acid-phosphatase and Periodontitis

Biomarkers of periodontal disease activity may be obtained from potential proteolytic and hydrolytic enzymes of inflammatory cell origin. Studies that have sought to correlate these enzymes with periodontal disease activity are reviewed with special consideration given to collagenases, cysteine, aspartate and serine proteinases, beta-glucuronidase, arylsulphate, alkaline and acid phosphatases, myeloperoxidase, lysozyme and lactoferrin.

Topics: Acid Phosphatase; Alkaline Phosphatase; Arylsulfatases; Aspartic Acid Endopeptidases; Biomarkers; Collagenases; Cysteine Endopeptidases; Glucuronidase; Humans; Hydrolases; Lactoferrin; Muramidase; Peptide Hydrolases; Periodontal Diseases; Periodontitis; Peroxidase; Serine Endopeptidases

The dysbiotic microbiota associated with aggressive periodontitis includes Aggregatibacter actinomycetemcomitans, the only oral species known to produce a cytolethal distending toxin (AaCDT). Give that CDT alters the cytokine profile in monocytic cells, we aimed to test the hypothesis that CDT plays a role in bone homeostasis by affecting the differentiation of precursor cells into osteoclasts. Recombinant AaCDT was added to murine bone marrow monocytes (BMMC) in the presence or absence of RANKL and the cell viability and cytokine profile of osteoclast precursor cells were determined. Multinucleated TRAP(+) cell numbers, and relative transcription of genes related to osteoclastogenesis were also evaluated. The addition of AaCDT did not lead to loss in cell viability but promoted an increase in the average number of TRAP(+) cells with 1-2 nuclei in the absence or presence of RANKL (Tukey, p < 0.05). This increase was also observed for TRAP(+) cells with ≥3nuclei, although this difference was not significant. Levels of TGF-β, TNF-α, and IL-6, in the supernatant fraction of cells, were higher when in AaCDT exposed cells, whereas levels of IL-1β and IL-10 were lower than controls under the same conditions. After interaction with AaCDT, transcription of the rank (encoding the receptor RANK), nfatc1 (transcription factor), and ctpK (encoding cathepsin K) genes was downregulated in pre-osteoclastic cells. The data indicated that despite the presence of RANKL and M-CSF, AaCDT may inhibit osteoclast differentiation by altering cytokine profiles and repressing transcription of genes involved in osteoclastogenesis. Therefore, the CDT may impair host defense mechanisms in periodontitis.

Topics: Acid Phosphatase; Aggregatibacter actinomycetemcomitans; Animals; Bone Marrow Cells; Bone Resorption; Cathepsin K; Cell Differentiation; Cell Survival; Cells, Cultured; Interleukin-10; Interleukin-1beta; Interleukin-6; Isoenzymes; Macrophages; Male; Mice; Mice, Inbred C57BL; Monocytes; NFATC Transcription Factors; Osteoclasts; Pasteurellaceae Infections; Periodontitis; RANK Ligand; Tartrate-Resistant Acid Phosphatase; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The purpose of this study was to evaluate the influence of an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser in the absence or presence of manual scaling and root planning (SRP) for the treatment of induced periodontitis in rats. Ligatures were placed in the subgingival region of the maxillary first molar. After a 7-day period, the ligatures were removed, and 40 rats were randomly divided into four groups (G), as follows: (GI) no treatment, (GII) scaling and root planning (SRP) with curettes, (GIII) Er,Cr:YSGG laser irradiation and (GIV) SRP with curettes followed by Er,Cr:YSGG laser irradiation. Seven and 30 days after the treatment, the animals were sacrificed and histologic, histometric and immunohistochemistry analyses were performed. All groups showed similar histopathological characteristics during the evaluation period. The histometric analysis was confirmed using Bonferroni and paired t tests. At 7 and 30 days, groups II, III and IV exhibited greater bone formation in the furcation area when compared to group I (p < 0.0001; p < 0.05). During the 7-day period, the groups irradiated with the laser (III and IV) showed a statistically larger new bone area than the group treated with SRP (II) (p < 0.01). Immunohistochemistry analysis revealed that the control group exhibited a higher expression of tartrate-resistant acid phosphatase (TRAP) and the receptor activator of nuclear factor κΒ ligand (RANKL) when compared to groups II, III and IV (p < 0.05). All treatments were able to reduce the inflammatory processes, consequently enabling the repair of periodontal tissues. The results achieved with the application of the Er,Cr:YSGG laser suggest that this laser can stimulate greater bone formation, especially over a shorter period of time.

Topics: Acid Phosphatase; Animals; Dental Scaling; Isoenzymes; Laser Therapy; Lasers, Solid-State; Male; Molar; Periodontitis; Rats; Tartrate-Resistant Acid Phosphatase

Pycnogenol(®) (PYC) is a standardized bark extract from French maritime pine (Pinus pinaster Aiton). We examined the inhibitory effects of PYC on alveolar bone resorption, which is a characteristic feature of periodontitis, induced by Porphyromonas gingivalis (P. gingivalis) and osteoclast differentiation. In rat periodontitis model, rats were divided into four groups: group A served as the non-infected control, group B was infected orally with P. gingivalis ATCC 33277, group C was administered PYC in the diet (0.025%: w/w), and group D was infected with P. gingivalis and administered PYC. Administration of PYC along with P. gingivalis infection significantly reduced alveolar bone resorption. Treatment of P. gingivalis with 1 µg/ml PYC reduced the number of viable bacterial cells. Addition of PYC to epithelial cells inhibited adhesion and invasion by P. gingivalis. The effect of PYC on osteoclast formation was confirmed by tartrate-resistant acid phosphatase staining. PYC treatment significantly inhibited osteoclast formation. Addition of PYC (1-100 µg/ml) to purified osteoclasts culture induced cell apoptosis. These results suggest that PYC may prevent alveolar bone resorption through its antibacterial activity against P. gingivalis and by suppressing osteoclastogenesis. Therefore, PYC may be useful as a therapeutic and preventative agent for bone diseases such as periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Anti-Bacterial Agents; Bone and Bones; Cell Differentiation; Cell Line; Cells, Cultured; Epithelial Cells; Flavonoids; Gingiva; Humans; Isoenzymes; Male; Mice, Inbred BALB C; Osteoclasts; Periodontitis; Pinus; Plant Bark; Plant Extracts; Porphyromonas gingivalis; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

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

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

Acupuncture has shown the capability of modulating the immuno-inflammatory response of the host. This study aims to evaluate the effects of electroacupuncture (EA) on ligature-induced periodontitis in rats.. Thirty-two animals were divided into four groups: 1) control; 2) experimental periodontitis (EP); 3) sham-treated (EP/EA-sham); and 4) treated with EA (EP/EA). For the EP groups, a ligature was placed around the right mandibular first molars at day 1. Sessions of EA or EA-sham were assigned every other day. For EA treatment, large intestine meridian points LI4 and LI11 and stomach meridian points ST36 and ST44 were used. EA-sham was performed in off-meridian points. Animals were euthanized at day 11. Histomorphometric and microtomographic analyses were performed. Immunolabeling patterns for the receptor activator of nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), and tartrate-resistant acid phosphatase (TRAP) were assessed. Expressions of interleukin (IL)-1β, matrix metalloproteinase (MMP)-8, IL-6, and cyclooxygenase (COX)-2 messenger RNAs (mRNAs) were evaluated by quantitative reverse transcription-polymerase chain reaction. Data were analyzed statistically (P <0.05, analysis of variance).. Histomorphometric and microtomographic analyses demonstrated that group EP/EA presented reduced alveolar bone loss when compared to group EP (P <0.05). Reduced RANKL immunolabeling and fewer TRAP-positive multinucleated cells were observed in the EA-treated group in relation to group EP. No differences were observed in OPG expression among groups. EA treatment decreased the genic expression of IL-1β and MMP-8 (P <0.05), increased the mRNA expression of IL-6 (P <0.05), and did not modify the genic expression of COX-2 in animals with EP (P >0.05).. It can be concluded that EA reduced periodontal tissue breakdown and the expression of some proinflammatory mediators and a proresorptive factor in EP in rats.

