Micro‐computerized tomography analysis of alveolar bone loss in ligature‐ and nicotine‐induced experimental periodontitis in rats

Liu Y‐F, Wu L‐A, Wang J, Wen L‐Y, Wang X‐J. Micro‐computerized tomography analysis of alveolar bone loss in ligature‐ and nicotine‐induced experimental periodontitis in rats. J Periodont Res 2010; 45: 714–719. © 2010 John Wiley & Sons A/S Background and Objective: Nicotine reportedly is a risk factor for periodontitis, but accurate data regarding nicotine‐induced alveolar bone loss is lacking. The aim of this study was to quantitatively assess alveolar bone loss in ligature‐ and nicotine‐induced periodontitis in rats using micro‐computerized tomography (micro‐CT). Material and Methods: Thirty‐six adult male rats were treated by placing silk ligatures around the cervixes of the right second maxillary molar; the contralateral tooth was untreated. After ligation, the animals were randomly divided into three groups: group A received intraperitoneal injections of saline solution, group B received 0.83 mg of nicotine/kg/d, and group C received 1.67 mg of nicotine/kg/d. Six animals in each group were killed on days 14 and 28 after ligature placement, and then micro‐CT examinations were conducted. Results: In all groups, bone mineral density (BMD), bone volume fraction (BVF), trabecular number (Tb.N) and trabecular thickness (Tb.Th) values of the ligated sides were significantly lower than those of the unligated sides (p < 0.001), whereas alveolar bone height loss (ABHL) and trabecular separation (Tb.Sp) of the ligated sides were significantly higher than those of the unligated sides (p < 0.001). Compared with the control group, nicotine administration increased the ABHL value and decreased the BMD, BVF and Tb.Th values of both sides in a dose‐dependent manner (p < 0.05). Conclusion: Our results confirmed that ligature could cause significant loss in the trabecula of alveolar bone, and daily administration of nicotine resulted in further bone loss and microstructure deterioration.     

Parathyroid hormone administration may modulate periodontal tissue levels of interleukin-6, matrix metalloproteinase-2 and matrix metalloproteinase-9 in experimental periodontitis

Background and Objective: 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. Material and Methods: 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‐1beta, 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. Results: 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. Conclusion: 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.     

N‐acetylcysteine decreases alveolar bone loss on experimental periodontitis in streptozotocin‐induced diabetic rats

Toker H, Ozdemir H, Balc? H, Ozer H. N‐acetylcysteine decreases alveolar bone loss on experimental periodontitis in streptozotocin‐induced diabetic rats. J Periodont Res 2012; 47: 793–799. © 2012 John Wiley & Sons A/S Background and Objective: The purpose of this study was to evaluate the morphometric and histopathological changes associated with experimental periodontitis in diabetic rats in response to systemic administration of N‐acetylcysteine (NAC), a sulfhydryl‐containing thiol antioxidant. Material and methods: Sixty Wistar rats were divided into six experimental groups: nonligated (NL) group; ligature‐only (L) group; streptozotocin‐only (STZ) group; STZ and ligature (STZ + L) group; and systemic administration of NAC and ligature (70 and 100 mg/kg body weight per day, respectively) (NAC70 and NAC100 groups). Diabetes mellitus was induced by 60 mg/kg of streptozotocin. Silk ligatures were placed at the gingival margin of the lower first molars of the mandibular quadrant. The study duration was 30 d and the animals were killed at the end of this period. Changes in alveolar bone levels were clinically measured and tissues were histopathologically examined to assess the differences among the study groups. Results: At the end of the 30‐d study period, alveolar bone loss was significantly higher in the STZ + L group compared with the other groups (p < 0.05). Also, alveolar bone loss in all the NAC groups was significantly lower than in the STZ + L and L groups (p < 0.05). The osteoblastic activity in the NAC100 group was significantly higher than in the other groups (p < 0.05). Conclusion: Within the limits of this study, it can be suggested that NAC, when administered systemically, prevents alveolar bone loss in the diabetic rat model.     

