成骨细胞与镁合金表面硅酸盐-磷酸盐复合涂层的体外相容性

The compatibility study of osteoblast and the silicate-phosphate coating on the surface of the magnesium in vitro

目的评价成骨细胞在镁合金表面硅酸盐-磷酸盐复合涂层上的生长状况,探讨镁合金表面硅酸盐-磷酸盐复合涂层与成骨细胞的生物相容性。方法用胶原酶消化法分离培养大鼠成骨细胞,并用免疫荧光和茜素红染色法进行鉴定;然后将第4代大鼠的成骨细胞接种于材料表面,用扫描电镜观察成骨细胞在不同表面上黏附形态的变化,同时制备材料浸提液,用浸提液法培养细胞,并通过四甲基偶氮唑盐(MTT)法和碱性磷酸酶(ALP)活性法检测成骨细胞的增殖和分化情况。结果将成骨细胞与材料进行复合培养后,扫描电镜下可见,涂层组的成骨细胞生长活力旺盛、形态饱满、分布均匀,而单纯镁合金组未见细胞生长;MTT和ALP活性分析结果显示,硅含量为1%～5%涂层对大鼠成骨细胞的体外生长、增殖和分化的促进作用最佳。结论镁合金表面硅酸盐-磷酸盐涂层与成骨细胞具有良好的相容性,在体外培养的环境下,有利于细胞的生长、黏附、增殖和分化。

Objective To evaluate the growing condition of osteoblasts, which were cultured on the silicate- phosphate coating on the surface of the magnesium and the biocompatibility between the coating and osteoblasts. Methods The osteoblasts were harvested by collagenase digesting method, and identified by immunofiuorescence and Alizarin red staining method. Then the osteoblasts of the forth passage were inoculated on the material surface and the changes in form of osteoblasts on different surfaces were observed by scanning electron microscopy （SEM）. At the same time the material leaching liquor was prepared to culture osteoblasts and the viabilities and differentiation of osteoblasts were detected by MTT and ALP method respectively. Results SEM showed that cells on the coating magnesium alloy growed well, and proliferated significantly,but cells were hardly finded on the naked group. The MTT and ALP analysis showed that i % -5% of Si coating could promote the growth, proliferation and differentiation of osteoblast more obviously than other groups in vitro. Conclusion The silicate-phosphate coating on the surface of the magnesium had good compatibility with osteoblasts and was conducive to cell growth ,adhere, proliferation and differentiation in vitro.