0.05) total antioxidant capacity, femoral density, and biomechanical properties and increased (P < 0.05) alkaline phosphatase, acid phosphatase, and urinary excretion of hydroxyproline compared with the sham-control group. In contrast to orchidectomy, orchidectomy plus orange juice and orchidectomy plus grapefruit juice reversed (P < 0.05) orchidectomy-induced antioxidant suppression, decreased (P < 0.05) alkaline phosphatase and acid phosphatase activities, moderately restored (P = 0.07) femoral density, increased (P < 0.05) femoral strength, significantly delayed time-induced femoral fracture, and decreased (P < 0.05) urinary excretion of hydroxyproline.. The present study supports the supposition in that drinking citrus juice positively affects serum antioxidant status and bone strength.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Antioxidants; Beverages; Bone and Bones; Bone Density; Citrus; Disease Models, Animal; Hydroxyproline; Male; Orchiectomy; Osteoporosis; Random Allocation; Rats; Rats, Sprague-Dawley

Localised bone loss in the form of bone erosions and peri-articular osteopenia constitutes an important criteria for the diagnosis of rheumatoid arthritis. In the present study, the effect of Semecarpus anacardium Linn. nut milk extract (SA) on the metabolism of bone turn over has been studied by analyzing various markers of bone turnover and by histological and radiological analysis of the joints in adjuvant arthritis in rats. Arthritis was induced in rats by injecting Freund's complete adjuvant containing 10mg of heat killed mycobacterium tuberculosis in 1 ml paraffin oil (0.1 ml) into the left hind paw of the rat intradermally. After 14 days of induction, SA (150 mg/kg body weight/day) was administered orally by gastric intubations for 14 days. SA significantly reverted the alterations in the bone turnover observed in arthritic animals by modulating the levels of calcium, phosphorus and the activities of the enzymes names tartrate resistant acid phosphatase, acid phosphatase and alkaline phosphatase. The drug increased the bone weights that were found to be decreased during arthritis. Protective effect of SA was also observed by the decrease in the levels and expression of tumour necrosis factor alpha (TNF-alpha) as well as the histopathological and radiological observations. From all these observations it can be concluded that SA possesses strong anti-arthritic property by regulating bone turnover.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Antirheumatic Agents; Arthritis, Experimental; Bone and Bones; Calcium; Disease Models, Animal; Hot Temperature; Immunohistochemistry; Nuts; Phosphorus; Plants, Medicinal; Radiology; Rats; Rats, Wistar; Semecarpus; Tumor Necrosis Factor-alpha

To investigate the effect of gallium salts on bone metabolism in osteoporosis rats caused by tretinoin.. After duplicating osteoporotic model of rats by using tretinoin, we observed the metabolism of blood, urine and bone in vivo. The rats were divided into control group, osteoporosis group, estrogen treated and gallium chloride treated group. The indexes of bone metabolism and related indexes in serum and urine were observed after 60 days of treatment. In the same time, we observed the dynamic effect of 30 days and 60 days of treatment.. After duplicating osteoporotic model, rats' bone structures were injured, the bone mineral density and the organic matrix decreased, the contents of tartrate-resistant acid phosphatase (TRAP) in serum were higher. After gallium salt treating, the above damages were inhibited or retarded. Trabecular width and thickness of bone cortex were significantly increased. AKP and TRAP activities were decreased to the level close to that of the control group.. Gallium salt is effective in treating osteoporosis.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Density; Disease Models, Animal; Female; Gallium; Isoenzymes; Osteoporosis; Random Allocation; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Tretinoin

In the current study, we investigated whether the systemic administration of alendronate, a third-generation bisphosphonate, suppressed the loosening of screws at the bone-screw interface. We systemically administered alendronate to rats fitted with external fixators. External fixators with two half pins were applied to the right femurs of rats, and alendronate was administrated once a week during a 5-week postoperative period. Radiographic, histologic, and immunohistochemical findings subsequently were analyzed. Treatment with alendronate reduced the width of the fibrous loosening membrane and the number of osteoclasts at the bone-screw interface. These findings indicate that systemic treatment with alendronate exerts an inhibitory effect on local bone resorption at the bone-screw interface.

Topics: Acid Phosphatase; Alendronate; Animals; Bone Resorption; Bone Screws; Cathepsin K; Cathepsins; Disease Models, Animal; External Fixators; Femur; Isoenzymes; Male; Prosthesis Failure; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Recent data suggest that the spleen is a crucial component of the immune system in the development of experimental autoimmune encephalomyelitis (EAE) in marmoset monkeys. Using immunohistochemistry, we investigated changes in the distribution of leukocytes in the spleen associated with clinical symptoms of EAE. Animals without EAE displayed well-developed T- and B-cell areas, germinal centers and red pulp. In contrast, a marked depletion of periarteriolar T cells with preservation of other elements was found in animals with clinical EAE. These findings suggest that immune responses within the spleen are impaired during a paralysing inflammatory process in the central nervous system.

Topics: Acid Phosphatase; Animals; Antigens, CD; Callithrix; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Humans; Immunoglobulin G; Immunoglobulin M; Immunohistochemistry; Lymphocyte Activation; Lymphocyte Depletion; Lymphocytes; Macrophages; Male; Membrane Glycoproteins; Microscopy, Electron, Transmission; Myelin Sheath; Nuclear Proteins; Plasma Cells; ran GTP-Binding Protein; Receptors, Immunologic; Sialic Acid Binding Ig-like Lectin 1; Spleen; T-Lymphocytes

We developed a rodent model that mimics the osteoblastic and osteolytic changes associated with human metastatic prostate cancer. Microarray analysis identified MMP-7, cathepsin-K, and apolipoprotein D as being upregulated at the tumor-bone interface. MMP-7, which was produced by osteoclasts at the tumor-bone interface, was capable of processing RANKL to a soluble form that promoted osteoclast activation. MMP-7-deficient mice demonstrated reduced prostate tumor-induced osteolysis and RANKL processing. This study suggests that inhibition of MMP-7 will have therapeutic benefit in the treatment of prostate cancer-induced osteolysis.

Topics: Acid Phosphatase; Actins; Animals; Carrier Proteins; Disease Models, Animal; Down-Regulation; Gene Expression; Gene Expression Profiling; Glycoproteins; Humans; Isoenzymes; Male; Matrix Metalloproteinase 7; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Models, Biological; Monocytes; Osteoclasts; Osteolysis; Osteoprotegerin; Parathyroid Hormone-Related Protein; Prostatic Neoplasms; RANK Ligand; Rats; Rats, Inbred F344; Receptor Activator of Nuclear Factor-kappa B; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Skull; Tartrate-Resistant Acid Phosphatase; Up-Regulation

This study was to develop a suitable model for the investigation for food allergy.. BALB/c mice were dosed by intraperitoneally with Ovalbumin, Beef serum albumin, Trypsin inhibitor and Potato acid phosphatase respectively (0.25ml 20mg/mnl) on day 0 and again on day 7. Control group was dosed with PBS. Sera form individual animals were analysed for specific IgE and Passive cutaneous anaphylaxis tests. Additionally, the level of histamine in plasma were detected.. The high titres of specific IgE (1: 32) could be provoked in test groups compared with control group. In addition, the level of histamine in plasma of test groups was higher than that in the control group. But there was no statistical significance between group food allergen and group Potato acid phosphatase.. Although allergic action of BALB/c mice could be provoked, the situation of the allergic action of BALB/c mice to the proteins was very different with the human being. The BALB/c mice could not be a suitable model for the investigation for food allergy.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Female; Food Hypersensitivity; Histamine; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Serum Albumin, Bovine; Trypsin Inhibitors

To develop a Brown Norway (BN) rat model to determine the potential allergenicity of novel proteins in genetically modified food.. The allergenicity of different proteins were compared, including ovalbumin (OVA), a potent respiratory and food allergen, bovine serum albumin (BSA), a protein that is considered to have a lesser allergenic potential, and potato acid phosphatase (PAP), a non-allergenic protein when administered to BN rats via different routes of exposure (intraperitoneally or by gavage). IgG and IgE antibody responses were determined by ELISA and PCA, respectively. An immunoassay kit was used to determine the plasma histamine level. In addition, possible systemic effect of allergens was investigated by monitoring blood pressure.. OVA provoked very vigorous protein-specific IgG and IgE responses, low grade protein-specific IgG and IgE responses were elicited by BSA, while by neither route did PAP elicit anything. In either routes of exposure, plasma histamine level in BN rats sensitized with OVA was higher than that of BSA or PAP. In addition, an oral challenge with BSA and PAP did not induce any effect on blood pressure, while a temporary drop in systolic blood pressure in few animals of each routes of exposure was found by an oral challenge with OVA.. BN rat model might be a useful and predictive animal model to study the potential allergenicity of novel food proteins.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Food Hypersensitivity; Food, Genetically Modified; Male; Ovalbumin; Rats; Rats, Inbred BN; Serum Albumin, Bovine; Solanum tuberosum

Abatacept is the first in a new class of agents that selectively modulates T-cell activation by attenuating CD28-mediated co-stimulation. This study examined the effects of abatacept on disease development in a rat model of collagen-induced arthritis (CIA). The rats were treated with either abatacept (1mg/kg) or control IgG beginning at the time of induction of CIA. By day 16, significant paw swelling was observed in IgG-treated control animals that continued to increase, reaching a plateau on day 21. Prophylactic treatment with abatacept completely abrogated paw swelling throughout the study. Histopathology demonstrated a significant reduction in inflammation, cartilage destruction, bone resorption and pannus formation. Abatacept treatment resulted in 90% inhibition of circulating collagen-specific antibodies and decreased the serum expression of many cytokines and chemokines that were upregulated in diseased animals. Immunohistochemical analysis of the ankle joints demonstrated that interleukin-6 production was reduced in the tissues and the numbers of osteoclasts present in the joints were also decreased. Ankle microcomputer tomography (micro-CT) analyses dramatically demonstrated the protective effects of abatacept on bone destruction in these animals. Data presented here demonstrate that prophylactic administration of abatacept significantly inhibits the onset and progression of disease in a rat CIA model, with reductions in inflammation, inflammatory mediators, and bone and joint destruction.

Topics: Abatacept; Acid Phosphatase; Animals; Arthritis, Experimental; Autoantibodies; Bone and Bones; Bone Resorption; Collagen; Disease Models, Animal; Female; Immunoconjugates; Inflammation; Isoenzymes; Osteoclasts; Rats; Tarsal Joints; Tartrate-Resistant Acid Phosphatase

We show that mice lacking beta3 integrin are protected from OVX-induced bone loss. Using a lentiviral-based strategy to express beta3 mutants in beta3(-/-) mice, we also show that beta3(S752), but not beta3(Y747/Y759), is important for osteoclastic bone resorption in vivo.. Mice lacking the beta3 integrin have dysfunctional osteoclasts and therefore accumulate bone mass with age. Thus, the alphavbeta3 integrin is a potential anti-osteoporosis target. Identifying components of the beta3 integrin that determine its function in vivo is essential for therapeutically exploiting the antiresorptive properties of alphavbeta3.. We used DXA and histomorphometry to assess bone loss after ovariectomy in wildtype and beta3 integrin null mice. We used lentiviral vectors carrying various human beta3 (hbeta3) integrin constructs to transduce beta3(-/-) bone marrow and reconstituted lethally irradiated beta3(-/-) mice with the transduced marrow. The expressed constructs include the intact integrin and two mutants, namely hbeta3(Y747F/Y759F) and hbeta3(S752P), each of which induces the bleeding dyscrasia, Glanzmann's thrombasthenia, in humans. Two months after transplantation, the expression of hbeta3 was measured by flow cytometry of marrow-derived macrophages. Osteoclast differentiation and function were assessed ex vivo by TRACP and actin-ring staining, respectively. Reconstituted mice were ovariectomized, and bone loss was assessed by DXA, histomorphometry, and serum TRACP5b assay.. beta3(-/-) mice are protected from ovariectomy-induced bone loss, showing no difference in BMD compared with sham-operated controls. We successfully expressed hbeta3 integrins in beta3(-/-) hosts using lentiviral transduction of bone marrow. Two months after transplantation, 25-35% of marrow-derived macrophages expressed the hbeta3 constructs. Similar to its effect in vitro, hbeta3(WT) completely rescued the osteoclast and platelet phenotype of beta3(-/-) mice. Whereas platelet function remained deranged in beta3(-/-) mice overexpressing hbeta3(Y747F/Y759F), osteoclast function was fully restored. In contrast, beta3(-/-) mice expressing hbeta3(S752P) continued to exhibit prolonged bleeding times and dysfunctional osteoclasts in vitro and ex vivo. Most importantly, hbeta3(WT) and hbeta3(Y747F/Y759F) transplanted mice underwent equivalent ovariectomy-induced bone loss, whereas, like those bearing the control vector, hbeta3(S752P) transplanted mice were protected.. Functional beta3 integrin is required for ovariectomy-induced bone loss. beta3(S752), but not beta3(Y747/Y759), is critical for osteoclast function in vivo.

Topics: Acid Phosphatase; Actins; Animals; Bleeding Time; Blood Platelets; Bone Density; Bone Marrow Cells; Bone Marrow Transplantation; Bone Resorption; Carrier Proteins; Cell Differentiation; Disease Models, Animal; Female; Femur Head; Genetic Vectors; Humans; Integrin beta3; Isoenzymes; Lentivirus; Lumbar Vertebrae; Macrophage Colony-Stimulating Factor; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Mutation; Osteoclasts; Osteoporosis, Postmenopausal; Ovariectomy; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; Tibia; Transfection

Osteocyte apoptosis caused by load-induced microdamage is followed by osteoclastic bone remodeling, and a causal link between apoptosis and repair has been suggested. The objectives of the present study were to use a chick model to examine the incidence of osteocyte apoptosis and the presence of osteoclasts during the first 96 hours following an osteotomy, prior to extensive callus mineralization. Osteotomies were performed on the right radii of 24 chicks at 23-24 days of age. The left radii served as controls. Radii were collected and processed at six time points following surgery (0, 12, 24, 48, 72, and 96 hours). Decalcified bone tissue sections were stained either for apoptosis using a modified TUNEL procedure or for tartrate-resistant acid phosphatase to identify osteoclasts in the intracortical and periosteal envelopes. The percentage of apoptotic osteocytes, as well as osteoclast counts (n/mm or n/mm2) were quantified in four regions (0-1, 1-2, 2-4, and 4-8 mm from the site of the osteotomy; regions 1-4, respectively) in the osteotomized radii and in the same measured areas in the control radii. Data for osteocyte apoptosis and osteoclasts in the control limb were subtracted from the osteotomized limb data to identify differences due to surgical influence. The incidence of osteocyte apoptosis was significantly higher at 12, 24, 48, and 72 hours versus 0 hours following osteotomy, and the response was highest in region 1; however, there was no interaction between time and region. Intracortical osteoclast counts (n/mm2) were elevated after 48 hours, and the response was similar in all regions. The data demonstrate that osteocyte apoptosis occurs within 24 hours in response to an osteotomy and temporally precedes an increase in osteoclast presence. Hence, osteocyte apoptosis may play a role in signaling during the bone healing process.

Topics: Acid Phosphatase; Animals; Apoptosis; Chickens; Disease Models, Animal; Fracture Healing; In Situ Nick-End Labeling; Isoenzymes; Male; Osteoclasts; Osteocytes; Osteogenesis; Osteotomy; Radius; Tartrate-Resistant Acid Phosphatase

Early gestational malaria is found to be more fatal than late gestational infection but the pathophysiology of early gestational placenta, the maternofoetal organ responsible for maintenance of pregnancy, remains unexplored. Present study dealing with hydrolytic enzymes in early gestational placenta of rhesus monkeys during Plasmodium cynomolgi infection was anticipated to provide a better insight into the functional impairment of this organ during early gestational maternal malaria.. Experimental monkeys (Macaca multtta) at 2-2 1/2 months of pregnancy were inoculated with P. cynomolgi bastianelli. After attaining first peak of parasitaemia the animals were anesthetised and placentae were collected for histochemical studies. The snap-frozen, cryostat sections were subjected to histochemical reactions for acid phosphatase and alkaline phosphatase.. The placental syncytiotrophoblast showed a loss in alkaline phosphatase activity, while the trophoblast layers and phagocytic cells of the maternal blood showed increased acid phosphatase activity during early gestational malarial infection. Morphological damage to the placental tissue whenever occurred was associated with altered Alk pase activity.. The altered distribution of Ac pase and Alk pase in malaria infected early gestational placenta has been discussed in the light of placental function. It could be concluded by present studies that these malaria induced changes in hydrolytic enzyme activities in monkey placenta have a direct bearing on functional and morphological integrity of the placental tissue. These changes are apparently responsible for early gestational foetal death and abortions as reported in literature.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Female; Immunohistochemistry; Macaca mulatta; Malaria; Placenta; Plasmodium cynomolgi; Pregnancy; Pregnancy Complications, Parasitic

We determined the effect of baicalein on prostatic hyperplasia in experimental animal models. Prostatic hyperplasia was induced by testosterone propionate in mice and castrated rats and by transplantation of homologous strain fetal mice urogenital sinus in mice. With the histopathological examination, the efficacy of baicalein on prostate hyperplasia in experimental animals was evaluated by the activity of serum acid phosphatase (ACP) and the following norm of the prostate gland: the volume, wet weight, wet weight index, dry weight index, DNA contents and prostatic epithelial height and cavity diameter. Results showed that baicalein at doses of 260 and 130 mg/kg administrated intragastrically (i.g.) significantly inhibited prostatic hyperplasia in castrated rats induced by testosterone propionate compared with the negative control group (p<0.01). Baicalein at doses of 520 and 260 mg/kg (i.g.) also significantly inhibited prostatic hyperplasia in mice induced by transplantation of homologous strain fetal mouse urogenital sinus and by testosterone propionate (p<0.01). These results suggested that baicalein has an inhibitory effect on prostatic hyperplasia in experimental animals.

Topics: Acid Phosphatase; Animals; Castration; Cell Division; Depression, Chemical; Disease Models, Animal; Dose-Response Relationship, Drug; Flavanones; Male; Mice; Organ Size; Prostatic Hyperplasia; Rats; Rats, Sprague-Dawley; Testosterone Propionate

It has been suggested that subchondral bone remodeling plays a role in the progression of osteoarthritis (OA). To test this hypothesis, we characterized the changes in the rat anterior cruciate ligament transection (ACLT) model of OA and evaluated the effects of alendronate (ALN), a potent inhibitor of bone resorption, on cartilage degradation and on osteophyte formation.. Male Sprague-Dawley rats underwent ACLT or sham operation of the right knee. Animals were then treated with ALN (0.03 and 0.24 microg/kg/week subcutaneously) and necropsied at 2 or 10 weeks postsurgery. OA changes were evaluated. Subchondral bone volume and osteophyte area were measured by histomorphometric analysis. Coimmunostaining for transforming growth factor beta (TGF beta), matrix metalloproteinase 9 (MMP-9), and MMP-13 was performed to investigate the effect of ALN on local activation of TGF beta.. ALN was chondroprotective at both dosages, as determined by histologic criteria and collagen degradation markers. ALN suppressed subchondral bone resorption, which was markedly increased 2 weeks postsurgery, and prevented the subsequent increase in bone formation 10 weeks postsurgery, in the untreated tibial plateau of ACLT joints. Furthermore, ALN reduced the incidence and area of osteophytes in a dose-dependent manner. ALN also inhibited vascular invasion into the calcified cartilage in rats with OA and blocked osteoclast recruitment to subchondral bone and osteophytes. ALN treatment reduced the local release of active TGF beta, possibly via inhibition of MMP-13 expression in articular cartilage and MMP-9 expression in subchondral bone.. Subchondral bone remodeling plays an important role in the pathogenesis of OA. ALN or other inhibitors of bone resorption could potentially be used as disease-modifying agents in the treatment of OA.