Topics: Acid Phosphatase; Acupuncture Points; Alveolar Bone Loss; Animals; Bone Density; Cyclooxygenase 2; Electroacupuncture; Giant Cells; Image Processing, Computer-Assisted; Interleukin-1beta; Interleukin-6; Isoenzymes; Male; Matrix Metalloproteinase 8; Osteoprotegerin; Periodontal Ligament; Periodontitis; Rats; Rats, Wistar; Receptor Activator of Nuclear Factor-kappa B; Tartrate-Resistant Acid Phosphatase; X-Ray Microtomography

Periodontitis is an inflammatory disease of tooth-supporting tissue leading to bone destruction and tooth loss. Periodontitis affects almost 50% of adults greater than 30 years of age.. This study evaluated the association between biomarkers linked to bone formation and resorption with the occurrence and progression of periodontal disease in older men (≥ 65 y).. The Osteoporotic Fractures in Men (MrOS) study is a prospective, observational study among men 65 years of age and older.. This ancillary study, Oral and Skeletal Bone Loss in Older Men, was conducted at two of the six MrOS study sites (Birmingham, AL and Portland, OR).. Patients underwent medical and dental evaluation. Diagnoses of periodontitis were based on clinical attachment loss, pocket depth, calculus, plaque, and bleeding on a random half-mouth. Bone metabolism biomarkers included serum levels of calcium, phosphate (Pi), alkaline phosphatase, albumin, carboxy-terminal collagen crosslinks (CTX), N-terminal propeptides of type I procollagen, isoform 5b of tartrate-resistant acid phosphatase, and urine alpha- carboxy-terminal collagen crosslinks (alpha-CTX) and beta-CTX and serum levels of calciotropic hormones vitamin D (25(OH)D) and PTH.. The aim of this study is to correlate bone metabolism biomarkers with prevalence and progression of periodontal disease in older men.. Patients with more severe periodontitis had significantly higher levels of PTH (P trend = .0004), whereas 25(OH)D was lower (P trend = .001). In a subset of men reevaluated at a second dental visit, improvement of periodontitis was associated with lower alpha-CTX, beta-CTX, and CTX levels at baseline after adjusting for age, site, and body mass index.. This study suggests that a distinct set of biomarkers of bone metabolism are associated with more severe periodontal disease (PTH, 25(OH)D) and periodontal progression (alpha-CTX, beta-CTX, and CTX) over time.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Biomarkers; Bone and Bones; Bone Diseases, Metabolic; Collagen Type I; Humans; Isoenzymes; Male; Osteoporotic Fractures; Peptides; Periodontal Diseases; Periodontitis; Tartrate-Resistant Acid Phosphatase; Vitamin D

Periodontitis is an inflammatory disease characterized by loss of connective tissue and alveolar bone, and osteoporosis is a common disease characterized by a systemic impairment of bone mass and microarchitecture. To date, the association between periodontitis and osteoporosis has remained to be fully elucidated. In the present study, an experimental rat model of periodontitis was used to explore the effects of oestrogen deficiency‑induced osteoporosis on the maxillary alveolar bone. Forty‑four female, six‑month‑old Sprague‑Dawley rats were randomly divided into four groups: Control, ligature, ovariectomized (OVX), and OVX + ligature. One month after ovariectomy, rats in the ligature and OVX + ligature groups received ligatures on their first and second maxillary molars for 1 month. Fluorescent labelling was performed prior to sacrificing the animals. At the end of the experiment, the maxillae and serum were collected and subjected to micro‑computed tomography analysis, confocal laser‑scanning microscopic observation, Van Gieson's fuchsin staining, tartrate‑resistant acid phosphatase staining and ELISA. Ligatures slightly reduced the alveolar bone mineral density (BMD) and bone formation rate, but significantly reduced alveolar crest height (ACH). Ovariectomy reduced the alveolar BMD, impaired the trabecular structure, reduced the bone formation rate and increased the serum levels of bone resorption markers. Animals in the OVX + ligature group exhibited a lower alveolar BMD, a poorer trabecular structure, a reduced ACH, a lower bone formation rate and higher serum levels of bone resorption markers compared with those in the control group. The results of the present study showed that ovariectomy enhanced alveolar bone loss and reduced the ACH of rats with experimental periodontitis. Thus, post‑menopausal osteoporosis may influence the progression of periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Biomarkers; Collagen Type I; Estrogens; Female; Humans; Isoenzymes; Osteoporosis, Postmenopausal; Peptides; Periodontitis; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride (NaF) on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin (IL)-1β, matrix metalloproteinase 9 (MMP9), and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL (receptor activator of nuclear factor κB ligand) and macrophage colony-stimulating factor (M-CSF). The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells (NFAT) c1 was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATc1, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents; Bacteroidaceae Infections; Bone Density Conservation Agents; Cathepsin K; Interleukin-1beta; Interleukin-6; Interleukin-8; Isoenzymes; Macrophage Colony-Stimulating Factor; Male; Matrix Metalloproteinase 9; Osteoclasts; Periodontitis; Porphyromonas gingivalis; RANK Ligand; Rats; Rats, Sprague-Dawley; Sodium Fluoride; Tartrate-Resistant Acid Phosphatase; Transcription Factors; X-Ray Microtomography

Porphyromonas gingivalis (Pg) may cause an immune-inflammatory response in host cells leading to bone degradation by osteoclasts. We investigated the osteoclast-inducing capacity of periodontal ligament fibroblasts from periodontitis patients and non-periodontitis donors after a challenge with viable Pg.. PDLFs from periodontitis patients (n = 8) and non-periodontitis donors (n = 7) were incubated for 6 h with or without viable Pg and subsequently co-cultured with osteoclast precursors from peripheral blood mononuclear cells (PBMCs). The number of multinucleated tartrate-resistant acid phosphatase-positive cells was determined at 21 days. Expression of osteoclastogenesis-associated genes was assessed after infection of PDLFs mono-cultures and in PDLFs-PBMCs co-cultures. Resorption activity was analysed on bone slices.. Pg induced the expression of osteoclastogenesis-associated genes by PDLFs. After bacterial challenge the formation of osteoclast-like cell was decreased in co-cultures of PBMCs with non-periodontitis PDLFs, but not with PDLFs from periodontitis patients.. PDLFs from sites free of periodontitis respond to an infection with Pg by tempering formation of osteoclast-like cells, probably promoting clearance of the infection. PDLFs from periodontitis sites are desensitized to a Pg challenge in terms of their osteoclast-inducing capacity.

Topics: Acid Phosphatase; Actins; Bone Resorption; Carbonic Anhydrase II; Cell Culture Techniques; Cell Differentiation; Coculture Techniques; Female; Fibroblasts; Giant Cells; Humans; Isoenzymes; Leukocytes, Mononuclear; Male; Middle Aged; Osteoclasts; Osteoprotegerin; Periodontal Ligament; Periodontitis; Porphyromonas gingivalis; RANK Ligand; Tartrate-Resistant Acid Phosphatase; Time Factors

The aim of the present study was to assess the impact of chronic consumption of Cachaça on alveolar bone loss (BL) induced by ligature and on alveolar bone density (BD) in peripubertal rats.. Male Wistar rats were assigned into one of the following groups:. non-ingestion of Cachaça (n=15); Cachaça: ingestion of ascending concentrations of Cachaça during 100 days (n=15). 70th day after the beginning of Cachaça ingestion, one first mandibular molar received a ligature while the contralateral tooth was left unligated. After 30 days, the rats were killed. BL, BD, the positive cells for tartrate-resistant acid phosphatase (TRAP), receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) were analyzed in the furcation area of the ligated and unligated mandibular molars.. The Cachaça group presented greater BL (0.75±0.1mm(2) for Cachaça and 0.66±0.1mm(2) for control group, respectively) and number of RANKL and OPG+ cells and lower BD (60.3±4.2% for Cachaça and 76.8±3.8% for control group, respectively) and number of TRAP+ cells around ligated teeth (p<0.05), when compared to the control group. The Cachaça group (0.42±0.02mm(2)) also presented a higher BL around unligated teeth when compared to control group (0.31±0.05mm(2)).. Cachaça consumption per se and in the presence of ligature negatively affects alveolar bone by increasing the alveolar BL and reducing BD.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Bone Density; Ethanol; Isoenzymes; Ligation; Male; Mandible; Osteoprotegerin; Periodontitis; RANK Ligand; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase

