生物复合材料体外诱导分化SD大鼠乳鼠骨髓间充质干细胞为成骨细胞分析  

Analysis of biological composites inducing SD rat bone marrow mesenchymal stem cells to differentiate into osteoblasts in vitro

作　　者：卞铁荣[1] 王莉丽[2] 邢宏运[1] 唐利[1] 

机构地区：[1]泸州医学院附属医院,四川泸州646000 [2]陕西科技大学,西安715400

出　　处：《中国骨质疏松杂志》2014年第8期885-889,共5页Chinese Journal of Osteoporosis

基　　金：四川泸州医学院青年基金项目(NO12050);四川泸州医学院附属医院人才基金(NO11195)

摘　　要：目的探讨生物复合材料HA-玻璃涂层/多孔ZrO2体外诱导分化SD大鼠乳鼠骨髓间充质干细胞为成骨细胞的能力,找到骨髓间充质干细胞理想的复合载体及移植后的安全性,为骨缺损的临床快速恢复治疗奠定基础。方法通过生物材料体外诱导SD大鼠骨髓间充质干细胞,培养至14天、21天,经Gomori钙钴法染色、Von Kossa染色及Ⅰ型胶原免疫组化分析比较诱导前后其碱性磷酸酶、钙结节、Ⅰ型胶原的表达变化;流式细胞技术检测诱导前后细胞CD45、CD44及CD90的表达变化。结果 SD大鼠骨髓间充质干细胞体外诱导14天,出现了三角形或不规则细胞,细胞之间界限清晰,碱性磷酸酶呈阳性表达;诱导至21天,细胞中出现致密圆形的矿化结节及Ⅰ型胶原呈阳性表达,细胞之间界限模糊;流式技术检测诱导前后细胞表面CD44仍为阳性表达,CD45仍为阴性表达,而CD90由原来的阳性表达转为阴性表达,细胞表达阳性率由99.7%下降为0.29%。结论该生物复合材料将骨髓间充质干细胞成功诱导为成骨细胞,且该材料又能与骨形成很强的化学结合,用作骨缺损的填充材料,为新骨的形成提供支架,发挥骨传导作用。Objective To explore the ability of the biological composite materials,HA-glass coating /porous ZrO2,inducing SD rat bone marrow mesenchymal stem cells to differentiate into osteoblasts in vitro,to find the ideal composite carrier of transplantated bone marrow mesenchymal stem cells and to investigate the security after transplantation,and to lay foundation for the fast recovery treatment of bone defect. Methods SD rat bone marrow mesenchymal stem cells were induced to differentiate by biological materials in vitro. Then cells were cultured for 14 days and 21 days. After dying with calcium-cobalt according to Gomori,Von Kossa staining,and immunohistochemical technology,the expression of alkaline phosphatase,calcium nodules,and type I collagen was detected. The changes of the expression of CD45,CD44,and CD90 in the cells before and after the induction were detected using flow cytometry. Results After 14-day induction in vitro,the cell morphology of SD rat bone marrow mesenchymal stem cells changed from long shuttle deformation cells to triangle or polygon cells,and there were clear boundaries between the cells. The expression of alkaline phosphatase was positive. After 21-day induction,dense and circular mineralized nodules appeared in cells,and the expression of type I collagen was positive. The boundaries between cells were blurred. According to the results of flow cytometry technology,the expression of CD44 was still positive,and the expression of CD45 was still negative. But the expression of CD90 transformed from positive to negative. The positive expression rate of cells decreased from 99. 7% to 0. 29%. Conclusion Biological composite materials,HA-glass coating /porous ZrO2 scaffolds,successfully induce SD rat bone marrow mesenchymal stem cells to osteoblasts. These materials can combine with bone formation through strong chemical bond,which can be used as bone defect filling materials,and provide support for the formation of new bone,and play the conduction role in bone.

关 键 词：HA-玻璃涂层/多孔ZrO2支架 诱导剂 骨髓间充质干细胞 成骨细胞 

分 类 号：R318.08[医药卫生—生物医学工程] Q813.11[医药卫生—基础医学]