acid-phosphatase and fluorexon

The purposes of the present study were to evaluate the effects of frequent applications of low-level laser therapy (LLLT) on corticotomy-assisted tooth movement in a beagle dog model and to compare the effects in the mandible and maxilla.. In 4 male beagles, the maxillary and mandibular second premolars were extracted. The third premolars were corticotomized and then protracted from the canines with a continuous force of 200 g. Daily LLLT (using an aluminum gallium indium phosphide [AlGaInP] diode) was applied at the buccal mucosa of the corticotomized premolars on 1 side only. The tooth movement was measured for 8 weeks. Fluorochromes were injected intravenously at the start of the experiment (T0) and after 2 (T2), 4 (T4), and 8 (T8) weeks to evaluate new bone formation on the tension sides. Histomorphometric and immunohistologic evaluations were performed.. In the mandible, the movement of the corticotomized premolars in the LLLT plus corticotomy group was less than that in the corticotomy-only group, although the difference was not statistically significant. In the maxilla, no significant differences between the 2 groups were found. Osteoclastic and proliferating cell activities and the amount of new bone formation were greater in the mandibular LLLT plus corticotomy group than in the corticotomy-only group.. The frequent application of LLLT showed no significant effect on the corticotomized tooth movement.

Topics: Acid Phosphatase; Alveolar Process; Animals; Anthraquinones; Bicuspid; Bone Resorption; Cell Proliferation; Dogs; Fluoresceins; Fluorescent Dyes; Isoenzymes; Lasers, Semiconductor; Low-Level Light Therapy; Male; Mandible; Maxilla; Models, Animal; Orthodontic Wires; Osteoclasts; Osteogenesis; Pilot Projects; Proliferating Cell Nuclear Antigen; Root Resorption; Tartrate-Resistant Acid Phosphatase; Tetracycline; Time Factors; Tooth Movement Techniques

The aim of the present study was to evaluate the feasibility of a gelatin hydrogel system to enhance recombinant human fibroblast growth factor-2 (rhFGF-2)-induced osteogenic effects during rat mandibular distraction.. Mandibular distraction was performed in 28 male Wistar rats. Then, the rats were divided into 5 groups in which the designated gel mix was inserted into the distracted area: group 1, rhFGF-2 alone (n = 5); group 2, collagen alone (n = 6); group 3, collagen incorporating rhFGF-2 (n = 6); group 4, gelatin hydrogel alone (n = 5); and group 5, gelatin hydrogel incorporating rhFGF-2 (n = 6). The mandibles were excised 29 days after surgery and the newly formed bone was analyzed radiologically and histologically. The experimental groups were compared using the Fisher post hoc test (95% statistical significance threshold; P < .05).. Peripheral quantitative computed tomographic analysis, von Kossa staining, and calcein staining showed that using gelatin hydrogel with rhFGF-2 (group 5) significantly increased cortical bone mineral density, the domain area of hard tissue, the domain area of cortical bone area, total bone mineral content, cortical bone mineral content, the von Kossa-stained area, and the calcein-stained area compared with the collagen carrier (group 3). Group 5 also had a significantly larger number of cells positive for tartrate-resistant acid phosphatase compared with group 3 and radiopaque areas were observed more frequently.. The present findings suggest that gelatin hydrogel is a feasible delivery system for rhFGF-2, and when used together perform better in regard to hard tissue healing and treatment time after surgery.

