Yu Xiao-Fei, Wang Xin-Yang, Wei Peng-Gong, Li Bin, Chen Yao-Zhong
Department of Stomatology, Affiliated Hospital of Southeast University. Nanjing 210029, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2023 Apr;32(2):137-142.
To investigate the effects of chitosan oligosaccharide on bone metabolism and IKK/NF-κB pathway in mice with osteoporosis and periodontitis.
Thirty rats were randomly divided into 3 groups, with 10 rats in each group. They were divided into control group, ovariectomized periodontitis group and chitosan oligosaccharide treatment group. Except for the control group, the other two groups were ovariectomized and smeared with Porphyromonas gingivalis fluid to establish the model of osteoporosis with periodontitis. Four weeks after ligation, the rats in chitosan oligosaccharide treatment group were gavaged with 200 mg/kg chitosan oligosaccharide, and the other two groups were gavaged with equal volume of normal saline once a day for 90 days. The periodontal tissues of each group were observed before administration, and the bone mineral density of rats was detected by dual energy X-ray animal bone mineral density and body composition analysis system. After 90 days of administration, the bone mineral density was detected again. After administration, blood was collected from tail vein, and the contents of serum alkaline phosphatase (ALP), bone Gla protein (BGP) and tartrate resistant acid phosphatase 5b (TRACP5b) were measured by enzyme-linked immunodeficient assay. The gingival index and periodontal attachment loss of rats in each group were obtained by visual examination and exploratory examination. The maxilla was removed, and the distance from the enamel cementum boundary to the alveolar crest was measured to obtain alveolar bone absorption value. H-E staining was used to observe the pathology of maxilla in each group. RT-PCR and Western blot were used to detect the nuclear factors in periodontal tissue of rats in each group. SPSS 22.0 software package was used for statistical analysis.
Before administration, the gums of the control group were pink without bleeding, and the gums of the other two groups were red and swollen with slight bleeding. After administration, compared with the control group, the bone mineral density, serum ALP, BGP of ovariectomized periodontitis group decreased significantly(P<0.05); while TRACP5b, gingival index, loss of periodontal attachment and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue increased significantly(P<0.05). Compared with the ovariectomized periodontitis group, the bone mineral density, serum ALP, BGP were significantly increased(P<0.05); while TRACP5b, gingival index, periodontal attachment loss and alveolar bone resorption, NF-κB and IKK mRNA and protein expression in periodontal tissue were significantly decreased (P<0.05). In the ovariectomized periodontitis group, the periodontal tissue combined with epithelium was separated from the tooth surface, the dental pocket was obvious and deep, and the height of alveolar bone decreased. Although dental pocket could be observed in the periodontal tissue of rats treated with chitosan oligosaccharide, it was not obvious, and new bone appeared around the alveolar bone.
Chitosan oligosaccharide can induce biochemical indexes of bone metabolism to become normal, alleviate the symptoms of periodontitis, this may be related to the inhibition of IKK/NF-κB pathway by chitosan oligosaccharide.
探讨壳寡糖对骨质疏松伴牙周炎小鼠骨代谢及IKK/NF-κB信号通路的影响。
将30只大鼠随机分为3组,每组10只。分为对照组、去卵巢牙周炎组和壳寡糖治疗组。除对照组外,其余两组行去卵巢手术并涂抹牙龈卟啉单胞菌菌液,建立骨质疏松伴牙周炎模型。结扎4周后,壳寡糖治疗组大鼠灌胃给予200mg/kg壳寡糖,其余两组灌胃等体积生理盐水,每天1次,共90天。给药前观察各组大鼠牙周组织情况,并用双能X线动物骨密度及体成分分析系统检测大鼠骨密度。给药90天后,再次检测骨密度。给药后,尾静脉采血,采用酶联免疫法检测血清碱性磷酸酶(ALP)、骨钙素(BGP)和抗酒石酸酸性磷酸酶5b(TRACP5b)含量。通过肉眼观察和探诊检查获得各组大鼠牙龈指数和牙周附着丧失情况。取下颌骨,测量釉牙骨质界至牙槽嵴的距离,获得牙槽骨吸收值。采用苏木精-伊红(H-E)染色观察各组上颌骨病理变化。采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法(Western blot)检测各组大鼠牙周组织中核因子表达。采用SPSS 22.0软件包进行统计学分析。
给药前,对照组牙龈呈粉红色,无出血,其余两组牙龈红肿,有少量出血。给药后,与对照组相比,去卵巢牙周炎组骨密度、血清ALP、BGP显著降低(P<0.05);而TRACP5b、牙龈指数、牙周附着丧失和牙槽骨吸收、牙周组织中NF-κB和IKK mRNA及蛋白表达显著升高(P<0.05)。与去卵巢牙周炎组相比,壳寡糖治疗组骨密度、血清ALP、BGP显著升高(P<0.05);而TRACP5b、牙龈指数、牙周附着丧失和牙槽骨吸收、牙周组织中NF-κB和IKK mRNA及蛋白表达显著降低(P<0.05)。去卵巢牙周炎组牙周组织结合上皮与牙面分离,牙周袋明显且深,牙槽骨高度降低。壳寡糖治疗组大鼠牙周组织虽可见牙周袋,但不明显,牙槽骨周围有新骨形成。
壳寡糖可使骨代谢生化指标恢复正常,缓解牙周炎症状,这可能与壳寡糖抑制IKK/NF-κB信号通路有关。