Topical and intermittent application of parathyroid hormone recovers alveolar bone loss in rat experimental periodontitis

Tokunaga K, Seto H, Ohba H, Mihara C, Hama H, Horibe M, Yoneda S, Nagata T. Topical and intermittent application of parathyroid hormone recovers alveolar bone loss in rat experimental periodontitis. J Periodont Res 2011; 46: 655–662. © 2011 John Wiley & Sons A/S Background and Objective: Periodontitis is characterized by periodontal tissue inflammation and alveolar bone loss. The intermittent administration of parathyroid hormone (PTH), a major regulator of bone remodeling, has been demonstrated to stimulate osteoblastic activity. Although the systemic administration of PTH has been reported to protect against periodontitis‐associated bone loss, the effect of the topical administration of PTH is unclear. In this study, the effect of intermittent administration of PTH on osteoblastic differentiation was examined in cultured calvaria cells and then the effect of topical and intermittent administration of PTH was determined by measuring the recovery of alveolar bone loss after inducing experimental periodontitis in rats. Material and Methods: Alkaline phosphatase activity and bone nodule formation were measured in fetal rat calvaria cells. Experimental periodontitis was induced by placing nylon ligature around rat maxillary molars for 20 d. After ligature removal (day 0), PTH was topically injected into buccal gingiva three times a week for 10 wk. Micro‐computed tomography analysis and histological examination were performed on days 35 and 70. Results: Intermittent exposure of PTH in calvaria cells increased alkaline phosphatase activity and bone nodule formation by 1.4‐ and 2.4‐fold, respectively. Ligature procedures induced marked alveolar bone loss around the molars on day 0 and greater bone recovery was observed in the PTH‐treated rats on day 70. An increase in osteoid formation on the surface of alveolar bone was detected in the PTH‐treated rats. Conclusion: Intermittent treatment with PTH stimulated osteoblastic differentiation in fetal rat calvaria cell cultures, and topical and intermittent administration of PTH recovered alveolar bone loss in rat experimental periodontitis.     

Effects of Polycan,a β‐glucan,on experimental periodontitis and alveolar bone loss in Sprague‐Dawley rats

Kim YS, Kang SJ, Kim JW, Cho HR, Moon SB, Kim KY, Lee HS, Han CH, Ku SK, Lee YJ. Effects of Polycan, a β‐glucan, on experimental periodontitis and alveolar bone loss in Sprague‐Dawley rats. J Periodont Res 2012; 47: 800–810. © 2012 John Wiley & Sons A/S Background and Objective: Polycan is a promising candidate for the treatment of periodontal disease. This study was undertaken to examine whether Polycan, a type of β‐glucan, has a protective effect on ligature‐induced experimental periodontitis and related alveolar bone loss in Sprague‐Dawley rats. Material and Methods: Polycan was orally administered, daily, for 10 d, at 21.25, 42.5 or 85 mg/kg, beginning 1 d after ligation. Changes in body weight and alveolar bone loss were monitored, and the anti‐inflammatory effects of Polycan were determined by measuring the levels of myeloperoxidase (MPO), interleukin‐1beta (IL‐1β) and tumor necrosis factor‐alpha (TNF‐α) in gingival tissue. We also evaluated inducible nitric oxide synthase (iNOS) activity and malondialdehyde (MDA) concentrations as a measure of the antioxidant effect. Results: Ligature placement led to a marked decrease in body weight, increased alveolar bone loss and increased concentrations of MPO, IL‐1β, TNF‐α and MDA, as well as increased iNOS activity and inflammatory cell infiltration and decreased collagen‐fiber content. Histological examination revealed increases in the number and activity of osteoclast cells, decreases in alveolar bone volume and elevated percentages of osteclasts on the alveolar bone surface. Daily oral treatment with 42.5 or 85 mg/kg of Polycan for 10 d led to significant, dose‐dependent inhibition of the effect of ligature placement. Conclusion: Taken together, these results suggest that 10 d of oral treatment with Polycan effectively inhibits ligature placement‐induced periodontitis and related alveolar bone loss via an antioxidant effect.     

Effects of Locally Administered Tiludronic Acid on Experimental Periodontitis in Rats

Background: 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. Methods: 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). Results: 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). Conclusion: 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.     