Antimicrobial therapy can suppress periodontal pathogens and increase the effectiveness of conventional mechanical treatment. The aim of this study was to assess bone loss and the immune inflammatory response of rats under the influence of two photosensitizing agents (MB and TBO) at two different concentrations in antimicrobial photodynamic therapy (aPDT), used as an adjuvant therapy in the treatment of periodontitis.. Periodontitis was induced in the mandibular first molars of 162 rats. The animals were divided into nine groups: G1 - scaling and root planing (SRP); G2 - SRP plus 100 μg/mL of methylene blue (MB); G3 - SRP plus 10 mg/mL of MB; G4 - SRP plus 100 μg/mL of toluidine blue (TBO); G5 - SRP plus 10 mg/mL of TBO; G6 - SRP plus 100 μg/mL of MB and laser; G7 - SRP plus 10 mg/mL of MB and laser; G8 - SRP plus 100 μg/mL of TBO and laser; and G9 - SRP plus 10 mg/mL of TBO and laser. Six animals from each group were euthanized 7, 15, or 30 d after treatment. Bone loss (BL) in the furcation region was evaluated using histomorphometric and immunohistochemical analyses to detect the receptor activator of nuclear factor-Κappa B ligand (RANKL), osteoprotegerin (OPG) and tartrate-resistant acid phosphatase (TRAP).. There was significantly less BL in animals treated with aPDT using low concentrations of MB and TBO at 7, 15 and 30 d. Immunohistochemical analysis revealed decreased RANKL and increased OPG in the aPDT groups and decreased TRAP-positive cells in G6 and G8.. aPDT, using low concentrations of MB and TBO, was the most effective adjuvant therapy to SRP, acting indirectly as a downregulator of the molecular mechanisms that control bone resorption in periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Combined Modality Therapy; Connective Tissue; Dental Scaling; Isoenzymes; Low-Level Light Therapy; Lymphocytes; Male; Methylene Blue; Neutrophils; Osteoclasts; Osteoprotegerin; Periodontitis; Phenothiazines; Photochemotherapy; Photosensitizing Agents; RANK Ligand; Rats; Rats, Wistar; Root Planing; Tartrate-Resistant Acid Phosphatase; Time Factors; Tolonium Chloride

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

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

BET proteins are a group of epigenetic regulators controlling transcription through reading acetylated histone tails and recruiting transcription complexes. They are considered as potential therapeutic targets in many distinct diseases. A novel synthetic bromodomain and extraterminal domain (BET) inhibitor, JQ1, was proved to suppress oncogene transcription and inflammatory responses. The present study was aimed to investigate the effects of JQ1 on inflammatory response and bone destruction in experimental periodontitis. We found that JQ1 significantly suppressed lipopolysaccharide (LPS)-stimulated inflammatory cytokine transcription, including interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α), as well as receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast markers, such as c-Fos, nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 1 (NFATc1), tartrate-resistant acid phosphatase (TRAP) and cathepsin K in vitro. JQ1 also inhibited toll-like receptors 2/4 (TLR2/4) expression and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) phosphorylation and nuclear translocation. Chromatin immunoprecipitation and quantitative polymerase chain reaction (ChIP-qPCR) revealed that JQ1 neutralized BRD4 enrichment at several gene promoter regions, including NF-κB, TNF-α, c-Fos, and NFATc1. In a murine periodontitis model, systemic administration of JQ1 significantly inhibited inflammatory cytokine expression in diseased gingival tissues. Alveolar bone loss was alleviated in JQ1-treated mice because of reduced osteoclasts in periodontal tissues. These unprecedented results suggest the BET inhibitor JQ1 as a prospective new approach for treating periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Cathepsin K; Cell Differentiation; Cell Line; Interleukin-1beta; Isoenzymes; Lipopolysaccharides; Mice; NF-kappa B; NFATC Transcription Factors; Nuclear Proteins; Osteoclasts; Periodontitis; Promoter Regions, Genetic; Proto-Oncogene Proteins c-ets; Proto-Oncogene Proteins c-fos; RANK Ligand; Tartrate-Resistant Acid Phosphatase; Toll-Like Receptor 2; Toll-Like Receptor 4; Transcription Factor AP-1; Transcription Factors; Tumor Necrosis Factor-alpha

Osteocytes are increasingly recognized as significant sources of osteoclast differentiation factor, receptor activator of nuclear factor-κB ligand (RANKL), and osteoblast differentiation inhibitory factor, sclerostin. In this study, RANKL and sclerostin expression of osteocytes is investigated in rats with ligature-induced periodontitis.. Rats were divided into control and periodontitis groups, and periodontitis was induced by ligature on the mandibular first molars. At 1, 3, 10, and 20 days after ligature, histologic analyses of alveolar bone (AB) and osteoid areas in the molar furcation were performed. The numbers of osteoclasts and RANKL- and sclerostin-positive osteocytes were estimated by tartrate-resistant acid phosphatase staining and immunohistochemistry, respectively.. The AB area gradually decreased at day 10 after ligature and increased at day 20. The number of osteoclasts markedly increased at day 3 and then decreased. Conversely, osteoid formation was suppressed up to day 3 and then showed a remarkable increase above control level at day 20. The number of RANKL-positive osteocytes increased at days 1 and 3 and then decreased. Sclerostin-positive osteocytes markedly increased at days 3 and 10 but decreased below control level at day 20.. These results show that AB loss is accompanied by enhanced osteoclast formation and suppressed osteoid formation. Osteocytes express RANKL when osteoclast formation increases, and they express sclerostin when osteoid formation is suppressed. Conversely, osteocytic sclerostin expression decreases when osteoid formation increases. These findings suggest that osteocytes may be important in AB loss via RANKL and sclerostin expression in periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Apoptosis; Bone Matrix; Bone Morphogenetic Proteins; Genetic Markers; Isoenzymes; Leukocytes, Mononuclear; Male; Mandibular Diseases; Neutrophils; Osteoclasts; Osteocytes; Periodontitis; Random Allocation; RANK Ligand; Rats; Rats, Inbred F344; Tartrate-Resistant Acid Phosphatase; Time Factors

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

Epigallocatechin-3-gallate (EGCG) is known for its beneficial properties, including anti-inflammatory and anti-oxidative activities. Recently, reports have suggested that EGCG plays a pivotal role in regulating cytokine expression and osteoclastic activity. In the present study, we investigated whether orally administered EGCG has a therapeutic effect on ligature-induced periodontitis.. Forty-eight Sprague-Dawley rats were treated with EGCG or phosphate-buffered saline. Periodontitis was induced by tying a ligature for 7 d. After removing ligation, EGCG (200 mg/kg) or phosphate-buffered saline was administered via oral gavage on a daily basis. Rats were killed after 1, 2 and 4 wk of administration. Histologic and histomorphometric analyses, tartrate resistant acid phosphatase staining and immunohistochemistry were carried out.. In the control group, bone loss did not recover even after the causative factor of periodontitis was eliminated. On the other hand, distance from cemento-enamel junction to alveolar bone crest, long junctional epithelium and collagen destruction were reduced in the EGCG group. Decreased interleukin (IL)-6 expression was shown from the early stage of EGCG administration, followed by reduced tumor necrosis factor (TNF) expression at week 4 EGCG group. The CT area showed a higher decrease of IL-6 expression between the control and EGCG group than alveolar bone area. Downregulation of TNF and IL-6 expression led to a decrease in osteoclast number and activity, which resulted in reduced bone loss.. Systemic administration of EGCG could have a therapeutic effect on damaged periodontal tissue. Inhibited cytokine expression, including TNF and IL-6 is responsible for the reduction in osteoclast formation, osteoclastic activity and collagen destruction.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Anti-Inflammatory Agents; Antioxidants; Biomarkers; Catechin; Collagen; Down-Regulation; Epithelial Attachment; Immunohistochemistry; Interleukin-6; Isoenzymes; Male; Osteoclasts; Periodontitis; Random Allocation; Rats; Rats, Sprague-Dawley; Tartrate-Resistant Acid Phosphatase; Time Factors; Tooth Cervix; Tumor Necrosis Factor-alpha