Topics: Acid Phosphatase; Alendronate; Animals; Anterior Cruciate Ligament; Bone Remodeling; Calcinosis; Cartilage, Articular; Collagen; Collagen Type I; Collagen Type II; Disease Models, Animal; Disease Progression; Extracellular Matrix Proteins; Glycoproteins; Isoenzymes; Male; Matrilin Proteins; Osteoarthritis, Knee; Osteoclasts; Peptides; Rats; Sclerosis; Severity of Illness Index; Tartrate-Resistant Acid Phosphatase; Transforming Growth Factor beta

Etodolac, a cyclooxygenase-2 inhibitor, may alleviate nociceptive pain and inhibit the activation of osteoclasts. The aim of the present study was to determine whether etodolac can alleviate heat-evoked hyperalgesia and investigate its possible protective effects on osteoporosis induced by chronic constriction injury (CCI) in rats. A CCI to the sciatic nerve was performed, after which the rats received etodolac orally in a volume of 2 ml at 0, 1, and 10 mg/kg/day for 1 to 5 weeks following surgery (experiment 1); at 0 and 10 mg/kg/day for 1 day to 5 weeks following surgery (experiment 2); and at 0 mg/kg/day for 1 to 5 weeks, 10 mg/kg/day for 1 to 2 weeks after surgery, or 10 mg/kg/day for 1 to 3 weeks after surgery (experiment 3). Paw withdrawal latency after exposure to heat, bone mineral content (BMC) and bone mineral density (BMD) in the whole tibial bone, and the number of tartrate resistant acid phosphate (TRAP)-positive multinucleated osteoclasts were measured. Etodolac alleviated heat-evoked hyperalgesia in the CCI rats and the increase in number of TRAP-positive multinucleated osteoclasts on the CCI-side was abrogated, however, it did not inhibit the decrease of BMC and BMD on the CCI-side. Our results suggest that etodolac is useful for treatment of neuropathic pain.

Topics: Acid Phosphatase; Animals; Bone Density; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; Disease Models, Animal; Etodolac; Hyperalgesia; Isoenzymes; Ligation; Neuralgia; Osteoclasts; Osteoporosis; Pain Measurement; Peripheral Nerves; Peripheral Nervous System Diseases; Prostaglandin-Endoperoxide Synthases; Rats; Rats, Sprague-Dawley; Reaction Time; Sciatic Neuropathy; Tartrate-Resistant Acid Phosphatase; Up-Regulation

Bone loss was previously shown to be induced in the femoral tissue of regucalcin transgenic (TG) rats. Regucalcin is expressed in rat bone marrow cells and its expression is enhanced in regucalcin TG rats. This study was undertaken to determine the change in osteoclastic bone resorption in regucalcin TG rats with increasing age. Femoral-diaphyseal and -metaphyseal tissues were obtained from normal (wild-type) and regucalcin TG rats aged 5, 14, 25 or 50 weeks. Calcium content in the femoral-diaphyseal and -metaphyseal tissues was significantly decreased in regucalcin TG male and female rats aged 5, 14, 25 or 50 weeks as compared with the value obtained from normal rats with each age. When the marrow cells obtained from normal or regucalcin TG rats were cultured for 7 days, the number of tartrate-resistant acid phosphatase (TRACP), a marker of osteoclasts, positive multinucleated cells (MNCs) were significantly increased in the marrow culture of regucalcin TG male and female rats aged 5, 14, 25 or 50 weeks. The effect of parathyroid hormone [human PTH (1-34); 10(-7) M] or 1,25-dihydroxyvitamin D3 [1,25(OH)2D3; 10(-7) M] in stimulating TRACP-positive MNC formation was significantly enhanced in regucalcin TG male and female rats aged 14 or 25 weeks. This study demonstrates that osteoclastic bone resorption is stimulated in regucalcin TG male and female rats with increasing age.

Topics: Acid Phosphatase; Aging; Animals; Animals, Genetically Modified; Biomarkers; Bone Marrow Cells; Calcium; Calcium-Binding Proteins; Carboxylic Ester Hydrolases; Cell Lineage; Cells, Cultured; Disease Models, Animal; Female; Gene Expression Regulation; Humans; Intracellular Signaling Peptides and Proteins; Isoenzymes; Male; Osteoclasts; Osteoporosis; Parathyroid Hormone; Rats; Rats, Sprague-Dawley; Sulfotransferases; Tartrate-Resistant Acid Phosphatase; Time Factors; Vitamin D

Oxalate induced renal calculi formation and the associated renal injury is thought to be caused by free radical mediated mechanisms. An in vivo model was used to investigate the effect of phycocyanin (from Spirulina platensis), a known antioxidant, against calcium oxalate urolithiasis. Male Wistar rats were divided into four groups. Hyperoxaluria was induced in two of these groups by intraperitoneal infusion of sodium oxalate (70 mg/kg) and a pretreatment of phycocyanin (100 mg/kg) as a single oral dosage was given, 1h prior to sodium oxalate infusion. An untreated control and drug control (phycocyanin alone) were also included in the study. We observed that phycocyanin significantly controlled the early biochemical changes in calcium oxalate stone formation. The antiurolithic nature of the drug was evaluated by the assessment of urinary risk factors and light microscopic observation of urinary crystals. Renal tubular damage as divulged by urinary marker enzymes (alkaline phosphatase, acid phosphatase and gamma-glutamyl transferase) and histopathological observations such as decreased tubulointerstitial, tubular dilatation and mononuclear inflammatory cells, indicated that renal damage was minimised in drug-pretreated group. Oxalate levels (P < 0.001) and lipid peroxidation (P < 0.001) in kidney tissue were significantly controlled by drug pretreatment, suggesting the ability of phycocyanin to quench the free radicals, thereby preventing the lipid peroxidation mediated tissue damage and oxalate entry. This accounts for the prevention of CaOx stones. Thus, the present analysis revealed the antioxidant and antiurolithic potential of phycocyanin thereby projecting it as a promising therapeutic agent against renal cell injury associated kidney stone formation.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Antioxidants; Bacterial Proteins; Biomarkers; Cyanobacteria; Disease Models, Animal; gamma-Glutamyltransferase; Hyperoxaluria; Kidney; Kidney Calculi; Lipid Peroxidation; Male; Oxalates; Phycocyanin; Rats; Rats, Wistar; Spirulina

Bisphosphonates induce major increases in strength of callus in distraction osteogenesis in the short term. Poor understanding of the underlying mechanism, however, raises concerns about long-term consequences. In this long-term study in 32 rabbits, zoledronic acid transiently increased trabeculae by delayed temporal progression of endochondral bone remodeling but did not prevent radiographic completion of bone repair.. We hypothesized that bisphosphonate inhibition of osteoclast-mediated resorption would retain bone during repair, producing a larger callus in the short term. However, if remodeling was not restored, completion of the bone repair process in the long term could be jeopardized.. Juvenile rabbits underwent right tibial osteotomy and 2 weeks of distraction, followed by a period of consolidation. Animals received saline (controls) or zoledronic acid (ZA; 0.1 mg/kg at surgery and again 2 weeks later), and distracted tibias were examined by radiograph, DXA, histology, and histomorphometry at 2, 4, 6, 18, and 44 weeks after surgery.. Regenerated bone in ZA-treated animals was denser than controls on radiographs at 6 weeks and had more distinct radiodense trabeculae and retention of original cortices at 18 weeks. By 44 weeks, controls and ZA-treated animals were radiographically healed and indistinguishable. Regenerate BMD and BMC increased between 2 and 4 weeks in all animals, with a greater effect in ZA. At 6 weeks, BMD and BMC in ZA-treated animals were 1.6- and 2-fold greater, respectively, than controls (p < 0.01). From 6 to 44 weeks, the control values gradually increased and approached the ZA-treated values. Regenerate bone volume and trabecular number by histomorphometry were from 1.6- to 2-fold greater in ZA-treated animals at 6 and 18 weeks (p < 0.05). Endochondral cartilaginous matrix volume was up to 2.4-fold greater in ZA-treated animals at 2 and 4 weeks (p < 0.05). TRACP+ cells in ZA-treated animals were larger with more nuclei. Mineral apposition rate and osteoblast number and surface were lower in ZA-treated animals at 6 weeks (p < 0.01) but not at later times.. Disruption of TRACP+ cell function by ZA during bone regeneration seems to lead to an accretion of cancellous bone built on a larger endochondral cartilaginous matrix and increased bone mass, consistent with reported increases in short-term callus strength. This increase in bone mass, caused by a delay in remodeling, provided a transient advantage without preventing radiographic completion of the bone repair process in the long term. Noncontinuous treatment with nitrogen-containing bisphosphonates thus can have short-term beneficial effects without preventing long-term bone repair.

Topics: Acid Phosphatase; Animals; Bone Density; Bone Regeneration; Bony Callus; Cartilage; Diphosphonates; Disease Models, Animal; Imidazoles; Isoenzymes; Male; Osteoblasts; Osteoclasts; Osteogenesis, Distraction; Rabbits; Tartrate-Resistant Acid Phosphatase; Tibia; Time Factors; Zoledronic Acid

Osteoclasts secrete tartrate-resistant acid phosphatase 5b (TRACP 5b) into the circulation. We have developed an immunoassay for the determination of rat TRACP 5b activity. Intra-assay variation of the immunoassay was 4.5%, interassay variation was 3.8%, dilution linearity was 104.6 +/- 7.6%, and recovery of recombinant rat TRACP was 99.1 +/- 5.8%. We studied serum TRACP 5b as a marker of bone resorption using orchidectomized (ORC) rats as a model for osteoporosis and age-matched sham-operated rats as controls in a 6-month study. After the operation, trabecular bone mineral density decreased significantly more in the ORC group than in the sham group, whereas cortical bone mineral density increased similarly in both groups. Serum TRACP 5b activity was significantly elevated within the first week after ORC, returned to the control level in the third week, and was not increased above the sham level at any of the later time points. At 6 months, trabecular bone volume was 80% lower in ORC rats than in controls. Osteoclast number per trabecular bone perimeter was slightly increased, but the absolute number of osteoclasts in trabecular bone was significantly decreased. These results suggest that absolute bone resorption is increased within the first week after ORC. Later, it is decreased because there is less bone to be resorbed. However, relative bone resorption (compared with the amount of remaining bone) is still increased, leading to further bone loss. We conclude that serum TRACP 5b is a useful marker for monitoring changes in the bone resorption rate in rat ORC model.

Topics: Acid Phosphatase; Animals; Blood Chemical Analysis; Bone Density; Bone Resorption; Disease Models, Animal; Immunoassay; Isoenzymes; Male; Orchiectomy; Osteoporosis; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Time Factors

Lipopolysaccharide (LPS) was recognized by CD14, which may be an important mediator in the deleterious effects of LPS on the periodontal destruction. To investigate the roles of CD14 molecules on LPS-induced soft tissue inflammation and bone destruction, the tissues of CD14-deficient mice were examined histopathologically following a local injection of either Salmonella minnesota or Porphyromonas gingivalis LPS. In the first group, 12 mice received a local injection of 500 microg of purified P. gingivalis LPS and six mice were injected with saline to the calvaria as controls. In the second group 13 mice were injected subcutaneously on the laterally abdominal skin with 50 microg of S. minnesota LPS and three mice were injected with PBS. Mice were sacrificed at day 5. After histological preparation, the tissue sections of calvaria and soft tissue specimen were stained with tartrate-resistant acid phosphatase (TRAP) marker for osteoclast and macrophage. The soft tissue sections were also stained with hematoxylin & eosin (H&E). Resorption surface and osteoclast index were measured to quantify bone resorption. Necrotic area and inflammatory cell numbers were estimated to assess the situation of local inflammation. Our results indicated that LPS-induced bone resorption is inhibited in CD14-deficient mice. An increase in the number of total inflammatory cells was noticed in both CD14-deficient mice and wild-type mice; however, the cell numbers were less in CD14-deficient mice than those in wild-type mice (two- to three-fold decrease). Therefore, we conclude that the LPS-stimulated bone resorption is mainly via CD14 receptor but the LPS-induced soft tissue inflammation appears to be partially dependent on the receptor.

Topics: Acid Phosphatase; Analysis of Variance; Animals; Biomarkers; Bone Resorption; Cell Count; Coloring Agents; Disease Models, Animal; Isoenzymes; Lipopolysaccharide Receptors; Lipopolysaccharides; Macrophages; Male; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Knockout; Necrosis; Osteoclasts; Porphyromonas gingivalis; Salmonella; Skin; Skull; Tartrate-Resistant Acid Phosphatase

The effect of ascorbic acid deficiency on bone metabolism was evaluated using the ascorbate-requiring Osteogenic Disorder Shionogi (ODS) rat model. Ascorbic acid (Asc)-deficient rats gained body weight in a manner similar to Asc-supplemented rats (control) during 3 weeks, but began to lose weight during the 4th week of Asc deficiency. The tartrate-resistant acid phosphatase (TRAP) activity in serum increased to about 2-fold the control value in the rats fed the Asc-free diet for 2, 3, and 4 weeks (AscD2, AscD3, and AscD4), while a decrease in the alkaline phosphatase (ALP) activity was observed only in AscD4 rats. The serum pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) level significantly increased to 1.3-, 1.4-, and 1.9-fold of that in the controls in AscD2, D3, and D4, respectively. The ALP activity in the distal femur was unchanged in AscD1, D2, and D3, but decreased to 50% of the control level in AscD4 rats. The TRAP activity in the distal femur increased to about 2-fold of that in the controls in the AscD2 and D3 and decreased to the control level in the AscD4 rats. The amount of hydroxyproline in the distal femur significantly decreased to about 80%, 70%, and 60% of the control in AscD2, D3, and D4 rats, respectively. These decreases were associated with a similar reduction in the calcium content of the distal femur. Histochemical analysis of the distal femur showed an increase in TRAP-positive cells in AscD2 and AscD3 rats and a decrease in the trabecular bone in AscD2, D3, and D4 rats. These results suggested that a deficiency of Asc stimulated bone resorption at an early stage, followed by a decrease in bone formation in mature ODS rats which already had a well-developed collagen matrix and fully differentiated osteoblasts.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Ascorbic Acid Deficiency; Body Weight; Bone Resorption; Calcium; Collagen Type I; Disease Models, Animal; Female; Femur; Hydroxyproline; Immunohistochemistry; Isoenzymes; Oligopeptides; Peptide Fragments; Peptides; Procollagen; Rats; Tartrate-Resistant Acid Phosphatase; Time Factors

First molars fail to erupt in the incisor-absent (ia/ia) rat because of a defect in osteoclast function. Growth factors that regulate local bone metabolism include growth hormone (GH), insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF) and interleukin-1 alpha (IL-1alpha). Since osteoclast function may be affected by these factors, the aim of this study was to determine the distribution of GH receptor (GHr), IGF-I, EGF and IL-1alpha, in osteoclasts located occlusal to the erupting first molar, in the 'eruption pathway', in normal and ia/ia rats. Sagittal sections of the first molar and adjacent bone from 3- and 9-d-old animals were examined. Osteoclasts were identified using tartrate-resistant acid phosphatase (TRAP). The TRAP-positive osteoclast cell numbers were higher in ia/ia animals at 3 and 9 days-of-age. In the ia/ia group, fewer osteoclasts were GHr- and IGF-I-positive at 3 d of age, and at 9 d of age fewer osteoclasts were GHr-positive. In the ia/ia rat, defective osteoclast function failed to resorb bone to provide an eruption pathway for the lower first molar. The expression of GHr, and to some degree IGF-I, by these osteoclasts was reduced, which may be related to their ability to differentiate and function.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Immunohistochemistry; Incisor; Isoenzymes; Mandible; Molar; Osteoclasts; Osteopetrosis; Rats; Rats, Mutant Strains; Receptors, Somatotropin; Tartrate-Resistant Acid Phosphatase; Tissue Distribution; Tooth Eruption; Tooth Germ

Chronic destructive periodontal disease is characterized by gingival inflammation, periodontal pocket formation, and bacterial plaque that lead to alveolar bone destruction. Polymorphonuclear neutrophil leukocytes (PMNs) are the first line of defense against infection caused by dental plaque bacteria. Renal patients present functional abnormalities of PMN, including impaired chemotaxis, phagocytosis, and intracellular killing of bacteria. In view of the above, the aim of this work was to evaluate the effect of renal failure on bone damaged by periodontal disease using histomorphometric and histochemical parameters.. Twenty male Wistar rats weighing 250 g were assigned to one of the following four groups: 1) control (no treatment); 2) renal failure (RF); 3) periodontal disease (PD); and 4) renal failure plus periodontal disease (RF+PD). All the animals were sacrificed 31 days after the onset of the experiment. Mesio-distally oriented sections of the first lower molar were obtained for histomorphometric and histochemical evaluation.. Total erosion, active erosion, and total number of tartrate-resistant acid phosphatase-positive (TRAP+) osteoclasts were found to be increased in the RF+PD group compared with the PD group.. Our results demonstrate increased bone resorption in animals with untreated renal failure and periodontal disease, and thus indicate that the release of different factors by inflammatory cells is magnified, accelerating the progression of the disease in this animal model.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Coloring Agents; Disease Models, Animal; Isoenzymes; Male; Osteoblasts; Osteoclasts; Rats; Rats, Wistar; Renal Insufficiency; Tartrate-Resistant Acid Phosphatase

To investigate the protective effect of pSVPoMcat (myelin basic protein microgene) modifying Schwann cell on injured spinal neurons.. A model of rat spinal cord injured by hemisection was used. One hundred and twenty healthy SD rats of both sexes weighing 250-300 g were divided into three groups: Group A (n=40, treated with implantation of pSVPoMcat modifying Schwann cell), Group B (n= 40, treated with implantation of Schwann cell only) and Group C (n=400, treated with sham operation as the control). One week after operation the rat functional recovery was observed dynamically by using combined behavioral score (CBS) and cortical somatasensory evoked potentials, the spinal cord sections were stained by Nissl, acid phosphatase enzyme histochemistry and cell apoptosis was examined by methye green, terminal deoxynucleotidyl and the dUTP Nick end labeling technique. Quantitative analysis was done by computer image analysis system.. In Group A the injured neurons recovered well morphologically. The imaging analysis showed a result of Group A

Topics: Acid Phosphatase; Animals; Apoptosis; Cell Transplantation; Disease Models, Animal; Evoked Potentials, Somatosensory; Female; Gene Transfer Techniques; Male; Methyl Green; Myelin Basic Protein; Nerve Regeneration; Rats; Rosaniline Dyes; Schwann Cells; Spinal Cord Injuries

Administration of 0.01 and 0.1 mg of lead nitrate for 4 and 7 days and infection of Ancylostoma caninum larvae orally altered the activation of alkaline phosphatase and acid phosphatase in the hearts of mice when compared to infected animals and controls. Alkaline phosphatase activity increased significantly in all drug-treated + infected mice. The level of acid phosphatase decreased significantly in mice exposed to chronic doses of lead. The altered levels of alkaline phosphatase and acid phosphatase suggest that administration of lead could cause toxicity in the heart, disturbing the cellular metabolism; infection alone could not cause any significant changes in enzymes of heart.

Topics: Acid Phosphatase; Alkaline Phosphatase; Ancylostoma; Ancylostomiasis; Animals; Disease Models, Animal; Female; Lead Poisoning; Mice; Myocardium

Unspecific inflammatory lesions of the urinary tract are known to be a vital question in urology and nephrology of our country and in foreign ones. Used in experiments were rats of mixed population. Lysosomal enzymes were studied in homogenates of the liver and contralateral kidney tissues and in the blood serum as well 1, 3, 6 days after simulation of the pathology experimental model. Activities were measured of such lysosomal enzymes as acid DNA-ase and RNA-ase, cathepsin, and acid phosphatase. Results of the studies made showed that in unspecific purulent affection of the kidneys there occurs an activation of acid blood hydrolases followed by their activation in the liver. The contralateral kidney, because of it being linked in functioning with the liver, remains relatively "inert" with respect to activation of lysosomal enzymes.

Topics: Acid Phosphatase; Animals; Cathepsin D; Deoxyribonucleases; Disease Models, Animal; Female; Kidney; Liver; Lysosomes; Male; Nephritis; Rats; Ribonucleases; Suppuration; Time Factors

Aphanamixis polystachya is a traditional medicinal plant of the Meliaceae family in India. A crude ethanolic extract of the leaf of this plant shows a beneficial effect on toxic liver injury. Its antihepatotoxic activity was evaluated on carbon tetrachloride (CCl4)-induced liver injury in a rat model. The assessment of hepatoprotective activity was evaluated by measuring the activities of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), acid phosphatase (ACP) and lactate dehydrogenase (LDH), serum total bilirubin and albumin and histology of the liver. The crude leaf extract significantly inhibits the enhanced ASAT, ALAT, ALP, ACP and LDH activities released from the CCl4-intoxicated animals. It also ameliorated the depressed value of serum albumin and the enhanced value of total bilirubin in plasma caused by CCl4 intoxication. The study showed that the crude ethanolic extract from A. polystachya leaves provided protection against acute carbon tetrachloride-induced liver damage.

