实验性糖尿病牙周炎骨丧失动物模型研究

目的:建立实验性糖尿病牙周炎骨丧失动物模型,以进一步揭示糖尿病加重牙周炎骨丧失的细胞学机制.方法:选用6周龄雄性SD大鼠62只,随机分为糖尿病牙周炎组(DP)、牙周炎组(P)以及正常对照组(N).采用一次性腹腔注射链脲佐菌素(streptozotocin,STZ)的方法诱导大鼠糖尿病模型,采用丝线结扎联合口内接种细菌的方法建立牙周炎模型.动物分别于丝线结扎后3周和6周分批处死,进行HE染色、TRAP染色.观测指标包括:牙槽骨丧失,组织病理学比较,炎症区破骨细胞计数等.资料采用单因素方差分析统计学处理.结果:丝线结扎后3周和6周,大鼠牙槽骨丧失在N组与P组、N组与DP组、P组与DP组不同,组间两两比较均有统计学差异(P＜0.05),牙槽骨丧失DP组＞P组＞N组.炎症区单位长度破骨细胞数在N组、P组、DP组不同,N组与P组比较,N组与DP组比较、P组与DP组比较均有统计学差异(P＜0.05),其炎症区单位长度破骨细胞数DP组＞P组＞N组.结论:糖尿病可加重牙周炎牙周组织破坏,糖尿病条件下牙周炎骨丧失明显增加.糖尿病可能通过促进炎症部位破骨细胞生成,增强骨吸收,促进牙周炎骨丧失.     

实验性糖尿病牙周炎诱导骨细胞凋亡的初步研究

目的初步探讨糖尿病牙周炎条件下骨细胞的凋亡情况。方法选用6wk雄性SD大鼠62只,随机分为糖尿病牙周炎组（DP,n=22）、牙周炎（P,n=20）以及正常对照组（N,n=20）。采用一次性腹腔注射STZ（55mg/kg）的方法建立大鼠糖尿病模型,注射STZ后1wk检测血糖,血糖≥16.65mmol/L者定为糖尿病大鼠。采用丝线结扎大鼠上颌第二磨牙联合口内接种细菌的方法建立牙周炎模型。动物分别于丝线结扎后3wk和6wk分批处死,进行HE染色和原位细胞凋亡检测。观察指标包括：牙槽骨丧失（ABL）,骨细胞计数,骨细胞凋亡百分率。资料采用单因素方差分析统计学处理。结果丝线结扎后3周和6周,大鼠牙槽骨丧失在N组与P组、N组与DP组、P组与DP组不同,组间两两比较均有统计学意义（P〈0.05）,牙槽骨丧DP组〉P组〉N组。与N组比较,P组和DP组单位面积骨细胞数均减少,与P组比较,DP组单位面积骨细胞数亦显著减少（P〈0.05）。在丝线结扎后3周和6周,糖尿病牙周炎组（DP）骨细胞凋亡百分率均达到牙周炎组（P）的2倍左右。结论糖尿病条件下牙周炎骨丧失明显增加,糖尿病可加强牙周炎条件下牙周组织中骨细胞的凋亡,降低骨细胞的数量。     

应激对豚鼠牙周炎模型的影响

目的探讨应激对豚鼠牙周炎模型的影响。方法龈沟接种伴放线放线杆菌和牙龈卟啉单胞菌,制备豚鼠牙周炎模型24只,随机分为应激组12只(冷刺激和惊吓刺激)和对照组12只(无刺激),于1、2、4、6周后分批处死,进行临床参数、病理切片、破骨细胞和成骨细胞计数以及血清皮质醇浓度检测。结果应激组第1、2、4周皮质醇浓度升高,第2、4周牙周袋深度与对照组比较有明显差异(P<0.05或P <0.01)。病理切片显示:与对照组相比,应激组牙周破坏更明显,骨修复不活跃,应激组第1、4、6周破骨细胞计数明显增高(P<0.05)。结论应激加重致病菌感染后牙周组织的破坏,延缓组织修复,是牙周炎的重要危险因子。     