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

Progression of diabetes-associated periodontal destruction and the roles of advanced glycation end products (AGEs) are investigated.. Diabetes was induced by streptozocotin injection, and periodontitis was induced via silk ligature placement with Porphyromonas gingivalis lipopolysaccharide injection in 64 Sprague-Dawley rats for 7 to 21 days. The quality of alveolar bone and attachment loss (AL) were measured by microcomputed tomography and histology. Destruction profiles were evaluated by histology, histochemistry, immunohistochemistry, and quantitative assessments of inflammatory cells, expression of receptors for AGEs (RAGE), tartrate-resistant acid phosphatase, and proliferating cell nuclear antigen.. Without periodontitis induction, there was no obvious morphologic change in the periodontium, although slight elevations of AGEs and RAGE levels were noted in animals with diabetes. In the group with experimental periodontitis, significant periodontal bone loss was noted in animals both with and without diabetes from day 7, with more progressive bone loss in animals with diabetes during days 14 to 21. Histologically, the disruption of attachment and inflammation were observed from day 7, but subsequently subsided in animals without diabetes. A stronger and more prolonged response with significant AL was observed in animals with diabetes. Stronger inflammation, attenuated and persistent resorptive activity, and weaker proliferating potential were demonstrated by animals with diabetes. AGE deposition and RAGE expression were noted in animals without diabetes but with periodontitis, although levels were considerably elevated in the later stages in animals with diabetes.. Diabetes augments periodontal destruction by reducing the proliferating capability and activating resorptive activities. Presence of the AGE-RAGE axis without diabetes implies that it is involved in the regulation of inflammation.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Collagen; Diabetes Mellitus, Experimental; Disease Progression; Glycation End Products, Advanced; Isoenzymes; Lipopolysaccharides; Male; Periodontitis; Porphyromonas gingivalis; Proliferating Cell Nuclear Antigen; Random Allocation; Rats; Rats, Sprague-Dawley; Receptor for Advanced Glycation End Products; Receptors, Immunologic; 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

Bovine lactoferrin (bLF) modulates the production of tumor necrosis factor-alpha (TNF-α) and inhibits alveolar bone breakdown associated with periodontitis. This study is designed to examine the effects of orally administered liposomal bLF (LbLF) on orthodontic force (OF)-induced alveolar bone remodeling during experimental tooth movement.. Two groups of male Wistar rats were treated with either LbLF or control solution in drinking water 7 days before OF application. Lipopolysaccharide (LPS) was injected into the gingival sulcus in half the rats in each group. Thus, four groups: OF, OF+LbLF, OF+LPS, and OF+LPS+LbLF were established.. Orally administered LbLF significantly reduced apical migration of junctional epithelium in the OF and OF+LPS groups. In OF+LPS, osteoclast number in the alveolar crestal area was increased by LPS treatment, whereas osteoclast number was significantly reduced in OF+LPS+LbLF through suppression of TNF-α production. Osteoclastic induction in the middle part, mainly from OF application, was not affected by LbLF administration. Inhibition of tooth movement was not induced by LbLF.. Orally administered LbLF significantly inhibits LPS-induced alveolar bone resorption but not OF-induced bone remodeling. LbLF could be a potent therapeutic and preventive agent to control periodontal inflammation in patients undergoing orthodontic treatment.

Topics: Acid Phosphatase; Administration, Oral; Alveolar Bone Loss; Alveolar Process; Animals; Anti-Infective Agents; Bone Remodeling; Cattle; Cell Count; Epithelial Attachment; Escherichia coli; Isoenzymes; Lactoferrin; Lipopolysaccharides; Liposomes; Male; Osteoclasts; Periodontitis; Pilot Projects; Random Allocation; RANK Ligand; Rats; Rats, Wistar; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Tooth Cervix; Tooth Movement Techniques; Tumor Necrosis Factor-alpha

Periodontitis is generally accepted to relate to gram-negative bacteria, and the host defense system influences its onset and progression. However, little is known about the relation between gram-positive bacteria and periodontitis. In this study, we topically applied gram-positive and gram-negative bacterial suspensions to the gingival sulcus in rats after immunization, and then histopathologically examined their influence on periodontal destruction.. Rats previously immunized with heat-treated and sonicated Staphylococcus aureus or Aggregatibacter actinomycetemcomitans were used as immunized groups. The non-immunized group received only sterile phosphate-buffered saline. In each animal, S. aureus or A. actinomycetemcomitans suspension was applied topically to the palatal gingival sulcus of first molars every 24 h for 10 d. Blood samples were collected and the serum level of anti-S. aureus or anti-A. actinomycetemcomitans immunoglobulin G (IgG) antibodies was determined by enzyme-linked immunosorbent assay. The first molar regions were resected and observed histopathologically. Osteoclasts were stained with tartrate-resistant acid phosphatase (TRAP). The formation of immune complexes was confirmed by immunohistological staining of C1qB.. Serum levels of anti-S. aureus and anti-A. actinomycetemcomitans IgG antibodies in the immunized groups were significantly higher than those in the non-immunized groups were. The loss of attachment, increase in apical migration of the junctional epithelium, and decreases in alveolar bone level and number of TRAP-positive multinuclear cells in each immunized group were significantly greater than in each non-immunized group. The presence of C1qB was observed in the junctional epithelium and adjacent connective tissue in the immunized groups.. Heat-treated and sonicated S. aureus and A. actinomycetemcomitans induced attachment loss in rats immunized with their suspensions. Our results suggest that not only gram-negative but also gram-positive bacteria are able to induce periodontal destruction.

Topics: Acid Phosphatase; Administration, Topical; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Antibodies, Bacterial; Antigen-Antibody Complex; Antigens, Bacterial; Biomarkers; Connective Tissue; Epithelial Attachment; Gingiva; Hyaluronan Receptors; Immunization; Immunoglobulin G; Isoenzymes; Male; Mitochondrial Proteins; Molar; Osteoclasts; Periodontal Attachment Loss; Periodontitis; Rats; Rats, Inbred Lew; Specific Pathogen-Free Organisms; Staphylococcus aureus; Tartrate-Resistant Acid Phosphatase

Diabetes is known to impair wound healing and deteriorate the periodontal condition. There is limited information about the patterns and events associated with periodontal wound repair. In this study, we evaluate the dynamics of periodontal wound repair using micro-computed tomography (microCT) and immunohistochemistry.. Thirty-six male rats were used, and diabetes was induced by streptozotocin. The maxillary first molars were extracted, and a tooth-associated osseous defect was created in the extraction area. Animals were sacrificed after 7, 14, and 21 days. Volumetry and distribution of bone trabeculae were evaluated by microCT imaging. The patterns of healing and collagen alignment were evaluated by histology. Advanced glycation end-product (AGE) deposition and expression of the receptor for AGEs (RAGE), tartrate-resistant acid phosphatase, and proliferating cell nuclear antigen were evaluated by histochemical and immunohistochemical staining.. Diabetic animals demonstrated a significantly reduced bone volume and trabecular number as well as thinner trabeculae and more trabecular separation in osseous defects. The early stage was characterized by significantly reduced cellular proliferation and prolonged active inflammation without evident bone resorption, whereas delayed recovery of collagen realignment, matrix deposition, and bone turnover was noted in later stages. Although AGEs and RAGE were present during healing in diabetes and controls, a stronger and more persistent level of expression was observed in the group with diabetes. Diabetes significantly delayed osseous defect healing by augmenting inflammation, impairing proliferation, and delaying bone resorption. The AGE-RAGE axis can be activated under metabolic disturbance and inflammation.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Bone Remodeling; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Fibrillar Collagens; Glycation End Products, Advanced; Immunohistochemistry; Isoenzymes; Male; Periodontitis; Proliferating Cell Nuclear Antigen; Rats; Rats, Sprague-Dawley; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Tartrate-Resistant Acid Phosphatase; Wound Healing; X-Ray Microtomography