Topics: Acid Phosphatase; Alanine Transaminase; Alkaline Phosphatase; Animals; Aspartate Aminotransferases; Bilirubin; Carbon Tetrachloride; Carbon Tetrachloride Poisoning; Disease Models, Animal; L-Lactate Dehydrogenase; Liver; Male; Plant Extracts; Plants, Medicinal; Rats; Serum Albumin

We examined whether topical administration of a bisphosphonate clodronate could prevent alveolar bone loss in rats with experimental periodontitis.. On day 0, elastic rings were placed around the cervix of the right and left maxillary first molars (M1) to induce inflammatory periodontitis. Fifty microl of clodronate solution at a concentration of either 0 (0.9% NaCl), 20, 40, or 60 mM was injected into the subperiosteal palatal area adjacent to the interdental area between M1 and M2 on either the left or right (experimental) side on days 0, 2, 4, and 6. The contralateral side served as a control and received 0.9% NaCl solution without clodronate. The animals were sacrificed on day 7.. Histological examination and determination of bone mineral density in the interdental alveolar bone area between M1 and M2 revealed that placement of an elastic ring caused severe vertical and horizontal bone resorption on the control side, while the topical administration of clodronate significantly prevented such alveolar bone loss. The number of osteoclasts on the experimental side was decreased compared with the control side. Furthermore, many of the osteoclasts on the experimental side were detached from the surface of the alveolar bone and had degenerated appearances, such as rounded shapes and a loss of polarity.. These results suggest that topical administration of clodronate may be effective in preventing osteoclastic bone resorption in periodontitis.

Topics: Acid Phosphatase; Administration, Topical; Alveolar Bone Loss; Analysis of Variance; Animals; Antimetabolites; Azo Compounds; Biomarkers; Bone Density; Bone Resorption; Cell Count; Clodronic Acid; Coloring Agents; Disease Models, Animal; Eosine Yellowish-(YS); Fluorescent Dyes; Injections; Isoenzymes; Male; Maxillary Diseases; Methyl Green; Molar; Osteoclasts; Periodontitis; Radiography; Rats; Rats, Wistar; Statistics as Topic; Tartrate-Resistant Acid Phosphatase

The present study was undertaken to determine the frequency and extent of apical root resorption associated with induced periradicular lesions in mice.. Bone and root resorption was quantified by using two- and three-dimensional micro-computed tomography (mu-CT) in the lower first molars of mice subjected to pulp exposure and infection.. mu-CT measurements showed significant apical resorption in exposed and infected teeth, resulting in an average distal root shortening of 12.7% (P <.001 vs unexposed). These findings were confirmed with three-dimensional reconstituted images that showed thinning and shortening of the distal root. Tartrate-resistant acid phosphatase clastic cells were associated with resorption lacunae on the cementum of root apices, as well as on bone at the periphery of the periradicular lesions. Brown and Brenn staining showed the presence of bacteria in dentinal tubules adjacent to resorbed cementum.. Apical root resorption is a prominent and consistent finding associated with periradicular infection in the mouse. This species represents a convenient model for studying the pathogenesis of inflammatory root resorption in vivo.

Topics: Acid Phosphatase; Animals; Biomarkers; Bone Resorption; Coloring Agents; Dental Cementum; Dental Pulp Diseases; Dental Pulp Exposure; Dentin; Disease Models, Animal; Gram-Negative Bacteria; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Isoenzymes; Male; Mandibular Diseases; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Molar; Periapical Diseases; Regression Analysis; Root Resorption; Statistics as Topic; Tartrate-Resistant Acid Phosphatase; Tomography, X-Ray Computed; Tooth Apex; Tooth Root

The osteopetrotic grey-lethal (gl) mouse mutant displays many similarities to the human malignant autosomal-recessive form of osteopetrosis. In this study, we show that the gl osteopetrotic bone phenotype is characterized by the presence of numerous differentiated multinucleated osteoclasts. A significant increase in the number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts was detected in vivo, suggesting induction of differentiation in the osteoclast lineage as a compensatory mechanism. These gl osteoclast cells demonstrated a defective cytoskeletal reorganization and an underdeveloped ruffled border, a membrane structure essential for active bone resorption. Accordingly, resorption activity of these cells is markedly impaired by four- to tenfold as evaluated with the pit formation assay. This low bone resorption in gl osteoclasts is highly reminiscent of the loss in key enzymes, V-ATPase or cathepsin-K, and in signaling factors, Src or TRAF-6, which were shown not to be significantly altered in gl osteoclasts. Thus, independently of a deficiency in V-ATPase, Src, cathepsin-K, and TRAF-6, the gl mutation results in increased number of osteoclasts, characterized by a disrupted cytoskeleton and an underdeveloped ruffled border.

Topics: Acid Phosphatase; Animals; Bone and Bones; Bone Resorption; Cell Differentiation; Cells, Cultured; Coculture Techniques; Cytoskeletal Proteins; Cytoskeleton; Disease Models, Animal; Genes, Lethal; Immunohistochemistry; Isoenzymes; Mice; Mice, Mutant Strains; Microscopy, Electron; Mutation; Osteoclasts; Osteopetrosis; Phenotype; Proteins; Proto-Oncogene Proteins pp60(c-src); Proton-Translocating ATPases; Tartrate-Resistant Acid Phosphatase; TNF Receptor-Associated Factor 6; Vacuolar Proton-Translocating ATPases

The aim of this study was to assess the skeletal metabolism in a murine model of systemic lupus erythematosus (SLE). MRL/n and MRL/l mice (respectively representing a benign and a malignant form of the disease) were observed from 1.5 to 6.5 months of life. The monthly follow-up included: biochemical and histomorphometrical studies of the femoral bone, serum biochemistry, immunoglobulins and osteocalcin, and histological evaluation of the kidney tissue. The results showed a higher femoral weight (+11.5%), calcium (+4.4%) and protein bone content (+11.4%) and a significantly higher (+77%) phosphorus bone content in the MRL/n group; significantly lower (-48.9%) bone alkaline phosphatase enzymatic activity, lower bone alkaline/acid phosphatase enzymatic activities ratio (-40.8%) and lower (-38.4%) serum osteocalcin values in the MRL/l group (which might suggest reduced bone formation in these animals); markedly smaller trabecular bone volume (BV/TV) in the femoral head (-36.2%) and femoral neck (-39.8%), and smaller cortical and femoral areas in the mid-femoral shaft (-38.8% and -38.1% respectively) in the MRL/l group; higher serum immunoglobulins, increased serum blood urea nitrogen (BUN) and creatinine and a higher index of activity in the kidney histology in the MRL/l group, indicating increased activity of the disease in this substrain. The MRL mice, through their two substrains, may serve as a valuable laboratory animal model for study of the skeletal changes in SLE and of the influence of the disease activity on the skeletal metabolism.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Calcium; Disease Models, Animal; Femur; Lupus Erythematosus, Systemic; Magnesium; Mice; Mice, Inbred MRL lpr; Organ Size; Osteocalcin; Osteoporosis; Phosphorus

The treatment for chronic nonbacterial prostatitis (NBP) has not been established. Cernitin pollen-extract (CN-009) is reported to have therapeutic effects for NBP. The effects and mechanisms of CN-009 were investigated.. Ten-month-old rats were used with administration of estradiol after castration, which were similar to human NBP histologically. Since CN-009 consists of T-60 and GBX, these drugs were administered, respectively. The prostate was evaluated histopathologically including glandular damage (epithelial score), stromal ratio and immunohistochemical assays for epithelial function (PAP), stromal evaluation (Vimentin), cell proliferation (PCNA) and apoptosis (deoxyuridine triphosphate biotin nick end-labeling (TUNEL)).. Controls revealed severe acinar gland atrophy and stromal proliferation. CN-009 showed diminished these damages. Epithelial score was better (P < 0.01) and PAP positive materials were more abundant in CN-009 and GBX than in Controls. The stromal ratio was lower in CN-009 (P < 0.01) and T-60 (P < 0.05). There was no difference for PCNA positive cells in the epithelium and stroma, and TUNEL positive cells in epithelium. While, the number of TUNEL positive cells in the stroma of CN-009 and T-60 increased (P < 0.01).. These findings suggest that CN-009 protects acinar epithelial cells mainly by GBX and also inhibits stromal proliferation in association with enhanced apoptosis mainly by T-60.

Topics: Acid Phosphatase; Animals; Apoptosis; Disease Models, Animal; Epithelial Cells; Estradiol; Immunohistochemistry; In Situ Nick-End Labeling; Male; Orchiectomy; Phytotherapy; Plant Extracts; Pollen; Proliferating Cell Nuclear Antigen; Prostatitis; Rats; Rats, Wistar; Secale; Statistics, Nonparametric; Stromal Cells; Vimentin

It was studying the effect of fluorinecontain pinacidil analogs (PF-5 and PF-10), which are the K(+)-channel openers, on morphofunctional lysosomes state at lung and heart tissues. The investigation was made on white pubertal rat-males under acute (30 min) hypoxic hypoxia (7% O2 [symbol: see text] N2). The application of PF-5 and PF-10 under acute hypoxic hypoxia leads in lung and myocardium tissues to morphofunctional changes of lysosome system in investigated cells, which were connected with decreasing of enzyemia. It, partly, may be explain by increasing of connected enzyme forms synthesis, because such forms are the functional latent, so the output of enzymes in blood under unfavourable conditions increased.

Topics: Acid Phosphatase; Acute Disease; Animals; Cathepsin D; Disease Models, Animal; Heart; Hypoxia; Lung; Lysosomes; Male; Myocardium; Pinacidil; Potassium Channels; Rats; Rats, Wistar

The TRAMP-C1 (C1) and TRAMP-C2 (C2) cell lines were derived from a prostate tumor that arose in a mouse from the transgenic adenocarcinoma mouse prostate (TRAMP) model. However, their similarity to primary prostate tumors and therefore their usefulness in immunotherapy studies has not been clearly defined. We showed using RT-PCR that these cell lines exhibited a variety of prostate-specific genes expressed by human prostate tumors that may be used as tumor-associated antigens for immunotherapy. Interestingly, several of these genes are also expressed in cell lines that are not prostatic in origin. The prostate cell lines were also shown to grow in an androgen-independent manner, to be capable of expressing MHC class I and to be susceptible to specific lysis by cytotoxic T lymphocytes. Therefore, these cell lines will provide us with the ability to evaluate immune responses to and tolerance of prostate-specific protein peptides in an animal model.

Topics: Acid Phosphatase; Adenocarcinoma; Animals; Antibody Specificity; Antigens, Neoplasm; Carboxypeptidases; Disease Models, Animal; Gene Expression; Genes, Tumor Suppressor; Glutamate Carboxypeptidase II; GPI-Linked Proteins; Homeodomain Proteins; Immunotherapy; In Vitro Techniques; Male; Membrane Glycoproteins; Mice; Mice, Transgenic; Neoplasm Proteins; Prostatic Neoplasms; Prostatic Secretory Proteins; Protein Tyrosine Phosphatases; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors; Tumor Cells, Cultured

To investigate the cellular mechanism of bone destruction in collagen-induced arthritis (CIA).. After induction of CIA in DA rats, a histologic study of the advanced arthritic lesion was carried out on whole, decalcified joints from the hindpaws of affected animals. To conclusively identify osteoclasts, joint tissue sections were stained for tartrate-resistant acid phosphatase (TRAP) enzyme activity, and calcitonin receptors (CTR) were identified using a specific rabbit polyclonal antibody. The expression of messenger RNA (mRNA) for the osteoclast differentiation factor (also known as receptor activator of nuclear factor kappaB ligand [RANKL]) was investigated using in situ hybridization with a specific riboprobe.. TRAP-positive and CTR-positive multinucleated cells were invariably detected in arthritic lesions that were characterized by bone destruction. Osteoclasts were identified at the pannus-bone and pannus-subchondral bone junctions of arthritic joints, where they formed erosive pits in the bone. TRAP-positive multinucleated cells were detected within synovium and at the bone erosive front; however, CTR-positive multinucleated cells were present only at sites adjacent to bone. RANKL mRNA was highly expressed in the synovial cell infiltrate in arthritic joints, as well as by osteoclasts at sites of bone erosion.. Focal bone erosion in CIA is attributed to cells expressing definitive features of osteoclasts, including CTR. The expression of RANKL by cells within inflamed synovium suggests a mechanism for osteoclast differentiation and activation at sites of bone erosion. Inhibitors of RANKL may represent a novel approach to treatment of bone loss in rheumatoid arthritis.

Topics: Acid Phosphatase; Animals; Arthritis, Rheumatoid; Biomarkers; Bone Diseases; Carrier Proteins; Collagen; Disease Models, Animal; Female; Histocytochemistry; In Situ Hybridization; Isoenzymes; Membrane Glycoproteins; RANK Ligand; Rats; Receptors, Calcitonin; Tartrate-Resistant Acid Phosphatase

Guided bone regeneration (GBR) has been widely utilized for the promotion of bone augmentation in bone loss areas. However, little information has been available regarding chronological changes in newly formed bone and alterations in the nature of newly formed bone after removal of a barrier membrane. The present study attempted to establish a GBR model for rat maxillae. We also examined the effects of membrane application periods on newly formed bone and its remodeling process after removal of the membrane in this experimental model.. Thirty-five Wistar rats were divided into 2 groups: a membrane application group and a membrane removal group. The chronological changes of newly formed bone were evaluated histologically and statistically.. At 2 weeks after the GBR procedure, bony cavities had completely filled the newly formed bone in the experimental side. In the control side, corticalization on the surface of the newly formed bone proceeded with a decrease in the bone marrow cavity, whereas the bone marrow space had enlarged by 12 weeks post-surgery in the experimental side. In the membrane removal group, the osteoblasts appeared on newly formed bone at 1 week after membrane removal. Comparatively thick compact bone had formed on the surface of the newly formed bone at 4 weeks after membrane removal, and corticalization proceeded later.. The long-term application of a barrier membrane induces the enlargement of the bone marrow spaces. We suggest that PTFE membrane removal in adequate time promotes the corticalization and maturation of the newly formed bone by the GBR technique.

Topics: Acid Phosphatase; Alkaline Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Bone Marrow; Bone Remodeling; Disease Models, Animal; Guided Tissue Regeneration, Periodontal; Histocytochemistry; Isoenzymes; Male; Maxilla; Membranes, Artificial; Osteoblasts; Osteogenesis; Polytetrafluoroethylene; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Socket

Live mycobacteria secrete a number of unique proteins early in their multiplication which are important for both the pathogenesis and the stimulation of specific host responses. We have investigated the mechanisms by which the host mounts immune response against tuberculosis after vaccination with secretory proteins (SP) of a vaccine candidate Mycobacterium habana TMC 5135. Mice vaccinated with SP of 10th day growth of M. habana, either alone or emulsified in Freund's incomplete adjuvant (FIA) possessed antituberculous resistance and cellular immune responses against M. tuberculosis H37Rv. These proteins induced a significant cutaneous delayed type hypersensitivity response in guinea pigs vaccinated with heat killed M. tuberculosis H37Rv, which was equivalent to that observed with a standard purified protein derivative (PPD). The splenocytes of these guinea pigs have shown higher proliferative response after stimulation with SP than with PPD. The SP + FIA immunization has been found to exert maximum prophylactic effect by potentiating both the oxygen dependent arms and enzymatic activities of macrophages. Macrophages from mice vaccinated with SP of M. habana produced enhanced levels of interleukin(IL)-2, interleukin-12 and interferon(IFN)-gamma. The protective as well as cell mediated immune responses were upregulated in SP immunized animals when compared to whole cell (M. habana) vaccinated animals. SDS-PAGE of SP from M. habana showed the prominent bands of 60, 32, 31 and 30 kDa. Furthermore, the western analysis of SP with pulmonary tuberculosis patient's serum has revealed the presence of immunoreactive antigens of 36, 35, 33/32 kDa. Overall study demonstrated that the secretory antigens released by actively growing M. habana bacilli could activate different arms of effective immune response.

Topics: Acid Phosphatase; Animals; Bacterial Proteins; Blotting, Western; Disease Models, Animal; Electrophoresis, Polyacrylamide Gel; Glucuronidase; Interferon-gamma; Interleukin-12; Interleukin-2; Mice; Mice, Inbred AKR; Muramidase; Mycobacterium; Reactive Oxygen Species; Tuberculosis

We studied the activity of lysosomal enzymes (acid phosphatase and cathepsine D) and the status of lysosomal membranes of the liver, lung, heart and brain tissues of the male adult rats under acute hypoxia. We found that disturbances of lysosomal membranes permeability in these organs depend on the tissues hypoxia development, not yet on the genesis of hypoxic state. The lysosomal enzymes activation depends on the heaviness of the hypoxic influence.

Topics: Acid Phosphatase; Acute Disease; Animals; Brain; Cathepsin D; Disease Models, Animal; Heart; Hypoxia; Intracellular Membranes; Liver; Lung; Lysosomes; Male; Myocardium; Rats; Rats, Wistar

The dependent of DIC development on lysosomal apparatus neutrophils activity was founded. The most severe changes in hemostasis system were registered in the period of maximum acid phosphatus activity.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Disseminated Intravascular Coagulation; Female; Hemostasis; Lysosomes; Male; Neutrophils; Rabbits; Time Factors

The goal of this study was to determine the effects of chronically elevated blood androstenedione and estrone levels on the quality and quantity of both cancellous (trabecular) and cortical bone in a young (mean age 9.4 years) female primate model (M. fascicularis). Thirteen intact female monkeys received continuous androstenedione/estrone supplementation via subcutaneous implants over a 24-month period to simulate the human condition known as polycystic ovarian disease (PCOD). A group of 16 untreated intact age-matched female monkeys served as controls. Lumbar spine and whole body bone mineral density (BMD) status was determined mid-study by dual photon absorptiometry (DPA); subsequent analysis of the bone related to data obtained following the 2-year treatment period without further BMD measurement. Bone markers, including serum acid phosphatase, total bone alkaline phosphatase, bone gla protein and tartrate-resistant acid phosphatase were measured at the end of the study. At necropsy, the lumbar vertebrae and femora were recovered in order to analyze the bone mineral quality and quantity of cancellous and cortical bone respectively and to compare these with the control group. Mineralization profiles of the vertebrae and femora were obtained using the density fractionation technique. Chemical analysis of the three largest fractions retrieved by density fractionation was performed to evaluate differences in %Ca, %P, Ca/P ratio and mineral content (%Ca + %PO4) between control and experimental groups. In addition, unfractionated bone powder was examined by X-ray diffraction to identify any changes in crystal size. Coronal sections of vertebrae were analyzed for structural parameters using histomorphometry and image analysis. Cross-sections taken at the midshaft diaphyseal femora were analyzed for structural macroscopic and intracortical parameters. There was a significant increase in BMD at the L2-L4 region in the treatment group compared with the control groups (p < 0.005) as measured at 1 year into the trial. Serum acid phosphatase was significantly lower (p < 0.05) in the treatment group compared with the controls near study termination. A nonsignificant shift in the mineralization profile of the vertebrae towards less dense bone was observed in the treatment group, while there was a significant shift in the mineralization profile towards more dense bone in the treated femora compared with controls (p < 0.05) after a 2-year period. There was no differen

Topics: Absorptiometry, Photon; Acid Phosphatase; Alkaline Phosphatase; Androstenedione; Animals; Biomarkers; Bone and Bones; Disease Models, Animal; Estrone; Female; Image Processing, Computer-Assisted; Macaca fascicularis; Osteocalcin; Photography; Polycystic Ovary Syndrome

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Electromagnetic Phenomena; Glucuronidase; Humans; Lysosomes; Osteoarthritis; Rabbits

We have used a rabbit leg-lengthening model for detailed studies of the histology of distraction osteogenesis. Some unusual features of the endochondral ossification that occurs during the rapid transition of cartilage to bone in the regenerate were observed. Histological staining techniques together with immunohistochemistry and nonradioactive in situ mRNA hybridization for cartilage and bone-related molecules have been used to document the presence of an overlapping cartilage-bone phenotype in cells of the cartilage-bone transitional region. In those particular areas, some chondrocytes appeared to be directly transformed into newly formed bone trabeculae which are surrounded by bone matrix. Acid phosphatases were found within the cartilage matrix in some of the cartilage/bone transitional regions and type I collagen mRNA and type II collagen protein were found together in some of the marginal hypertrophic chondrocytes. This study indicates an unusual role of chondrocytes in the process of ossification at a distraction rate of 1.3 mm/day in the rabbit. Further direct evidence is required to prove the hypothesis that the hypertrophic chondrocytes may transdifferentiate into bone cells in this model.

