珊瑚羟基磷灰石负载含BMP-2纳米缓释微球体系促进人间充质干细胞骨形成的研究

Study on the bone formation of human mesenchymal stem cells by coral hydroxyapatite loaded with BMP-2 nanoscale microspheres

目的探讨珊瑚羟基磷灰石负载含骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)纳米缓释微球体系在促进人间充质干细胞(human mesenchymal stem cells,hMSCs)骨形成中的作用。方法从骨移植患者中收集hMSCs,分离培养后使用BMP-2纳米微球作为载体,装载到珊瑚羟基磷灰石(coral hydroxyapatite,CHA)支架上。将CHA-BMP-2-hMSCs与CHA-hMSCs分别植入两组小鼠的L4和L5横向软组织中,10周后检测小鼠碱性磷酸酶(alkaline phosphatase,ALP)活性,通过Western blot检测Runx2蛋白与骨桥蛋白表达水平,通过显微镜观察骨质生长情况。结果 CHA-BMP-2-hMSCs小鼠的支架上骨组织覆盖面积显著大于CHA-hMSCs小鼠,ALP活性显著高于非缓释组小鼠,骨钙素、Runx2蛋白与骨桥蛋白表达水平高于非缓释组小鼠。结论 CHA-BMP-2-hMSCs缓释系统有利于在较长时间内诱导骨形成。

Objective To investigate the role of coral-hydroxyapatite-loaded BMP-2 nano-release microspheres in promoting bone formation of human mesenchymal stem cells. Methods Human mesenchymal stem cells(MSCs) were collected from bone transplant recipients. After isolation and culture, bone morphogenetic protein-2(BMP-2) was loaded onto coral hydroxyapatite(CHA) scaffolds. The CHA-BMP2-hMSCs and CHA-hMSCs were implanted into L4 and L5 transverse soft tissues of mice, respectively. After 10 weeks, alkaline phosphatase(ALP) activity in mice was detected, Runx2 protein and osteogenic protein expression were detected by Western blot, and bone growth was observed using microscope. Results Compared with CHA-hMSCs, the ALP activity of the sustained-release mice was significantly higher than that of the non-sustained-release mice, and the osteocalcin, Runx2 and osteopontin levels were higher than those of the non-sustained release group mice. Conclusion CHA-BMP2-hMSCs sustained release system is conducive to induce bone formation in a long time.