The influence of loss of bone mass on induced periodontal disease: A radiographic and densitometric study of female rats

Background: Changes in mineral density in the mandibular and femoral bones (BMD) after estrogen deficiency caused by ovariectomy (OVX) and the influence of these changes on induced periodontal disease were evaluated in female rats.Methods: Forty-eight female Holtzman rats (90 days old) were randomly divided into five groups: 0: control (N = 9); 1: SHAM without induced periodontal disease (N = 11); 2: SHAM with induced disease (N = 10); 3: OVX without induced disease (N = 9); and 4: OVX with induced disease (N = 9). In groups 2 and 4, the first lower molars were tied with ligatures for 30 days 120 days after surgery. After 5 months the animals were sacrificed to measure global mineral density (BMD) and that of the sub-regions of the mandible and femur by dual energy x-ray absorptiometry (DXA). The extent of vertical bone loss was evaluated with digital radiography by measuring the distance from the bone crest to the cemento-enamel junction at the mesial of the first lower molar.Results: Results of the femur (Kruskal-Wallis test) showed a significant difference (P < 0.001) between the groups SHAM and OVX in bone density values for all regions. Comparison between the groups in relation to the BMD of the mandible, both in the sub-regions and global revealed no differences (P < 0.05). The vertical bone loss measured for the groups with induced disease was similar (P= 0.713).Conclusions: Differences between the groups were found in the bone mineral density BMD of the femur but not of the mandible. OVX had no influence on induced periodontal disease.

Curcumin modulates the immune response associated with LPS-induced periodontal disease in rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Differential regulation of MMP-13 expression in two models of experimentally induced periodontal disease in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of the period after ovariectomy on femoral and mandibular bone density and on induced periodontal disease

Background: This study investigated the influence of the period after ovariectomy on femoral and mandibular bone mineral density (BMD) and on induced periodontal disease.Methods: One hundred and twenty-six female Holtzman rats were divided into nine groups: control, sham surgery (SHAM) with and without induction of periodontal disease for 51 and 150 days, and ovariectomy (OVX) with and without induction of periodontal disease for 51 and 150 days. Periodontal disease was induced by placing ligatures on the first lower molars during the last 30 days of each period. BMD was measured by dual-energy x-ray absorptiometry. Vertical bone loss was determined by measuring the distance from the alveolar bone crest to the cemento-enamel junction on the mesial side of the first lower molar.Results: Statistical analyses (Kruskal-Wallis test) revealed a significant difference between the OVX and SHAM groups' global and femoral proximal epiphysis BMD (P < 0.001) for 150 days and in the global evaluation for 51 days. For mandibular BMD, no difference was found between the groups of each period. Influence of the period on femoral BMD was found only for the SHAM groups, with lower BMD for the 51-day period compared to the 150-day period (P < 0.05). In the global evaluation of the mandible, a lower BMD was found after 51 days. The period was a contributing factor for the vertical bone loss, and it resulted in higher values for the 51-day period (P < 0.05).Conclusion: the period influenced the femoral BMD and the vertical bone loss in induced periodontal disease.

Energy Expenditure and Oxygen Consumption as Novel Biomarkers of Obesity-induced Cardiac Disease in Rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of induced diabetes mellitus on alveolar bone loss after 30 days of ligature-induced periodontal disease

Several studies have shown that diabetics are more susceptible to the development of severe periodontal disease. Currently, the use of animal models can be considered a feasible alternative in radiographic assessments of these two pathologies. The purpose of this radiographic study was to evaluate the effect of induced diabetes mellitus on alveolar bone loss after 30 days of ligature-induced periodontal disease. Sixty-four Wistar rats were randomly distributed into four experimental groups. Diabetes was induced in Groups II and IV, while periodontal disease was induced in Groups III and IV; Group I was used as control. In order to perform the radiographic assessment of the specimens, the rats were killed on the 3rd and 30th days of the study. Radiographic measurements were assessed with ANOVA and Tukey's test to determine statistically significant differences (p < 0.05). It was observed that Groups III and IV featured greater bone loss when compared to Groups I and II. Only the diabetic group with periodontal disease (Group IV) featured statistically significant greater bone loss when compared to the other groups. These results suggested that the alveolar bone loss resulting from the periodontal disease installation is greater when associated to the diabetes mellitus.