生命早期负性应激对大鼠实验性牙周炎的影响

目的:探究生命早期负性应激对大鼠成年后实验性牙周炎进展的影响。方法:在3周龄雄性SD大鼠建立慢性温和不可预知应激(unpredictable chronic mild stress, UCMS)模型,以正常饲养大鼠为对照。应激5周后对所有大鼠右上第二磨牙进行实验性牙周炎处理。通过行为学实验评估大鼠情绪状态;ELISA检测血清皮质酮、IL-1β和TNF-α水平;HE染色观察破骨细胞数目,测量牙槽骨丧失量。结果:UCMS大鼠出现糖水偏好下降(P<0.001)、强迫游泳实验中不动时间增加(P<0.01)等抑郁样行为;血清皮质酮及炎性细胞因子水平升高(P<0.05)。HE染色可见UCMS大鼠牙周炎侧牙槽骨吸收量最大,其次为对照组牙周炎侧,再次为对照组正常侧(P<0.001);破骨细胞数目变化趋势与之类似(P<0.001)。血清皮质酮含量与牙周破坏程度呈正相关关系(P<0.05)。结论:生命早期负性应激可能通过HPA轴增强体液免疫,加剧大鼠实验性牙周炎病变程度。     

Toll样受体⁃4抑制剂TAK⁃242对大鼠重度牙周炎骨吸收的影响

目的探讨Toll样受体?4(Toll like receptor?4,TLR?4)抑制剂TAK?242对大鼠重度牙周炎骨质吸收的影响,为重度牙周炎寻找辅助治疗手段提供实验基础。方法18只3周龄雄性Wistar大鼠随机分为3组(n=6),其中1组为正常对照组,另外2组以含有牙龈卟啉单胞菌(P.gingivalis)ATCC33277的5?0丝线结扎大鼠双侧上颌磨牙行重度牙周炎建模,分为牙周炎组、TAK?242组;TAK?242组从丝线结扎第1天起,通过尾静脉隔天注射1次溶于DMSO的TAK?242(2 mg/kg),另外两组注射相同体质量比例的DMSO溶剂,连续8周;第8周末处死3组大鼠,获取大鼠上颌骨标本,采用micro?CT扫描后三维重建,测量特定位点釉牙骨质界?牙槽嵴顶的距离评估骨丧失量,并对牙槽骨骨质相关参数和骨质微结构进行分析;组织学切片苏木精?伊(HE)染色观察牙周组织病理改变;甲基绿染色观察牙槽骨吸收情况;抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞分布情况。结果Micro?CT定量分析显示:牙周炎组与TAK?242组牙槽骨吸收显著高于对照组;与牙周炎组相比,TAK?242组大鼠上颌第一磨牙近、远中根吸收位点的骨丧失均显著减轻(P<0.001),骨密度(P<0.05)与骨体积/总体积分数(P<0.01)显著增高,骨小梁数目与骨小梁厚度(P<0.01)相对增多,骨小梁分离度(P<0.01)和骨小梁结构模式指数显著降低。牙周炎组骨质呈现疏松多孔的蜂窝状结构,骨小梁结构恶化,向杆状结构转变;TAK?242组骨质微结构改善,骨量改善,骨小梁分布相对更致密,骨小梁结构与对照组更相似。HE染色发现牙周炎组与TAK?242组牙周附着丧失与牙槽骨吸收较对照组显著;与牙周炎组相比,甲基绿染色表明TAK?242组骨吸收减轻,TRAP染色显示破骨细胞浸润减少(P<0.001)。结论TLR?4抑制剂TAK?242能缓解大鼠重度牙周炎骨吸收,改善其多孔、稀疏、排列紊乱的炎症性骨小梁结构。     

