重组人骨形成蛋白2诱导成肌细胞成骨分化中的矿化反应(英文)  

Mineralization reaction during osteogenic differentiation of myoblasts stimulated by bone morphogenetic protein 2

作　　者：张力[1,2] 王伟[2] 

机构地区：[1]辽宁中医药大学,辽宁省沈阳市110032 [2]锦州市中心医院骨科,辽宁省锦州市121000

出　　处：《中国组织工程研究》2012年第20期3620-3625,共6页Chinese Journal of Tissue Engineering Research

基　　金：the National Natural Science Foundation of China,No.30772190;Postdoctorate Foundation Program of China,No.2005038055;Science and Technology Development Program of Liaoning Province,No.2005225003-14~~

摘　　要：背景:近年来成肌细胞在重组人骨形态发生蛋白2诱导下向成骨细胞分化已经通过多种方式得到了证实。目的:探讨成肌细胞在重组人骨形态发生蛋白2诱导下向成骨分化过程的矿化反应及其体外诱导表达成骨表型的可行性。方法:采用差速贴壁法和酶消化法获取Wistar乳鼠成肌细胞并体外培养,纯化鉴定后取第3代培养成肌细胞加入含重组人骨形成蛋白2的培养液进行诱导,对照组不加入重组人骨形成蛋白2,体外培养21d。结果与结论:成肌细胞经重组人骨形成蛋白2诱导后,细胞增殖减缓,相邻细胞聚集呈层状排列,诱导8d可见胞浆有少量不透光结节,诱导14d结节增多,21d时胞浆内可见大量不透光结节,未经重组人骨形成蛋白2诱导的成肌细胞融合成有收缩性的肌管。诱导后的成肌细胞碱性磷酸酶活性随时间延长而增强,诱导21d时细胞经碱性磷酸酶染色、Ⅰ型胶原免疫组化染色和钙结节染色均呈阳性反应。表明大鼠成肌细胞经重组人骨形成蛋白2诱导可以出现矿化反应,可在体外一定条件下诱导能够转化为成骨细胞。BACKGROUND： In recent years, it has been confirmed by a variety of ways that myoblasts can differentiate into osteoblasts under the induction of recombinant human bone morphogenetic protein 2 （rhBMP-2）. OBJECTIVE： To explore the mineralization reaction during the osteogenic differentiation of myoblasts under the induction of recombinant rhBMP-2 and the feasibility of osteogenic phenotype expression by in vitro induction. METHODS： Myoblasts were isolated and harvested from neonatal Wistar rats using differential velocity adherent technique and trypsinization method. After in vitro culture, purification and identification, myoblasts at passage 3 were induced by a medium containing rhBMP-2 for 21 days. Myoblasts in the control group were cultured in vitro in complete medium without rhBMP-2 for 21 days. RESULTS AND CONCLUSION： After rhBMP-2 induction, myoblast proliferation gradually slowed down. A small quantity of opaque secretory granules were found in the cytoplasm on day 8 after induction; the number of opaque secretory granules increased on day 14 after induction; and a great quantity of opaque secretory granules were found in the cytoplasm on day 21 after induction while the myoblasts without induction fused into contractile myotubes. The alkaline phosphatase activity of the induced myoblasts increased as time extended; myoblasts reacted positively in the alkaline phosphatase staining, immunochemical staining for type Ⅰ collagen and calcium node staining on day 21 after induction. These findings suggest that mineralization reaction is found in rat myoblasts by rhBMP-2 induction and myoblasts can differentiate into osteoblasts under certain inducing conditions in vitro.

关 键 词：成肌细胞 成骨细胞 骨形态发生 蛋白 分化 矿化 

分 类 号：R318[医药卫生—生物医学工程]