The influence of short-term diabetes mellitus and insulin therapy on alveolar bone loss in rats

BACKGROUND: It is well known that the multiple direct and indirect consequences of hyperglycemia in diabetic individuals have been linked to a number of abnormal host effector mechanisms that could lead to an increased risk of developing periodontal disease. OBJECTIVE: The aim of this study was to investigate the effect of short-term experimental diabetes and insulin therapy on the severity of alveolar bone loss in rats, and the effect of experimental periodontitis on glycemic control. METHODS: Seventy-two male Wistar rats were divided into four groups: group I animals were submitted to dental ligature around lower right first molars (ligated); group II consisted of streptozotocin (STZ)-diabetic, ligated rats; group III represented STZ-diabetic, unligated rats; and group IV consisted of insulin-treated (6 U/day), STZ-diabetic, ligated rats. Blood glucose of all diabetic rats was monitored at regular intervals. Standardized digital radiographs were taken after killing at 7, 15 and 30 days to measure the amount of bone loss about the mesial root surface of the first molar tooth in each rat. Results: No significant (p < 0.05) changes in plasma glucose levels of insulin-treated diabetic rats were found among the different examinations after the beginning of insulin therapy. Rats from group II showed significantly greater increases in mean plasma glucose levels at 15 and 30 days after ligature placement compared with rats from group III (p < 0.05). Furthermore, in spite of the significant alveolar bone loss progression that was observed in groups I, II and IV (p < 0.00001; two-way anova), no significant differences among these groups regarding the severity of bone loss (p = 0.77) and no significant interaction between treatment group and time (p = 0.81) were found. CONCLUSIONS: Within the limits of this study, it can be suggested that the severity of periodontal disease was not affected by short-term diabetes, and that experimental periodontitis increased blood glucose levels in uncontrolled diabetic rats.     

iNOS-derived nitric oxide stimulates osteoclast activity and alveolar bone loss in ligature-induced periodontitis in rats

Background: Inflammatory stimuli activate inducible nitric oxide synthase (iNOS) in a variety of cell types, including osteoclasts (OC) and osteoblasts, resulting in sustained NO production. In this study, we evaluate the alveolar bone loss in rats with periodontitis under long‐term iNOS inhibition, and the differentiation and activity of OC from iNOS‐knockout (KO) mice in vitro. Methods: Oral aminoguanidine (an iNOS inhibitor) or water treatment was started 2 weeks before induction of periodontitis. Rats were sacrificed 3, 7, or 14 days after ligature placement, and alveolar bone loss was evaluated. In vitro OC culture experiments were also performed to study the differentiation of freshly isolated bone marrow cells from both iNOS KO and wild‐type C57BL/6 mice. OC were counted 6 days later after tartrate‐resistant acid phosphatase staining (a marker of osteoclast identity), and bone resorption activity was assessed by counting the number of resorption pits on dentin disks. Results: Rats with ligature showed progressive and significant alveolar bone loss compared to sham animals, and aminoguanidine treatment significantly inhibited ligature‐induced bone loss at 7 and 14 days after the induction. In comparison to bone marrow cells from wild‐type mice, cells from iNOS KO mice showed decreased OC growth and the resulting OC covered a smaller culture dish area and generated fewer resorption pit counts. Conclusion: Our results demonstrate that iNOS inhibition prevents alveolar bone loss in a rat model of ligature‐induced periodontitis, thus confirming that iNOS‐derived NO plays a crucial role in the pathogenesis of periodontitis, probably by stimulating OC differentiation and activity.     

Involvement of toll‐like receptor 4 in alveolar bone loss and glucose homeostasis in experimental periodontitis

