微管仿生人工骨对犬骨髓基质细胞生物学行为的影响

目的]研究不同微管结构仿生人T骨对犬骨髓基质细胞（BMSCs）增殖和分化的影响。方法]仿生人工骨分为正交结构（A组）和同心结构（B组），以磷酸钙骨水泥（CPC）／重组人骨形成蛋白2（rhBMP-2）（C组）、单纯CPC（D组）为对照组：梯度密度离心法获取犬骨髓单个核细胞，取第3代与人工骨复合培养。通过荧光显微镜、扫描电镜观察细胞在人工骨表面附壁和生长情况；以MTT实验检测各组细胞增殖能力；测定碱性磷酸酶（ALP）检测BMSCs的成骨活性：结果]细胞在各组人工骨表面黏附、生长，以A、B组数量较多；MTT法检测显示A、B、C组吸光度（A）值大于D组（P〈0．05）、A、B、C组A值相似（P〉0．05）；ALP测定显示ALP活性A、B组〉C组〉D组（P〈0．05），A、B组间差异无统计学意义（P〉0．05）。结论]微管结构仿生人工骨与犬BMSCs有良好的生物相容性，是BMP的良好缓释载体，可以促进犬骨髓基质细胞向成骨方向分化.

Effects of bionic microstructure artificial bone on dog bone marrow stromal cells: an in vitro study

Objective]To study the effects of different bionic microstructure of artificial bones on the growth and osteogenesis characteristics of dog bone marrow stromal cells(BMSCs).Method]The artificial bones were divided into 4 groups in the experiments:perpendicular structure(experimental group,A),concentric structure(experimental group,B),CPC/ rhBMP-2(calcimrt phosphate cement composited with recombinant human bone morphogenetic protein 2)(control group,C)and pure CPC(control group,D).Dog bone marrow mononuclear cells were isolated from the bone marrow by gradient centrifugation and then cultured in non-induced condition.BMSCs at 3~(rd) passage were harvested and inoculated onto the artificial bones of the 4 groups.The implanted cells and artificial bones were retrieved and observed at different times with fluorescence microscope,scanning electron microscope.The proliferation of BMSCs was evaluated by MTT method,and the osteogenic differentiation was evaluated by alkaline phosphatase(ALP)activity.Result]The cultured BMSCs adhered well to the surface of all artificial bones and it was better to A and B groups than to C and D groups.By MTT method,the A values of group A,B,C were significantly higher compared with group D(all P<0.05),while there were no significant differences between group A,group B and group C(P>0.05);The ALP activities of BMSCs in group A,B,C and D all increased significantly from the 4~(th) day to the 7~(th) day(P<0.05),quantitative analysis of the ALP activities by the four groups showed significant differences between group A,B and group C(P<0.05),and between group C and group D(P<0.05),but there was no significant difference between group A and group B(P>0.05).Conclusion]The bionic microstrueture artificial bones are satisfactory carriers of BMP and possess good cellular biocompatibility with dog bone marrow stromal cells and have the ability to enhance the osteogenic differentiation of BMSCs.