海藻酸钠凝胶对骨髓间充质干细胞生物学效应的初步研究

目的：初步探讨海藻酸钠凝胶对骨髓间充质干细胞（bone mesenchymal stem cells, bMSCs）的生物学效应。方法：将在海藻酸钠凝胶上培养的细胞设为实验组，在塑料培养板上的细胞设为对照组。以MTT法检测细胞在两种材料上的增殖；采用甲苯胺蓝染色观察细胞在凝胶上的生长形态；RT-PCR检测bMSCs生长6天后的成骨相关基因。结果：两组的bMSCs均能迅速贴附、铺展进入增殖期；在实验组，细胞成集落样生长较明显，且形态发生改变，伸出较多的伪足，细胞间接触更为紧密；培养至第6天，进行成骨相关基因检测，发现对照组和实验组均有Ⅰ型胶原和碱性磷酸酶表达，但它们在实验组的表达量高于对照组；同时，实验组骨连接蛋白为阳性表达，而对照组为阴性。结论：bMSCs可能具有自发向成骨细胞分化的趋势；海藻酸钠凝胶具有促进干细胞向成骨细胞分化的生物学活性。提示海藻酸钠凝胶可能是比较适合的骨组织工程支架材料。

The effects of alginate gels on bone mesenchymal stem cells

Objective: To examine the effects of alginate on bone mesenchymal stem cells(bMSCs).Methods: bMSCs were cultured on the alginate gels(experimental group) and tissue culture plastics(control group);the morphous of bMSCs was observed by microscope after toluidine blue staining;the growth curves were analyzed by MTT;and the expressions of osteoblast related genes(alkaline phosphatase,collagen I and osteonectin) were detected by RT-PCR when cultured for 6 days.Results: There were no difference in the cell proliferation between the experimental group and control group.The morphous of bMSCs in the experimental group was induced to be similar to that of osteoblasts and form colony proliferation obviously.The alkaline phosphatase and collagen I genes were positive in the two groups,but their expressions in the experimental group were higher;at the same time,osteonectin gene was positive in the experimental group and negative in the control group.Conclusion: bMSCs may differentiate into osteoblasts automatically when cultured for longer time and that alginate gels could promote this differentiation and may be suitable biomaterial of scaffold in bone tissue engineering.