Watanabe K, Iizuka T, Adeleke A, Pham L, Shlimon AE, Yasin M, Horvath P, Unterman TG. Involvement of toll‐like receptor 4 in alveolar bone loss and glucose homeostasis in experimental periodontitis. J Periodont Res 2011; 46: 21–30. © 2010 John Wiley & Sons A/S Background and Objective: There is general agreement that certain fatty acids and lipopolysaccharides (LPS) promote inflammation through toll‐like receptor 4 (TLR4), and that inflammation promotes insulin resistance. We therefore hypothesized that mice with periodontitis and a TLR4 loss‐of‐function (LOF) mutation fed a high‐fat (HF) diet would develop improved glucose homeostasis compared with wild‐type (WT) animals with periodontitis fed a HF diet. Material and Methods: Wild‐type and TLR4 mutant mice fed a HF diet were divided into four groups (n = 6/group): WT; WT with periodontitis (WT/P); mutant (Mut); and mutant with periodontitis (Mut/P). Periodontitis was induced by placing LPS soaked ligatures around maxillary second molars. Fasting insulin and glucose levels were measured weekly for 10 wk. Glucose tolerance was evaluated at baseline (week 1) and at 9 wk. Insulin signaling (phosphorylation of Akt) and tumor necrosis factor‐α (TNF‐α) mRNA levels in liver were determined when the mice were killed at week 10. Results: Mut/P mice developed less alveolar bone loss compared with WT/P mice (p < 0.05). Fasting glucose levels were improved after 8 wk of feeding a HF diet (weeks 9 and 10) in Mut/P mice compared with Mut, WT and WT/P mice (p < 0.05). Glucose tolerance was impaired in all groups compared with baseline (p < 0.05), except for the Mut/P group. Insulin signaling was improved (p < 0.05), and expression of TNF‐α was decreased (p < 0.05) in the liver of Mut/P mice compared with the liver of WT/P mice. Conclusion: The TLR4 LOF mutation partially protects against alveolar bone loss and improves glucose homeostasis in mice with periodontitis fed a HF diet.     

Preventive effects of thymoquinone in a rat periodontitis model: a morphometric and histopathological study

Ozdemir H, Kara MI, Erciyas K, Ozer H, Ay S. Preventive effects of thymoquinone in a rat periodontitis model: a morphometric and histopathological study. J Periodont Res 2012; 47: 74–80. © 2011 John Wiley & Sons A/S Background and Objective: Thymoquinone has a variety of pharmacologic properties, including antihistaminic, antibacterial, antihypertensive, hypoglycemic, anti‐inflammatory and anti‐oxidative activities. Through its anti‐inflammatory and antioxidant properties, thymoquinone may play an important role in preventing periodontal diseases. The aim of this study was to evaluate the effectiveness of thymoquinone in preventing the initiation and progression of periodontitis in a rat periodontitis model. Material and Methods: Twenty‐four rats were randomly divided into three experimental groups: a nonligated (NL) treatment group (n = 8), a ligature‐only (LO) treatment group (n = 8) and a ligature plus thymoquinone (10 mg/kg, daily for 11 d) (TQ) treatment group. In order to induce experimental periodontitis, a 4/0 silk suture was placed at the gingival margin of the right‐mandibular first molars of the rats. Thymoquinone was administered by gastric feeding until the animals were killed on day 11. Changes in the alveolar bone levels of rats in each group were measured clinically, and tissues of rats in each group were examined histopathologically to determine inflammatory cell infiltration (ICI), osteoblast and osteoclast activities, and osteoclast morphology. Results: Alveolar bone loss around the mandibular molar tooth was significantly higher in the LO group compared with NL and TQ groups (p < 0.05). The ratio of the presence of ICI and osteoclast numbers was significantly higher in the LO group than in the NL and TQ groups (p < 0.05). Osteoblastic activity was significantly lower in the LO group than in the NL and TQ groups (p < 0.05). Conclusion: The present study showed that the oral administration of thymoquinone diminishes alveolar bone resorption in a rat periodontitis model.     

Effects of low-dose doxycycline and bisphosphonate clodronate on alveolar bone loss and gingival levels of matrix metalloproteinase-9 and interleukin-1β in rats with diabetes: a histomorphometric and immunohistochemical study

