TiO2纳米管负载BMP-2指节肽对骨髓间充质干细胞增殖与分化的影响

摘要

Objective To investigate the effect of the complex structure of titanium dioxide (TiO2) nanotubes loaded with BMP-2 knuckle peptide on the proliferation and differentiation of bone marrow mesenchymal stem cells. Methods The TiO2nanotube arrays were manufactured by the method of anodic oxidation. The MSCs were isolated from the bone marrow of Wistar rats by all bone marrow adherence method. In the experimental group,the TiO2nanotubes titanium plate was loading BMP-2,while in the control group,only TiO2nanotubes titanium plate was used. The titanium plate of the two groups was inoculated with MSCs. Samples were assayed by MTT and ALP. OCN and OPN RNA expression were examined. Results After 1,3,5 days of culture,the cell proliferation in the experimental group was greater tnat that in the control. After 7,10,15 days,the alkaline phosphatase was more active in the experimental group than that in the control. After 21 days,the fold change of OCN and OPN was greater in the experimental group than that in the control. Conclusion The promoting effect of TiO2nanotubes loaded with BMP-2 knuckle peptide on the proliferation and early osteogenetic differentiation of bone marrow mesenchymal stem cells was stronger than that of pure TiO2nanotubes.%目的 检测负载骨形态发生蛋白2(bone morphogenetic protein 2, BMP-2)指节肽的二氧化钛(TiO2)纳米管复合结构对骨髓间充质干细胞增殖与成骨分化的影响.方法 通过阳极氧化法在钛片表面制备TiO2纳米管阵列.全骨髓差速贴壁法分离培养Wistar大鼠骨髓间充质干细胞.实验组负载BMP-2指节肽TiO2纳米管;对照组只采用TiO2纳米管.2组纳米管钛片接种间充质干细胞培养,测定细胞增殖、碱性磷酸酶活性、骨钙素(osteocalcin,OCN)及骨桥蛋白(osteopontin,OPN)RNA表达,并进行统计学分析.结果 2组细胞培养1、3、5天,细胞增殖实验组>对照组(P<0.05);2组细胞培养7、10、15天,碱性磷酸酶活性实验组>对照组(P<0.01);2组细胞培养21天后,骨钙素和骨桥蛋白的差异倍数(fold change)显示为实验组>对照组.结论 TiO2纳米管负载BMP-2指节肽对骨髓间充质干细胞增殖和早期成骨分化的促进作用强于单纯TiO2纳米管结构.