骨形成蛋白-2和牙胚细胞联合作用诱导人牙周膜干细胞表达成牙骨质细胞的表型研究  

Periodontal ligament stem cells co-cultured with tooth germ cells containing bone morphogenetic protein-2 acquired cementoblast feature in vitro

作　　者：童晓洁 兰泽栋[1] 郑雅心 陈瑾[1] 谢跃强 陈良娇[1] 

机构地区：[1]广州医科大学附属口腔医院正畸科,广东广州510140 [2]厦门市口腔医院正畸科

出　　处：《中国美容医学》2014年第16期1343-1348,共6页Chinese Journal of Aesthetic Medicine

基　　金：广州医科大学课题(骨形成蛋白-2对人牙周膜干细胞成牙骨质特性的影响:课题编号:2010A31)

摘　　要：目的:探讨大鼠牙胚细胞(Tooth germ cells,TGC)与骨形成蛋白-2(Bone morphogenetic proteins-2,BMP-2)联合作用对人牙周膜干细胞(Human periodontal ligament stem cells,hPDLSCs)向成牙骨质细胞表型分化的作用。方法:将免疫磁珠法分离的hPDLSCs与TGC共培养,并加入50ng/mL的BMP-2,通过RT-PCR,茜素红染色和碱性磷酸酶(Alkaline Phosphatase,ALP)活性检测等方法分析其相关成牙骨质基因及蛋白的表达变化。结果:诱导后hPDLSCs的ALP活性明显升高(P<0.001),牙骨质细胞相关基因如牙骨质附着蛋白(Cementum attachment protein,CAP)、牙骨质蛋白(Cementum protein1,CEMP-1)、ALP,骨钙素(Osteocalcin,OCN)的转录水平表达量均有不同程度的增高(P<0.001)。矿化诱导14d后,诱导组形成大量矿化结节,与对照组相比具有统计学意义(P<0.001)。结论:TGC与BMP-2联合作用能够诱导hPDLSCs与向成牙骨质细胞的表型分化。Objective To investigated the role of Bone morphogenetic protein-2 （BMP-2） as a factor to induce human periodontal ligament stem cells （hPDLSCs） differentiate into cementoblast or cementum like cells when co-cultured with tooth germ cells （TGC）.Methods hPDLSCs isolated by magneticactivated method.The cells were co-cluture with tooth germ cells （control group） or co-cultured with tooth germ cells containing BMP-2 （test group）.Then RT-PCR,Alkaline Phosphatase （ALP） analyses and mineralization assay were used to identify the cementoblast markers,such as CAP（Cementum attachment protein）,CEMP-1（Cementum protein1）,ALP and OCN （Osteocalcin）.Results The induced cells showed high ALP activity （P〈0.001）,increased calcified nodules formation （P〈0.001） and a significantly increase cementum-related genes expression such as CEMP-1,CAP,ALP and OCN （P〈 0.001）.Conclusion hPDLSCs co-culture with TGC containing bone morphogenetic proteins-2 acquired more cementoblast features.

关 键 词：牙周膜干细胞 骨形成蛋白-2 成牙骨质细胞 牙胚细胞 

分 类 号：R246.83[医药卫生—针灸推拿学]