龟板对大鼠骨髓间充质干细胞向成骨分化的影响

Effects of Plastrum Testudinis on Os teogenesis of Cultured Rat Mesenchymal Stem Cells

目的探讨益肾中药龟板对大鼠骨髓间充质干细胞(mesenchymalstemcells,MSCs)体外向成骨分化的影响。方法使用密度梯度法分离成年大鼠骨髓间充质干细胞,观察在培养液中添加不同浓度龟板血清条件下MSCs的成骨变化。应用形态学、碱性磷酸酶组织化学染色、VonKossa染色、骨钙素测定等方法观察细胞成骨活性。结果形态学表明,MSCs贴壁细胞呈集落生长,有成纤维细胞外观。龟板组碱性磷酸酶、钙化结节、骨钙素明显升高,与对照组比较具有显著性差异(P<0.05)。结论MSCs受龟板诱导高效地向成骨细胞分化,可以为体内骨组织工程提供种子细胞。

Objective To investigate the effects of kidney-tonifying Plastrum Testudinis on o steogenesis of cultured rat mesenchymal stem cells(MSCs ).Methods MSCs were isolated from adult rats wi th density gradient separation meth od.Osteogenesis of MSCs in the culture a dded with different concentrations of serum containing Plastrum Testudinis was evalu-ated by detecting bone glaprotein(BGP)level and observing the morphologic al features of MSCs and by alkaline ph os-phatase(ALP)and Von Kossa immunohistochemical m ethods.Results Morphological examination showed t hat the MSCs attachment formed soon after seedin g ,and grew into colonies with the appearance of fibroblastic cells.Seru m containing Plastrum Testudinis promoted the expression of alkaline phosphatase(ALP)in MSCs ,and increased BGP level and the number of calcified deposition in dose -dependant manner,the difference being significant in comparison wi th the control groups(P<0.05).Conclusion Induced by the serum containing Plastrum Testudinis,MSCs can different iate into os-teoblast effectively,and this can b e used to provide seed cells for in -vivo bone tissue engineer.