This study aimed to investigate whether chronic antigen-induced arthritis (AIA) influences infection-induced periodontitis (PD) in mice and whether PD modifies the clinical course of AIA. The contribution of anti-TNF-α therapy was also evaluated.. The PD was induced in C57BL/6 mice by oral infection with Aggregatibacter actinomycetemcomitans. AIA was induced after infection. Anti-TNF-α and chlorhexidine therapies were used to investigate the role of TNF-α and oral infection on PD and AIA interaction. Maxillae, knee joints, lymph nodes and serum samples were used for histomorphometric, immunoenzymatic and/or real time-PCR analyses.. Antigen-induced arthritis exacerbated alveolar bone loss triggered by PD infection. In contrast, PD did not influence AIA in the evaluated time-points. PD exacerbation was associated with enhanced production of IFN-γ in maxillae and expression of the Th1 transcription factor tBET in submandibular lymph nodes. Increased serum levels of IL-6 and C-reactive protein were also detected. Anti-TNF-α and antiseptic therapies prevented the development and exacerbation of infectious-PD. Anti-TNF-α therapy also resulted in reduced expression of IFN-γ, TNF-α and IL-17 in maxillae.. Altogether, the current results indicate that the exacerbation of infection-induced PD by arthritis is associated with an alteration in lymphocyte polarization pattern and increased systemic immunoreactivity. This process was ameliorated by anti-TNF-α and antiseptic therapies.

Topics: Acid Phosphatase; Actinobacillus Infections; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Anti-Infective Agents, Local; Arthritis, Experimental; C-Reactive Protein; Chlorhexidine; Collagen Type I; Immunoglobulin G; Interferon-gamma; Interleukin-17; Interleukin-6; Isoenzymes; Lymph Nodes; Male; Maxilla; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Osteoclasts; Periodontitis; T-Box Domain Proteins; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

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

Recent studies have pointed to potentially periodontal risk indicators, however no information is available on the impact of changes in thyroid hormone levels on the progression of periodontitis and on the quality of alveolar bone. Thus, the aim of the present study was to evaluate histologically, in rats, the influence of thyroid hormones on the rate of periodontal bone loss resulting from ligature placement and on the quality of tooth-supporting alveolar bone.. Thirty-six male Wistar rats were randomly assigned to the following groups: healthy (control, n = 12), hypothyroidism (n = 12) and hyperthyroidism (n = 12). Once alterations were confirmed by total serum levels of triiodothyronine and thyroxine, ligatures were randomly placed around one of the first mandibular molars. Thirty days later, the animals were killed and specimens routinely processed for serial decalcified sections. The parameters assessed were periodontitis-related bone loss, quality of tooth-supporting alveolar bone and the number of cells positive for tartrate-resistant acid phosphatase (TRAP), a marker of bone resorption.. At the ligated sites, intergroup analysis revealed that hypothyroidism significantly increased the bone loss resulting from ligature-induced periodontitis (p = 0.02) and the number of TRAP-positive cells on the linear surface of bone crest (p = 0.01). In addition, no significant differences were detected regarding the quality of the bone (p = 0.24) or the number of TRAP-positive cells in the area of the interradicular bone for ligated teeth among the groups (p = 0.17).. It may be concluded that decreased serum levels of thyroid hormones may enhance periodontitis-related bone loss, as a function of an increased number of resorbing cells, whereas the tooth-supporting alveolar bone seems to be less sensitive to alterations in hormone levels.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Animals; Biomarkers; Bone Density; Disease Progression; Furcation Defects; Gingivitis; Hyperthyroidism; Hypothyroidism; Isoenzymes; Male; Periodontitis; Random Allocation; Rats; Rats, Wistar; Tartrate-Resistant Acid Phosphatase; Thyroid Hormones; Thyroxine; Triiodothyronine

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

Rosiglitazone (RGZ), an oral anti-hyperglycemic agent used for non-insulin-dependent diabetes mellitus, is a high-affinity synthetic agonist for peroxisome proliferator-activated receptor-gamma (PPAR-gamma). Both in vitro and in vivo experiments have also revealed that RGZ possesses anti-inflammatory properties. Therefore, in the present study, we investigated the anti-inflammatory effects of RGZ in a rat model of periodontal disease induced by ligature placed around the mandible first molars of each animal. Male Wister rats were divided into four groups: 1) animals without ligature placement receiving administration of empty vehicle (control); 2) animals with ligature receiving administration of empty vehicle; 3) animals with ligature receiving administration with oral RGZ (10 mg/kg/day); and 4) animals with ligature receiving administration of subcutaneous RGZ (10 mg/kg/day). Thirty days after induction of periodontal disease, the animals were sacrificed, and mandibles and gingival tissues were removed for further analysis. An in vitro assay was also employed to test the inhibitory effects of RGZ on osteoclastogenesis. Histomorphological and immunohistochemical analyses of periodontal tissue demonstrated that RGZ-treated animals presented decreased bone resorption, along with reduced RANKL expression, compared to those animals with ligature, but treated with empty vehicle. Corresponding to such results obtained from in vivo experiments, RGZ also suppressed in vitro osteoclast differentiation in the presence of RANKL in MOCP-5 osteoclast precursor cells, along with the down-regulation of the expression of RANKL-induced TRAP mRNA. These data indicated that RGZ may suppress the bone resorption by inhibiting RANKL-mediated osteoclastogenesis elicited during the course of experimental periodontitis in rats.

Topics: Acid Phosphatase; Administration, Oral; Alveolar Bone Loss; Animals; Cell Differentiation; Cell Line; Hypoglycemic Agents; Immunohistochemistry; Isoenzymes; Male; Osteoclasts; Osteogenesis; Periodontitis; PPAR gamma; RANK Ligand; Rats; Rats, Wistar; Rosiglitazone; Tartrate-Resistant Acid Phosphatase; Thiazolidinediones

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

The mitogen-activated protein (MAP) kinase phosphatase (MKP) family plays an important function in regulating the pro-inflammatory cytokines by deactivating MAP kinases. MKP-1 is essential for the dephosphorylation of p38 MAP kinase that regulates expression of IL-6, TNF-alpha, and IL-1 beta. We hypothesized that MKP-1 regulates inflammatory bone loss in experimental periodontitis. Wild-type and Mkp-1(-/-) mice received A. actinomycetemcomitans LPS injection in the palatal region or PBS control 3 times/wk for 30 days. Mice were killed, and maxillae were assessed by microcomputed tomography, histological analysis, and TRAP staining for measurement of bone loss, extent of inflammation, and degree of osteoclastogenesis. Results indicated that, in LPS-injected Mkp-1(-/-) mice, significantly greater bone loss occurred with more inflammatory infiltrate and a significant increase in osteoclastogenesis compared with Mkp-1(-/-) control sites or either wild-type group. Analysis of these data indicates that MKP-1 plays a key role in the regulation of inflammatory bone loss.

Topics: Acid Phosphatase; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Biomarkers; Cell Count; Cell Line; Cone-Beam Computed Tomography; Dual Specificity Phosphatase 1; Imaging, Three-Dimensional; Immunity, Innate; Interleukin-6; Isoenzymes; Leukocytes; Lipopolysaccharides; Maxillary Diseases; Mice; Mice, Knockout; Osteoclasts; p38 Mitogen-Activated Protein Kinases; Palate; Periodontitis; Phosphorylation; Tartrate-Resistant Acid Phosphatase; Time Factors; X-Ray Microtomography

To determine whether peripheral blood mononuclear cells (PBMCs) from chronic periodontitis patients differ from PBMCs from matched control patients in their capacity to form osteoclast-like cells.. PBMCs from 10 subjects with severe chronic periodontitis and their matched controls were cultured on plastic or on bone slices without or with macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-kappaB ligand (RANKL). The number of tartrate-resistant acid phosphatase-positive (TRACP(+)) multinucleated cells (MNCs) and bone resorption were assessed.. TRACP(+) MNCs were formed under all culture conditions, in patient and control cultures. In periodontitis patients, the formation of TRACP(+) MNC was similar for all three culture conditions; thus supplementation of the cytokines was not needed to induce MNC formation. In control cultures, however, M-CSF or M-CSF/RANKL resulted in higher numbers compared with cultures without cytokines. Upregulations of osteoclast marker mRNA cathepsin K and carbonic anhydrase II confirmed the osteoclastic character. Bone resorption was only observed when PBMCs were cultured in the presence of M-CSF and RANKL.. Our data indicate that PBMCs from periodontitis patients do not need priming by M-CSF to become osteoclast-like cells, suggesting that PBMCs from periodontitis patients are present in the circulation in a different state of activity.