人骨保护素质粒转染抑制大鼠实验性牙周炎骨吸收的研究

目的:将自行构建的质粒载体pcDNA3.1-h OPG,通过体内转染,评价OPG直接基因转染疗法对大鼠实验性牙周炎牙槽骨吸收的影响,为牙周炎以及种植体周炎的生物治疗提供实验依据.方法:将30只SD大鼠随机分为3组,即I组生理盐水组(n=10,100μg/只)、Ⅱ组pcDNA3.1(-)组(n=10,100μg/只)、Ⅲ组pcDNA3.1-hOPG组(n=10,100μg/只).通过丝线结扎、接种牙周炎可疑致病菌、喂高糖软食诱发实验性牙周炎.结扎28d后处死,通过大体标本、组织学等观察牙槽骨吸收、OPG及破骨细胞变化.结果:Ⅲ组结扎侧OPG表达强度增加,牙槽骨吸收量减少(P＜0.05),活化破骨细胞数降低(P＜0.05).结论:OPG重组质粒转染,减少破骨细胞数量,有效减缓实验性牙周炎引起的牙槽骨吸收破坏.     

云南白药对实验性牙周炎的影响

目的:观察云南白药对大鼠实验性牙周炎的影响并探讨其治疗牙周炎的机制.方法:6周龄雄性SD大鼠30只,建立牙周炎模型,随机分为用药组(A组)和对照组(B组),每组15只.A组:云南白药治疗组;B组:牙周炎对照组,均于第2、4、6周处死.取上颌骨标本,测量探诊深度值、牙槽骨丧失值进行统计学分析,作切片光镜观察牙周组织病理学变化.结果:A组在灌药第4周和第6周时牙周探诊深度小于B组(P<0.05);在第6周时牙槽骨丧失少于B组(P<0.05)并且牙周膜纤维排列较B组整齐,牙槽嵴顶有新骨形成.结论:云南白药可减轻大鼠牙周组织炎症、抑制牙槽骨吸收并促进新骨形成,具有治疗牙周炎的作用.     

三种中药提取物抑制牙周炎牙槽骨吸收的实验研究

目的:观察三种中药提取物淫羊藿苷、大黄素和黄芩苷联合应用对实验性牙周炎形成的影响。方法:将3 6只SD大鼠随机分为三组:正常对照组(N)、空白对照组(C)和实验组(E)。用大肠杆菌内毒素脂多糖(E LPS)牙龈局部注射法建立大鼠实验性牙周炎模型,E组造模期间在相同部位注射三种中药提取物混合液,C组注射生理盐水。光学显微镜下观察牙周组织病理学变化,并进行破骨细胞计数,测量釉牙本质界到牙槽嵴顶的距离。结果:E组大鼠牙周组织炎症反应明显轻于C组,破骨细胞数少(P <0 .0 1) ,牙槽骨吸收量小(P <0 .0 5 )。结论:淫羊藿苷、大黄素和黄芩苷联合应用可抑制实验性牙周炎牙槽骨吸收。     

亚抗菌剂量的米诺环素对大鼠实验性牙周炎治疗作用的初步观察

目的:初步观察亚抗菌剂量的米诺环素在大鼠实验性牙周炎中的治疗作用。方法:由24只成年雄性SD大鼠组成3个实验组:1组是模型组:仅诱导牙周炎;2组是治疗组:诱导牙周炎且用亚抗菌剂量的米诺环素治疗;3组是阴性对照组:假手术(仅腹腔麻醉)。牙周炎的诱导采用丝线结扎法并辅以100g/L蔗糖水为饮料。动物于实验的28d和58d处死,观察指标为:(1)视觉指标:牙松动度(MT),牙龈指数(GI)和牙槽骨丧失(ABL);(2)组织学指标:牙周组织中单核细胞的渗出数、破骨细胞数和牙周组织胶原的含量。资料采用方差分析统计学处理。结果:与模型组相比,28d和56d指标均显示亚抗菌剂量的米诺环素能显著抑制GI,MT,ABL和破骨细胞的形成,56d指标表明亚抗菌剂量的米诺环素能显著抑制牙周组织中单核细胞的渗出和胶原组织的降解。结论:亚抗菌剂量的米诺环素能有效抑制牙槽骨的吸收和牙周胶原纤维的降解,减缓大鼠实验性牙周炎的发展进程。     

