转化生长因子β3与骨形成蛋白2对牙髓干细胞增殖和成骨分化的影响  

Effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on proliferation and osteogenic differentiation of dental pulp stem cells

作　　者：艾力麦尔旦·艾尼瓦尔 木合塔尔·霍加 王玲[1] Ailimaierdan·Ainiwaer;Muhetaer·Huojia;Wang Ling(Department of Oral Surgery,First Affiliated Hospital(Affiliated Stomatological Hospital)of Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Oral Center,Shenzhen Luohu People’s Hospital,The Third Affiliated Hospital of Shenzhen University,Shenzhen 518001,Guangdong Province,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)口腔外科门诊,新疆维吾尔自治区乌鲁木齐市830011 [2]深圳市罗湖区人民医院·深圳大学附属第三医院口腔中心,广东省深圳市518001

出　　处：《中国组织工程研究》2022年第30期4862-4866,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81560180),项目负责人:木合塔尔·霍加。

摘　　要：背景:转化生长因子β3与骨形成蛋白2对牙髓干细胞的增殖和成骨向分化能力对比相关研究甚少。目的:探讨相同质量浓度转化生长因子β3与骨形成蛋白2对牙髓干细胞成骨向分化的作用及其初步机制。方法:采用酶解组织块法从新西兰大白兔牙髓组织中分离培养牙髓干细胞;将第3代牙髓干细胞分为空白组、骨形成蛋白2(80μg/L)组和转化生长因子β3(80μg/L)组,采用MTT法检测各组细胞的增殖活力;培养第7,14天行碱性磷酸酶活性定量检测,Runt相关转录因子2、骨涎蛋白免疫细胞化学染色,RT-qPCR法检测成骨相关基因的表达;培养第14,21天进行茜素红染色观察矿化结节形成能力。结果与结论:①骨形成蛋白2组和转化生长因子β3组的牙髓干细胞增殖活性、碱性磷酸酶活性均高于空白组(P<0.05);转化生长因子β3组第7天碱性磷酸酶活性高于骨形成蛋白2组(P<0.05);②转化生长因子β3组与骨形成蛋白2组Runt相关转录因子2、骨涎蛋白表达均高于空白组(P<0.05),其中转化生长因子β3组第7天Runt相关转录因子2蛋白表达高于骨形成蛋白2组(P<0.05);③转化生长因子β3组与骨形成蛋白2组碱性磷酸酶、Ⅰ型胶原A1、Runt相关转录因子2、骨钙蛋白、骨桥蛋白和Osx基因表达量高于空白组(P<0.05),第7天转化因子β3组碱性磷酸酶、Ⅰ型胶原A1、Runt相关转录因子2基因的表达高于骨形成蛋白2组(P<0.05);④转化生长因子β3组与骨形成蛋白2组矿化结节较空白组明显;⑤结果表明,转化生长因子β3与骨形成蛋白2均能促进牙髓干细胞增殖和成骨向分化;转化生长因子β3对牙髓干细胞成骨早期促进作用优于骨形成蛋白2。BACKGROUND:Less is reported about the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the proliferation and osteogenic differentiation of dental pulp stem cells.OBJECTIVE:To explore the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the osteogenic differentiation of dental pulp stem cells and its preliminary mechanism.METHODS:Dental pulp stem cells were isolated from the pulp tissue of New Zealand rabbit by the enzymatic digestion method.Dental pulp stem cells at passage 3 were divided into the blank group,bone morphogenetic protein-2 group(80μg/L)and transforming growth factor-beta 3 group(80μg/L).The cell proliferation activity in each group was tested by MTT assay.After 7 and 14 days of culture,alkaline phosphatase activities were measured.Runt-related transcription factor 2 and bone sialoprotein protein expression levels were conducted by immunocytochemical staining.Osteogenesis-related gene expression was tested through RT-qPCR method.Alizarin red S staining was conducted after 14 and 21 days of culture to observe mineralized nodules.RESULTS AND CONCLUSION:(1)Dental pulp stem cell proliferation activity and alkaline phosphatase activity were higher in the bone morphogenetic protein-2 and transforming growth factor-beta 3 groups than those in the blank group(P<0.05).The alkaline phosphatase activity of the transforming growth factor-beta 3 group on day 7 was higher than that of the bone morphogenetic protein-2 group(P<0.05).(2)The expression levels of Runt-related transcription factor 2 and bone sialoprotein in the transforming growth factor-beta 3 group and the bone morphogenetic protein-2 group were higher than those in the blank group(P<0.05).The protein expression of Runt-related transcription factor 2 in the transforming growth factor-beta 3 group on day 7 was higher than that in the bone morphogenetic protein-2 group(P<0.05).(3)The expression levels of alkaline phosphatase,type I collagen A1,Runt-related transcription factor 2,osteocalci

关 键 词：转化生长因子Β3 骨形成蛋白2 牙髓干细胞 增殖 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]