Topics: Acid Phosphatase; Adult; Case-Control Studies; Cell Differentiation; Chronic Disease; Female; Humans; Isoenzymes; Leukocytes, Mononuclear; Macrophage Colony-Stimulating Factor; Male; Matched-Pair Analysis; Middle Aged; Osteoclasts; Periodontitis; RANK Ligand; Tartrate-Resistant Acid Phosphatase

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

Periodontitis is an inflammatory disease that often leads to destruction of alveolar bone; a number of bacteria in subgingival plaque are associated with bone destruction in periodontitis. To understand the mechanism of how periodontopathogens induce osteoclastogenesis, we determined which mediators are involved in the osteoclastogenesis.. We investigated effects of sonicates from three periodontopathic bacteria, Porphyromonas gingivalis, Treponema denticola, and Treponema socranskii, on osteoclast formation in a co-culture system of mouse calvaria-derived osteoblasts and bone marrow cells. The osteoclast formation was determined by tartrate resistant acid phosphatase (TRAP) staining. The expression of the receptor activator of nuclear factor-kappa B ligand (RANKL), prostaglandin E(2) (PGE(2)) and osteoprotegerin (OPG) in mouse calvaria-derived osteoblasts was determined by immunoassay.. Each bacterial sonicate induced the osteoclast formation in the co-culture system. These bacterial sonicates increased the expression of RANKL and PGE(2), and decreased the expression of OPG in osteoblasts. The addition of OPG, an inhibitor of RANKL, in the co-culture completely suppressed the osteoclastogenesis that was stimulated by each bacterial sonicate. Indomethacin, which is an inhibitor of PGE(2) synthesis, reduced more than 88% of the osteoclast formation induced by each bacterial sonicate. Indomethacin inhibited more than 80% of RANKL expression in osteoblasts induced by T. denticola and T. socranskii, and 59% by P. gingivalis. Indomethacin completely recovered the depression of OPG expression in osteoblasts by T. denticola and T. socranskii to the level of the untreated osteoblasts. Indomethacin recovered the reduction of OPG expression by P. gingivalis to 67%.. These findings suggest that the osteoclastogenesis by P. gingivalis, T. denticola, and T. socranskii is mediated by a RANKL-dependent pathway and that PGE(2) is a main factor in the pathway by the enhancing of RANKL expression and the depression of osteoprotegerin, a RANKL inhibitor.

Topics: Acid Phosphatase; Animals; Carrier Proteins; Cattle; Cell Differentiation; Dinoprostone; Glycoproteins; Indomethacin; Isoenzymes; Membrane Glycoproteins; Mice; NF-kappa B; Osteoblasts; Osteoclasts; Osteoprotegerin; Periodontitis; Porphyromonas gingivalis; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Skull; Statistics, Nonparametric; Tartrate-Resistant Acid Phosphatase; Treponema; Treponema denticola

The effects of the potassium channel (Kv1.3) blocker kaliotoxin on T-cell-mediated periodontal bone resorption were examined in rats. Systemic administration of kaliotoxin abrogated the bone resorption in conjunction with decreased RANKL mRNA expression by T-cells in gingival tissue. This study suggests a plausible therapeutic approach for inflammatory bone resorption by targeting Kv1.3.. Kv1.3 is a critical potassium channel to counterbalance calcium influx at T-cell receptor activation. It is not known if Kv1.3 also regulates RANKL expression by antigen-activated T-cells, and consequently affects in vivo bone resorption mediated by activated T-cells.. Actinobacillus actinomycetemcomitans 29-kDa outer membrane protein-specific Th1-clone cells were used to evaluate the expression of Kv1.3 (using reverse transcriptase-polymerase chain reaction [RT-PCR] and Western blot analyses) and the effects of the potassium channel blocker kaliotoxin (0-100 nM) on T-cell activation parameters ([3H]thymidine incorporation assays and ELISA) and expression of RANKL and osteoprotegerin (OPG; flow cytometry, Western blot, and RT-PCR analyses). A rat periodontal disease model based on the adoptive transfer of activated 29-kDa outer membrane protein-specific Th1 clone cells was used to analyze the effects of kaliotoxin in T-cell-mediated alveolar bone resorption and RANKL and OPG mRNA expression by gingival T-cells. Stimulated 29-kDa outer membrane protein-specific Th1 clone cells were transferred intravenously on day 0 to all animals used in the study (n = 7 animals per group). Ten micrograms of kaliotoxin were injected subcutaneously twice per day on days 0, 1, 2, and 3, after adoptive transfer of the T-cells. The control group of rats was injected with saline as placebo on the same days as injections for the kaliotoxin-treated group. The MOCP-5 osteoclast precursor cell line was used in co-culture studies with fixed 29-kDa outer membrane protein-specific Th1-clone cells to measure T-cell-derived RANKL-mediated effects on osteoclastogenesis and resorption pit formation assays in vitro. Statistical significance was evaluated by Student's t-test.. Kaliotoxin decreased T-cell activation parameters of 29-kDa outer membrane protein-specific Th1 clone cells in vitro and in vivo. Most importantly, kaliotoxin administration resulted in an 84% decrease of the bone resorption induced in the saline-treated control group. T-cells recovered from the gingival tissue of kaliotoxin-treated rats displayed lower ratios of RANKL and OPG mRNA expression than those recovered from the control group. The ratio of RANKL and osteoprotegerin protein expression and induction of RANKL-dependent osteoclastogenesis by the activated T-cells were also markedly decreased after kaliotoxin treatments in vitro.. The use of kaliotoxin or other means to block Kv1.3 may constitute a potential intervention therapy to prevent alveolar bone loss in periodontal disease.

Topics: Acid Phosphatase; Adoptive Transfer; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Antigen Presentation; Bacterial Outer Membrane Proteins; Blotting, Western; Carrier Proteins; CD3 Complex; Cell Differentiation; Coculture Techniques; Down-Regulation; Female; Gene Expression; Glycoproteins; Immunoglobulin G; Interferon-gamma; Isoenzymes; Kv1.3 Potassium Channel; Lipopolysaccharides; Lymphocyte Activation; Maxilla; Membrane Glycoproteins; Osteoclasts; Osteoprotegerin; Periodontitis; Potassium Channels; Potassium Channels, Voltage-Gated; RANK Ligand; Rats; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Scorpion Venoms; Spleen; T-Lymphocytes; Tartrate-Resistant Acid Phosphatase

This study investigated the expression of key mediators that regulate differentiation of osteoclasts, receptor activator of nuclear factor kappaB ligand (RANKL), and its natural inhibitor, osteoprotegerin (OPG), in periodontitis. We aimed to compare the levels of the RANKL and OPG in the granulomatous tissue adjacent to areas of alveolar bone loss from patients with periodontitis to that present in tissue from patients without periodontitis. In addition, we aimed to determine the types of cells expressing these factors in these tissues and to demonstrate the expression of the osteoclastic markers, RANK and tartrate-resistant acid phosphatase (TRAP), in periodontitis.. Frozen biopsy specimens were analysed using specific monoclonal antibodies and were evaluated by semiquantitative analysis and digital image analysis to compare levels of RANKL and OPG protein expression. Double labelling of frozen sections with antibodies to different cell lineage specific markers was used to determine the types of cells expressing these proteins. In situ hybridization was used to detect cells expressing RANK mRNA.. Semiquantitative image analysis demonstrated that significantly higher levels of RANKL protein (P < 0.05) were expressed in the periodontitis tissue. Conversely, OPG protein was significantly lower (P < 0.05) in the periodontitis tissues. RANKL protein was associated with lymphocytes and macrophages. OPG protein was associated with endothelial cells in both tissues. Many leukocytes expressing RANK mRNA and TRAP were observed in periodontitis tissues.. The change in the levels of these key regulators of osteoclast differentiation may play a major role in the bone loss seen in periodontitis.

