微弧氧化陶瓷膜表面骨髓间充质干细胞生物相容性实验

目的观察微弧氧化（micro-arc,MAO）陶瓷膜表面骨髓间充质干细胞（mesenchymal stem cell,MSC）的黏附和增殖生长情况,通过MSC在三组不同表面处理材料表面的黏附和增殖情况评价微弧氧化陶瓷膜表面的生物相容性。方法贴壁法培养大鼠骨髓间充质干细胞,骨诱导培养后,做碱性磷酸酶染色和茜素红钙结节染色,鉴定其成骨潜能。取生长良好的第三代MSC,调整细胞密度为5×10^4/mL,接种到三组钛片表面。在接种细胞后第1、3、5、7天每组分别取出5枚钛片,一个做电镜扫描,另外4个细胞计数。观察MSC在不同材料表面的黏附增殖情况。电镜扫描MAO陶瓷膜表面形貌特征,EDX分析其表面主要元素含量。结果MSC具有良好的成骨特性,在MAO陶瓷膜表面的增殖优于光滑组和喷沙组。电镜下微弧氧化陶瓷膜表面有无数2～10m的微孔,并含有钙、磷成分。结论MAO陶瓷膜具有良好的生物相容性,大鼠骨髓间充质干细胞在其多孔,含有钙、磷元素表面的黏附及增殖均优于其它组。

The experiment of the micor-arc oxidation film''''s biocompatibility with mesenchymal stem cell

Objective The biocompatibility of the micro-arc oxidation （MAO） film by seeding rat bone marrow mesenchymal stem cell （MSC） on it. Methods Rat bone marrow mesenchymal stem cells were cultivated by cell attachment method. Its bone formarion ability was tested through alkaline phosphatase（ALP） stain and calcium tubercle alizarin Bordeaux stain. We put the three groups titanium pieces in twenty-four wells cultivate plate, twenty pieces per plate. The MSCs at the density of 5× 10^4/mL was seeded on the surface of titanium. On the first, third, fifth and seventh day after seeding the MSC, we take out five pieces of titanium per group every time, One of them for electronic microscopy scanning, the others for cell counting. The surface feature of MAO film was scanned by Scanning Electronic Microscopy（SEM） and the element contained with in the MAO film was analyzed by Energy Dispersion X Ray（EDX）. Results The mesenchymal cells showed it＇ s good bone forming ability. The attachment and proliferation of MSCs on MAO surface were better than the other groups indicated by the cell counting result. SEM photo exhibited that there were numerous 2 - 10μm pores on the MAO surface. EDX results indicated that the main element on the MAO surface is calcium and phosphonium.Conclusion the MAO film showed its excellent biocompatibility with MSCs. The attachment and proliferation of MSCs on MAO surface contained with calcium and phosphonium were excelled other groups.