Topics: Acid Phosphatase; Age Factors; Animals; Cartilage; Cell Differentiation; Chondrocytes; Collagen; Disease Models, Animal; Isoenzymes; Osteogenesis; Osteogenesis, Distraction; Rabbits; RNA, Messenger; Tartrate-Resistant Acid Phosphatase

Development of experimental the syndrome of disseminated intravascular coagulation, DIC syndrome, in organism laboratory animals from affect preparation "EFA-2" by accompanied increase quantity of neutrophiles circulations, modifications of the lysosomal apparatus of neutrophiles the activity of the serum acid phosphatase increased, characteristic damage on hemostasis system and typical alteration in some organs. As a result limitation of number of neutrophiles was achieved of suppression, of granulocytopoiesis by means "Myelosan" pharmacy, did not increase activity in blood serum solution lysosomal enzyme of neutrophils and not development in organism DIC syndrome. Made conclusion, the neutrophils have influence of promotion generalization DIC syndrome.

Topics: Acid Phosphatase; Animals; Biomarkers; Busulfan; Disease Models, Animal; Disseminated Intravascular Coagulation; Female; Hematopoiesis; Hemostasis; Lysosomes; Male; Neutrophils; Rabbits

The aim of this study was to investigate the effect of heparin on the rat bronchial mucosa changes induced by sulphur dioxide (SO2) inhalation. Sixty five rats were used in this experiment. Five of them constituted a control group, while 60 were exposed to SO2. Forty of the latter subgroup were additionally treated with low molecular weight heparin (LMWH), either during or after terminating exposure to SO2. In all animals exposed to SO2 inflammatory cells were found in broncho-alveolar lavage fluid (BALf) in numbers significantly higher from those observed in healthy controls. The rats exposed to SO2 and treated with LMWH showed intermediate cell pattern in the bronchi between healthy and SO2- exposed animals. When comparing histological picture of the bronchi, we noted extensive changes in irritated rats. These changes were either less expressed or totally absent in animals treated with heparin. The activity of enzymes: acid phosphatase (ACP), alkaline phosphatase (AP) and lactate dehydrogenase (LDH) rose in BALf, although the rise was not parallel and did not correlate with the magnitude of cellular influx or histological changes. Heparin did not influence this changes.

Topics: Acid Phosphatase; Animals; Bronchi; Bronchitis; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Heparin; L-Lactate Dehydrogenase; Male; Mucous Membrane; Partial Thromboplastin Time; Rats; Sulfur Dioxide

To investigate the participation of osteoclast-like bone resorbing cells in the joint destruction of murine collagen induced arthritis (CIA).. After induction of CIA in DBA/1J mice, a histological time course study was conducted on paw sections stained for tartrate resistant acid phosphatase (TRAP), a marker of osteoclasts. Cells from arthritic paws were cultured in vitro with or without indomethacin (IM) or anti-interleukin 6 neutralizing antibody (anti-IL-6), and stained for TRAP. Levels of prostaglandin E2 (PGE2), IL-1beta, IL-6, and tumor necrosis factor-alpha in the culture supernatants were determined by ELISA. The bone resorbing ability of these cells was examined on dentine slices. In control experiments, cells of normal paws or of arthritic tibiae were cultured in the same manner.. TRAP positive osteoclast-like cells were detected late in the development of bone lesions at every eroded front in the pannus-bone and the pannus-subchondral bone junctions of arthritic joints. In vitro, cells of arthritic paws formed bone resorbing osteoclast-like cells spontaneously. However, the control culture failed to form these cells. PGE2 and IL-6 were detected at higher levels in arthritic culture than in control culture. Although both indomethacin and anti-IL-6 reduced osteoclast-like cell formation and indomethacin inhibited PGE2 synthesis, indomethacin failed to reduce IL-6.. These findings suggest the direct participation of osteoclast-like cells in the joint destruction of CIA, the locally enhanced activity of osteoclast-like cell differentiation in arthritic paws, and the participation of prostaglandins and prostaglandin-independent IL-6 in this differentiation.

Topics: Acid Phosphatase; Animals; Arthritis, Experimental; Bone Resorption; Cell Count; Cells, Cultured; Collagen; Dinoprostone; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hindlimb; Indomethacin; Interleukin-1; Interleukin-6; Isoenzymes; Male; Mice; Mice, Inbred DBA; Neutralization Tests; Osteoclasts; Tarsal Joints; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

Accessory molecules and cytokines are involved in the immunopathogenesis of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) in rodent models, and are potential targets for immunotherapy. Evaluation of such experimental therapies requires appropriate animal models. Therefore, we analysed the expression of selected accessory molecules and cytokines in the brain of marmoset monkeys (Callithrix jacchus) with acute EAE, a newly described non-human primate model for MS. All animals experienced active disease clinically and histopathologically with strong resemblance to MS. Perivascular infiltrates of mononuclear cells showed abundant expression of CD40. CD40 was expressed on macrophages, indicating that T cell priming and macrophage effector functions may result from local CD40-CD40L interactions. CD40 ligand (CD40L) and B7-2 (CD86) were also expressed, but to a lower extent, while B7-1 (CD80) expression was limited. Both pro-inflammatory and anti-inflammatory cytokines were produced within individual lesions during active disease (IFN-alpha, IFN-gamma, TNF-alpha, IL-1alpha, IL-1beta, IL-2, IL-4, IL-10 and IL-12). This suggests that relative levels rather than sequential expression of Th1- and Th2-type cytokines determine disease activity. These findings demonstrate the value of EAE in marmoset monkeys as a model to assess the role of accessory molecules and cytokines in multiple sclerosis, and to evaluate targeted intervention.

Topics: Acid Phosphatase; Animals; Antigens, CD; B7-1 Antigen; B7-2 Antigen; Brain; Brain Chemistry; Callithrix; CD40 Antigens; Cytokines; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Histocompatibility Antigens Class II; HLA-DR Antigens; Interferon-alpha; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-2; Interleukin-4; Macrophages; Male; Membrane Glycoproteins; Multiple Sclerosis; Tumor Necrosis Factor-alpha

A tibial lengthening scheme in the mouse was used to study the molecular and cellular events regulating tissue regeneration during distraction osteogenesis. Here, we report on the surgical technique and frame design and describe the histochemical and molecular aspects of distraction during different phases of treatment. A total of 26 mice were used in this study. The treatment protocol was divided into a latency period of 7 days, a phase of active distraction that lasted 10 days with a distraction rate of 0.42 mm/day, and a maturation phase of 9 days. During latency, the distraction site resembled a stabilized fracture callus on both a histochemical and a molecular level. During active distraction, the gap was characterized by a central fibrous interzone bordered by primary matrix fronts, regenerate bone aligned with the distraction force, parallel columns of vascular sinusoids, and a medullary cavity. Alkaline phosphatase activity was detected in the endosteal and periosteal surfaces of the bone ends. Tartrate resistant acid phosphatase staining revealed that osteoclasts remodeled the bone regenerate as it formed. Collagen type I was expressed in the periosteum and the primary matrix front during distraction, whereas collagen type-II transcripts were localized to discrete regions on the periosteal surfaces, immediately adjacent to the osteotomy ends. Collagen type-II transcripts were not detected in the fibrous interzone. During the maturation phase, cells within the fibrous interzone expressed collagen type I and exhibited abundant alkaline phosphatase activity, suggesting that they had begun to terminally differentiate. Collectively, these data demonstrate the utility of a mouse model to study the molecular and cellular bases for the regeneration and remodeling of tissue.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Diseases; Bone Regeneration; Bony Callus; Collagen; Disease Models, Animal; External Fixators; Mice; Osteogenesis, Distraction; Radiography

We examined the effects of nandrolone decanoate (25 mg im every 3 weeks) on bone mass, serum biomarkers, and bone histomorphometric endpoints in 52 female cynomolgus macaques randomized into four treatment groups: (1) sham-ovariectomized (sham); (2) ovariectomized + placebo for 2 years (ovx); (3) ovx + nandrolone decanoate for 2 years (Nan); and (4) ovx + nandrolone decanoate beginning 1 year after ovx (dNan). Serum alkaline phosphatase (ALP), osteocalcin, and tartrate-resistant acid phosphatase (TRAP) were assayed every 3 months, and X-ray densitometry of the lumbar spine was done every 6 months. Fluorochrome-labeled iliac biopsies collected at baseline and 1 year, and lumbar vertebrae and midshaft femur collected at 2 years, were evaluated histomorphometrically. Body weight increased over 50% with administration of nandrolone. After 2 years, ovx animals had lower spinal BMC and BMD than all other groups. Ovx animals also had higher bone turnover rates than all other groups, as indicated by higher levels of the serum and urine biomarkers, and by at least twofold higher label-based bone formation rates in the femur diaphysis and in both cancellous and cortical bone of the ilium and vertebral bodies. Nandrolone-treated animals had similar serum estradiol levels as the sham animals, presumably due to conversion of endogenous or exogenous androgens. The effects of nandrolone on bone in this experiment are consistent with estradiol action and may be attributable to the increased serum estradiol. Despite >50% higher body weight, nandrolone-treated, ovariectomized animals did not have higher bone mass than sham animals.

Topics: Absorptiometry, Photon; Acid Phosphatase; Alkaline Phosphatase; Amino Acids; Anabolic Agents; Animals; Biomarkers; Body Composition; Bone Density; Bone Development; Bone Diseases, Metabolic; Disease Models, Animal; Estradiol; Female; Femur; Humans; Ilium; Isoenzymes; Lumbar Vertebrae; Macaca fascicularis; Nandrolone; Nandrolone Decanoate; Osteocalcin; Osteoporosis, Postmenopausal; Ovariectomy; Random Allocation; Tartrate-Resistant Acid Phosphatase

The anti-inflammatory activity of Salacia oblonga rootbark powder and Azima tetracantha leaf powder was assayed in male albino rats using carrageenan-induced rat paw oedema (acute inflammation) and cotton pellet granuloma (chronic inflammation) methods. Both the crude drugs were maximally active at a dose of 1000 mg/kg. In the cotton pellet granuloma assay, these drugs were able to suppress the transudative, exudative and proliferative components of chronic inflammation. Furthermore, these drugs were able to lower the lipid peroxide content of exudate and liver, gamma-glutamyl transpeptidase activity in the exudate of cotton pellet granuloma. The increased acid and alkaline phosphatase activity and decreased serum albumin in cotton pellet granulomatous rats were normalised after treatment with these drugs. It is likely that these drugs may exert their activity by antiproliferative, antioxidative and lysosomal membrane stabilization.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Anti-Inflammatory Agents; Antioxidants; Blood Chemical Analysis; Carrageenan; Disease Models, Animal; Edema; gamma-Glutamyltransferase; Gossypium; Granuloma, Foreign-Body; Lipid Peroxidation; Liver; Lysosomes; Male; Plant Extracts; Plant Leaves; Plants, Medicinal; Random Allocation; Rats; Rats, Wistar; Serum Albumin

Menkes disease is an X-linked disorder of copper metabolism. Excess amounts of copper in the kidney of Macular mice, a model for this disease, were found as copper-metallothionein (Cu-MT) from kidney of the mice. Histochemical studies of Cu-MT based on its autofluorescent emission properties showed that the protein was predominant in the proximal convoluted tubule (PCT) cells of the cortex. PCT cells are known to be the primary site of the nephrotoxicity caused by heavy metals. MT mRNA was also observed in the cortex, indicating that the protein was biosynthesized in this region. On the basis of these results, we suggest that biosynthesis and degradation of Cu-MT occur repeatedly in the PCT cells of the cortex. We also compared the histochemical localization of Cu-MT in Macular mice and Long-Evans cinnamon rats, a model for Wilson's disease. The significance of this comparison is discussed.

Topics: Acid Phosphatase; Animals; Carrier Proteins; Copper; Disease Models, Animal; Histocytochemistry; Injections, Subcutaneous; Kidney; Kidney Cortex; Kidney Tubules, Proximal; Lysosomes; Male; Menkes Kinky Hair Syndrome; Metallothionein; Mice; Mice, Inbred C3H; Mice, Inbred Strains; Rats; RNA, Messenger

In addition to various biological activities, interferon-gamma (IFN-gamma) inhibits bone resorption and collagen synthesis. We produced a transgenic mouse line expressing the murine IFN-gamma gene and protein in the bone marrow and thymus. Forty-five transgenic FVB/NCr mice, 23 days-9 months of age, were studied for anomalies in the skeletal system. The transgenic mice had short, wide, and deformed long bones. Young transgenic mice had epiphyseal plates severely thickened with zones of hypertrophy and degeneration with irregular metaphyseal borders. Cartilagenous masses were also observed in the metadiaphyseal marrow cavities. These lesions were primarily seen in long bones and ribs. Adult transgenic mice had residues of degenerated cartilagenous masses in the diaphyses. Many osteoclasts with well-developed ruffled borders were present on the metaphyseal cartilagenous masses in young transgenic mice. Adult transgenic mice had less prominent primary spongiosa with fewer osteoclasts at the metaphysis as compared with nontransgenic controls. The cortical bones of the transgenic mice were thinner and more immature compared with controls. Transgenic mice also had fractures, disruption of the epiphyseal plate, and degeneration of articular cartilage. Thus, the IFN-gamma transgenic mice developed a complex chondro-osseous lesion that was diagnosed as osteochondrodysplasia. The lesions may originate from primarily decreased matrix synthesis in bone and cartilage and also possible osteoclast-related changes caused by IFN-gamma overexpression in the bone marrow. Our IFN-gamma transgenic mouse will be a useful model to investigate the role of IFN-gamma in bone metabolism.

Topics: Acid Phosphatase; Animals; Antigens, Differentiation; Bone and Bones; Bone Marrow; Collagen; Disease Models, Animal; Female; Galectin 3; Gene Expression Regulation; Growth Plate; Immunohistochemistry; Incidence; Interferon-gamma; Isoenzymes; Male; Mice; Mice, Transgenic; Microscopy, Electron; Osteochondrodysplasias; Osteoclasts; Radiography; Tartrate-Resistant Acid Phosphatase; Thymus Gland

Pyelonephritis is the most common urinary tract infection affecting females of all age groups. Despite concerted efforts the mechanism of renal injury in pyelonephritis is not clearly understood. In the present study we have made an attempt to characterise the mediators of inflammatory insult in an experimental model of ascending pyelonephritis. Mice infected with Escherichia coli O6:K13:H1 were sacrificed at 2, 7 and 14 days post-infection. Luminol-dependent chemiluminescence response, NADPH oxidase, acid phosphatase, beta-glucuronidase and N-acetyl-beta-D-glucosaminidase activities were monitored in circulating as well as renal phagocytic cells in order to determine the role of reactive oxygen species and lysosomal enzymes in genesis of renal injury. We have demonstrated that reactive oxygen species are generated at the initiation of infection and the levels increase progressively during the course of infection. While intracellular release of lysosomal enzymes was seen in all groups, extracellular release was primarily observed at 7 and 14 days post-infection only. The results indicate that while reactive oxygen species play a significant role in tissue injury during all stages of infection, lysosomal enzyme release in extracellular milieu augments tissue destruction at later stages only.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Disease Models, Animal; Female; Glucuronidase; Kidney; Luminescent Measurements; Lysosomes; Mice; NADPH Oxidases; Pyelonephritis; Reactive Oxygen Species

Recent evidence has implicated canine distemper virus (CDV) as a possible aetiologic agent in Paget's disease of bone and the canine bone disorder, metaphyseal osteopathy. We have therefore examined the effects of CDV on the formation of multinucleated osteoclast-like cells in cultures of canine bone marrow mononuclear cells. Marrow cells from a distemper-infected dog and from five uninfected dogs were cultured in the presence of 1 alpha, 25-(OH)2 vitamin D3 and the number of tartrate resistant acid phosphatase positive multinucleated cells (MNCs) was determined. The presence of calcitonin (CT) receptors was confirmed by autoradiography with 125I-labeled human CT. Cultures from the distemper-infected dog contained a higher level of MNCs than those from the normal dogs. The in vitro addition of CDV to the cultures from all the dogs produced a dose-dependent increase in the number of MNCs, and an increase in size of these cells in the cultures from the infected dog. Cells infected with CDV were hyperresponsive to 1 alpha,25-(OH)2 vitamin D3. The presence of the virus in the relevant samples was confirmed using molecular techniques. In situ hybridization studies also revealed a significant increase in the level of infection following in vitro addition of the virus to the culture from the distemper-infected dog, suggesting that further infection had taken place. Resorption pits were formed on bone slices, although the number of pits was not significantly altered by viral infection.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acid Phosphatase; Animals; Autoradiography; Bone Marrow; Bone Marrow Cells; Bone Resorption; Calcitriol; Cells, Cultured; Disease Models, Animal; Distemper Virus, Canine; Dogs; Giant Cells; Humans; In Situ Hybridization; Isoenzymes; Osteitis Deformans; Osteoclasts; Polymerase Chain Reaction; Receptors, Calcitonin; Tartrate-Resistant Acid Phosphatase

Tartrate-resistant acid phosphatase (TRACP) and carbonic anhydrase II (CA II) are key enzymes responsible for osteoclastic bone resorption. In this study, we proposed that estrogen loss in postmenopausal osteoporosis may enhance gene expression of TRACP and CA II, and subsequently increase osteoclastic bone resorption. We have, therefore, used the ovariectomized rat model of postmenopausal bone loss to investigate changes at the gene transcriptional level in osteoclastic bone-resorbing enzymes in ovariectomized (OVX) rats, sham ovariectomized (S-OVX) rats, and estrogen-treated ovariectomized (E-OVX) rats. We have demonstrated for the first time that ovariectomy in rats enhances gene expression of TRACP, and CA II. The mRNA levels in OVX were approximately three- and four-fold higher, respectively, than those in S-OVX. Enhancement was observed 1 week after ovariectomy and transcripts remain high during the experimental period of 8 weeks. Administration of 17 beta-estradiol to OVX (E-OVX) reduced gene expression of these osteoclastic bone-resorbing enzymes 18 hours after injection. It appeared that the suppression of the osteoclastic bone-resorbing enzymes by 17 beta-estradiol was most effective during the first 1-2 weeks but the degree of suppression was reduced at 8 weeks after ovariectomy. In conclusion, our results suggest that estrogen prevents bone loss by reducing the mRNA levels of osteoclastic bone-resorbing enzymes in bone tissue.

Topics: Acid Phosphatase; Animals; Bone Resorption; Carbonic Anhydrases; Disease Models, Animal; Estradiol; Female; Gene Expression Regulation, Enzymologic; Humans; Osteoclasts; Osteoporosis, Postmenopausal; Ovariectomy; Rats; Rats, Sprague-Dawley; Tartrates

Bone resorption resulting from the metastatic human melanoma cell line (A375) was investigated morphologically using an experimental model of bone metastases in nude mice. An injection of A375 (1 x 10(5)) in the left ventricle produced multiple osteolytic lesions. Many TRAPase-positive multinucleated cells, identified by EM as osteoclasts, were observed on the bone surface at the site of metastases. The findings suggest that bone resorption was caused by osteoclasts developed in the presence of tumor cells. Even where tumor cells were juxtaposed to bone surface, small and flat TRAPase-positive cells were shown to exist on the bone surface. Thus, bone resorption was mainly associated with the occurrence of osteoclasts. A large number of osteoclast progenitor cells were also observed adjacent to tumor cells and/or stromal cells located apart from bone, indicating possible participation of tumor cells and/or stromal cells in the differentiation of osteoclasts. Ultrastructurally, stromal cells and/or extracellular matrices were present between tumor cells and osteoclast progenitor cells. Immunohistochemical observation clarified the localization of heparan sulfate proteoglycan (HSPG) and fibronectin (FN) around osteoclast progenitor cells. These findings suggest that they play an important role in providing a microenvironment favorable for osteoclast differentiation and activation. The immunohistochemical localization of IL-6, PGE2, and TGF-alpha also indicates that they are involved in osteoclast differentiation and activation. In conclusion, bone resorption at the metastatic sites of A375 is mediated via osteoclasts and A375 cells may be involved in the differentiation and activation of osteoclasts in association with stromal cells, extracellular matrices (HSPG, FN) and osteotropic cytokines (IL-6, PGE2, TGF-alpha).