Topics: Acid Phosphatase; Adolescent; Adult; Aged; Alveolar Bone Loss; Biomarkers; Carrier Proteins; Cell Differentiation; Endothelium, Vascular; Female; Gene Expression Regulation; Glycoproteins; Humans; Isoenzymes; Leukocytes, Mononuclear; Ligands; Male; Membrane Glycoproteins; Middle Aged; NF-kappa B; Osteoclasts; Osteoprotegerin; Periodontitis; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor; Tartrate-Resistant Acid Phosphatase

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 CD28 costimulation at TCR signaling plays a pivotal role in the regulation of the T cell response. To elucidate the role of T cells in periodontal disease, a system of cell transfer with TCR/CD28-dependent Th1 or Th2 clones was developed in rats. Gingival injection of specific Ag, Actinobacillus actinomycetemcomitans 29-kDa outer membrane protein, and LPS could induce local bone resorption 10 days after the transfer of Ag-specific Th1 clone cells, but not after transfer of Th2 clone cells. Interestingly, the presence of LPS was required not only for the induction of bone resorption but also for Ag-specific IgG2a production. LPS injection elicited the induction of expression of both B7-1 and B7-2 expression on gingival macrophages, which otherwise expressed only MHC class II when animals were injected with Ag alone. The expression of B7 molecules was observed for up to 3 days, which corresponded to the duration of retention of T clone cells in gingival tissues. Either local or systemic administration of CTLA4Ig, a functional antagonist of CD28 binding to B7, could abrogate the bone resorption induced by Th1 clone cells combined with gingival challenge with both Ag and LPS. These results suggest that local Ag-specific activation of Th1-type T cells by B7 costimulation appeared to trigger inflammatory bone resorption, whereas inhibition of B7 expression by CTLA4Ig might be a therapeutic approach for intervention with inflammatory bone resorption.

Topics: Abatacept; Acid Phosphatase; Alveolar Bone Loss; Animals; Antigens, CD; Antigens, Differentiation; B7-1 Antigen; B7-2 Antigen; Bone Resorption; Clone Cells; CTLA-4 Antigen; Female; Gingiva; Histocompatibility Antigens Class II; Humans; Immunoconjugates; Immunoglobulin G; Injections, Intravenous; Isoenzymes; Kinetics; Lipopolysaccharides; Lymph Nodes; Macrophages; Membrane Glycoproteins; Microinjections; Neck; Periodontitis; Rats; Rats, Inbred Strains; RNA, Messenger; Spleen; Tartrate-Resistant Acid Phosphatase; Th1 Cells; Th2 Cells

This study was undertaken to observe osteoclast differentiation related to inflammatory progression in aggressive periodontitis induced in beagle dogs by ligature of the gingival sulcus. To monitor osteoclastic activity, we used histochemical methods (staining for tartrate-resistant acid phosphatase [TRAP]) to visualize osteoclasts and their TRAP-positive precursors and biochemical methods (ELISA assay of pyridinium crosslinks) to detect bone matrix degradation products in gingival crevicular fluid (GCF), serum, and urine. For histochemical study, tissue specimens were prepared from 3 adult female beagle dogs induced with experimental periodontitis by silk ligature placement below the gingival margin of mandibular molars ligated for 3, 7, and 21 days. For biochemical study for pyridinoline measurement, the 24 mandibular molars of 4 male beagle dogs were ligated. GCF, urine, and serum were collected at day 0 and at 3, 7, 14, and 21 days after ligation. In the early inflammatory phase of ligature-induced periodontitis (day 3), TRAP+ mononuclear and TRAP+ multinucleated cells were present in the gingival connective tissue, and active bone-resorbing cells were found in excavated lacunae at the alveolar crest, but osteoclasts were not infiltrating the periodontal ligament during this early phase. During later stages of the inflammatory process (7 and 21 days), osteoclasts appeared at both the gingival and ligament side of the alveolar bone. Osteoclastic bone resorption appeared to be more severe on the bone surface at the gingival side than on the bone surface of the periodontal ligament side. Measurement of pyridinoline significantly increased in GCF and urine 3 days after ligation. The results suggested that bone at the crest of the alveolar bone is rapidly resorbed within 3 days of inducing experimental periodontitis.

Topics: Acid Phosphatase; Alveolar Bone Loss; Alveolar Process; Amino Acids; Animals; Biomarkers; Cell Differentiation; Coloring Agents; Connective Tissue; Cross-Linking Reagents; Disease Progression; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Giant Cells; Gingiva; Gingival Crevicular Fluid; Histocytochemistry; Isoenzymes; Leukocytes, Mononuclear; Male; Osteoclasts; Periodontal Ligament; Periodontitis; Pyridinium Compounds; Tartrate-Resistant Acid Phosphatase

The effects of recombinant human interleukin-1 beta (rhIL-1 beta) on alveolar bone resorptive activity in rats were examined. Continuous administration of rhIL-1 beta or phosphate-buffered saline (PBS) was given via osmotic pumps for 3, 7 and 14 days to rats with silk ligatures around second maxillary molars. Other animals without ligatures received insertion of pumps containing rhIL-1 beta or remained untreated. Sections were subject to three different stains:--hematoxylin and eosin (H-E) for histology, acid phosphatase (ACPase) activity for osteoclast detection, and immunohistochemistry using anti-rat monocyte/macrophage monoclonal antibody (ED 1). In addition, body weight, plasma calcium and phosphorus levels were monitored. The mean body weight of rats receiving rhIL-1 beta was significantly lower (P < 0.05 to P < 0.01) compared with untreated rats throughout the experimental period. On Day 7, plasma calcium and phosphorus levels were significantly lower in rats receiving rhIL-1 beta than in rats receiving PBS only (P < 0.05). Sections revealed a moderate inflammatory cell infiltrate reaching near the alveolar crest in both groups with ligatures on Day 3. Only rats receiving rhIL-1 beta exhibited enhancement of inflammatory cell invasion on Days 7 and 14. In rats receiving rhIL-1 beta with ligatures, numerous resorption lacunae containing ACPase-positive multinucleated giant cells (MNGCs), coinciding with ED1-positive cells, were located on the mesial side of the septum where extensive bone resorption had occurred throughout the experimental period. In animals receiving rhIL-1 beta without ligatures, compared with untreated rats, increased ACPase-positive cells were observed on the mesial side of the septum on Day 3. In animals receiving PBS only, a few ACPase-positive cells were observed confined to the mesial regions where slight bone resorption occurred on Days 7 and 14. These results indicate that the administration of rhIL-1 beta accelerated alveolar bone destruction in ligature-induced periodontal tissue inflammation over a two-week period.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Antibodies, Monoclonal; Body Weight; Bone Resorption; Calcium; Coloring Agents; Giant Cells; Humans; Immunohistochemistry; Infusion Pumps; Insect Proteins; Interleukin-1; Ligation; Macrophages; Male; Maxillary Diseases; Molar; Monocytes; Osteoclasts; Periodontitis; Phosphorus; Proteins; Rats; Rats, Wistar; Recombinant Proteins; Silk; Sutures