大黄素治疗实验性牙周炎的骨计量学研究

目的:观察不同浓度大黄素对实验性牙周炎大鼠附着丧失和牙槽骨吸收的治疗作用.方法:选取纯种雌性SD大鼠随机分为4 组,各30 只: 正常对照组(N组)、牙周炎组(P组)、低浓度大黄素治疗组(PL组)及高浓度大黄素治疗组(PH组).建立牙周炎动物模型并按分组用药,分别于4、 6、 8 周时处死动物,取上颌骨标本进行骨计量学观察.结果:PL组和PH组结缔组织附着丧失量、牙槽骨嵴高度丧失量和破骨细胞个数均明显小于P组,成骨细胞个数均明显大于P组(P<0.05).结论:大黄素可抑制牙周附着丧失和牙槽骨吸收,并能促进牙槽骨形成.     

雌激素缺乏对大鼠牙槽骨吸收影响的实验研究

目的观察雌激素缺乏对大鼠牙槽骨吸收的影响。方法34只雌性SD大鼠随机分为4组。第1组假手术(n=8),第2组卵巢切除(n=9),第3组卵巢切除加牙周结扎(n=9),第4组卵巢切除、牙周结扎加雌激素治疗(n=8)。适应性喂养7天后行假手术或双侧卵巢切除术。第4组于术后第二天起皮下注射苯甲酸雌二醇．20μg／kg体重／次,三天一次。第3、4两组于卵巢切除术后28天,结扎丝结扎上颌第一磨牙诱导牙周炎。第63天处死全部大鼠。常规取材。观察牙用组织组织学改变。测量牙用骨丧失值(PBL)。比较牙用骨支持率(PBS)。检测血清碱性磷酸酶(ALP)。结果采用成组f检验,第1、2两组的PBL分别为0．398±O．147mm,0．663±0．132哪。PBS分别为O．588±O．058。0．440±0．197,组间差异均有统计学意义(P<0．05);第2、3两组的PBL、PBS组间差异均有统计学意义(P<0．05)。第3组的PBL为0．875±0．197mm,PBS为0．336±O．087;第3、4两组的PBL、PBS组间差别没有统计学意义(P>0．05),第4组的PBL为O．823±0．119mm,PBS为0．360±0．950。结论雌激素缺乏促进牙槽骨吸收,茵斑刺激加剧骨质疏松大鼠牙槽骨的吸收,雌激素替代治疗不能预防骨质疏松大鼠因茵斑刺激引发的牙槽骨吸收。     

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.     

The effect of indomethacin on alveolar bone loss in experimental periodontitis

This study was undertaken to determine if prostaglandins play a role in the events leading to loss of bone in the ligature model of periodontitis. Periodontitis was induced by placement of the ligatures around mandibular teeth on one side of the jaw of squirrel monkeys ( Saimiri sciureus ). From one day prior to ligature placement, half the animals were administered indomethacin (5 mg/kg/day), a potent inhibitor of prostaglandin synthesis. Animals were sacrificed after one and two weeks of experimental periodontitis. It was found that indomethacin treatment abolished the significant losses of alveolar bone height and bone mass seen in non-indomethacin-treated (NIT) animals following ligature placement. Indomethacin also depressed the large increase in osteoclast density measured at one week in the NIT animals. The results support the hypothesis that prostaglandins are an important mediator of bone loss in the ligature model of periodontitis. Evidence is also presented for the coupling of bone resorption with osteoblastic neo-osteogenesis on both periodontal ligament and endosteal bone surfaces.     

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.     