Topics: Acid Phosphatase; Animals; Bone Neoplasms; Bone Resorption; Cell Differentiation; Dinoprostone; Disease Models, Animal; Fibronectins; Giant Cells; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Immunohistochemistry; Interleukin-6; Male; Melanoma, Amelanotic; Mice; Mice, Inbred BALB C; Mice, Nude; Microscopy, Electron; Osteoclasts; Proteoglycans; Stem Cells; Stromal Cells; Transforming Growth Factor alpha; Tumor Cells, Cultured

Adult male albino rats, maintained on normal or protein deficient diets from weanling, were exposed to repeated doses of MIC vapour (0.32 mg/L for 8 min for 5 consecutive days) under static conditions. Histopathology and the activities of alkaline and acid phosphatases and GSH content of lung were studied upto day 14 after exposure. Mild but repeated exposures of MIC vapour caused severe pulmonary lesions like denudation of bronchiolar epithelial lining tissue, cellular infiltration, edema, emphysema followed by hyperplasia, hypertrophy, fibrosis and intraluminal fibroplasia. The activities of alkaline and acid phosphatases were increased at earlier intervals while GSH content decreased significantly and remained low throughout the experimental duration. Protein deficiency was found to aggravate the toxic potentials of MIC in present condition.

Topics: Acid Phosphatase; Administration, Inhalation; Aerosols; Alkaline Phosphatase; Animals; Antisickling Agents; Disease Models, Animal; Glutathione; Isocyanates; Lung; Male; Occupational Exposure; Protein Deficiency; Random Allocation; Rats

A detailed investigation on the anti-inflammatory activity of the butanol fraction of a methanol extract (BMEL) of the defatted leaves of Paederia foetida was undertaken to find the pharmacological basis for the ethnomedical use of the plant. This fraction produced a significant inhibition of granulation tissue formation in cotton-pellet implanted rats. It decreased liver aspartate transaminase activity without affecting serum aspartate transaminase activity. It did not, however, affect adrenal weight and ascorbic acid content significantly, thus ruling out a stimulation of the adrenal-pituitary axis. BMEL antagonised hyposaline-induced haemolysis of human red blood cells and an elevation of rat serum acid phosphatase activity, indicating the presence of a membrane stabilising activity. It also inhibited the elevation of serum orosomucoid levels in rats, suggesting the possibility of the presence of disease-modifying antirheumatic activity. The results indicate that there is some rationale behind the ethnomedical use of the plant for treating inflammatory disorders.

Topics: 1-Butanol; Acid Phosphatase; Adrenal Glands; Animals; Anti-Inflammatory Agents; Ascorbic Acid; Aspartate Aminotransferases; Blood Proteins; Butanols; Chemical Fractionation; Disease Models, Animal; Erythrocyte Membrane; Female; Granuloma; Hemolysis; Humans; Liver; Male; Medicine, Traditional; Methanol; Organ Size; Orosomucoid; Plant Extracts; Rats; Spleen

Several parameters of bone mass and function were investigated in three experiments involving intact, ovariectomized, or hormone-supplemented ovariectomized female cynomolgus monkeys. Ovariectomized animals had increased serum levels of alkaline phosphatase and acid phosphatase compared with intact and hormone-supplemented animals. Vertebral bone mass measured ex vivo by dual-photon absorptiometry was reduced by 11-19% in ovariectomized animals compared with intact and hormone-supplemented animals. The most dramatic effects observed with ovariectomy were markedly increased (30-60%) bone formation rates in vertebral cancellous bone, primarily caused by higher activation frequency of basic multicellular units of bone. In addition, combined resorption and reversal periods were decreased and formation period increased in untreated ovariectomized animals. Changes in static histomorphometry parameters were less dramatic, cancellous bone volume being 1-14% lower in ovariectomized animals compared with intact or ovariectomized hormone-supplemented animals. The data indicate that changes in bone resorption are primarily responsible for the lower bone mass of estrogen deficiency and increased bone mass in hormone-supplemented animals. Bone changes in ovariectomized cynomolgus monkeys resemble those in women after menopause and similarly respond positively to hormone supplementation. As such, cynomolgus monkeys are an excellent model for studying the basic mechanisms of osteoporosis and for the development of suitable therapeutic regimens.

Topics: Acid Phosphatase; Age Factors; Alkaline Phosphatase; Animals; Biomarkers; Bone and Bones; Bone Density; Diet; Disease Models, Animal; Estrogen Replacement Therapy; Female; Humans; Macaca fascicularis; Osteocalcin; Osteoporosis, Postmenopausal; Ovariectomy

Our previous study reported the rich existence of multinucleated giant cells in an autoimmune myocarditis experimentally induced in rats. The present study investigated the histochemical and ultrastructural characteristics of these giant cells. Histochemistry for an acid phosphatase clearly demonstrated multinucleated giant cells dispersed at the inflammatory foci. Ultrastructurally, the giant cells were shown to be single cells, but not clustered cells. Their ultrastructural characteristics were very similar to the basic features of macrophages, except that the giant cells were poor in lysosomes and phagosomes. It was noticeable that some macrophages possessed three or more nuclei, displaying an intermediate form between mononuclear macrophages and multinucleated giant cells. These findings suggest that the giant cell in the experimental autoimmune myocarditis is a single multinucleated cell, and possibly derived from macrophages by cell-to-cell fusion.

Topics: Acid Phosphatase; Animals; Autoimmune Diseases; Cell Fusion; Disease Models, Animal; Giant Cells; Macrophages; Male; Myocarditis; Rats; Rats, Inbred Lew

The main purpose of this study was to investigate the influence of early total parenteral nutrition on acute sodium-taurocholate-induced pancreatitis in rats. Total parenteral nutrition did not change the survival rate, serum amylase, calcium or liver transaminase level on the degree of pancreatic damage, but reduced serum acid phosphatase and lactate dehydrogenase levels. Hyperglycemia occurred during the use of total parenteral nutrition. Total parenteral nutrition is not harmful in the course of acute experimental pancreatitis, and could be used with few side effects.

Topics: Acid Phosphatase; Acute Disease; Amylases; Animals; Calcium; Disease Models, Animal; Hyperglycemia; L-Lactate Dehydrogenase; Liver; Male; Pancreatitis; Parenteral Nutrition, Total; Rats; Rats, Sprague-Dawley; Transaminases

The influence of hypothyroidism in the adult rat on brain biochemistry was investigated. Hypothyroidism was induced in 6-month-old male rats by partial thyroidectomy coupled with the administration of 6-n-propyl-2-thiouracil (0.005%, w/v) in the drinking water. Age-matched euthyroid males served as the controls. Hypothyroidism resulted in brain region-specific changes in certain catabolic enzyme activities. Acid phosphatase activity was reduced in the cerebellum (by 34%) and the medulla (by 38%), whereas alkaline phosphatase activity was decreased in the midbrain (by 37%) and the subcortex (by 49%). A differential response was also observed in the case of aryl sulphatase activity: aryl sulphatase A (myelin-degradative activity) was diminished in the cerebellum (by 56%), whereas aryl sulphatase B remained unchanged in all regions. Acetylcholine esterase activity was reduced in the cerebellum (by 45%), the medulla (by 34%) and the subcortex (by 45%), whereas monoamine oxidase activity was affected in only one region, the cerebellum, where it was increased by (61%). The compromise of myelin and neurotransmitter degradative enzyme activities may place severe restrictions on normal brain function. The vulnerability of the adult rat cerebellum to the effects of thyroidectomy is commensurate with the known clinical signs of cerebellar dysfunction in adult hypothyroid man. These findings raise the possibility of an important role for the thyroid hormones in the mature brain.

Topics: Acetylcholinesterase; Acid Phosphatase; Alkaline Phosphatase; Animals; Brain; Cerebellum; Cerebroside-Sulfatase; Chondro-4-Sulfatase; Disease Models, Animal; Hypothyroidism; Male; Medulla Oblongata; Rats; Rats, Sprague-Dawley; Thyroidectomy

C2 mouse myogenic cells carrying the lacZ gene coding for beta-galactosidase (beta-gal) were injected into the tibialis anterior muscle of dystrophin-deficient mdx mice. Introduced cells were shown to have been incorporated into fibres of the injected muscle by virtue of the colocalization of beta-gal and dystrophin within them. Synthetic Nuclepore membrane inserted between the injected tibialis anterior and adjacent extensor digitorum longus muscle permitted the visualization of cells migrating between the two muscles through the pores of the membrane. Although the exact nature of the cells passing through the Nuclepore could not be determined by this method, they were thought to include implanted myogenic cells. Evidence for this was gained by the presence of beta-gal/dystrophin positive fibres within the extensor digitorum longus. Incorporation of cells into the adjacent extensor digitorum longus was greater in animals where this muscle had been autografted by the cutting and resuturing of the distal tendon. Autografted extensor digitorum longi differed from those which had not been subject to this procedure, by undergoing extensive fibre degeneration followed by regeneration, and further by the stripping of their surrounding epimysial covering. Implanted cells substantially participated in extensor digitorum longus fibre formation in these mice, up to 31% of their fibres 3 weeks after implantation coexpressing both the introduced lacZ gene product and the dystrophin gene product, the latter not normally expressed within the fibres of this myopathic recipient.

Topics: Acid Phosphatase; Animals; beta-Galactosidase; Cell Line; Cell Movement; Disease Models, Animal; Genetic Linkage; Glucose-6-Phosphate Isomerase; Lac Operon; Mice; Mice, Mutant Strains; Mice, Nude; Muscles; Muscular Dystrophy, Animal; Transfection; X Chromosome

Young and adult guinea pigs had been put in a chamber containing an 4-5 per cent addition of CO in the air. After 7 days of intoxication the authors performed a histopathological and histochemical examination of the retina. They discovered disturbances of the histological structure of the retina, a decrease of the content of nucleic acids and alkaline phosphatase and an increase of the acid phosphatase. The more pronounced intensification of the pathological changes was seen in the retina of younger individuals.

Topics: Acid Phosphatase; Age Factors; Air Pollutants; Alkaline Phosphatase; Animals; Carbon Monoxide Poisoning; Disease Models, Animal; Guinea Pigs; Nucleic Acids; Retina

We examined the effect of the amino bisphosphonate alendronate, administered IV every 2 weeks at 0.05 and 0.25 mg/kg for 1 year, on bone loss and parameters related to bone metabolism in ovariectomized baboons. Relative to non-OVX animals, the OVX baboons experienced increased bone turnover, reflected in biochemical and histomorphometric measurements, and bone loss assessed by dual-beam absorptiometry in the lumbar spine, which was similar to changes observed in ovariectomized women. Alendronate treatment maintained all parameters of bone turnover at control (nonovariectomized) levels and prevented the bone loss in a dose-dependent manner. We concluded that ovariectomized baboons offer a suitable model for the bone changes observed in ovariectomized women and that these changes can be prevented by sustained administration of an appropriate dose of this aminobisphosphonate.

Topics: Acid Phosphatase; Alendronate; Analysis of Variance; Animals; Bone and Bones; Bone Density; Calcium; Diphosphonates; Disease Models, Animal; Dose-Response Relationship, Drug; Estradiol; Female; Humans; Osteocalcin; Osteoporosis, Postmenopausal; Ovariectomy; Papio; Parathyroid Hormone; Radioimmunoassay

After a single intraperitoneal injection of cell wall fragments of Eubacterium aerofaciens, a main resident from the human intestinal flora, an acute arthritis develops within 2 days which is followed by a chronic arthritis that lasts at least 90 days. In an earlier report the histological appearance of the joint inflammation during this period has been described. In this study we investigated in more detail the cell types that are involved in the development of arthritis by using cell-type-specific monoclonal antibodies in an immunohistological assay. In the acute phase of arthritis, T-helper cells appeared in the synovial tissue together with ED1-positive (ED1+) and ED3-positive (ED3+) macrophages. After a temporary decline at day 12 all macrophage subsets, as well as T-helper cells, reappeared or increased again at day 33. Later, in the chronic phase (days 47-90), an increased number of ED1-positive (ED1+) cells in the synovial tissue and a decreased number of ED2-positive (ED2+) cells in the synovial lining was the most prominent finding when compared with control rats. These results indicate that, apart from T lymphocytes, macrophages also play an important role in the development and continuation of chronic arthritis in this model.

Topics: Acid Phosphatase; Animals; Antibodies, Monoclonal; Antibody Formation; Arthritis, Reactive; Cell Wall; Disease Models, Animal; Eubacterium; Female; Immunohistochemistry; Injections, Intraperitoneal; Rats; Rats, Inbred Lew; Synovial Membrane

Diabetes mellitus caused significant reduction in serum testosterone and accessory sex glands weight. The sperm content of epididymal regions also decreased. Among the epididymal regions, the cauda epididymidal tissue alone showed significant reduction in Na(+)-K+ ATPase activity. However, Mg2+ ATPase activity was lowered in caput epididymidis only. Specific activity of Ca2+ ATPase significantly decreased in caput and cauda epididymides. All three ATPases decreased significantly in caput epididymidal spermatozoa leaving cauda epididymidal spermatozoa unaffected. Specific activity of alkaline phosphatase was suppressed in caput epididymidis and in the spermatozoa collected from caput and cauda epididymides, while the acid phosphatase was unaffected. In general, the results are suggestive of definite influence of diabetes on epididymal phosphatases which is region specific. Diabetes induced decrease in phosphatases may have an impact on secretory and absorptive functions of epididymis and thus on sperm maturation.

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Alloxan; Animals; Ca(2+) Mg(2+)-ATPase; Cation Transport Proteins; Diabetes Mellitus, Experimental; Disease Models, Animal; Epididymis; Male; Organ Size; Prostate; Rats; Rats, Inbred Strains; Seminal Vesicles; Sodium-Potassium-Exchanging ATPase; Sperm Count; Testosterone

We have previously demonstrated the effects of various inhibitors of arachidonic acid metabolism on experimental lens-induced granulomatous uveitis. In the present study, we investigated the effect of these same inhibitors on the expression of lysosomal enzymes at different stages of choroidal inflammation in experimental lens-induced granulomatous uveitis and compared this to the inflammation observed at each stage examined. Lysosomal enzymes such as acid phosphatase, beta-glucuronidase and succinate dehydrogenase are known to be liberated during the maturation of mononuclear phagocytes to epithelioid cell granulomas. Although animals treated with nordihydroguaiaretic acid showed less severe inflammation than did indomethacin-treated or control animals, none of these agents appeared to affect the expression of acid phosphatase and beta-glucuronidase, as determined histochemically. Succinate dehydrogenase could not be detected in any of the eyes examined, even though sections of liver and kidney from these same animals were positive for this enzyme.

Topics: Acid Phosphatase; Animals; Anti-Inflammatory Agents, Non-Steroidal; Crystallins; Disease Models, Animal; Glucuronidase; Granuloma; Histocytochemistry; Indomethacin; Masoprocol; Rats; Rats, Inbred BN; Succinate Dehydrogenase; Uveitis, Posterior

The autosomal recessive disease Chediak-Higashi syndrome (CHS) is a progressive and generally fatal disease of humans. The underlying genetic defect in CHS is unknown and prenatal diagnostic methods have not been applied to this disease. The purpose of this study was to determine if CHS chorionic cells expressed a characteristic of CHS--enlarged lysosomes--that would permit the prenatal diagnosis of the disease. Cats with CHS, which have been shown to be homologous with human CHS, were used as the model system in this study. Chorionic tissue samples were obtained from CHS and control cat fetuses and cultures of cells were established. Acid phosphatase was utilized as a marker of lysosomes and cultures of chorionic fibroblasts from CHS and control fetuses were stained histochemically for acid phosphatase. The diameter of the largest lysosomes in 150 cells of each fetus was determined. The mean (+/- SD) diameter (in microns) of the largest lysosomes of normal fetuses was 0.9 +/- 0.13 (range 0.5-7.0 microns), whereas the mean diameter of lysosomes in CHS chorionic cells was 3.9 +/- 0.65 microns (range 0.5-25 microns). These means were significantly different (P less than 0.0001). These data suggest that it should be possible to diagnose human CHS in the first trimester by chorionic villus sampling.

Topics: Acid Phosphatase; Animals; Cats; Cells, Cultured; Chediak-Higashi Syndrome; Chorion; Disease Models, Animal; Lysosomes; Prenatal Diagnosis; Statistics as Topic

The activities of leucine amino peptidase (LAP) and acid phosphatase (Acid-P), conceivable markers of acute appendicitis, were determined in the portal blood of rabbits with acute appendicitis. An experimental model of acute appendicitis was established using No.-O silk ties to block the base of the appendix. The clinical and histopathological picture of acute appendicitis was seen after 12 hr in all the rabbits in the model group (9/9) and in none of the control group. Catheterization of the superior mesenteric vein was performed in rabbits with acute appendicitis, and portal blood samples were taken at 0, 6, and 12 hr for assay of LAP and Acid-P activities. No statistically significant difference between the experimental group and the control group, in the activities of LAP and Acid-P, was found at any time interval. The experimental model of acute appendicitis in the rabbit which is described here is simple and carries a high rate of success. This is probably the first report of using continuous catheterization and repeated sampling of portal blood, for the measurement of enzyme activities, in an experimental model of acute appendicitis. It was concluded that serum LAP and Acid-P activities cannot be used as markers for acute appendicitis.

Topics: Acid Phosphatase; Acute Disease; Animals; Appendicitis; Disease Models, Animal; Female; Leucyl Aminopeptidase; Male; Portal Vein; Rabbits

Chediak-Higashi syndrome (CHS) is an autosomal recessive disease in humans, cats, and 8 other species. The homology of CHS in humans and cats has been demonstrated. Since human CHS is a progressive, serious, and eventually fatal disease, a method for prenatal diagnosis would be desirable. This study was designed to determine whether CHS could be diagnosed prenatally by examination of amniotic fluid cells. The amniotic fluid samples were obtained from CHS and control cat fetuses on the 45th day of gestation and cultures of cells were established. Because the underlying enzyme deficiency in CHS has not been identified, it was necessary to use a secondary manifestation of the syndrome in these studies. The secondary manifestation used was the characteristic enlargement of lysosomes associated with the disease. The lysosomes of these cells were stained by acid phosphatase histochemistry and the diameter of the largest lysosome in each cell was measured by light microscopy with a calibrated ocular micrometer. The diameters of the largest lysosomes in cells of normal fetuses ranged from 0.5 to 7.0 micron (means ranged from 0.9 to 1.8 micron), whereas the diameter of the largest lysosomes in the cells of CHS fetuses ranged from 0.5 to 30 microns (means ranged from 6.4 to 12.8 microns). The approximate t-test for independent samples with unequal variances disclosed that the largest acid phosphatase-positive lysosomes in amniotic fluid cells of CHS cat fetuses were significantly larger than the lysosomes in the cells of normal cat fetuses (P less than 0.0001). This information should, by extrapolation, provide the basis for the prenatal diagnosis of human CHS by amniocentesis.

Topics: Acid Phosphatase; Amniocentesis; Amniotic Fluid; Animals; Cats; Cells, Cultured; Chediak-Higashi Syndrome; Disease Models, Animal; Female; Fetal Diseases; Lysosomes; Male; Phenotype

120 d after the experimental administration of ethanol to laboratory animals (mature male white rats), the changes manifesting themselves by the steatosis and the disorders of the carbohydrate balance and of the activity of the respiratory and the hydrolytic enzymes were observed in the animals' livers. There were no symptoms of a liver fibrosis.

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alcoholism; Alkaline Phosphatase; Animals; Disease Models, Animal; Ethanol; Histocytochemistry; L-Lactate Dehydrogenase; Liver; Male; Rats; Rats, Inbred Strains; Reference Values; Succinate Dehydrogenase

The Chediak-Higashi syndrome (CHS) is an autosomal recessive genetic disease of humans, and clinically similar diseases occur in cats, mink, cattle, mice, killer whales, blue foxes, and silver foxes. It is characterized by incomplete albinism, increased susceptibility to infection, and the most distinctive hallmark, the presence of enlarged cytoplasmic granules in many cell types. The acid phosphatase-positive granules, lysosomes, of fibroblasts from control and CHS humans, cats, mink, cattle, and mice were examined. These studies represent the initial characterization of the lesions in fibroblasts of CHS cats, mink, and cattle. Fibroblasts from each species and genotype were stained histochemically for acid phosphatase, and morphometric analysis of the distribution of acid phosphatase-positive granules was performed. The lysosomes in the CHS fibroblasts tended to be restricted to the perinuclear area of the cytoplasm, whereas the lysosomes in the normal fibroblasts were generally more widely distributed in the cytoplasm. The lysosomes in the CHS fibroblasts of all species examined were also more enlarged and heterogeneous than those in the control fibroblasts.

