›› 2015, Vol. 35 ›› Issue (6): 425-429.

• 基础研究 • 上一篇    下一篇

骨形态发生蛋白9和矿化液对牙髓干细胞成骨向诱导分化的对比研究

邵元元1,袁梦桐2,刘明月2,史欣2,胡伟平2   

  1. 1. 哈尔滨医科大学附属第二临床医院
    2. 哈尔滨医科大学附属第二医院
  • 收稿日期:2015-01-16 修回日期:2015-03-20 出版日期:2015-06-28 发布日期:2015-04-27
  • 通讯作者: 胡伟平 E-mail:huweiping1963@163.com
  • 基金资助:
    骨桥蛋白与骨涎蛋白促进人牙髓干细胞成骨分化的旁分泌作用

Comparative study in the osteogenic differentiation of human dental pulp stem cells by bone morphogenetic proteins-9 and mineralization medium

  • Received:2015-01-16 Revised:2015-03-20 Online:2015-06-28 Published:2015-04-27

摘要: 目的 比较骨形态发生蛋白9(Bone Morphogenetic Protein 9,BMP-9)和矿化液对牙髓干细胞(dental pulp stem cells, DPSCs)向成骨细胞分化的作用,选择理想的诱导方法。方法:矿化液培养DPSCs作为矿化液组,添加适宜浓度BMP-9作为BMP-9组,在倒置相差显微镜下观察诱导后的DPSCs的形态变化;用MTT法检测细胞的生长曲线;用茜素红染色法检测矿化结节的形成;采用逆转录-聚合酶链式反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)方法分别检测DPSCs的骨涎蛋白 (Bone Sialoprotein, BPS)、Runt相关转录因子-2 (Runx-2) 以及Ⅰ型胶原酶 (Collagen-1, Col-1) 及骨钙蛋白(Osteocalcin ,OCN)等骨源性基因的表达情况。结果:MTT法检测结果表明,BMP-9和矿化液能促进DPSCs增殖且促增殖作用接近;茜素红染色显示诱导培养28d后两组出现的矿化结节数量及大小相近;诱导培养14d后两组DPSCs的骨源性基因表达均成阳性且表达水平接近。结论:BMP-9和矿化液对DPSCs的成骨向诱导能力相近。

关键词: 牙髓干细胞, 骨形态发生蛋白-9, 分化

Abstract: Objective To compare the osteogenic differentiation of dental pulp stem cells (DPSCs) induced by bone morphogenetic protein-9 (BMP-9) and mineralizing culture medium, and to find out a relatively ideal inducing method. Methods DPSCs cultured with appropriate concentration of BMP-9 were made as BMP-9 group, and those cultured in mineralizing culture medium were made as mineralization medium group. The morphological changes of DPSCs were observed under the inverted phase contrast microscope; MTT was used to test the cell growth curves; a Alizarin red staining was used for detecting the mineralization nodules; reverse transcription polymerase chain reaction (RT-RCR) was used for detecting the expression of osteogenesis related genes including bone sialoprotein (BSP), Runt-related transcription factor 2 (Runx-2), collagen-1 (Col-1) and osteocalcin (OCN). Results The results of MTT showed that BMP-9 and mineralizing culture medium could promote the proliferation of DPSCs, and the promotion of both groups was close to each other. The result of Alizarin red staining showed that mineralization nodules were formed after being induced by BMP-9 or mineralizing culture medium for 28 days, and the number as well as sizes of mineralization nodules in both groups were similar. The expression of osteogenesis related genes in both groups was positive after 14 days’ inducing cultivation, and the expression levels of both groups were close. Conclusion The capabilities of BMP-9 and mineralizing culture medium in inducing DPSCs’ osteogenic differentiation were similar.