镁合金表面含硅涂层对成骨细胞的黏附、形态和细胞周期的影响

摘要

目的:评价成骨细胞在镁合金表面含硅涂层上的生长状况,为研究镁合金表面含硅涂层与成骨细胞的生物相容性提供实验依据.方法:用胶原酶消化法分离培养大鼠成骨细胞,并用免疫荧光和碱性磷酸酶染色法进行鉴定;然后将处于对数生长期的大鼠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粘附形态的变化,同时制备材料浸提液,用浸提液法培养细胞,并通过倒置相差显微镜观察细胞形态;用流式细胞仪检测成骨细胞的细胞周期变化.结果:分离出了大鼠成骨细胞,I型胶原免疫荧光显色及碱性磷酸酶显色阳性;成骨细胞与材料进行复合培养后,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;硅含量为1%～5%涂层组的成骨细胞G2期+S期所占百分比最高.结论:镁合金表面含硅涂层与成骨细胞具有良好的相容性,有利于细胞的黏附、生长和增殖.%Objective: To evaluate the growing condition of osteoblast, which were cultured on the silicate 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 immunofluorescence and alkaline phosphatase staining method. Then the osteoblasts of log phase growth were inoculated on the material surface and the changes in form of osteoblasts on different surfaces were observed by scanning electron microscopy. At the same time the material leaching liquor was prepared to culture osteoblasts and the changes in form of osteoblasts were observed under an inverted phase contrast microscope and the cell cycle were detected by flow cytometry. Results: The osteoblasts were successfully separated, and immunofluorescence staining of collagen Ⅰ and alkaline phosphatase staining was positive. After being cultured with the materials, cells of the coating group grew well and proliferated significantly, but cells were hardly found on the magnesium without coating group.The cell cycle analysis showed that the percentage of G2 phase+S phase in the 1％- 5％ Si content group was highest. Conclusion: The silicate coating on the surface of the magnesium have good compatibility of osteoblasts, which is conducive to cell adhesion, growth and proliferation in uitro.