Topics: Acid Phosphatase; Anatomy, Cross-Sectional; Animals; Biomarkers; Bone Density; Collagen; Coloring Agents; Drug Carriers; Feasibility Studies; Fibroblast Growth Factor 2; Fluoresceins; Fluorescent Dyes; Gelatin; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate; Isoenzymes; Male; Mandible; Osteogenesis; Osteogenesis, Distraction; Rats, Wistar; Recombinant Proteins; Tartrate-Resistant Acid Phosphatase; Tomography, X-Ray Computed; X-Ray Microtomography

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

Calcitonin gene related peptide (CGRP) is a neuropeptide that is abundant in the sensory neurons which innervate bone. The effects of CGRP on isolated bone cells have been widely studied, and CGRP is currently considered to be an osteoanabolic peptide that has effects on both osteoclasts and osteoblasts. However, relatively little is known about the physiological role of CGRP in-vivo in the skeletal responses to bone loading, particularly fatigue loading.. We used the rat ulna end-loading model to induce fatigue damage in the ulna unilaterally during cyclic loading. We postulated that CGRP would influence skeletal responses to cyclic fatigue loading. Rats were fatigue loaded and groups of rats were infused systemically with 0.9% saline, CGRP, or the receptor antagonist, CGRP(8-37), for a 10 day study period. Ten days after fatigue loading, bone and serum CGRP concentrations, serum tartrate-resistant acid phosphatase 5b (TRAP5b) concentrations, and fatigue-induced skeletal responses were quantified. We found that cyclic fatigue loading led to increased CGRP concentrations in both loaded and contralateral ulnae. Administration of CGRP(8-37) was associated with increased targeted remodeling in the fatigue-loaded ulna. Administration of CGRP or CGRP(8-37) both increased reparative bone formation over the study period. Plasma concentration of TRAP5b was not significantly influenced by either CGRP or CGRP(8-37) administration.. CGRP signaling modulates targeted remodeling of microdamage and reparative new bone formation after bone fatigue, and may be part of a neuronal signaling pathway which has regulatory effects on load-induced repair responses within the skeleton.

Topics: Acid Phosphatase; Animals; Bone Remodeling; Calcitonin Gene-Related Peptide; Fluoresceins; Isoenzymes; Male; Osteogenesis; Peptide Fragments; Rats; Rats, Sprague-Dawley; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Ulna; Weight-Bearing; Wound Healing

Modifying the balance between resorption and apposition through selectively injuring the cortical plate of the alveolus has been an approach to speed tooth movement and is referred to as periodontally accelerated osteogenic orthodontics. The aim of this study was to investigate the alveolar response to corticotomy as a function of time and proximity to the surgical injury in a rat model.. Maxillary buccal and lingual cortical plates were injured in 36 healthy adult rats adjacent to the upper left first molars. Twenty-four animals were euthanized at 3, 7, or 11 weeks. In one group, the maxillae were removed and stripped of soft tissues, and histomorphometric analysis was performed to study alveolar spongiosa and periodontal ligament (PDL) modeling dynamics. Catabolic activity was analyzed with tartrate-resistant acid phosphatase-positive osteoclasts and preosteoclasts. Anabolic actions were measured using a fluorescent vital bone stain series followed by sacrifice at 30 and 51 days. To further analyze the new bone formation, a separate group of animals were fed with calcein fluorescent stain and processed for non-decalcified fluorescent stain histology.. At 3 weeks, the surgery group had significantly (P <0.05) less calcified spongiosa bone surface, greater periodontal ligament surface, higher osteoclast number, and greater lamina dura apposition width. The catabolic activity (osteoclast count) and anabolic activity (apposition rate) were three-fold greater, calcified spongiosa decreased by two-fold, and PDL surface increased by two-fold. Surgical injury to the alveolus that induced a significant increase in tissue turnover by week 3 dissipated to a steady state by postoperative week 11. The impact of the injury was localized to the area immediately adjacent to the decortication injury.. Selective alveolar decortication induced increased turnover of alveolar spongiosa, and the activity was localized; dramatic escalation of demineralization-remineralization dynamics is the likely biologic mechanism underlying rapid tooth movement following selective alveolar decortication.