Background: Bisphosphonates (BPs) and low‐dose doxycycline (LDD) have been shown to inhibit bone resorption and to improve the levels of proinflammatory mediators and destructive enzymes in gingival tissues, respectively. The purpose of this study is to evaluate the effect of mono and combined BP clodronate and LDD therapies in reducing gingival levels of matrix metalloproteinase‐9 (MMP‐9), interleukin‐1β (IL‐1β), and alveolar bone loss in rats with diabetes. Methods: Fifty adult Wistar rats were divided into five study groups as follows: 1) group 1 = diabetes control; 2) group 2 = diabetes + periodontitis; 3) group 3 = diabetes + periodontitis + LDD; 4) group 4 = diabetes + periodontitis + clodronate; and 5) group 5 = diabetes + periodontitis + LDD + clodronate. LDD and clodronate were given as a single agent or as combination therapy during the 7 days of the post‐experimental periodontitis period. On day 7, the rats were sacrificed, the mobility of the tooth was recorded, and block biopsies were removed. The gingival tissues were analyzed histologically and immunohistochemically for expression of MMP‐9 and IL‐1β. Alveolar bone loss was evaluated morphometrically under a light microscope. Data analysis was performed statistically by Kruskal‐Wallis and post hoc Tukey and Spearman correlation tests. Results: Alveolar bone loss was significantly greater in groups 2 through 5 than group 1 (P <0.05) but was not significantly different among groups 2 through 5 (P >0.05). Animals with periodontitis (group 2) expressed significantly higher levels of MMP‐9 and IL‐1β compared with those without periodontitis (group 1) (P <0.05). MMP‐9 expression was significantly lower in group 3 than groups 1, 2, and 5 (P <0.05). IL‐1β expression was significantly lower in the groups 1, 3, 4, and 5 than 2 (P <0.01) but was not significantly different among groups 1, 3, 4, and 5. Positive correlations were found between alveolar bone loss and density of inflammation (ρ = 0.319, P = 0.021) and between MMP‐9 and IL‐1β (ρ = 0.418, P = 0.002), respectively. Conclusion: Our findings suggest that ligature‐induced periodontitis in animals with diabetes results in significantly higher levels of MMP‐9 and IL‐1β expression in gingiva. The use of mono and combined clodronate and LDD administrations may significantly reduce levels of MMP‐9 and IL‐1β expression. However, drug administration did not affect alveolar bone levels during the study period.     

Curcumin inhibits inflammatory response and bone loss during experimental periodontitis in rats

Abstract

Objective. Curcumin, an active ingredient of turmeric, is proved to be a potential candidate of controlling inflammation and bone resorption, but few reports are on the periodontitis. The purpose of this study was to evaluate whether the intra-gastric administration of curcumin could inhibit the in?ammation and alveolar bone resorption in rats following ligature-induced experimental periodontitis. Materials and method. Male Wistar rats were randomly divided into three groups: no ligature placement and administration of vehicle, ligature placement and administration of vehicle, ligature placement and administration of curcumin. After the animals were sacrificed, their mandibles were collected for morphological, histological and immunohistochemical analysis; their gingival tissues were collected for cytokine measurements. Results. Bone resorption was significantly higher in the experimental periodontitis animals treated with vehicle compared with the curcumin-treated group or the control group. Furthermore, receptor activator of nuclear factor-κB ligand (RANKL), receptor activator of nuclear factor-κB (RANK), osteoprotegerin (OPG), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) expression levels were higher in the experimental periodontitis animals treated with vehicle compared with the curcumin treated group or the control group. Conclusions. Curcumin may decrease alveolar bone loss in the experimental periodontitis rats via suppressing the expression of RANKL/RANK/OPG and its anti-inflammatory properties.     

Effects of Estrogen Deficiency and/or Caffeine Intake on Alveolar Bone Loss,Density, and Healing: A Study in Rats

Background: The aim of this study is to evaluate the effects of caffeine and/or estrogen deficiency on ligature‐induced bone loss (BL), trabecular bone area (TBA), and postextraction bone healing (BH). Methods: Rats were assigned into one of the following groups (15 each): 1) control = non‐ingestion of caffeine/sham surgery; 2) caffeine = ingestion of caffeine/sham surgery); 3) ovariectomized (OVX) = non‐ingestion of caffeine/ovariectomy; or 4) caffeine/OVX = ingestion of caffeine/ovariectomy. The rats were under caffeine administration for 65 days and/or estrogen deficiency for 51 days. On day 21 after ovariectomy, one first mandibular molar received a ligature and the contralateral tooth was not ligated. The first maxillary molars were extracted 8 days before sacrifice. BL, TBA, the positive cells for tartrate‐resistant acid phosphatase (TRAP), receptor activator of nuclear factor‐κB ligand (RANKL), and osteoprotegerin (OPG) were analyzed in the furcation area of mandibular molars. Histometric BH and gene expression of bone morphogenetic protein (BMP)‐2, BMP‐7, osteopontin, and bone sialoprotein were evaluated in alveolar sockets. Results: The caffeine group presented the greatest BL and the OVX group the highest number of TRAP‐positive (TRAP⁺) cells around ligated teeth (P <0.05). The control group presented higher TBA and BH than the other groups (P <0.05). All test groups presented higher RANKL/OPG⁺ cells than the control group around ligated/unligated teeth. The OVX and caffeine/OVX groups presented a greater number of TRAP⁺ cells around unligated teeth than the control group (P <0.05). There were no differences among groups for gene expression (P >0.05). Conclusions: Caffeine increased BL in ligated teeth. Caffeine and/or estrogen deficiency decreased TBA in the unligated teeth and reduced BH after tooth extraction.     