Evaluation of effect of cyclosporine A on the bone tissue with induced periodontal disease to ligature in rats

The administration of cyclosporine A (CsA) has been associated with significant bone loss and increased bone remodeling. The present investigation was designed to evaluate the effects of CsA on alveolar bone of rats subjected to experimental periodontitis, using histomorphometric and histological analysis. Twenty-four rats were divided into groups with 6 animals each: 1, control; 2, rats with ligature around the lower first molars; 3, rats with ligature around the lower first molars and that were treated with 10 mg CsA/kg of body weight/d; and 4, rats treated with 10 mg CsA/kg of body weight/d. At the end of 30 days, rats were humanely killed and subjected to a histological processing, with analysis of the distance cemento-enamel junction and alveolar bone crest, bone area, eroded bone area, and cemento surface. All of them were assessed at the mesial region of the alveolar bone. The CsA therapy combined with ligature placement decreased bone area and increased the eroded bone area around the tooth surface. The results at the histological analysis showed the same combination and changes. Therefore, in spite of the lack of a direct effect on the alveolar bone height, the CsA therapy intensified the imbalance of the alveolar bone homeostasia in a rat model of experimental periodontitis. © 2013 Elsevier Inc.

Evaluation of the Host Response in Various Models of Induced Periodontal Disease in Mice

Background: The aim of this study is to characterize and evaluate the host response caused by three different models of experimental periodontitis in mice.Methods: C57BL/6 wild-type female mice were distributed into six experimental groups and sacrificed at 7, 15, and 30 days after the induction of periodontal disease: 1) group C: no treatment control group; 2) group L: periodontal disease induced by ligature; 3) group G-Pg: oral gavage with Porphyromonas gingivalis (Pg); 4) group G-PgFn: oral gavage with Fusobacterium nucleatum + Pg; 5) group I-Pg: heat-killed Pg injected into the palatal mucosa between the molars; and 6) group I-V: phosphatebuffered saline injected into the palatal mucosa. The samples were used to analyze the immune-inflammatory process in the gingival tissue via descriptive histologic and real-time polymerase chain reaction analyses. The alveolar bone loss was evaluated using microcomputed tomography. The data were analyzed using the Kruskal-Wallis test, followed by a post hoc Dunn test and analysis of variance, followed by a Tukey test using a 5% significance level.Results: Only the ligature model displayed significant alveolar bone loss in the initial period (7 days), which was maintained with time. The group injected with heat-killed Pg displayed significant alveolar bone loss starting from day 15, which continued to progress with time (P < 0.05). A significant increase (P < 0.05) in the gene expression of proinflammatory cytokines (interleukin-6 and -1b) and proteins involved in osteoclastogenesis (receptor activator of nuclear factor-kB ligand and osteoprotegerin) was observed in the ligature group on day 7.Conclusion: The ligature and injection of heat-killed Pg models were the most representative of periodontal disease in humans, whereas the oral gavage models were not effective at inducing the disease under the experimental conditions.

Alloxan-Induced Diabetes Triggers the Development of Periodontal Disease in Rats

Background. Periodontal disease in diabetic patients presents higher severity and prevalence; and increased severity of ligature-induced periodontal disease has been verified in diabetic rats. However, in absence of aggressive stimuli such as ligatures, the influence of diabetes on rat periodontal tissues is incompletely explored. The aim of this study was to evaluate the establishment and progression of periodontal diseases in rats only with diabetes induction. Methodology/Principal Findings. Diabetes was induced in Wistar rats (n = 25) by intravenous administration of alloxan (42 mg/kg) and were analyzed at 1, 3, 6, 9 and 12 months after diabetes induction. The hemimandibles were removed and submitted to radiographical and histopathological procedures. A significant reduction was observed in height of bone crest in diabetic animals at 3, 6, 9 and 12 months, which was associated with increased numbers of osteoclasts and inflammatory cells. The histopathological analyses of diabetic rats also showed a reduction in density of collagen fibers, fibroblasts and blood vessels. Severe caries were also detected in the diabetic group. Conclusions/Significance. The results demonstrate that diabetes induction triggers, or even co-induces the onset of alterations which are typical of periodontal diseases even in the absence of aggressive factors such as ligatures. Therefore, diabetes induction renders a previously resistant host into a susceptible phenotype, and hence diabetes can be considered a very important risk factor to the development of periodontal disease.

Periodontal Disease Decreases Insulin Sensitivity and Insulin Signaling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fluoxetine Inhibits Inflammatory Response and Bone Loss in a Rat Model of Ligature-Induced Periodontitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of photodynamic therapy on the development of ligature-induced periodontitis in rats

Background: the purpose of this study was to evaluate, histologically and radiographically, the effect of photodynamic therapy on the progression of experimentally induced periodontal disease in rats.Methods: Ligatures were placed at the first mandibular molar in rats. The animals were divided into four groups: group 1 (C) received no treatment; group 2 was treated topically with methylene blue (MB; 100 mu g/ml); group 3 was treated with low-level laser therapy (LLLT); and group 4 was treated topically with methylene blue followed by LLLT (4.5 J/cm(2)) (photodynamic therapy; PDT). Rats were sacrificed 5, 15, or 30 days postoperatively. Standardized radiographs were taken to measure bone loss around the mesial root surface of the first molar. Data were analyzed statistically (analysis of variance and Tukey test; P < 0.05). A scoring system was used to evaluate the connective tissue, periodontal ligament, and alveolar bone histologically. Data were analyzed statistically (Kruskal-Wallis test; P < 0.05).Results: Radiographic examination showed that there was significantly less bone loss in Group PDT compared to Group C at 5 and 15 days postoperatively. There was no significant difference in bone loss at 30 days. At 15 days, the histologic results showed significant differences in the extent of inflammatory reaction in the gingival tissue, with a greater extent of chronic inflammatory reaction in Group LLLT.Conclusion: PDT transiently reduced the periodontal tissue destruction.

