BMP-2促进人炎症牙髓干细胞骨向诱导分化的体外研究

›› 2015, Vol. 6 ›› Issue (2): 71-77.

BMP-2促进人炎症牙髓干细胞骨向诱导分化的体外研究

邓云贞,李珏丹,魏虹,石建峰,李昂,苟建重

西安交通大学口腔医院

收稿日期:2014-11-17 修回日期:2015-03-10 出版日期:2015-06-25 发布日期:2015-07-13
通讯作者: 邓云贞 E-mail:812247448@qq.com
基金资助:
陕西省科技统筹创新工程计划资源主导型产业关键技术（链）项目;陕西省自然科学基础研究计划项目

Studies of BMP-2 promote osteogenic differentiation of human inflammatory dental pulp stem cells in vitro

Received:2014-11-17 Revised:2015-03-10 Online:2015-06-25 Published:2015-07-13
Contact: Yun-Zhen DENG E-mail:812247448@qq.com

摘要/Abstract

摘要： 目的：研究人骨形态发生蛋白-2（bone morphogenetic protein，BMP-2）对人炎症牙髓组织来源的牙髓干细胞（dental pulp stem cells from inflamed pulps，DPSCs-IPs）体外骨向诱导分化的影响。方法：取第三代DPSCs-IPs，按是否于培养基中加入BMP-2分成：BMP-2+DPSCs-IPs组（实验组）和DPSCs-IPs组（对照组）。无成骨诱导条件培养1周后，对各组分泌的胶原基质行Trichrome染色，观察并比较实验组和对照组之间的骨向诱导分化情况；实时定量RT-PCR检测二者的Ⅰ型胶原蛋白（collagenⅠ，COL-Ⅰ）mRNA表达情况。在成骨诱导条件下，培养3周后对各组分泌的钙化基质行Von Kossa染色，通过实时定量RT-PCR检测转录因子及成骨相关基因的表达。结果：无成骨诱导培养1周后，实验组DPSCs-IPs分泌的胶原基质Trichrome染色较对照组深，COL-ⅠmRNA的表达水平显著上调（P<0.05），说明BMP-2可诱导DPSCs-IPs沉积更多的胶原基质；成骨诱导培养3周后，相对于对照组，实验组DPSCs-IPs沉积了更多的钙化基质，Nanog、八聚体结合转录因子4（octamer-binding transcription factor 4，Oct4）、性别决定区因子（SRY-related high-mobility group box 2，Sox2）等转录因子表达升高，碱性磷酸酶（alkaline phosphatase，ALP）、骨钙素（osteocalcin，OCN）、骨涎蛋白（bone sialoprotein，BSP），牙骨质蛋白1（cementum protein 1，CEMP-1）、COL-Ⅰ等成骨相关基因表达水平显著上调（P<0.05）。结论：BMP-2在成骨诱导条件下可促进DPSCs-IPs进行体外骨向诱导分化。

Abstract: Objective：To investigate the influence of bone morphogenetic protein-2(BMP-2) on osteogenic differentiation of human inflammatory dental pulp stem cells(DPSCs-IPs) in vitro. Methods：Passage 3 cells dental pulp stem cells from inflamed teeth in medium with or without osteogenic matter. According to whether 100 μg/L BMP-2 was added they were divided into four groups: group 1，DPSCs-IPs culturing with 100 μg/L BMP-2 and osteogenic medium containing; group 2, DPSCs-IPs culturing with 100 μg/L BMP-2 but without osteogenic medium containing; group 3, DPSCs-IPs culturing without 100 μg/L BMP-2 but with osteogenic medium containing; group 4, DPSCs-IPs culturing without 100 μg/L BMP-2 and osteogenic medium containing. After one week of their culture in nonosteogenic condition, we used Trichrome to stain the secretion of collagen matrix, and compared the difference of osteogenic differentiation between them. Real-time RT-PCR was used to test the expression of COL-ⅠmRNA. Three weeks after their culture in osteogenic condition, the cell differentiation was examined by von Kossa staining. Real-time RT-PCR was used to test nuclear transcription factor Nanog, Oct4, SOX2 and osteogenesis related molecules ALP, OCN, BSP, CEMP-1 and COL-Ⅰ gene expression. Results：One week later, Trichrome staining matrix secretion of DPSCs-IPs with BMP-2 group is more obvious than the control group in the nonosteogenic medium. Real-time RT-PCR tests showed that expression of COL-ⅠmRNA increased (P<0.05). The results suggested that BMP-2 could induce DPSCs-IPs deposit more collagen matrix; 3 weeks later, more mineralized matrix secretion of DPSCs-IPs can be observed in the osteogenic medium. In addition, real-time RT-PCR tests showed that experimental group cells express more Nanog, Oct4 and SOX2. And the osteogenesis related molecules ALP, OCN, BSP, CEMP-1 and COL-Ⅰ expression increased significantly compared with the control group (P<0.05). Conclusions：BMP-2 can promote osteogenic differentiation of human inflammatory dental pulp stem cells in vitro in osteogenic condition.

邓云贞李珏丹魏虹石建峰李昂苟建重. BMP-2促进人炎症牙髓干细胞骨向诱导分化的体外研究[J]. , 2015, 6(2): 71-77.

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