Impact of cannabis sativa (marijuana) smoke on alveolar bone loss: a histometric study in rats

Background: Cannabis sativa (marijuana) can interfere with bone physiopathology because of its effect on osteoblast and osteoclast activity. However, its impact on periodontal tissues is still controversial. The present study evaluates whether marijuana smoke affects bone loss (BL) on ligature‐induced periodontitis in rats. Methods: Thirty male Wistar rats were used in the study. A ligature was placed around one of the mandible first molars (ligated teeth) of each animal, and they were then randomly assigned to one of two groups: control (n = 15) or marijuana smoke inhalation ([MSI] for 8 minutes per day; n = 15). Urine samples were obtained to detect the presence of tetrahydrocannabinol. After 30 days, the animals were sacrificed and decalcified sections of the furcation area were obtained and evaluated according to the following histometric parameters: bone area (BA), bone density (BD), and BL. Results: Tetrahydrocannabinol was positive in urine samples only for the rats of the MSI group. Non‐significant differences were observed for unligated teeth from both groups regarding BL, BA, and BD (P >0.05). However, intragroup analysis showed that all ligated teeth presented BL and a lower BA and BD compared to unligated teeth (P <0.05). The intergroup evaluation of the ligated teeth showed that the MSI group presented higher BL and lower BD (P <0.05) compared to ligated teeth from the control group. Conclusion: Considering the limitations of this animal study, cannabis smoke may impact alveolar bone by increasing BL resulting from ligature‐induced periodontitis.     

Therapeutic Effects of Melatonin on Alveolar Bone Resorption After Experimental Periodontitis in Rats: A Biochemical and Immunohistochemical Study

Background: The present study aims to investigate the effects of systemic melatonin administration on alveolar bone resorption in experimental periodontitis in rats. Methods: Twenty‐four male Sprague‐Dawley rats were divided into three groups (control, experimental periodontitis [Ped], and experimental periodontitis treated with melatonin [Mel‐Ped]). For periodontitis induction, first molars were ligatured submarginally for 4 weeks. After ligature removal, rats in the Mel‐Ped group were treated with a daily single dose of 10 mg/kg body weight melatonin for 15 consecutive days. At the end of the study, intracardiac blood samples and mandible tissues were obtained for histologic, biochemical, and radiographic analysis. Serum markers related to bone turnover, calcium, phosphorus, bone alkaline phosphatase (b‐ALP), and terminal C telopeptide of collagen Type I (CTX) were analyzed. Myeloperoxidase levels were determined in gingival tissue homogenates, and receptor activator of nuclear factor‐kappa B ligand (RANKL) activation was analyzed in the mandible samples stereologically. Alveolar bone loss was also evaluated radiographically in the mandible samples of each group. Results: Melatonin treatment decreased serum CTX levels and increased b‐ALP levels. Serum calcium and phosphorus levels were not statistically different among groups (P >0.05). Alveolar bone resorption and myeloperoxidase activity were statistically higher in the Ped group compared to the Mel‐Ped group (P <0.05). Immunohistochemical staining of RANKL and osteoclast activity were significantly lower in the Mel‐Ped group compared to the Ped group (P <0.05). Conclusion: This study reveals that melatonin treatment significantly inhibits regional alveolar bone resorption and contributes to periodontal healing in an experimental periodontitis rat model.     