补肾坚骨汤对牙周炎大鼠细胞因子及牙周组织影响的研究

目的：通过检测牙周炎大鼠血清中白细胞介素-4(interleukin-4,IL-4)、白细胞介素-6(interleukin-6, IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)水平的变化,及牙周组织的形态改变,初步探讨补肾坚骨汤对牙周炎大鼠的治疗作用。方法雄性12月龄SD大鼠45只,其中8只为正常对照组,37只采用丝线结扎8周,建立大鼠牙周炎模型后,将建模成功大鼠(34只)随机分为牙周炎模型对照组10只,补肾坚骨汤高剂量组(16g/kg/d)8只,补肾坚骨汤中剂量组(8g/kg/d)8只,和补肾坚骨汤低剂量组(4g/kg/d)8只,给予补肾坚骨汤灌胃4周。各组大鼠末次给药后24 h处死,采用ELISA法检测外周血IL-4、IL-6和TNF-α的水平;取上颌骨制作牙体牙周组织联合切片,HE染色,光学显微镜下观察牙周的组织学改变。结果牙周炎模型对照组血清IL-6、TNF-α水平显著高于正常对照组,IL-4水平显著低于正常对照组。补肾坚骨汤各剂量治疗组血清IL-6、TNF-α水平显著低于牙周炎模型对照组,补肾坚骨汤高、中剂量治疗组血清IL-4水平显著高于牙周炎模型对照组(p<0．05)。牙周炎模型对照组牙周组织表现为明显的炎症破坏,牙槽骨吸收明显,而补肾坚骨汤高、中剂量治疗组牙周组织炎症均明显减轻,牙槽骨吸收减少且骨修复反应明显。结论补肾坚骨汤可能通过降低牙周炎大鼠血清中IL-6和TNF-α水平,升高IL-4水平,减轻牙周组织炎症,减缓牙周组织破坏。     

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.     

实验性牙周炎动物模型研究

目的：建立一种近似人类临床牙周炎动物模型。方法：将大鼠随机分成对照组及实验组，并分别给予生理盐水肌肉注射、左上颌第2磨牙钢丝结扎和醋酸泼尼松龙肌肉注射。于实验第6周及第8周分2批处死大鼠。结果：实验组动物激素注射第4天后便出现了进食减少、倦怠少动等表现，符合中医肾虚的症状，并且出现了牙槽骨疏松、牙槽嵴吸收、牙周袋形成及破骨细胞活跃等病理性改变。对照组大鼠则无上述表现。结论：牙间结扎加糖皮质激素注射的方法可成功地建立近似于人类临床牙周炎的动物模型，为研究牙周炎提供了很好的方法。     

补肾坚骨汤对牙周炎大鼠血清白细胞介素-6和骨钙素影响的实验研究

目的：通过检测牙周炎大鼠血清中白细胞介素-6（interleukin-6/IL-6）和骨钙素（O steocalcin/OC）水平的变化,观察光学显微镜下牙周组织的形态改变,探讨补肾坚骨汤对牙周炎大鼠的治疗作用及可能机制。方法：雄性12月龄SD大鼠24只,8只为正常对照组（N）,其余成功建立大鼠牙周炎模型之后,分为牙周炎模型对照组（D）、补肾坚骨汤治疗组16g/（kg·d）（H）,给予补肾坚骨汤灌胃4周,每日一次,每次5m L,各组大鼠末次给药后24 h处死,摘取上颌骨制作牙体牙周组织联合切片,H E染色,在光学显微镜下观察牙周的组织学改变;采用ELISA法检测外周血IL-6和OC的水平。结果：牙周炎模型对照组牙周组织表现为明显的炎症破坏,牙槽骨吸收明显,而补肾坚骨汤治疗组牙周组织炎症明显减轻,牙槽骨吸收减少且骨修复反应明显;牙周炎模型对照组血清IL-6水平显著高于正常对照组和补肾坚骨汤治疗组（P〈0.05）;牙周炎模型对照组血清OC水平显著低于正常对照组和补肾坚骨汤治疗组（P〈0.05）。结论：补肾坚骨汤可能通过降低牙周炎大鼠血清中IL-6水平,升高OC水平,减轻牙周组织炎症,促进牙周组织再生,其作用机制还需进一步深入研究。