血管内皮生长因子/纳米晶胶原基骨缓释系统与人骨髓间充质干细胞的体外生物相容性  

In vitro biocompatibility between vascular endothelial growth factor/nanohydroxyapatite collagen slow-release system and human bone marrow mesenchymal stem cells

作　　者：徐晓峰[1] 刘小平[2] 王明伟[2] 刘进炼[3] 陈静家[2] 李阳[2] 张志坚[2] 

机构地区：[1]江苏大学附属医院骨科,江苏省镇江市212001 [2]江苏大学临床医学院,江苏省镇江市212013 [3]上海交通大学医学院苏州九龙医院骨科,江苏省苏州市215021

出　　处：《中国组织工程研究与临床康复》2010年第25期4636-4640,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：镇江市社会发展基金资助项目(SH2002019)课题名称:骨髓间质干细胞体外扩增回植对骨缺损的作用~~

摘　　要：背景:利用载体或缓释系统负载生长因子,既能保护生长因子的生物活性,又可以使生长因子缓慢释放,从而持续促进细胞生长及组织修复再生,是目前控制释放载体材料应用研究的方向之一。目的:观察血管内皮生长因子/纳米晶胶原基骨缓释系统的体外缓释效果及与种子细胞的生物相容性。方法:人骨髓间充质干细胞经体外诱导培养、扩增后种植于血管内皮生长因子+肝素/纤维连接蛋白+纳米晶胶原基骨支架(实验组)和单纯的纳米晶胶原基骨支架(对照组)行体外培养。测定缓释支架材料上血管内皮生长因子的释放量和持续时间;种植细胞后细胞的黏附率;培养3,7,10,14d时支架材料中细胞数、碱性磷酸酶活性以及细胞在材料上的生长状况。结果与结论:①该缓释支架具有一定的血管内皮生长因子缓释效果,持续时间可达14d。②第3代人骨髓间充质干细胞经成骨诱导培养,可表达成骨细胞表型:碱性磷酸酶细胞化学染色、I型胶原免疫荧光染色均为阳性。③体外实验显示该缓释支架与人骨髓间充质干细胞具有优良的生物相容性:相同时间点实验组的细胞黏附率、支架上细胞数量及细胞碱性磷酸酶活性均明显高于对照组;扫描电镜发现两组材料上均有细胞生长,但实验组的细胞生长状况明显好于对照组。BACKGROUND：Using carrier or slow-release system to load growth factor can protect the biological activity of the growth factor and slow down its release,which enhances the cell growth and tissue repairing,regeneration constantly.This is one direction to the exploratory development of controlled release carrier.OBJECTIVE：To observe the effect of vascular endothelial growth factor（VEGF）/nano-hydroxyapatite collagen（nHAC） slow-release system and the in vitro biocompatibility with human bone marrow mesenchymal stem cells（MSCs）.METHODS：MSCs obtained from human bone marrow were induced,proliferated in vitro and seeded onto the nHAC scaffolds loaded with VEGF,heparin and fibronectin（experimental group） or single nHAC scaffolds（control group）.The quantity and the duration of VEGF release from the scaffolds which loaded with VEGF were determined;the attachment rate of cells on the scaffolds,number of cells in the scaffolds,and alkaline phosphatase（ALP） activity at different time points（3,7,10,14 days） were observed.The cell growth on scaffolds was also observed.RESULTS AND CONCLUSION：①The release of VEGF on the scaffolds of experimental group could last for 14 days.②The differentiation of MSCs into osteoblastic phenotype was demonstrated by the positive staining of alkaline phosphatase（ALP） and collagen type I.③The adhesive rates,cell numbers and the expression of the ALP at the same time point in the scaffolds of experimental group were significantly greater compared with control group（P 0.05）.④Cells could be observed on every scaffold by scanning electron microscopy,and the cells in experimental group grew better than the control group.

关 键 词：血管内皮生长因子 肝素 纤维连接蛋白 纳米晶胶原基骨 人骨髓间充质干细胞 

分 类 号：R318[医药卫生—生物医学工程]