Drinking green tea alleviates alveolar bone resorption in ligature-induced periodontitis in mice

ObjectivesIt has been reported that green tea exerts antibacterial, anti-inflammatory, and antioxidant effects. The purpose of the present study was to evaluate the effects of drinking green tea on bone resorption in ligature-induced periodontitis in mice.MethodsSixty C57BL/6 eight-week-old male mice were used. To induce periodontitis, a ligature was placed for 7 days around the upper left second maxillary molar. After ligature removal, the animals were administered different concentrations of green tea (1.5 g/60 mL, 3 g/60 mL, or 6 g/60 mL) or distilled water. At 1 and 2 weeks of administration, the animals were sacrificed and micro-CT images of the maxillae were taken. Next, the depth and area of alveolar bone loss in the buccal and palatal sides were measured. The number of inflammatory cells and osteoclasts in histological sections were counted.ResultsThe result showed ligature-induced alveolar bone loss. Green tea inhibited ligature-induced bone loss in the buccal side in a dose-dependent manner. Histologically, ligature increased the number of inflammatory cells and osteoclasts, but this effect was alleviated by green tea.ConclusionsEvidence from this animal experiment suggested that drinking green tea would be potentially beneficial to reduce alveolar bone loss in ligature-induced periodontitis.     

Effects of Probiotic Therapy on Metabolic and Inflammatory Parameters of Rats With Ligature‐Induced Periodontitis Associated With Restraint Stress

Background: This study evaluates the effects of probiotic therapy (PT) in rats with ligature‐induced periodontitis associated with restraint stress. Methods: Sixty‐four rats were divided into control, stress (STR), probiotic (PROB), periodontal disease (PD), STR‐PROB, STR‐PD, STR‐PROB‐PD, and PROB‐PD groups. The probiotic was added to the drinking water for 44 days. PD was induced by a ligature. In STR groups, the animals were subjected to restraint stress for 2.5 hours per day for 30 days. Results: Rats with PD exhibited increased alveolar bone loss (P <0.05), as well as increased levels of cyclooxygenase‐2, serum C‐terminal telopeptide (CTX), p38 mitogen‐activated protein kinase (p38), and receptor activator of nuclear factor‐κB ligand and decreased levels of osteoprotegerin (OPG). Stressed rats presented high levels of C‐peptide, corticosterone, and glucose (P <0.05). In general, the presence of stress reduced the expression of CTX and p38 (P <0.05). PT reduced alveolar bone loss in unstressed animals. It also decreased expression of CTX and induced increased expression of OPG in unstressed animals with PD. However, PT was not effective in preventing bone loss or altering the expression of inflammatory markers in stressed animals. PT decreased the number of inflammatory cells in the periodontal tissue (P <0.05). Groups with stress and PD showed decreased villous height and crypt depth. Stress seemed to prevent part of the probiotic beneficial effects on the small intestine. Conclusions: Based on the methodology used, PT may reduce tissue breakdown resulting from PD in unstressed rats. The protocol used for restraint stress influenced the immunomodulatory effects of PT in intestinal and periodontal tissues.     

2种给药方式下辛伐他汀对牙周炎骨质疏松大鼠上颌骨骨丢失的影响

目的比较辛伐他汀(Simvastatin,SIM)在不同给药方式下抑制牙周炎骨质疏松大鼠上颌骨骨丢失的作用效果。方法 24只4月龄雌性SD(Sprague Dawley)大鼠,随机分成4组:假手术组(SHAM),去势+绑线组(OVX+LIG),去势+绑线+辛伐他汀灌胃给药组(OVX+LIG+Oral SIM),去势+绑线+辛伐他汀局部用药组(OVX+LIG+Local SIM)。适应性喂养1周后,进行骨质疏松造模手术——双侧卵巢切除术(OVX),4周后,进行牙周炎造模手术——分别于双侧上颌骨第一、第二磨牙进行8字结扎绑线术(LIG),4周后拆除绑线,用药组开始辛伐他汀给药。8周后处死所有大鼠,收集双侧上颌骨和血清,分别进行进一步检测。检测包括micro-CT扫描,硬组织切片观察、染色,酶联免疫吸附实验(ELISA)。结果相比于对照OVX+LIG组,OVX+LIG+Local SIM组可见明显釉牙骨质界-牙槽嵴顶(CEJ-ABC)距离的减少(P<0.05),OVX+LIG+Oral SIM组与OVX+LIG+Local SIM组可见明显牙槽骨的骨密度(BMD)和骨体积分数(BVF)升高,OVX+LIG+Oral SIM组可见明显骨钙素(OC)上升,抗酒石酸酸性磷酸酶5b(TRAP5b)下降。结论局部或全身系统应用辛伐他汀能减缓牙周炎伴骨质疏松大鼠上颌骨的骨丢失,其中局部注射辛伐他汀对牙槽嵴顶骨形成具有促进作用。     

