【2016年】无定形磷酸钙负载rhBMP2纳米缓释体的制备及细胞毒性实验【药学论文】.doc

药学论文-无定形磷酸钙负载rhBMP2纳米缓释体的制备及细胞毒性实验 【摘要】? 制备无定形磷酸钙(ACP)负载重组人骨形态发生蛋白2(rhBMP2／ACP)纳米缓释体并观察其细胞毒性，为进一步体内植入提供实验依据。 ［方法］采用湿化学法合成rhBMP2／ACP纳米缓释体；兔间充质干细胞(BMSCs)与rhBMP2／ACP纳米缓释体进行体外复合培养，观察细胞在材料表面的黏附、增殖及菜单达，检测材料的细胞毒性。［结果］材料浸提液对兔BMSCs的生长无影响，其细胞相对增殖率在1002％～1025％之间，细胞毒性评级为0级；BMSCs于复合材料表面的黏附、生长速度、形态与对照组无明显差别。［结论］rhBMP2／ACP纳米缓释体无细胞毒性，复合培养不影响BMSCs的正常生理功能，细胞兼容性良好。 【关键词】? 重组人骨形态发生蛋白-2 无定形硫酸钙 细胞培养 骨髓间充质干细胞 细胞毒性 ??? Abstract:［Objeetive］ To develop the recombinant human bone morphogenetic protein2(rhBMP2)loaded amorphous calcium phosphate(ACP)delayed release nanosized material，investigate its cytotoxicity of cell，and provide a reference for the experiment of composite material in vivo．［Method］The rhBMP2／ACP delayed release nanosized material were prepared by chemical wet method and cultured on rabbit bone marrowderived mesenchymal stem cells(BMSCs)in vitro．Then the adhesion，proliferation，growth and functional expression of BMSCs were measured．［Result］Cytotoxicity test demonstrated that rhBMP2／ACP delayed release nanosized material had not affect the percentage of cell’s proliferation with materialextracted liquid cultured with BMSCs and the cytotoxicity was graded zero．The adhesion，proliferation，configuration of the cells on the surface of this material were identical to the control group．［Conclusion］It was suggested that rhBMP2／ACP delayed release nanosized material might have good cellular biocompatibility,no cytotoxicity and not effected the normal functional expression of BMSCs in vitro． ??? Key words:rhBMP2；amorphous calcium phosphate；cell culture；BMSCs；cytotoxicity ??? 目前，骨修复替代材料的研究在生物取材、细胞的体外培养、支架材料复合体等诸多方面积聚了丰厚的基础。无定形磷酸钙(amorphous calcium plosphate，ACP)是在采用湿化学法合成羟基磷灰石(hydroxyapatite，HA)时发现一种磷酸钙的无定形中间相，是一类X射线衍射为非晶态的磷酸钙材料的总称，具有典型的无定形物质的球形面貌。研究发现ACP的骨传导及细胞黏附性能优于HA〔1〕，生物降解速率高于磷酸三钙〔2〕，可降解可任意塑形，其化学组分、磷酸钙颗粒的表面形态、表面亲水性等，能满足药物分子和人体细胞对载体的多重要求〔3〕，但ACP不具备诱导成骨能力、降解速率不易控制等缺点。作者采用ACP包裹重组人骨形态发生蛋白2(recombinant human bone morphogenetic protein2，rhBMP2)，通过湿化学法合成技术，制备出rhBMP2