Topics: Acid Phosphatase; Animals; Cats; Cattle; Cells, Cultured; Chediak-Higashi Syndrome; Disease Models, Animal; Fibroblasts; Genotype; Histocytochemistry; Humans; Lysosomes; Mice; Mink

Malignant fibrous histiocytoma (MFH) was produced by injection of 9,10-dimethyl-1,2-benzanthracene (DMBA) into the rat knee joint. The tumor was observed in or around the knee in nearly all the animals 13 to 36 weeks after the initial DMBA administration. Histologically, these lesions were of the storiform-pleomorphic type (39/58, 67.2%), myxoid type (9/58, 15.5%), or giant cell type (8/58, 13.8%). Six cell types reported in human MFH were confirmed and phagocytosis of 0.81-micron latex particles by histiocyte-like cells was noted by electron microscopic examination. Acid phosphatase, beta-glucuronidase, and alpha-naphthyl acetate esterase were positive in enzyme histochemical examinations. Acid phosphatase activity was electron microscopically noted primarily in the lysosomes and the Golgi apparatus of the histiocyte-like cells. Cells from the storiform-pleomorphic (M1) and myxoid (M2) type tumors were serially transplanted subcutaneously in the back of the rats, and are now at the thirtieth and fortieth passage, respectively. They also were studied by enzyme histochemical and electron microscopic techniques. Our observations suggested an undifferentiated mesenchymal cell origin of MFH. Transplantable MFH can be produced in rats by intra-articular injection of DMBA, and lesions thus produced are a useful experimental model for the investigation of the histogenesis and the effect of chemotherapy of MFH.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Acid Phosphatase; Animals; Disease Models, Animal; Glucuronidase; Golgi Apparatus; Histiocytoma, Benign Fibrous; Histocytochemistry; Lysosomes; Male; Naphthol AS D Esterase; Neoplasm Transplantation; Phagocytosis; Rats; Rats, Inbred Strains

An experimental subcutaneous inflammation was produced in guinea pigs with peripheral blood eosinophilia. The eosinophilia resulted from two subsequent infections with Trichinella spiralis larvae. One group of guinea pigs served as non-infected control. Inflammation was induced by carrageenan, bradykinin, histamine, platelet activating factor and eosinophilotactic factors of lymphocytic or neutrophilic origin. Whereas in the control group no eosinophil granulocytic response was observed, this response was seen in the group with peripheral blood eosinophilia. The inflammatory substances and mediators (carrageenan, bradykinin, histamine, platelet activating factor) did not attract eosinophils alone, but also neutrophils. Under peripheral blood eosinophilia within the time course of the inflammatory reaction a second emigration with a shifted neutrophil/eosinophil ratio in favour of eosinophils was found. This could be due to a generation of chemoattractants by the injected substances themselves or more probably, by already emigrated granulocytes. Neither histamine, bradykinin, carrageenan, nor the eosinophilotactic factors (ECF's) in the concentrations used did release the cytotoxic major basic protein from eosinophils. Platelet activating factor exhibited a release of major basic protein from some eosinophils but no release of the peroxidase under our experimental conditions. The immigration of sufficient numbers of eosinophils into inflammatory areas might be one cause of the reduction of the inflammatory edema found in a previous investigation under similar conditions.

Topics: Acid Phosphatase; Animals; Arylsulfatases; Blood Proteins; Bradykinin; Carrageenan; Cell Movement; Chemotactic Factors; Chemotactic Factors, Eosinophil; Disease Models, Animal; Eosinophil Granule Proteins; Eosinophilia; Eosinophils; Female; Guinea Pigs; Histamine; Inflammation; Isoenzymes; Male; Neutrophils; Peroxidase; Peroxidases; Platelet Activating Factor; Ribonucleases; Time Factors; Trichinellosis

In osteoarthritis, despite increased matrix synthesis, there is a reduction of both major matrix components, proteoglycan and collagen. This study suggests that this is the result of enhanced degradative activity intrinsic to the cartilage. Because osteoarthritis is a focal disease, histologic controls were used to measure the severity in different areas of the cartilage and in different specimens. Neutral proteoglycanase and collagenase were both described in human cartilage, and their levels matched the severity of the disease as did acid phosphatase, a marker of lysosomal enzymes. Articular cartilage collagenase has an inhibitor in the cartilage and has negligible activity in normal cartilage. This was found not to be a lysosomal enzyme. A model of osteoarthritis was studied and found to have the same biochemical pattern as human disease. Using this method, inhibitors of degradative enzymes were used as a treatment. The chelator of metallic cations EDTA was found to have a significant effect on reduction of degradative enzyme activity and altered the arthritic process.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Cytidine; Disease Models, Animal; Edetic Acid; Egtazic Acid; Endopeptidases; Glycosaminoglycans; Humans; Hydrogen-Ion Concentration; Metalloendopeptidases; Microbial Collagenase; Osteoarthritis; Rabbits; Thymidine

The pathology of canine glycogen storage disease type II (acid alpha-glucosidase deficiency, GSD II) was studied in three genetically related Lapland dogs and compared to the pathology of human GSD II (McKusick 23230). Canine GSD II closely parallels the infantile form of the human disease, except for the presence of oesophageal dilatation. Generalized glycogen storage particularly affected muscular tissues (skeletal, oesophageal, cardiac and smooth muscle). The altered cells showed glycogen accumulation in the cytosol and in autophagic membrane-bound vacuoles (glycogenosomes). They also showed increased acid phosphatase activity consistent with the lysosomal nature of this storage disorder. The cytopathology in canine and human GSD II appears to evolve from segregation of glycogen during regular cellular autophagy, phagolysosomal accumulation of the undigested glycogen, and eventually rupture of distended glycogenosomes. This study indicates that the usefulness of canine GSD II as an animal model of human disease, extends to the area of pathogenesis.

Topics: Acid Phosphatase; Animals; Brain; Cytoplasm; Disease Models, Animal; Dogs; Esophagus; Female; Glycogen; Glycogen Storage Disease; Glycogen Storage Disease Type II; Humans; Kidney; Liver; Male; Microscopy, Electron; Muscle, Smooth; Muscles; Myocardium; Neurons; Spinal Cord; Vacuoles

Ligation of the common bile duct in pigs causes esophagogastric ulceration in 100 per cent of instances within seven days postoperatively. The results of previous studies have suggested a peptic cause for these ulcers. The possible role of lysosomal acid hydrolases in the primary pathogenesis was investigated herein. The total activities of two lysosomal acid hydrolases in mucosal specimens taken at biopsy from ulcerated pars esophagea and one in ulcerated gastric cardia were significantly lower in pigs undergoing ligation of the bile duct postoperatively, than in comparable pigs undergoing sham operations while there was evidence of increased extracellular activity. Mucosal deoxythymidine kinase activity was increased in gastric cardia suggesting attempted repair. Pigs with parakeratosis of the pars esophagea and edema of the gastric cardia, consistent with the preulcerative state also showed lowered enzyme activity. These results in intact pigs suggest that release of lysosomal acid hydrolases occurs with and may precede peptic ulceration.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; beta-Galactosidase; Cathepsin D; Disease Models, Animal; Gastric Mucosa; Hydrolases; Lysosomes; Peptic Ulcer; Swine; Thymidine Kinase; Time Factors

Monoarticular arthritis in the rabbit has been used to study the effect of intra-articular administration of cortisol-21-phosphate and alpha-1-proteinase inhibitor. The preparations were administered both separately and in combination. All treatments improved parameters associated with joint biochemistry and histopathology, but the greatest effect was found when steroid was combined with anti-proteinase. Cortisol-21-phosphate had both an anti-inflammatory and anti-arthritic action, whereas alpha-1-proteinase inhibitor showed little anti-inflammatory action but had some anti-arthritic effect. Alpha-1-proteinase inhibitor had no anti-inflammatory action against carrageenan induced oedema in the rat, but was anti-arthritic against adjuvant induced arthritis in the rat where it reduced both primary and secondary arthritis.

Topics: Acid Phosphatase; alpha 1-Antitrypsin; Animals; Arthritis; Arthritis, Experimental; Blood Proteins; Disease Models, Animal; Edema; Hydrocortisone; Injections, Intra-Articular; Proteins; Rabbits; Rats; Rats, Inbred Strains; Synovial Fluid

Majority of literature data support the significance of proteases activation in pathogenesis of acute pancreatitis. The ability of cathepsins to the activation of trypsinogen was shown and the labilization of lysosomes of pancreas in different models of acute experimental pancreatitis (AEP) was reported. In present work the dynamic of lysosomal changes during the course of AEP in dogs is evaluated. AEP was induced in 17 mongrel dogs by Elliot's method. Six healthy dogs served as a control group (I). Pancreatitic dogs were killed after 6 hr (G. II, n = 5), after 12 hrs (G. III, n = 5), and after 24 hrs (G. IV, n = 6 survivors). The pancreata were removed and divided into segments A (less advanced changes, [B] most advanced changes) and C (intermediate changes). The lysosomal enriched subfraction was isolated from the C segments at 15 000 X g for 20 min. The total (T) and free (F) activity of beta-glucuronidase (beta-G), acid phosphatase (AP), acid cathepsins (Cs) was estimated and the value F/T (relative free activity-r.f.a.) was calculated as an index of lysosomal stability. The progressive increase of r.f.a. of hydrolases in whole homogenate and in lysosomal enriched subfraction depending on time of AEP was observed suggesting labilization of pancreatic lysosomes. This labilization was more expressed in corresponding parts of organ with more advanced pathological changes. The differences between part A and B were most evident after 6 hrs of AEP. The labilization of lysosomes is more pronounced after 12 and 24 hrs than after 6 hrs in analogical parts of organ. These results indicate that labilization of lysosomes in pancreas correspond to the degree of pathological changes of pancreatic tissue.

Topics: Acid Phosphatase; Acute Disease; Animals; Cathepsins; Disease Models, Animal; Dogs; Female; Glucuronidase; Hydrolases; Lysosomes; Male; Pancreas; Pancreatitis; Reference Values; Time Factors

The ultrastructural alteration of Langerhans' islet cells as well as acinar structure in endotoxin shock was investigated in virtue of the electron histochemical procedures. One hour after administration of endotoxin, swelling and loss of cristae of the mitochondria, dilatation of the cisterna of the rough endoplasmic reticulum and swelling of the secretory granules in the B cell of Langerhans' islet cell were observed. On the contrary, the teleinsular acinar cells disclosed relatively intact ultrastructure, while the periinsular acinar cells showed in rather remarkable changes, such as the swollen mitochondria and obliteration of the rough endoplasmic reticulum. Four hours after administration of endotoxin, increase of the autophagic vacuoles in the cytoplasm of the acinar cells and in B cells of the Langerhans' islet cells were observed. Electron histochemical study disclosed the increase of rather small primary lysosomes in B cells of the Langerhans' islet cells, comparing to those of the acinar cells in normal condition. However, one hour after administration of endotoxin, acid phosphatase activity was activated and dispersed reactive substances were discernible in the cytoplasm of the B cells as well as intercellular and perivascular spaces.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Islets of Langerhans; Lysosomes; Male; Microscopy, Electron; Rats; Rats, Inbred Strains; Shock, Septic

A partial lateral meniscectomy procedure has been developed for the induction of a predictable and reproducible degenerative joint disease in knees of rabbits. The procedure adopted involves section of the fibular collateral and sesamoid ligaments and removal of 4-5 mm of the anterior lateral meniscus. In most experiments the animals are killed and tissues obtained for histologic examination at 6 weeks. Section of the ligaments alone (with or without penetration of the joint space) did not result in significant pathologic change. Significant degeneration was observed in tibial and femoral cartilage when the meniscus as well as the ligaments were cut, but the most extensive lesions were seen when a piece of the anterolateral meniscus was actually removed. These lesions included fibrillation, ulceration and erosion, "clone" and osteophyte formation, loss of chondrocytes, and loss of safraninophilic staining in the articular cartilage. The incidence and distribution of lesions with time following surgery were also investigated. Lesions were observed as early as 1-2 weeks post-surgery and increased in number and severity up to 12 weeks. A global scoring system has been devised to permit statistical comparisons of lesion incidence and severity in different groups of rabbits. This scoring system has enabled us to test drug efficacy in the rabbit lateral meniscectomy model of osteoarthritis.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Histocytochemistry; Knee Joint; Ligaments, Articular; Male; Menisci, Tibial; Osteoarthritis; Rabbits

Topics: Acid Phosphatase; Animals; beta-Glucosidase; Brain; Cerebrosides; Disease Models, Animal; Galactosylceramides; Gangliosides; Gaucher Disease; Humans; Inositol; Mice; Myelin Sheath; Spleen

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Dogs; Humans; Male; Mice; Mice, Nude; Prostatic Neoplasms; Rats; Rats, Inbred Strains; Substrate Specificity; Vitamin K

Essential fatty acid deficient (EFAD) rats are significantly more resistant to the lethal effects of S. enteritidis endotoxin (20 mg/kg, IV) than normal control rats. Compared to endotoxin-treated normal rats, EFAD rats also manifested less severe alterations of hepatic and lysosomal integrity and became less hypoglycemic. Administration of the ethyl ester of the essential fatty acid, arachidonic acid (100 mp, IP) two days prior to challenge with S. enteritidis endotoxin (20 mg/kg) in EFAD rats restored their sensitivity to endotoxin, as denoted by a 100% mortality compared to a 24% mortality (P less than 0.01) in EFAD rats. Treatment of EFAD rats with the fatty acid docosahexaenoic acid, a non-prostaglandin and thromboxane precursor, (100 mg, IP) produced significantly less (less than 0.01) mortality than ethyl-arachidonate-treated groups (ie, 40% vs 100%). The arachidonate metabolite, thromboxane B2 (TxB2), increased from nondetectable plasma levels (less than 200 pg/ml) to 2285 +/- 449 pg/ml (N = 10) at 30 min and remained elevated for 180 minutes after endotoxin administration in nondeficient rats. However, plasma TxB2 was not detectable in endotoxin-treated EFAD rats and was only slightly elevated in groups supplemented with docosahexaenoic acid (273 +/- 104 pg/ml, N = 6) after 30 minutes. In ethyl arachidonate (100 mg, IP) supplemented EFAD rats, plasma TxB2 rose to 873 +/- 204 pg/ml (N = 8), 30 min after endotoxin. Pretreatment of the ethyl-arachidonate-supplemented EFAD group with a specific thromboxane synthetase inhibitor, 7-(1-imidazolyl)-heptanoic acid (30 mg/kg, IV), significantly reduced mortality 100% to 50% (P less than 0.05) from endotoxic shock. These observations suggest a deleterious role for arachidonic acid and its conversion to TxA2 in the pathogenesis of endotoxic shock.

Topics: Acid Phosphatase; Animals; Arachidonic Acid; Arachidonic Acids; Blood Glucose; Disease Models, Animal; Fatty Acids, Essential; Female; Fibrin Fibrinogen Degradation Products; Male; Muridae; Pregnancy; Shock, Septic; Thromboxane B2

The effect of 3 alpha-androstanediol (3 alpha-diol), 17 beta-estradiol (E2) and cyproterone acetate (CA) on prostates in castrated beagle dogs were investigated by histological and histochemical examinations. A significant increase of prostatic weight occurred after 6 months' treatment with 3 alpha-diol alone and in combination with E2. Histologically and histochemically, two different types of prostatic enlargement were observed: first, administration of 3 alpha-diol resulted in diffuse glandular hyperplasia with replacement of functional activity monitored by strongly positive reactions for acid phosphatase, aminopeptidase and zinc. Second, 3 alpha-diol plus E2 produced stratified squamous metaplasia with cystic lumina and loss of the typical morphological structure. These glands showed negative reactions for acid phosphatase, aminopeptidase and zinc. In both types of prostatic hyperplasia CA abolished epithelial or metaplastic proliferation and induced atrophy of glandular epithelium. In estrogenized dogs activation of the fibromuscular stroma was obvious. CA prevented prostatic hyperplasia by atrophying epithelial effects.

Topics: Acid Phosphatase; Aminopeptidases; Androstanols; Animals; Castration; Cyproterone; Cyproterone Acetate; Disease Models, Animal; Dogs; Estradiol; Male; Prostatic Hyperplasia; Zinc

Topics: Acid Phosphatase; Adenocarcinoma; Animals; Antineoplastic Agents; Castration; Disease Models, Animal; Male; Neoplasms, Experimental; Prostatic Neoplasms; Rats

The efficacy of corticosteroid therapy in the treatment of shock remains controversial. In order to evaluate this question, the following controlled experimental study was undertaken. There were 44 puppies (2-6 kg) used to examine the effects of methylprednisolone (30 mg/kg) in both hemorrhagic and live Escherichia coli septic shock. In order to isolate the effects of steroid treatment, no volume or antibiotic therapy was given. Arterial, venous, and pulmonary artery catheterization allowed continuous hemodynamic and metabolic monitoring. One control group received steroid treatment and was not subjected to shock. Septic shock was achieved by a rapid bolus infusion of 10(9) live E. coli organisms. Hemorrhagic shock was produced by bleeding the puppy an average of 43% of his blood volume. Four septic and four hemorrhagic shock groups received either no treatment, steroids at the time of shock, or steroids 30 min before or after shock. Cardiac outputs of less than 50% of control values and significant lactic acidosis were documented in all of the shock animals. The septic groups showed more profound alterations in these parameters and a decreased overall survival. No statistically significant differences could be found, however, in the hemodynamic, metabolic or survival figures among the different septic shock groups, or among the various hemorrhagic shock animals. The anticipated preservation of cardiac output and decrease in leakage of lysosomal acid phosphatase were not seen with any treatment schedule. The theoretical benefits of corticosteroid treatment in shock could not be documented in these two models of severe septic and hemorrhagic shock in puppies.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Dogs; Escherichia coli; Fatty Acids, Nonesterified; Glucose; Hemodynamics; Lactates; Methylprednisolone; Methylprednisolone Hemisuccinate; Shock, Hemorrhagic; Shock, Septic; Triglycerides

Rabbit hearts perfused under hypoxic conditions underwent progressive subcellular damage, which becomes irreversible by one hour. During the first 20 minutes of perfusion, minor dilation of mitochondria and condensation of nuclear chromatin were the only salient features of cell injury. By 40 minutes moderate mitochondrial swelling was evident in hypoxic myocytes. Moreover, an increase in degenerating mitochondria and autophagic vacuoles was apparent. Reperfusion after either 20 or 40 minutes of hypoxia restored contractility, and injured myocytes underwent a cellular repair process that involved a dramatic increase in lysosomal autoplagy. One hour of hypoxia yielded irreversibly injured myocytes. Upon reoxygenation, some of these cells displayed typical changes of necrosis, but others apparently underwent an abortive repair process involving the formation of large, probably nonfunctional lysosomes. These observations suggest that lysosomal autophagy is important in the efforts at repair that cardiac cells initiate during and after hypoxia.

Topics: Acid Phosphatase; Animals; Autolysis; Coronary Disease; Disease Models, Animal; Hydrolases; Hypoxia; In Vitro Techniques; Lysosomes; Male; Mitochondria, Heart; Myocardium; Oxygen Consumption; Rabbits

Topics: Acid Phosphatase; Adenocarcinoma; Animals; Cell Line; Disease Models, Animal; Humans; Karyotyping; Lymphatic Metastasis; Male; Mice; Mice, Nude; Middle Aged; Neoplasms, Experimental; Prostatic Neoplasms

The Dunning rat prostatic adenocarcinoma models (R-3327 series) are providing a valuable system for elucidating new principles of prostatic tumor biology. Each of the new lines provides a new tool for these studies, but each line must be carefully characterized and monitored.

