组织工程骨膜的构建及重组人骨形态发生蛋白-2对其体外成骨能力的影响  

Construction of Tissue Engineered Periosteum and Effects of Recombinant Human Bone Morphogenetic Protein-2 on Its Osteogentic Capacity in Vitro

作　　者：任广铁[1] 张苍宇[1] 拓振合[1] 庾佳佳[1] 孙瑞[1] 汪静[1] 赵琳[1] 

机构地区：[1]兰州大学第二医院骨科/甘肃省骨与关节疾病研究重点实验室,兰州730030

出　　处：《世界科技研究与发展》2014年第1期80-84,共5页World Sci-Tech R&D

基　　金：国家自然科学基金(30973064)资助

摘　　要：目的探讨重组人骨形态发生蛋白-2对组织工程骨膜体外成骨能力的影响,以期构建性能更加优良的产品用于临床。方法兔骨髓间充质干细胞经成骨诱导培养,接种于猪小肠黏膜下层构建组织工程骨膜,后分别对其使用含有100 ng/ml、50 ng/ml、10 ng/ml及不含有重组人骨形态发生蛋白-2的成骨诱导培养基进行培养,7天后行电镜扫描,并对各组在多个时间点采用MTT法绘制生长曲线、测定碱性磷酸酶和骨钙素含量,分析重组人骨形态发生蛋白-2对组织工程骨膜成骨能力的影响。结果加入重组人骨形态发生蛋白-2培养的组织工程骨膜在细胞增殖速度、碱性磷酸酶和骨钙素素表达量均高于未加入重组人骨形态发生蛋白-2组,且与浓度成正相关。结论重组人骨形态发生蛋白-2可提高组织工程骨膜体外成骨能力。Objective To investigate the effects of recombinant human bone morphogenetic protein-2 on osteogentic capacity in vitro of tissue-engineered periosteum which were constructed by porcine small intestinal submucosa（SIS） and the bone marrow mesenchymal stem cells（BMSCs）. Methods 150 ul lxl05cell/nd New Zealand rabbits BMSCs cultured and osteo- genie induced in vitro were seeded on O. 8 era2 round SIS to manufacture different group tissue-engineered periosteum （TEP）. The TEPs were divided into four groups and cultured with rhBMP-2 of different concentrations （100 ng/ml,50 ng/ ml,10 ng/ml,and 0 ng/ml）. Then growth curves of BMSCs on TEP of each group were detected by MTT,and the micro - structure of TEP were scanned by electronic microscopy at 7 d. Moreover, the the ALP activity and osteocalcin content of each group TEP were determined by ELISA kit on several different time points. Result Growth curves,ALP activity, and os- teocalcin content show that the TEP with rhBMP-2 get significantly better performance than none, and there is a positive cor- relation between rhBMP-2 doses and the TEP osteogentic capacity. Conclusion rhBMP-2 can enhance the TEP＇s osteogentic capacity in vitro,so it can be used to improve the TEP.

关 键 词：骨髓间充质干细胞 猪小肠粘膜下层 组织工程骨膜 骨形态发生蛋白-2 

分 类 号：R318.08[医药卫生—生物医学工程]