Topics: Acid Phosphatase; Alveolar Process; Animals; Anthraquinones; Biomarkers; Bone Remodeling; Bone Resorption; Cell Count; Fluoresceins; Fluorescent Dyes; Isoenzymes; Maxilla; Models, Animal; Osteoclasts; Osteogenesis; Osteotomy; Periodontal Ligament; Rats; Tartrate-Resistant Acid Phosphatase; Tetracycline; Time Factors

Topics: Acid Phosphatase; Alcian Blue; Alkaline Phosphatase; Animals; Anthraquinones; Biomarkers; Bone and Bones; Bone Development; Bromodeoxyuridine; Cartilage; Chondrocytes; Dissection; Fluoresceins; Histological Techniques; Isoenzymes; Mutagenesis; Mutation; Osteoblasts; Osteoclasts; Phenotype; Radiography; Staining and Labeling; Tartrate-Resistant Acid Phosphatase; Zebrafish

Osteoblastic metastases are common in patients with advanced prostate cancer. The pathophysiology of the new bone formation at metastatic sites is not currently known, but it is hypothesized that growth factors secreted by the prostate may be involved. Unfortunately, most rodent models of prostate cancer with metastasis to bone are osteolytic and not osteoblastic. Significant osteolysis by tumor cells at metastatic sites also may lead to fractures or bone instability. Misinterpretation of new periosteal bone due to bone instability as tumor-cell osteo-induction is another disadvantage of the osteolytic models. To circumvent these problems, we have developed a model system of new bone formation in the calvaria of nude mice stimulated by normal canine prostate tissue. Collagenase-digested normal prostate tissue was implanted adjacent to the calvaria of nude mice. Calvaria were examined at 2 weeks post-implantation for changes in the bone microenvironment by histology, calcein uptake at sites of bone mineralization, and tartrate-resistant acid phosphatase staining for osteoclasts. The prostate tissue remained viable and induced abundant new woven bone formation on the adjacent periosteal surface. In some cases new bone formation also was induced on the distant or concave calvarial periosteum. The new bone stained intensely with calcein, which demonstrated mineralization of the bone matrix. The new bone formation on prostate-implanted calvaria significantly increased (1.7-fold) the thickness of the calvaria compared with control calvaria. New bone formation was not induced in calvaria of mice implanted with normal canine kidney, urinary bladder, spleen, or skeletal muscle tissue, or mice with surgically-induced disruption of the periosteum. Osteoclast numbers in the medullary spaces and periosteum of calvaria were mildly increased (61%) in mice with implanted prostate tissue. In conclusion, this animal model will be useful for investigating the roles of prostate-derived growth factors on new bone formation in vivo.

Topics: Acid Phosphatase; Animals; Calcification, Physiologic; Dogs; Fluoresceins; Humans; Isoenzymes; Male; Mice; Mice, Nude; Osteoclasts; Osteogenesis; Prostate; Skull; Tartrate-Resistant Acid Phosphatase; Tissue Transplantation; Transplantation, Heterologous

Prosthesis failure due to wear debris-induced osteolysis remains a major clinical problem and the greatest limitation for total joint arthroplasty. Based on our knowledge of osteoclast involvement in this process and the requirements of receptor activator of NF-kappaB (RANK) signaling in osteoclastogenesis and bone resorption, we investigated the efficacy of RANK blockade in preventing and ameliorating titanium (Ti)-induced osteolysis in a mouse calvaria model. Compared with placebo controls we found that all doses of RANK:Fc above 1 mg/kg intraperitoneally (ip) per 48 h significantly inhibited osteoclastogenesis and bone resorption in response to Ti implanted locally. Complete inhibition occurred at 10 mg/kg ip per 48 h, yielding results that were statistically equivalent to data obtained with Ti-treated RANK-/- mice. We also evaluated the effects of a single injection of RANK:Fc on day 5 on established osteolysis and found that Ti-treated were still depleted for multinucleated tartrate-resistant acid phosphatase-positive (TRAP+) cells 16 days later. More importantly, this osteoclast depletion did not affect bone formation because the bone lost from the osteolysis on day 5 was restored by day 21. An assessment of the quantity and quality of the newly formed bone in these calvariae by calcein labeling and infrared (IR) microscopy, respectively, showed no significant negative effect of RANK:Fc treatment. These studies indicate that osteoclast depletion via RANK blockade is an effective method to prevent and reverse wear debris-induced osteolysis without jeopardizing osteogenesis.