Topics: Acid Phosphatase; Adenocarcinoma; Animals; Castration; Diethylstilbestrol; Disease Models, Animal; Flutamide; Male; Neoplasm Metastasis; Prostatic Neoplasms; Rats; Steroids; Testosterone

Topics: Acid Phosphatase; Animals; Coronary Disease; Disease Models, Animal; Dogs; Indomethacin; Intracellular Membranes; Lysosomes; Male; Myocardium; Nucleotides, Cyclic; Prostaglandins

A histochemical enzyme profile has been used to determine the origin of the cells of glomerular crescents in rabbit and sheep models of rapidly progressive crescentic glomerulonephritis. Crescentic glomeruli were examined for beta-glucuronidase, esterase and acid phosphatase, the characteristic phagolysosomal enzymes of the monocyte-macrophage series and its transformed tissue counterpart, the epithelioid cell. More than 50% of the cells of the glomerular crescents of animals with experimental crescentic glomerulonephritis contained large amounts of all the enzymes, whereas the cells of normal glomeruli contained only trace amounts of acid phosphatase and esterase and no glucuronidase. These findings support the hypothesis that the major proportion of the cells of glomerular crescents are monocytes which have accumulated in Bowman's space rather than intrinsic glomerular cells which have proliferated.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Esterases; Glomerulonephritis; Glucuronidase; Histocytochemistry; Kidney Glomerulus; Macrophages; Monocytes; Rats; Sheep

A high incidence of spontaneous, non-acute, age-dependent prostatitis was observed in the lateral prostate of Copenhagen rats and Wistar rats. The lumen of infected acini was filled with polymorphonuclear leucocytes, shed epithelial cells and cell residues. Epithelial cells lining such acini showed degenerative changes. Lymphocytes and macrophages were seen in the stroma. A histochemically observed increase in acid phosphatase and beta-glucuronidase activity in affected epithelial cells may indicate an increased lysosomal activity. Some bacteriological cultures of infected lateral prostates were positive for Proteus vulgaris and diphtheroids. It is suggested that this spontaneous rat prostatitis may be a useful model for the study of the pathogenesis and treatment of human non-acute prostatitis.

Topics: Acid Phosphatase; Age Factors; Alkaline Phosphatase; Animals; Bacteria; Disease Models, Animal; Epithelium; Esterases; Glucuronidase; Leucyl Aminopeptidase; Male; Prostate; Prostatitis; Rats

A transplantable, metastasizing prostatic adenocarcinoma (Tumor I) in Lobund Wistar rats was examined for activity and distribution of five hydrolytic enzymes and for ability to accumulate radioactive zinc. The results suggest that the tumor had arisen in the ventral lobe of the prostate and that its growth was not affected by orchiectomy, adrenalectomy, or replacement treatment with exogenous androgen or corticosteroids. The androgen independency of the tumor was further shown by the low uptake of 3H-testosterone, in contrast to the high uptake in the ventral prostate. Tumor growth was retarded by Cytoxan but not by 5-fluorouracil, Estracyt, or streptozotocin, three agents clinically effective in the treatment of some patients with prostatic cancer resistant to endocrine therapy. It is concluded that this tumor in Lobund Wistar rats may be an adequate model for human prostatic cancers resistant to the agents mentioned above.

Topics: Acid Phosphatase; Adenocarcinoma; Adrenal Cortex Hormones; Adrenalectomy; Alkaline Phosphatase; Aminopeptidases; Animals; Antineoplastic Agents; Castration; Disease Models, Animal; Drug Resistance; Esterases; Glucuronidase; Male; Neoplasm Metastasis; Prostatic Neoplasms; Rats; Testosterone; Tritium; Zinc Radioisotopes

A model of chronic pancreatitis has been created in 13 mongrel dogs. The animals were withdrawn from the experiment 2 months after the beginning of it. According to the author's data, artifically created insufficiency of the constrictors of the common bile and pancreatic ducts in dogs is analogous to unfitness of the sphincter in man and results in the development of the changes in the abdominal cavity, specific for chronic pancreatitis.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Chronic Disease; Common Bile Duct; Disease Models, Animal; DNA; Dogs; Pancreas; Pancreatic Ducts; Pancreatitis; RNA; Sphincter of Oddi

Acid phosphatase and beta-glucuronidase were used as marker enzymes of lysosome, and their role in adjuvant-induced uveitis was studied. These enzyme activities in the iris-ciliary processes were increased in the inflamed tissues. Changes in these enzyme activities in the tissues paralleled the development of uveitis. While protein concentration in the aqueous humor as a parameter of vascular permeability was significantly correlated with these enzyme activities, there was no correlation in the leucocyte counts in the aqueous humor. Topically applied dexamethasone reduced the increase in the aqueous protein, the leucocyte migration and the swelling of the iris-ciliary processes, while topically applied indomethacin reduced only the leucocyte migration among these inflammatory parameters. Acid phosphatase activities in the inflamed tissues were reduced also by dexamethasone, but not by indomethacin.

Topics: Acid Phosphatase; Adjuvants, Immunologic; Animals; Anti-Inflammatory Agents; Dexamethasone; Disease Models, Animal; Glucuronidase; Indomethacin; Iris; Lysosomes; Male; Rabbits; Uveitis

Topics: Acid Phosphatase; Animals; Brain; Cerebrosides; Disease Models, Animal; Gangliosides; Gaucher Disease; Humans; Inositol; Liver; Mice; Spleen; Xylosidases

Lepromatous tissue from armadillos inoculated 24--36 months earlier with Mycobacterium leprae was obtained for electron microscopic studies. Cytochemically stained lepromas revealed a subpopulation of macrophages containing peroxisomes. These peroxidase reactive macrophages were not infected with bacilli. Acid phosphatase was present in macrophages and many of these were infected with bacilli and contained vacuoles and lipid globules. Within the membrane-bound vacuoles, acid phosphatase surrounded bacilli. However, the reaction product ended abruptly at a 15--40 millimicron thick zone of low electron density surrounding intact bacilli. Acid phosphatase was more intensely reactive and localized less precisely in heavily infected and vacuolated macrophages than in lightly and non-infected cells. The effectiveness of this bacillary barrier and the numerous infected macrophages with substantial acid phosphatase argue against the ability of acid phosphatase to protect host cells from leprosy bacilli. Evidence suggests a protective action of peroxidase or the rapid turnover of macrophages within lepromas. Granular and membranous debris were commonly seen within vacuoles of infected macrophages. A portion of the debris was ultrastructurally similar to bacillary matrix and was nonreactive for peroxidase and acid phosphatase. Following homogenization and centrifugation, similar materials banded with bacilli above 60% sucrose. Another portion of the debris was ultrastructurally similar to host lysosomal matrix and was reactive for acid phosphatase. Results support the concept of dual host and parasitic origins of the debris found in phagolysosomes of infected macrophages. Transparent, oval Epon defects remained eccentric to the majority of intact bacilli in centrifuged fractions. Apparently, an intrinsic property of leprosy produced these Epon defects.

Topics: Acid Phosphatase; Animals; Armadillos; Disease Models, Animal; Histocytochemistry; Leprosy; Macrophages; Mycobacterium leprae; Peroxidases; Vacuoles

Topics: Acid Phosphatase; Animals; Demyelinating Diseases; Disease Models, Animal; Macrophages; Peptide Hydrolases; Peripheral Nerves; Polyradiculoneuropathy; Rabbits; Spinal Nerve Roots

Occlusion of the circumflex coronary artery induced a profound redistribution in ischemic rabbit myocardium of several lysosomal acid hydrolases, including cathepsin D, B-acetylglycosaminidase, and acid phosphatase. 30-45 min after ligation non-sedimentable cathepsin D activity rose from 36% of the total activity to 42-48%, and in immunohistochemical preparations cathepsin D appeared to have diffused from lysosomes into the cytosol of injured cells. A pharmacologic dose of methylprednisolone (50mg/kg) significantly delayed the subcellular redistribution of cathepsin D and the other hydrolases in ischemic heart. Thus, in treated hearts the nonsedimentable activity of cathepsin D rose to only 38% after 30 min of ischemia and 42% after 45 min (P is less than 0.05 compared to untreated ischemia at each time). Similarly, unlike untreated hearts, noevidence of enzyme diffusion from lysosomes could be demonstrated immunohistochemically in corticosteroid-treated ischemic hearts for over 45 min. After 1-2 h of ischemia, however, steroid-protected myocytes deteriorated and the biochemical activity and anatomical distribution of cathepsin D were indistinguishable from untreated ischemic hearts. This study demonstrates that corticosteroid pretreatment does not prevent alterations in cardiac lysosomes during severe ischemia indefinitely, but does delay their development significantly.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Cathepsins; Coronary Disease; Disease Models, Animal; Lysosomes; Male; Methylprednisolone; Myocardium; Rabbits; Time Factors

A machine designed upon the principles of a crank lever mechanism was built to produce excessive use of the joints of small animals by means of continuous flexion-extension movements. Articular cartilage changes could be consistently produced after 5 to 10 days of exercise. Histochemical studies of the articular cartilage demonstrated a decrease in proteoglycan and an increase in acid phosphatase. These findings suggest that this animal model may be of value for the analysis of the earliest degenerative stages of the articular cartilage.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Male; Physical Exertion; Physiology; Proteoglycans; Rats; Synovial Fluid

Acute uremia was induced in male Swiss albino mice by complete urethral ligation and the animals were sacrificed 2, 4-6, 24, and 48 h after operation. Sham-opeated animals (without the urethral ligation) were similarly treated. The blood urea levels of animals with total urinary tract obstruction went up to 175 mg/100 ml at 4-6 h of urethral ligation and reached an average level of 827 mg/100 ml at 48 h, while the control group exhibited and average blood urea level of 37 mg/100 ml. Lysosomes obtained from livers of uremic mice sacrificed at different time intervals demonstrated a lability of the lysosomal membranes (as determined by the acid phosphatase activity in mU/mg) which was maximal at 4-6 h of urethral ligation, declining towards normal at 24 and 48 h, despite an increase in the animal's blood urea. In vitro studies exposing liver lysosomes to progressively higher urea concentrations (differences of as much as 100,000 times) did not reveal any effect of urea upon the stability of lysosomal membranes. The reason for the lability of lysosomal membranes in the uremic group was not apparent in the present study.

Topics: Acid Phosphatase; Acute Disease; Animals; Cell Fractionation; Disease Models, Animal; Liver; Lysosomes; Male; Membranes; Mice; Urea; Uremia

The Dunning R3327 prostate adenocarcinoma of the Copenhagen rat was developed as a suitable model of human prostate cancer. Inoculation of tumor tissue mince or cells sc in the flanks of recipient rats produced tumors that had the macroscopic and microscopic characteristics of the human disease. The histologic picture of these tumors was that of a well-differentiated adenocarcinoma with the formation of glands and acid secretions within the acini. Tumors were also produced in the dorsolateral lobe of the prostate by the injection of cells. The intraprostate tumor, although initially confined to the injected lobe, grew to involve the surrounding tissues and eventually metastasized to the lymph nodes and lungs. Occasional metastatic lesions were found in other organs also. Acid phosphatase could be domonstrated by histochemical staining of frozen tumor sections, and elevated levels of the enzyme were seen in the sera of rats bearing long-term subcutaneous tumors. During investigation of the tumor, a fast-growing tumor line arose that grew equally as well in females as in males. The histology of this tumor was that of an undifferentiated anaplastic tumor.

Topics: Acid Phosphatase; Adenocarcinoma; Anaplasia; Animals; Disease Models, Animal; Female; Male; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Prostatic Neoplasms; Rats; Rats, Inbred Strains; Transplantation, Isogeneic

Feeding of fructose for 7 days has been morphometrically shown to induce a SER-reduction and an accumulation of glycogen in rat liver cells. This hypothetical model "glycogenosis" is investigated with histochemical methods.. Rats are given a solution of 60% fructose in water as only nutritional source. Controls are given a solution of 60% glucose in water, an isocaloric Altromin-R-standard diet and an Altromin-R-standard diet ad libitum. Reversion of fructose induced metabolic changes is investigated by a 7 days fructose diet followed by an 1-4 days Altromin-R-standard diet ad libitum. Glycogen and glycogen metabolizing enzymes are demonstrated after a 7 days diet and in the course of an 1-7 days fructose diet.. Feeding of fructose leads to a high glycogen content, combined with a high activity of glycogen-phosphorylase and glucose-6-phosphatase in the liver parenchyma. Glycogen-synthetase activity increases during the first 4 days and then it drops to a low level. A pathological alteration of liver cell metabolism seems to be improbable, for all fructose induced changes are reversibel after 2 days of Altromin-R-standard diet. Glucose-6-phosphatase, as a marker-enzyme of the smooth endoplasmatic reticulum, is discussed to become activated by disruption of SER membranes due to fructose.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Endoplasmic Reticulum; Fructose; Glucose; Glucose-6-Phosphatase; Glycogen Storage Disease; Glycogen Storage Disease Type I; Glycogen Synthase; Histocytochemistry; Liver; Liver Glycogen; Male; Phosphorylases; Rats

Topics: Acid Phosphatase; Animals; Demyelinating Diseases; Disease Models, Animal; DNA; Encephalomyelitis, Autoimmune, Experimental; Glucosephosphate Dehydrogenase; Haplorhini; Humans; Hydrolases; L-Lactate Dehydrogenase; Mice; Multiple Sclerosis; Wallerian Degeneration

Topics: Acid Phosphatase; Animals; Cathepsins; Disease Models, Animal; Hymenolepiasis; Liver; Lysosomes; Male; Mice; Nitrogen

After inducing experimental ileus in albino rats, the authors found changed lysosome enzyme activities in liver homogenizate. In the same kind "free" activity of acid phosphatase and acid ribonuclease is elevated by strangulation ileus. According to literature, these alterations result from changed permeability of lysosome membranes, resp. from rupture of lysosomes. Ileus by obstruction causes no significant changes of the "free" lysosomes activities in liver homogenizate. Increase of acid phosphatase and ribonuclease in blood serum by strangulation or obstruction is equally considerable in both kinds of ileus. The results of these experiments suggest the developing of hepatic damage under both kinds of experimental ileus, the extent of which can be assessed by determination of lysosome enzyme activities.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Endonucleases; Intestinal Obstruction; Lysosomes; Rats; Ribonucleases

Topics: Acid Phosphatase; Acute Disease; Alkaline Phosphatase; Animals; Chronic Disease; Disease Models, Animal; Erysipelothrix Infections; Joints; Rats; Synovial Membrane

Myocardial contractility was evaluated in 8 out of 14 anaesthetized mongrel dogs during haemorrhagic shock and after volume replacement with Dextran 60 using the force-velocity relation of the contractile elements at zero load (Vmax). 7 animals received 50,000 or 20,000 KIE respectively of a proteinase inhibitor after bleeding and immediately before and one hour after the infusion of Dextran 60. The release of the lysosomal enzymes acid phosphatase and beta-glucuronidase was inhibited significantly (p less than 0.05) in the animals treated with Trasylol. However, the inhibition of the lysosomal enzymes seems not to have a decisive influence on the dynamic of the macro- and microcirculation and the myocardial contractility.

Topics: Acid Phosphatase; Animals; Aprotinin; Blood Pressure; Carbon Dioxide; Dextrans; Disease Models, Animal; Dogs; Enzyme Inhibitors; Female; Glucuronidase; Heart Ventricles; Hematocrit; Male; Microcirculation; Muscles; Myocardial Contraction; Oxygen; Partial Pressure; Plasma Substitutes; Shock, Hemorrhagic

Severe cardiac hypertrophy has been produced experimentally in rats by long-term, low-dose treatment with tri-iodothyroacetic acid. The dose used was insufficient to cause any apparent systemic or metabolic effect. It is suggested that similar iodinated substances in the blood in man, resulting from normal or abnormal thyroid hormone catabolism, may be causally related to some forms of cardiomyopathy.

Topics: Acetates; Acid Phosphatase; Alkaline Phosphatase; Animals; Cardiomegaly; Disease Models, Animal; Glycogen; Heart; Heart Ventricles; Mitochondria, Muscle; Myocardium; Organ Size; Rats; Succinate Dehydrogenase; Triiodothyronine

Local irradiation of the carotid artery of the hypercholesterolemic rabbit with 2000 rd of X-rays gives rise to infiltration of lipid droplets in the intima and media, becoming visible 3 days after the irradiation. At the same time, acid phosphatase and beta-glucuronidase become activated. These enhanced activities are localized in different cells of the arterial wall. Acid phosphatase activity is localized in the intima, while the beta-glucuronidase activation is found preferentially in the media. A functional heterogeneity of the lysosomal content of the different cells is suggested. A model for the development of the radiation-induced atheromatosis is presented.

Topics: Acid Phosphatase; Animals; Arteriosclerosis; Carotid Arteries; Disease Models, Animal; Enzyme Activation; Female; Glucuronidase; Glycosaminoglycans; Histocytochemistry; Hypercholesterolemia; Lipid Metabolism; Lysosomes; Male; Models, Biological; Rabbits; Radiation Injuries, Experimental; Radiotherapy

Unusual inclusions, which occurred in the reticuloendothelial cells intimately associated with fresh amyloid deposits, were analyzed by electron microscopy. The inclusions were located in the areas rich in the primary lysosome type of dense bodies and the cytoplasmic invaginations containing well-oriented amyloid fibrils. They were single-membrane-bounded, measured 0.3 to 0.8 mu in width and 0.5 to several microns in length, and showed considerable variation in the electron density of their contents. The latter consisted of two different ultrastructural elements: fibrillar profiles and a homogeneous or finely granular electron-dense substance. The fibrillar profiles were virtually identical in ulstrastructure to the amyloid fibrils and were well-oriented parallel to the long axis of the inclusion. The homogeneous or finely granular electron-dense substance appeared to be comparable to that composing the dense body matrix. The inclusions were usually acid phosphatase positive, but did not take up intravenously injected Thorotrast particles. These data led us to conclude that these inclusions were transitional forms from the usual dense bodies to the deep cytoplasmic invaginations containing well-oriented amyloid fibrils (which are accepted by most investigators as the sites of amyloid formation) and thus constitute direct evidence for the involvement of lysosomes in amyloid fibril formation.

Topics: Acid Phosphatase; Amyloid; Amyloidosis; Animals; Disease Models, Animal; Inclusion Bodies; Lysosomes; Membranes; Mice; Mononuclear Phagocyte System; Thorium Dioxide

A new experimental model for the study of two important aspects of ischemia, namely, oxygen and substrate deprivation, is proposed: the intact, beating fetal mouse heart in organ culture. This model offers long-term stability, ease and reproducibility of preparation, and the ability to manipulate experimental conditions. Hearts deprived of oxygen and glucose ceased beating immediately. After 3-4 hr of deprivation, biochemical and ultrastructural changes consistent with ischemic injury were evident. These include depletion of ATP and glycogen levels, loss of cytoplasmic enzymes, and extensive swelling and disruption of mitochondrial structure. Glucose and insulin partially protected against ATP depletion. Upon resupply of oxygen and glucose , beating resumed immediately, ATP levels rapidly increased to control levels and, consistent with this, mitochondrial structure returned toward normal. During the recovery phase autophagic vacuoles containing damaged mitochondria and myofibrils were seen, indicating that repair mechanisms were activated. Consistent with this, the proportion of lysosomal enzymes that were present in the nonsedimentable fraction of the tissue homogenate increased. We conclude that the cultured fetal mouse heart is a model useful for studying myocardial responses to anoxia and/or substrate deprivation and for assessing interventions designed to limit damage or to stimulate repair after ischemic injury.

Topics: Acetylglucosaminidase; Acid Phosphatase; Adenosine Triphosphate; Animals; Cathepsins; Coronary Disease; Disease Models, Animal; Female; Fetus; Gestational Age; Glucose; Lysosomes; Mice; Microscopy, Electron; Myocardium; Organ Culture Techniques; Oxygen Consumption; Pregnancy; Time Factors

The origin of giant granules in the retinal pigment epithelium of the beige mouse was investigated with electron microscopy and ultrastructural histochemistry. These granules were found to contain melanin and acid phosphatase. Apparently they arise from fusions of primary lysosomes with melanin granules which are already enlarged from multiple fusions among melanosomes. Therefore, the giant granules are not primary lysosomes, nor are they simply enlarged melanin granules as suspected from light microscopic studies. A deficiency of primary lysosomes in the pigment epithelium results, suggesting a defect in intracellular digestion similar to that found in the leukocytes of Chediak-Higashi patients and several animal models. Affected humans probably have defective digestion in their retinal pigment epithelium also; which could impair the renewal process for rod outer segments. Thus, Chediak-Higashi patients may show an increased susceptibility to light damage due not only to hypopigmentation, but to defective intracellular digestion, as well.