Apical periodontitis was surgically induced in the mandibular first molar of rats and chronological changes in the periapical bone tissue were observed by histochemistry and electron microscopy. On the second postoperative day (Day 2), tartrateresistant acid phosphatase (TRACPase)-positive cells emerged on the bone surface facing the inferior alveolar nerve, whereas alkaline phosphatase (ALPase)-positive cells proliferated on the bone marrow surface of the mandibular canal wall. On Day 3, the active resorption of the mandibular canal wall appeared on the surface facing the inferior alveolar nerve. The bone of the upper wall of the canal was completely resorbed. On Day 4, however, numerous ALPase-positive cells emerged over the bone surface facing the inferior alveolar nerve intermingled with TRACPase-positive cells. On Day 5, repair of the upper wall of the mandibular canal by new bone progressed. Bone formation was also observed on the bone surface facing the inferior alveolar nerve. On Day 6, the upper wall of the mandibular canal was remodeled by the new bone, whereas TRACPase-positive cells had already migrated over the bone surface in the vicinity of ALPase-positive cells. From Days 2 to 5, active trabecular bone formation continued in the bone marrow cavity close to the mandibular canal, while TRACPase-positive cells were found only on Day 6. These demonstrate that inflammatory stimuli activate bone formation coupled with bone resorption, as well as direct trabecular bone formation without a bone resorption phase. A rapid bone turnover in the early stage of apical periodontitis is also suggested. We conclude that bone defects in apical periodontitis are not the result of sole bone resorption but rather, active bone remodeling.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Remodeling; Male; Mandible; Periodontitis; Rats; Rats, Inbred Strains; Tartrates

Quantification of osteoclast resorption, a good index of periodontitis destruction, is primarily based on osteoclast identification. As their identification is sometimes dubious, we compared osteoclastic counts in hamster specimens processed for either routine histology or tartrate-resistant acid phosphatase (TRAP) staining. No difference was found between the two approaches concerning the number of osteoclasts. However the mean bone-osteoclast interface was higher in the TRAP-stained specimens (+30%, p less than 0.02). As osteoclast precursors are also TRAP+ cells, they were quantified too. Compared with controls, there was a dramatic increase (p less than 0.0001) in periodontitis-affected animals. Precursors were strongly correlated to active osteoclasts (r = 0.97). Our data suggest that precursors are recruited only when the disease is active in a given site.

Topics: Acid Phosphatase; Alveolar Bone Loss; Animals; Cell Count; Cricetinae; Histocytochemistry; Male; Mesocricetus; Osteoclasts; Periodontitis; Staining and Labeling; Stem Cells

The therapeutic effect of superoxide dismutase (SOD) and the role of O2- were assessed on 3 groups of Wistar rats (total 115). Fifty-four received injections of gingival bacteria or of anaerobically cultured rat dental plaque in their peritoneum, then received both intravenous (i.v.) and intraperitoneal (i.p.) injection of SOD. The rats were killed 48 hours later to collect their peritoneal exudate for cell count and for acid phosphatase activity assessment. Twenty-six received injections of bacteria in their footpads, after which SOD was administered intravenously. These rats were killed at 6 hours, 48 hours and 1 week respectively for histological examination. The gingiva of 26 rats were incised to create artificial lesions. The rats were killed at 24 or 48 hours and examined histologically. The nine remaining rats were used as controls (untreated) for the 3 experiments. The results of the 3 experiments showed that: Injection of SOD reduced exudation and acid phosphatase activity enhanced by the injection of B. gingivalis, at dosages of 1, 5 mg/kg i.p. and 5 mg/kg i.v., but 10 mg/kg i.p. had no apparent effect; i.v. injection of SOD had inhibitory effects on cell infiltration of B. gingivalis into the footpad, and the increase in fibrin and fibroblast formation through time was greater in SOD-administered rats; a decreased cell infiltration rate and increased fibrin network, fibroblast proliferation and gingival tissue regeneration occurred in specimens with artificial lesions given SOD. Apparently SOD has a curative effect on both inflammatory reaction induced by B. gingivalis and periodontal wound healing.

Topics: Acid Phosphatase; Animals; Dental Plaque; Gingival Crevicular Fluid; Periodontitis; Porphyromonas gingivalis; Rats; Rats, Inbred Strains; Superoxide Dismutase; Wound Healing

PMN leukocytes from circulating blood, were investigated using the following parameters: chemiluminescence, superoxide generation, exocytic degranulation, lactoferrin, myeloperoxydase and protein-kinase-C activities. The results did not allow to point out any significative discrepancy between samples collected from either patients suffering of periodontal disease or healthy individuals. However, the phagocytic ability (as measured by light emission and superoxide production) was found to be severely inhibited in both instances, when PMN leukocytes were incubated in the presence of succinic acid. Similarly, protein-kinase-C activity in these cells, collapsed when the pH was brought down to 5.5. These data and the available literature indicate that pH decrease as well as succinic acid release in the crevicular fluid, may play a role in the pathogeny of chronic periodontitis in adults.

Topics: Acid Phosphatase; Adult; Cell Migration Inhibition; Female; Humans; Hydrogen-Ion Concentration; L-Lactate Dehydrogenase; Luminescent Measurements; Male; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Periodontitis; Phagocytosis; Protein Kinase C; Succinates; Succinic Acid; Superoxides; Zymosan

Topics: Acid Phosphatase; Actinobacillus; Alkaline Phosphatase; Alveolar Process; Animals; Bone Resorption; Capnocytophaga; Extracellular Matrix; Male; Osteogenesis; Periodontitis; Rats; Rats, Inbred Strains

Acid phosphatase activity was examined ultracytochemically in gingival specimens to elucidate the response of plasma cells to Russell's bodies. The acid phosphatase activity was discernible in lysosomes of various morphology, some of which contained Russell's bodies. The acid phosphatase activity was stronger in the peripheries of such lysosomes, but weak activity was also found inside Russell's bodies. These findings indicated that at least some of Russell's bodies formed within the plasma cells were degraded in autophagolysosomes.

Topics: Acid Phosphatase; Adult; Endoplasmic Reticulum; Gingiva; Humans; Inclusion Bodies; Lysosomes; Microscopy, Electron; Middle Aged; Periodontitis; Plasma Cells

Fifteen patients, eight males and seven females, ranging from 30 to 88 years of age with advanced periodontal disease were selected for this study. Biopsies and blood samples were taken of both normal and inflamed gingival tissues, and processed for detection of nonspecific esterase and acid phosphatase activity in monocytes and macrophages. Activated macrophages, as indicated by their intense reaction to acid phosphatase and nonspecific esterase, were found in the gingival epithelium, lamina propria, perivascular tissues and in the blood vessels in human chronic periodontitis. Blood smears of monocytes showed variability of stain intensity suggesting that their activation occurred in blood vessels where they marginate and emigrate into the perivascular tissues in chronic periodontitis. They then appear as macrophages that migrate through the connective tissue, penetrate the basement membrane and continue through the epithelium. The nonspecific esterase stain identified T-cells, by a singular dot-like granule, and plasma cells by multiple granules in the cytoplasm. Lymphocytes containing multiple cytoplasmic nonspecific esterase positive granules commonly were found only in the perivascular connective tissue and may represent B-cell differentiation to plasma cells. The plasma cell predominance, the presence of T-cells and activated macrophages indicated both humoral and cell-mediated responses are operative in human chronic periodontitis.

Topics: Acid Phosphatase; Adult; Aged; Cell Differentiation; Cell Movement; Female; Humans; Macrophages; Male; Middle Aged; Monocytes; Naphthol AS D Esterase; Periodontitis; Staining and Labeling

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Dogs; Glucuronidase; Lysosomes; Periodic Acid-Schiff Reaction; Periodontal Pocket; Periodontitis

Topics: Acid Phosphatase; Adult; Alkaline Phosphatase; Cathepsins; Cytoplasm; Gingiva; Gingivitis; Glucuronidase; Humans; Hydrogen-Ion Concentration; Hydrolases; Kinetics; Lysosomes; Periodontitis

Topics: Acid Phosphatase; Densitometry; Dental Plaque; Epithelial Cells; Epithelium; Gingival Crevicular Fluid; Gingivitis; Humans; In Vitro Techniques; Kinetics; Periodontitis

Topics: Acid Phosphatase; Alkaline Phosphatase; Humans; Hydrogen Peroxide; Keratosis; Occlusive Dressings; Periodontitis; Succinate Dehydrogenase

Topics: Acid Phosphatase; Alkaline Phosphatase; Gingiva; Oxidoreductases; Periodontitis

Topics: Acid Phosphatase; Alkaline Phosphatase; L-Lactate Dehydrogenase; Periodontitis; Succinate Dehydrogenase