透明质酸钠对大鼠成肌细胞体外成骨分化的影响

摘要

目的 研究透明质酸钠对成肌细胞体外成骨诱导分化的影响.方法 运用差速贴壁法和(或)酶消化法分离纯化的大鼠成肌细胞,将培养至第3代的细胞分为:(1)实验组:采用含0.1%的透明质酸钠和500ng/mL的重组人骨形态发生蛋白2(rhBMP-2)的培养基进行培养;(2)对照组:以含500ng/mL rhBMP-2的培养基进行培养.观察诱导过程中成肌细胞形态变化.第3周时对其进行成骨指标检测(碱性磷酸酶活性,Ⅰ型胶原免疫组化,茜素红和碱性磷酸酶的钙结节染色).结果 实验组和对照组成肌细胞在诱导过程中形态学改变相似.实验组和对照组碱性磷酸酶活性均升高,2组间差异无统计学意义,Ⅰ型胶原免疫组化均呈阳性,茜素红和碱性磷酸酶染色均可见大量钙结节.结论 透明质酸钠不抑制成肌细胞体外成骨分化,与成肌细胞具有良好的生物相容性.%Objective To investigate the effects of sodium hyaluronate solution on the osteoplastic diferentation of rat's myoblasts. Methods The minced muscles were digested with mixed digestive juices,followed by the isolation,culture and purification of the myoblasts. The third subculture myoblasts were cultured in two growth media,in which experimental group containing 0.1％ sodium hyaluronate and 500 ng/mL rhBMP-2, the control group containing 500 ng/mL rhBMP-2, respectively. The morphological change of induced myoblasts in each medium was observed carefully. Three weeks later,a series of examination were performed,including:ALP activity,type Ⅰ collagen immunohistochemistry, and new calcifying nodules staining with Alizarin red S and ALP. Results The morphological change in each medium was similar during the induced process. There was a difference between the two groups on ALP activity,but the ALP activity increased and there was no statistic difference (P ＞ 0.05 ). The immunohistochemistry of type Ⅰ collagen was positive,a great quantity of new calcifying nodules was seen by the staining of Alizarin red S and ALP. Conclusion Sodium hyaluronate could not inhibit the osteoplastic differentiation of rat's myoblasts in vitro,it could be a proper media for myoblasts.