Topics: Acid Phosphatase; Animals; Chediak-Higashi Syndrome; Cytoplasmic Granules; Disease Models, Animal; Epithelium; Histocytochemistry; Lysosomes; Melanins; Mice; Mice, Inbred Strains; Microscopy, Electron; Mutation; Retina

Section of the medial collateral and both cruciate ligaments combined with resection of the medial meniscus in rabbit knees caused instability and during the ensuing six months these knees showed progressive histological changes similar to those of human osteoarthritis. Biochemical analysis of the cartilage from such knee joints demonstrated a decrease in proteoglycan, an increase in acid phosphatase, and increases in the rates of synthesis of protein and glycosaminoglycan. These findings, which are quite consistent with those in human osteoarthritis, suggest that this animal model may be of value in the study of the pathogenesis and treatment of human disease.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Femur; Glycine; Glycosaminoglycans; Hexosamines; Hydroxyproline; Knee Joint; Osteoarthritis; Protein Biosynthesis; Proteoglycans; Rabbits; Tritium

An experimental model of degenerative joint disease on chondromalacia consists of a surgically-scarified articular surface of the adult dog knee joint. In 52 dogs, evaluated by histologic and enzymatic assays over a period of 1 to 110 weeks post-surgery, the levels of acid hydrolase activity varied on various areas of articular cartilage within the same joint. There was a transient rise in most of the acid hydrolases in the synovium as a response to arthrotomy of the knee joint. All of the acid hydrolases studied did not respond uniformly to surgically created trauma. There was evidence of repair of the cartilage lacerations even when the subchondral zone was not breached. Lacerations in the central portion of the patella rarely showed healing in contrast to those placed more to the periphery of the articular surface. There was no gross or histologic evidence of progressive degenerative joint disease up to 2 years post-surgery. Thus an injury inflicted to the surface of the articular cartilage may be in itself insufficient in severity to produce destructive changes in the joint. This should not be too surprising, since, clinically, all joint surface injury does not lead to degenerative arthritis. The joint seems to have an injury threshold whereby chondrocytes are capable of repairing surface injury if the damage is not massive or repetitive. Insofar as lacerations in the center of the patella rarely healed, while the peripheral ones showed consistent signs of healing, the site of injury, as well as the magnitude of injury, may be critical.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Arylsulfatases; Cartilage Diseases; Cartilage, Articular; Cathepsins; Deoxyribonucleases; Disease Models, Animal; Dogs; Glucuronidase; Joint Diseases; Knee Joint; Patella; Synovial Membrane; Time Factors

Early changes in lysosomal enzymes must occur if their role is significant in irreversible myocardial injury. Therefore, we ligated the anterior descending coronary artery in 14 dogs and after 60 min excised epicardial and endocardial samples from the ischemic and adjacent normal heart. The collateral flow measured with radioactive microspheres in the endocardial samples averaged 19% of control. The muscle was disrupted and fractionated by ultracentrifugation into nuclear pellet (NP), heavy lysosomal pellet (HL), light lysosomal pellet (LL), microsomal pellet (M) and supernate (S). Electron microscopy demonstrated changes characteristic of sichemia in whole tissues and sedimented fractions. Acid phosphatase reaction product was present in residual bodies in the HL fraction and membrane-bound vesicles in the LL fraction and in the intact tissue. Significant decreases in the specific activity of N-acetyl-beta-glucosaminidase and beta-glucuronidase occurred in the endocardial LL fraction, while significant increases in both were found in the ts fraction (P less than 0.05). Losses of acid phosphatase occurred in both LL and S fractions. Moreover, decreases of total N-acetyl-beta-glucosaminidase in the HL fraction and of total beta-glucuronidase and acid phosphatase in the LL fraction were positively correlated (P less than 0.01) with the degree of ischemia measured with radioactive microspheres. Only insignificant enzymatic changes were found when the collateral flow was greater than 40%, and the differences were less significant in epicardial samples where the flow averaged 29%. The early loss of enzymes from the lysosomal fractions in severe ischemia suggests a role for lysosomal hydrolases in the necrosis that follows coronary occlusion.

Topics: Acetylglucosaminidase; Acid Phosphatase; Acute Disease; Anaerobiosis; Animals; Collateral Circulation; Coronary Circulation; Coronary Disease; Disease Models, Animal; Dogs; Endocardium; Hydrolases; Hydrolysis; Lysosomes; Myocardium; Polyethylene Glycols; Proteins

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Fibroblasts; Kidney Cortex; Kidney Medulla; Kidney Tubules; Macrophages; Microscopy, Electron; Pyelonephritis; Rabbits; Staining and Labeling; Time Factors

Topics: Acid Phosphatase; Animals; Cerebral Cortex; Cytoplasmic Granules; Disease Models, Animal; Endoplasmic Reticulum; Golgi Apparatus; Haplorhini; Ischemic Attack, Transient; Lysosomes; Microscopy, Electron; Microscopy, Fluorescence; Mitochondrial Swelling; Nissl Bodies; Ribosomes

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Disease Models, Animal; Esterases; Female; Histocytochemistry; Iliac Artery; Ischemia; Ligation; Malate Dehydrogenase; Muscles; Nucleotidases; Physical Exertion; Rats; Regeneration; Succinate Dehydrogenase; Time Factors

Topics: Acid Phosphatase; Animals; Body Weight; Cardiomegaly; Cathepsins; Creatine Kinase; Disease Models, Animal; Heart Ventricles; Hexosaminidases; Hyperthyroidism; Lysosomes; Male; Myocardium; Organ Size; Proteins; Rats; Remission, Spontaneous; Thyroxine

Topics: Acid Phosphatase; Adenosine Triphosphatases; Animals; Cats; Cerebrospinal Fluid; Demyelinating Diseases; Disease Models, Animal; Lysosomes; Microscopy, Electron; Neuroglia; Time Factors

Topics: Acid Phosphatase; Adenosine Monophosphate; Alkaline Phosphatase; Animals; Bone Development; Cartilage; Chickens; Disease Models, Animal; Epiphyses; Osteogenesis; Phosphates; Phosphoric Monoester Hydrolases; Pyrophosphatases; Vitamin A Deficiency

Topics: Acid Phosphatase; Animals; Body Weight; Calcium; Cell Fractionation; Cell Nucleus; Centrifugation; Diet; Disease Models, Animal; Electron Transport Complex IV; Glucosephosphate Dehydrogenase; Kidney; Magnesium; Magnesium Deficiency; Male; Mitochondria; Nephrocalcinosis; Parathyroid Hormone; Proteins; Rats; Tissue Extracts

Topics: Acid Phosphatase; Animals; Cell Movement; Cell Nucleus; Cytoplasm; Disease Models, Animal; Foreign Bodies; Gold Colloid, Radioactive; Intercellular Junctions; Leukocytes; Lysosomes; Mice; Mice, Inbred C57BL; Microscopy, Electron; Microscopy, Phase-Contrast; Motion Pictures; Organoids; Peroxidases; Pinocytosis; Plastics; Pseudopodia; Thorium Dioxide; Vacuoles

The beige mouse is considered to be a homologue of Chediak-Higashi syndrome (CHS). Cytochemical and electron microscopic studies have revealed an inherited lesion in the kidneys of these mice. The alteration was confined to the distal segments (S3) of the proximal tubules and was characterized by the accumulation of numerous massive polysaccharide-rich granules. These granules were reactive for acid phosphatase and beta-glucuronidase activities and were therefore considered to be lysosomes. Small numbers of lymphocytes were also observed in the perivascular spaces and within the tubules of the S3 segment. The fine structure of S3 cells was also markedly altered. In addition to the massive lysosomes, dense material was found associated with the brush border and was present in pinocytotic vesicles at the base of the brush border and between basal invaginations of the plasma membranes. Despite these changes, reabsorption of exogeneous peroxidase by S3 cells appeared to be normal. The presence of a congenital defect in the kidney of the beige mouse appears to provide a useful model for studying the morphology and function of the S3 region of the nephron.

Topics: Acid Phosphatase; Animals; Cell Membrane; Chediak-Higashi Syndrome; Colloids; Cytoplasm; Disease Models, Animal; Endoplasmic Reticulum; Glucuronidase; Histocytochemistry; Iron; Kidney; Kidney Tubules; Lysosomes; Mice; Microscopy, Electron; Peroxidases; Staining and Labeling

Topics: Acetylcholinesterase; Acid Phosphatase; Adenosine Triphosphatases; Animals; Central Nervous System; Cerebellar Cortex; Cerebellum; Disease Models, Animal; Esterases; Glucosephosphate Dehydrogenase; Haplorhini; Histocytochemistry; L-Lactate Dehydrogenase; Malate Dehydrogenase; Monoamine Oxidase; Motor Neurons; Phosphogluconate Dehydrogenase; Protein Deficiency; Pyrophosphatases; Spinal Cord; Succinate Dehydrogenase; Thiamine

Topics: Acid Phosphatase; Adenosine Triphosphatases; Animals; Disease Models, Animal; Hypoxia; Lysosomes; Male; Microelectrodes; Mitochondria, Liver; Muscles; Oxygen; Proteins; Rats; Shock, Hemorrhagic; Shock, Septic; Spectrophotometry

Proteus infection in the rat produces an acute bacterial pneumonia which resolves without necrosis or significant organization within 2 weeks. Ultrastructural changes included widespread damage to type 2 cells and endothelial cells and rapid proliferation of the alveolar epithelial cells, which were the predominant site of labeling with tritiated thymidine. Histochemical staining for lysosomal enzymes showed an initial reduction in type 2 cell reactivity. The majority of proliferating epithelial cells were also unreactive until the normal pattern of staining and morphology returned at 8 to 12 days after infection. The acute inflammatory exudate was reactive, but there was only minimal to moderate staining in the subsequent clusters of alveolar macrophages. These data suggest that resolution with the preservation of the normal architecture of the peripheral airspaces may be correlated with superficial injury and limited reactivity for digestive enzymes.

Topics: Acid Phosphatase; Animals; Autoradiography; Cell Nucleolus; Cell Nucleus; Cytoplasm; Disease Models, Animal; Epithelial Cells; Glucuronidase; Hyperplasia; Lung; Macrophages; Male; Mitochondria; Pneumonia; Proteus Infections; Proteus mirabilis; Pulmonary Alveoli; Rats; Ribosomes

Topics: Acid Phosphatase; Animals; Capillaries; Cerebellum; Cerebral Arteries; Chickens; Disease Models, Animal; Encephalomalacia; Histocytochemistry; Linoleic Acids; Microscopy, Electron; Microscopy, Fluorescence; Vitamin E Deficiency

On the basis of recent morpholgic and biochemical studies which suggested the possible involvement of reticuloendothelial (RE) cells and proteolytic enzymes in amyloidogenesis, the present study was undertaken to examine the ultrastructural interrelationship between lysosomes and amyloid fibrils at the sites of very early amyloid deposition. In the spleen, liver and kidney of the experimental mouse model, foci of amyloid deposits were closely associated with the RE cells. The lysosomal enzyme activity, as marked by cytochemical demonstration of acid glycerophosphatase activity, was localized in the primary type lysosomes (as defined by their electron microscopic appearance), in the Golgi complexes, in the small cytoplasmic vesicles and occasionally widespread in the cytoplasm. They showed an intimate relationship to the amyloid fibrils. The findings were interpreted as favoring the hypothesis that the hydrolases play a role in amyloid fibril formation. The enzyme activity was also demonstrated in the secondary type lysosomes which occasionally contained amyloid fibrils that appeared to be phagocytized.

Topics: Acid Phosphatase; Amyloid; Amyloidosis; Animals; Caseins; Cytoplasm; Disease Models, Animal; Female; Golgi Apparatus; Histocytochemistry; Kidney; Kupffer Cells; Liver; Lysosomes; Mice; Microscopy, Electron; Mononuclear Phagocyte System; Spleen

Topics: Acid Phosphatase; Animals; Chronic Disease; Disease Models, Animal; Kidney Glomerulus; Kidney Medulla; Kidney Tubules; Male; Pyelonephritis; Rabbits

Topics: Acid Phosphatase; Carrageenan; Disease Models, Animal; Galactosidases; Glucosidases; Golgi Apparatus; Granuloma; Histocytochemistry; Lysosomes; Macrophages

Topics: Acid Phosphatase; Animals; Animals, Newborn; Antilymphocyte Serum; Disease Models, Animal; Esterases; Humans; Immunosuppression Therapy; Male; Muscles; Phosphates; Prostate; Prostatic Hyperplasia; Rabbits; Rats; Succinate Dehydrogenase; Transplantation, Heterologous

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Burns; Disease Models, Animal; Esterases; Histocytochemistry; Liver; Methods; Necrosis; Rats

Topics: Acid Phosphatase; Analysis of Variance; Animals; Brain; Colorimetry; Disease Models, Animal; Enzyme Activation; Histological Techniques; Leucyl Aminopeptidase; Liver; Mice; Microscopy, Electron; Spectrophotometry; Time Factors; Yellow Fever; Yellow fever virus

Topics: Acid Phosphatase; Adenosine Diphosphate; Adenosine Triphosphate; Anabolic Agents; Animals; Disease Models, Animal; Dogs; Glucuronidase; Heart; Heart Ventricles; Lactates; Myocardial Contraction; Myocardial Infarction; Myocardium; Norepinephrine; Phosphocreatine; Proline; Pyruvates; Time Factors

Topics: Acid Phosphatase; Aminopeptidases; Animals; Antigens; Autoimmune Diseases; Cathepsins; Deoxyribonucleases; Disease Models, Animal; Glucuronidase; Guinea Pigs; Hyaluronoglucosaminidase; L-Lactate Dehydrogenase; Leucine; Lysosomes; Male; Organ Size; Oxidoreductases; Ribonucleases; Sorbitol; Spermatogenesis; Spermatozoa; Testis

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Aorta, Abdominal; Disease Models, Animal; Esterases; Female; Hot Temperature; L-Lactate Dehydrogenase; Malate Dehydrogenase; Rats; Rats, Inbred Strains; Succinate Dehydrogenase

Topics: Acid Phosphatase; Age Factors; Animals; Brain; Cerebral Cortex; Corpus Callosum; Demyelinating Diseases; Diffuse Cerebral Sclerosis of Schilder; Disease Models, Animal; Female; Male; Mice; Mice, Inbred Strains; Microscopy, Electron; Myelin Sheath; Neuroglia; Peripheral Nerves

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Disease Models, Animal; Dogs; Histocytochemistry; Methods; Pancreas; Pancreatitis; Phosphoric Monoester Hydrolases

Topics: Acid Phosphatase; Animals; Chelating Agents; Diabetes Mellitus; Diabetes Mellitus, Experimental; Disease Models, Animal; Histocytochemistry; Indicators and Reagents; Insulin; Islets of Langerhans; Methods; Microscopy, Fluorescence; Rabbits; Zinc

Topics: Acid Phosphatase; Animals; Brain; Cerebral Cortex; Cerebrovascular Disorders; Disease Models, Animal; Glucuronidase; Glycogen; Hemiplegia; Hydrolases; Hypoxia; Hypoxia, Brain; Ischemic Attack, Transient; Lysosomes; Male; Mitochondria; Rats

Topics: Acid Phosphatase; Amino Acids; Animals; Arthritis; Aspirin; Chronic Disease; Disease Models, Animal; Freund's Adjuvant; Glucuronidase; Liver; Lysosomes; Male; Membranes; Phenylbutazone; Rats

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Clinical Enzyme Tests; Disease Models, Animal; Esterases; Forensic Medicine; Histocytochemistry; Rats; Skin; Time Factors; Wounds and Injuries

Topics: Acid Phosphatase; Animals; Carbohydrate Metabolism; Cathepsins; Chemical and Drug Induced Liver Injury; Disease Models, Animal; Galactosidases; Lipid Metabolism; Liver; Lysosomes; Mushroom Poisoning; Peptides; Proteins; Rats; Sulfuric Acids

Topics: Acid Phosphatase; Animals; Atrophy; Cytoplasm; Disease Models, Animal; Histocytochemistry; Liver; Liver Diseases; Liver Glycogen; Male; Microscopy, Electron; Phagocytosis; Rats

Topics: Acid Phosphatase; Anesthesia; Animals; Disease Models, Animal; Dogs; Female; Heparin; Histocytochemistry; Kidney; Liver; Lysosomes; Male; Mitochondria; Mitochondria, Liver; Mononuclear Phagocyte System; Protamines; Shock, Hemorrhagic; Spleen

Topics: Acid Phosphatase; Aging; Alkaline Phosphatase; Animals; Central Nervous System; Central Nervous System Diseases; Cerebrosides; Cholesterol; Demyelinating Diseases; Disease Models, Animal; Female; Genes, Recessive; Genotype; Lipids; Male; Metabolism, Inborn Errors; Mice; Mutation; Phosphoric Monoester Hydrolases; Thyroid Function Tests; Time Factors

Topics: Acid Phosphatase; Ammonium Chloride; Animals; Animals, Newborn; Cerebellum; Copper; Culture Techniques; Disease Models, Animal; Glucosephosphate Dehydrogenase; Glutamate Dehydrogenase; Glycosaminoglycans; Hepatic Encephalopathy; Hepatolenticular Degeneration; Histocytochemistry; Humans; Neuroglia; Rats; Succinate Dehydrogenase

Topics: Acid Phosphatase; Adenosine Triphosphatases; Adult; Aged; Alkaline Phosphatase; Animals; Biopsy; Cerebral Cortex; Dementia; Disease Models, Animal; Female; Frontal Lobe; Histocytochemistry; Humans; Male; Middle Aged; Neurofibrils; Neurons; Oxidoreductases; Phosphoric Monoester Hydrolases; Rabbits; Succinate Dehydrogenase; Temporal Lobe

Topics: Acid Phosphatase; Adenine Nucleotides; Adenosine Triphosphatases; Animals; Biological Transport, Active; Cardiomyopathies; Cricetinae; Disease Models, Animal; Endoplasmic Reticulum; Female; Heart Valves; Lipid Metabolism; Male; Microscopy, Electron; Mitochondria, Muscle; Myocardium; Myofibrils; NAD; Oxidoreductases; Sarcolemma

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Dihydrolipoamide Dehydrogenase; Disease Models, Animal; DNA; Electron Transport Complex IV; Esterases; Histocytochemistry; L-Lactate Dehydrogenase; Lipase; Liver; Liver Cirrhosis; Liver Glycogen; Methods; Proteins; Rabbits; RNA; Succinate Dehydrogenase

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Aorta; Arteriosclerosis; Cholesterol; Diet, Atherogenic; Disease Models, Animal; Esterases; Female; Glucuronidase; Kidney; L-Lactate Dehydrogenase; Liver; Rabbits; Time Factors

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Histocytochemistry; Lung; Lung Diseases; Lung Injury; Phospholipids; Pulmonary Emphysema; Pulmonary Fibrosis; Radiation Injuries, Experimental; Rats

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Disease Models, Animal; Glycine; Histocytochemistry; Lung; Lung Diseases; Microscopy, Electron; Pulmonary Emphysema; Pulmonary Fibrosis; Radiation Injuries, Experimental; Rats; Tritium

Topics: Acid Phosphatase; Adenocarcinoma; Alkaline Phosphatase; Animals; Disease Models, Animal; Endometrial Hyperplasia; Endometrium; Estradiol; Female; Histocytochemistry; Microscopy, Electron; Neoplasms, Experimental; Rabbits; Uterine Neoplasms

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Brain Neoplasms; Disease Models, Animal; Enzymes; Esterases; Glioma; Glucuronidase; Histocytochemistry; Neoplasms, Experimental; Neurilemmoma; Nitroso Compounds; Oligodendroglioma; Rats

Topics: Acid Phosphatase; Animals; Buffers; Cartilage, Articular; Catechols; Cathepsins; Disease Models, Animal; DNA; Enzyme Activation; Glucuronidase; Glycoside Hydrolases; Hydrolases; In Vitro Techniques; Joint Diseases; Joints; Knee Joint; Organ Size; Pressure; Rabbits; Sucrose; Sulfatases; Synovial Membrane

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Arboviruses; Body Temperature; Disease Models, Animal; Female; Guinea Pigs; Hemorrhagic Fevers, Viral; Leukocyte Count; Male; Peptide Hydrolases

Topics: Acid Phosphatase; Adaptation, Physiological; Animals; Aortic Coarctation; Deoxyribonucleases; Disease Models, Animal; Heart; Heart Diseases; Hypoxia; Isoproterenol; Lysosomes; Male; Methods; Myocardium; Rats; Ribonucleases

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Eye Diseases; Inclusion Bodies, Viral; Inflammation; Lacrimal Apparatus; Lysosomes; Microscopy, Electron; Parotid Gland; Rats; Rodent Diseases; Salivary Gland Diseases; Submandibular Gland; Virus Diseases