Divergence of the systemic immune response following oral infection with distinct strains of Porphyromonas gingivalis

Periodontitis is a polymicrobial oral infection characterized by the destruction of tooth‐supporting structures that can be linked to systemic diseases such as cardiovascular disease, diabetes or rheumatoid arthritis. Porphyromonas gingivalis, a bacterium implicated in the etiology of periodontitis, has shown variation in inducing T‐cell responses among different strains. Therefore, in this study we investigated the strain‐specific immune response using a murine experimental model of periodontitis. Periodontitis was induced by P. gingivalis strains A7A1‐28, W83 and W50, and later confirmed by the presence of P. gingivalis in the oral microflora and by alveolar bone resorption. Splenocytes were evaluated for gene expression, cellular proteins and cytokine expression. Dendritic cells were stimulated in vitro for T helper cell–cytokine profiling. Results showed that P. gingivalis had the ability to alter the systemic immune response after bacterial exposure. Strains W50 and W83 were shown to induce alveolar bone loss, whereas the A7A1‐28 strain did not significantly promote bone resorption in mice. Splenocytes derived from mice infected with strains W50 and W83 induced expression of high levels of interleukin‐4 (IL‐4) but A7A1‐28 stimulated increased IL‐10. Stimulation of dendritic cells in vitro showed a similar pattern of cytokine expression of IL‐12p40, IL‐6 and transforming growth factor‐β among strains. A distinct systemic response in vivo was observed among different strains of P. gingivalis, with IL‐10 associated with the least amount of alveolar bone loss. Evaluation of pathogen‐driven systemic immune responses associated with periodontal disease pathogenesis may assist in defining how periodontitis may impact other diseases.     

Exenatide and Sitagliptin Decrease Interleukin 1β, Matrix Metalloproteinase 9, and Nitric Oxide Synthase 2 Gene Expression But Does Not Reduce Alveolar Bone Loss in Rats With Periodontitis

Background: New drugs for the treatment of diabetes, glucagon‐like peptide‐1 (GLP‐1) receptor agonists and inhibitors of dipeptidyl peptidase‐4 (DPP‐4) have shown pleiotropic effects on bone metabolism and anti‐inflammatory properties. The aim of this study is to evaluate the effects of exenatide (GLP‐1 agonist) and sitagliptin (DPP‐4 inhibitor) during periodontitis induction by ligature insertion in rats. Methods: Forty rats were divided into four groups: 1) animals with induced periodontitis that received exenatide (EG); 2) animals with induced periodontitis that received sitagliptin (SG); 3) animals with induced periodontitis and without drug treatment (LG); and 4) animals without induced periodontitis and without drug treatment (controls). The drugs were administered for 28 days. On the day the animals were sacrificed, blood was collected for analysis of glucose and DPP‐4 levels. The gene expressions of prostaglandin‐endoperoxide synthase 2, tissue inhibitor of metalloproteinase 1, Dpp4, nitric oxide synthase 2 (Nos2), interleukin 1β (Il1b), and matrix metalloproteinase 9 (Mmp9) in the gingiva; support and alveolar bone loss; connective tissue attachment; and the quantity of gingival collagen were evaluated. Results: Exenatide and sitagliptin treatments have led to a lower percentage of weight gain but did not influence glycemia. Sitagliptin reduced the serum concentration of DPP‐4. Interestingly, although the gene expression profile has revealed a downregulation of Mmp9, Nos2, and Il1b in both EG and SG compared to LG, a significant protective effect was not observed on alveolar bone and collagen tissue in this model. Conclusion: Regardless of the reduction of the expression of Il1b, Nos2, and Mmp9, the drugs were not effective in the stabilization or reduction of alveolar bone loss and collagen degradation in rats.