生物活性氧化锆对成骨细胞粘附、增殖及矿化的影响

The influence of bio-active zirconia on the adhesion,proliferation and mineralization of osteoblast

目的观察精氨酸-甘氨酸-天冬氨酸三肽复合体修饰的氧化锆材料对小鼠成骨细胞粘附、增殖及矿化的影响。方法采用等离子喷涂的方法在钛片表面制备氧化锆涂层,经羟基化、硅烷化后在其表面连接精氨酸-甘氨酸-天冬氨酸三肽复合体(Arg-Gly-Asp tripeptide complex,RGD),并培养小鼠成骨细胞。使用激光共聚焦显微镜观察材料表面成骨细胞的粘着斑分布,并在培养24、48、72 h后进行细胞计数评价细胞增殖水平,用蛋白质印迹法检测成骨指标:碱性磷酸酶及骨钙素评价细胞矿化水平。结果带有RGD的氧化锆涂层表面成骨细胞形成更多粘着斑,增殖活性高,并且碱性磷酸酶及骨钙素表达水平升高。结论带有RGD的生物活性氧化锆涂层对成骨细胞的粘附、增殖及矿化具有促进作用。

Objective To investigate the influence of Arg-Gly-Asp tripeptide complex connected zirconia on the adhesion, prolifera- tion and mineralization of osteohlast. Methods Plasma spray was employed to prepare zirconia coating on titanium plate, after hydrox- ylation and silanization, Arg-Gly-Asp tripeptide complex was connected on zirconia coating. The focal adhesion of osteoblasts was oh- served by confocaI microscopy and the stained cell nucleus were counted to evaluate the proliferative ability of osteoblasts. Alkaline phosphatase and osteoealcin were measured to evaluate the minerahzation of osteoblast by western blot. Results Osteobiasts on Arg- Gly-Asp tripeptide complex connected zirconia presented more focal adhesion and excellent proliferative ability. The levels of alkaline phosphatase and osteocalcin were higher in Arg-Gly-Asp connected zirconia than those of regular zirconia. Conclusion Arg-Gly-Asp tripeptide complex connected bio-active zirconia can promote the adhesion, proliferation and osteogenic mineralization of osteoblast.