Topics: Acid Phosphatase; Animals; Bone Matrix; Bone Remodeling; Bone Resorption; Female; Fluoresceins; Glycoproteins; Mice; Mice, Inbred CBA; Osteoclasts; Osteogenesis; Osteolysis; Osteoprotegerin; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Signal Transduction; Skull; Titanium

Although ovariectomized cynomolgus monkeys are used extensively for studies examining perimenopausal changes in the skeleton, very little is known about the changes that occur naturally during growth and aging in these animals. To evaluate the changes in bone during growth and aging in female cynomolgus monkeys, 29 monkeys ranging from 3 years to >22 years of age were given a bone fluorochrome label and iliac biopsies were collected. Serum and urine were collected at the time of biopsy to determine alkaline phosphatase (ALP), osteocalcin, tartrate-resistant acid phosphatase (TRAP), serum and urinary calcium, estradiol, urinary creatinine, and urinary CrossLaps levels. The biopsies from 8 young (3-4 years of age), 13 mature (11-16 years of age) and 8 old (>22 years of age) were analyzed histomorphometrically. ALP, TRAP, and urinary CrossLaps levels were lower in the mature and old monkeys when compared with the young group. Urinary calcium/urinary creatinine levels increased with age. Bone volume (BV/TV), trabecular number (Tb.N), bone formation rate (BFR), and activation frequency (Ac.f) were greatest in the young monkeys and declined in the older groups. The biomarker and histomorphometric indices in the young animals reflect the growth that was occurring in this group. The older monkeys do not appear to differ significantly between 10 and 22 years of age.

Topics: Acid Phosphatase; Aging; Alkaline Phosphatase; Animals; Biomarkers; Biopsy; Calcium; Collagen; Collagen Type I; Creatinine; Estradiol; Female; Fluoresceins; Fluorescent Dyes; Ilium; Isoenzymes; Macaca fascicularis; Osteocalcin; Peptides; Sexual Maturation; Tartrate-Resistant Acid Phosphatase

The most intense endogenous transforming growth factor (TGF)-beta 1 expression was detected in osteoblasts and fibroblasts in the mid-palatal suture 24 hours after the start of palatal expansion with an immunohistochemical technique. Based on these data, local effects of transforming growth factor-beta 1 (TGF-beta 1) in the expanding mid-palatal suture of the rat were examined. Single doses of human recombinant (rhu) TGF-beta 1 (40 and 200 ng, and 1 microgram) were injected into the expanding mid-palatal suture 24 hours after expansion started. Calcein was also injected immediately, and 6 and 12 days after expansion. The width of each calcein label was measured to evaluate bone formation along the suture with bone histomorphometry. In the experimental groups, bone formation during the 12-day expansion period was stimulated significantly (P < 0.05), in a dose-dependent manner, compared with that in the control group. Furthermore, bone formation during the later stage (6-12 days) of the 12-day expansion period, was still significantly higher than that in the control group. In addition, the amount of bone formation in response to a course of 3 injections of 200 ng rhu TGF-beta 1 on days 3, 6, and 9 was almost equal to that induced by a single injection of 200 ng rhu TGF-beta 1 on day 1. These results suggest that TGF-beta 1 may play an important role in bone formation at the active site of the suture in response to rapid palatal expansion and application of TGF-beta 1 during the early stages may induce rapid bone formation.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Cranial Sutures; Dose-Response Relationship, Drug; Fibroblasts; Fluoresceins; Indicators and Reagents; Male; Osteoblasts; Osteogenesis; Palatal Expansion Technique; Palate; Rats; Rats, Wistar; Recombinant Proteins; Transforming Growth Factor beta