2型糖尿病伴牙周炎患者牙周膜干细胞骨向分化研究  

An in vitro study on the osteogenic differentiation of human periodontal ligament stem cells of the patients with type 2 diabetes mellitus and periodontitis

作　　者：夏佳佳[1] 王岚[1] 刘琪[1] 金岩 邓超[1] 伍燕[1] 

机构地区：[1]遵义医学院附属口腔医院,563003 [2]第四军医大学口腔医院组织工程中心

出　　处：《实用口腔医学杂志》2011年第5期624-629,共6页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:30725042;81020108019);国家基础研究项目(973项目)(编号:2010C13944800;2011CB964700);贵州省省长基金(编号:C_397);贵州省科技厅自然基金(编号:C_393);遵义市科技基金(编号:E_063)

摘　　要：目的:比较牙周炎和2型糖尿病伴牙周炎患者牙周膜干细胞成骨分化能力。方法:体外组织块法和有限稀释法克隆化培养牙周炎患者牙周膜干细胞(P-PDLSCs组)和2型糖尿病伴牙周炎患者牙周膜干细胞(D-PDLSCs组),计算克隆形成率,免疫荧光检测细胞表型分子CD146、STRO-1进行干细胞鉴定,矿化诱导后茜素红染色观察矿化结节形成,实时定量聚合酶链反应(real time PCR)检测成骨相关基因表达。结果:P-PDLSCs组和D-PDLSCs组细胞的克隆形成率分别为(25.6±2.7)%和(17.9±1.7)%(P﹤0.05);P-PDLSCs组CD146和STRO-1表达明显高于D-PDLSCs组;成骨诱导21 d后茜素红染色,2组均出现不同程度的矿化结节,P-PDLSCs组的矿化能力较D-PDLSCs组强;成骨诱导1周后D-PDLSCs组碱性磷酸酶(alkaline phosphatase,ALP)、Runx-2和Ⅰ型胶原蛋白(type-Ⅰcollagen,Col-Ⅰ)的mRNA表达均明显低于P-PDLSCs组(P<0.05)。结论:糖尿病伴牙周炎患者牙周膜干细胞成骨分化能力低于单纯牙周炎患者牙周膜干细胞。Objective：To investigate the osteogenic differentiation potential of human periodontal ligament stem cells（PDLSCs） of the patients with type 2 diabetes mellitus and period ontitis.Methods：PDLSCs were isolated from the patients with periodontitis patients（P-PDLSCs group） and those with type 2 diabetes mellitus and periodontitis （D-PDLSCs group）.Cells were purified by single-colony selection.Cloning efficiency,immunofluorescence staining were used to identify the stem cells.The osteogenic differentiation capacity of PDLSCs was evaluated by alizarin red staining and real time PCR.Results：The colony formation rate of P-PDLSCs and D-PDLSCs was（25.6 ± 2.7） % and（17.9 ± 1.7） % separately（P0.05）.Expression of cell surface molecules CD146 and STRO-1 in P-PDLSCs was significantly higher than those in D-PDLSCs.Alizarin red staining showed varying degree of mineralization nodules in both groups after 21d induction,the area of mineralization in D-PDLSCs group was smaller than that in P-PDLSCs group;the expressions of osteogenic genes ALP,Runx-2 and Col-Ⅰ mRNA in D-PDLSCs were significantly lower than those in P-PDLSCs after 7day induction （all,P﹤0.05）.Conclusion：In patients with type 2 diabetes mellitus and periodontitis the severity of periodontitis is increased by reducing the osteogenic differentiation capacity of PDLSCs.

关 键 词：2型糖尿病 牙周炎 牙周膜干细胞 成骨诱导 

分 类 号：R781.4[医药卫生—口腔医学]