Bone regeneration in cranioplasty and clinical complications in rabbits with alloxan-induced diabetes

This research evaluated the bone repair process in surgical defects created on the parietal bones of diabetic rabbits using the guided bone regeneration technique to observe the effects of alloxan in the induction of diabetes mellitus. Twenty-four adult rabbits were divided into three study groups: control (C), diabetic (D) and diabetic associated to polytetrafluoroethylene (PTFE) membrane (D-PTFE). For diabetes induction the animals received one dose of monohydrated alloxan (90 mg/kg) by intravenous administration in the auricular or femoral vein. In group D-PTFE the membrane covered both the floor and the surface of the bone defect. In groups D and C, the bone defect was filled up with blood clot. The specimens were fixed in 10% formol and prepared for histomorphometric analysis. The results showed that the 90 mg/kg dose of monohydrate alloxan was sufficient to promote diabetes mellitus when administered in the auricular vein. Bone regeneration was slower in the diabetic group when compared with the control and diabetic-PTFE groups, but there was no significant statistical difference between the two experimental groups (D and D-PTFE). The oral and general clinical complications among the diabetics were weight loss, polyuria, polyphagia and severe chronic gingivitis.

Low caloric value of ethanol itself increases alveolar bone loss in ligature-induced periodontitis in male rats

This study aimed at morphometrically evaluating the influence of variable caloric values of ethanol consumption on alveolar bone loss in periodontitis in male rats. Thirty-six male rats were randomized into four groups of nine rats each, as follows: Test group A (low) - rats were fed an ethanol-containing liquid diet (ethanol representing 22% of total caloric value); Control group A - rats were fed a pair-fed control diet (ethanol replaced by isocaloric amounts of carbohydrate); Test group B (high) - rats were fed an ethanol-containing liquid diet (ethanol representing 36% of total caloric value); Control group B - rats were fed a pair-fed control diet for Test B. Following anesthesia, cotton ligatures were placed around the cervix of the right upper second molar. At eight weeks, the maxillary bones were removed and alveolar bone loss was analyzed by measuring the distance between the cementoenamel junction and the alveolar bone crest at buccal and palatal sites of the upper second molar. The unligated groups showed no significant differences between the bone loss values observed for the low and high caloric values of ethanol (p > 0.05). In the ligated groups, the rats receiving low caloric values of ethanol showed significantly greater bone loss compared to the isocaloric rats (p < 0.05); however, the rats receiving high caloric values of ethanol showed no significant differences compared to the controls. Analysis of the results demonstrated that, in male rats, ethanol itself affected ligature-induced bone loss when representing a low value in the total caloric value.

SOCS3 expression correlates with severity of inflammation, expression of proinflammatory cytokines, and activation of STAT3 and p38 MAPK in LPS-induced inflammation in vivo

SOCS3 is an inducible endogenous negative regulator of JAK/STAT pathway, which is relevant in inflammatory conditions. We used a model of LPS-induced periodontal disease in rats to correlate SOCS3 expression with the inflammatory status. In vitro we used a murine macrophage cell line to assess the physical interaction between SOCS3 and STAT3 by coimmunoprecipitation. 30 ug of LPS from Escherichia coli were injected in the gingival tissues on the palatal aspect of first molars of the animals 3x/week for up to 4 weeks. Control animals were injected with the vehicle (PBS). The rats were sacrificed at 7, 15, and 30 days. Inflammation and gene expression were assessed by stereometric analysis, immunohistochemistry, RT-qPCR, and western blot. LPS injections increased inflammation, paralleled by an upregulation of SOCS3, of the proinflammatory cytokines IL-1β, IL-6, and TNF-and increased phosphorylation of STAT3 and p38 MAPK. SOCS3 expression accompanied the severity of inflammation and the expression of proinflammatory cytokines, as well as the activation status of STAT3 and p38 MAPK. LPS stimulation in a macrophage cell line in vitro induced transient STAT3 activation, which was inversely correlated with a dynamic physical interaction with SOCS3, suggesting that this may be a mechanism for SOCS3 regulatory function. © 2013 João Antônio Chaves de Souza et al.