表面矿化修饰煅烧骨/P24活性多肽仿生骨材料对MC3T3-E1细胞黏附、增殖和向成骨方向分化的影响

Effects of Sintered Bone Modified with Surface Mineralization/P24 Peptide Composite Biomaterial on the Adhesion,Proliferation and Osteodifferentiation of MC3T3-E1 Cells

本研究目的是评价表面矿化修饰煅烧骨/P24活性多肽复合仿生骨材料对小鼠MC3T3-E1细胞黏附、增殖和诱导成骨分化的影响。实验分三组：A组为表面矿化修饰煅烧骨加骨形态发生蛋白-2（BMP-2）活性多肽P24修饰的复合仿生骨材料,B组为表面矿化修饰煅烧骨材料,C组为单纯的煅烧骨材料。三组材料在细胞实验前行电镜扫描观察。然后将小鼠MC3T3-E1细胞分别种植在三种材料表面,测定细胞的黏附率,MTT法检测细胞体外增殖活性,同时通过碱性磷酸酶（ALP）染色和ALP活性检测,了解小鼠MC3T3-E1细胞成骨分化情况。电镜观察显示三组材料均保留了天然的孔隙结构,孔径为200-850μm。细胞黏附率和MTT实验显示,A组MC3T3-E1细胞在材料表面的黏附性能和增殖能力明显高于B组和C组,B组则高于C组,组间差异均有统计学意义（P〈0.05）。ALP染色和ALP活性检测显示：A组细胞ALP活性明显高于B组和C组,B组则高于C组,组间差异均有统计学意义（P〈0.05）。研究表明表面矿化修饰煅烧骨/P24活性多肽仿生骨材料能促进小鼠MC3T3-E1细胞在骨基质材料表面的黏附、增殖与分化,并能较好地保持细胞的形态。

In the present research,the effects of sintered bone modified with surface mineralization/P24 peptide composite biomaterials on the adhesion,proliferation and osteodifferentiation of MC3T3-E1 cells were investigated.The experiments were divided into three groups due to biomaterials used：Group A（composite materials of sintered bone modified with surface mineralization and P24,apeptide of bone morphogenetic protein-2）;Group B（sintered bone modified with surface mineralization）and Group C（sintered bone only）.The three groups were observed by scanning electron microscopy（SEM）before the experiments,respectively.Then MC3T3-E1 cells were cultured on the surfaces of the three kinds of material,respectively.The cell adhesion rate was assessed by precipitation method.The proliferative ability of MC3T3-E1 cells were measured with MTT assay.And the ALP staining and measurement of alkaline phosphatase（ALP）activity were performed to assess the differentiation of cells into osteoblasts.The SEM results showed that the materials in the three groups retained the natural pore structure and the pore sizes were in the range between 200-850μm.The adhesive ratio measurements and MTT assay suggested that adhesion and proliferation of MC3T3-E1 cells in Group A were much higher than those in Group B and Group C（P〈0.05）.The ALP staining and ALP activity of MC3T3-E1 cells in Group A were significantly higher than those in Group B and Group C（P〈0.05）.The sintered bone modified with surface mineralization/P24 composite material was confirmed to improve the adhesion rate and proliferation and osteodifferentiation of MC3T3-E1 cells,and maintained their morphology.