摘要
背景:骨形态发生蛋白2是目前发现的唯一的一种可单独诱导骨形成的生长因子,但是研究发现骨形态发生蛋白2单一基因的促血管生成作用较弱,无法在新生骨组织处产生足量的毛细血管,所以在诱导骨形成过程中还需要与其他诱导因子协同作用。低氧诱导因子1α在促进新生血管的形成方面有重要作用。目的:观察腺病毒介导的人骨形态发生蛋白2联合突变型低氧诱导因子1α(Ad-BMP-2-IRES-HIF-1α^(mu))转染骨髓间充质干细胞对激素性股骨头缺血性坏死的修复作用。方法:(1)转染Ad-BMP-2-IRES-HIF-1α^(mu)到骨髓间充质干细胞,检测碱性磷酸酶活性、钙结节茜素红染色检测其成骨能力;(2)建立激素性股骨头缺血性坏死兔模型,随机分为3组:实验组股骨坏死部位移植转染Ad-BMP-2-IRES-HIF-1α^(mu)的骨髓间充质干细胞;对照组移植未转染的骨髓间充质干细胞;空白组移植细胞培养液。移植8周后,Westen blot法检测骨形态发生蛋白2和低氧诱导因子1α蛋白表达;苏木精-伊红染色组织学观察股骨头缺血性坏死修复情况;墨汁灌注透明切片血管形态学观察股骨头缺损区域血管生成情况。结果与结论:(1)转染Ad-BMP-2-IRES-HIF-1α^(mu)的骨髓间充质干细胞的碱性磷酸酶活性和钙结节数量均高于未转染的细胞,说明转染Ad-BMP-2-IRES-HIF-1α^(mu)能促进骨髓间充质干细胞的成骨分化;(2)实验组骨形态发生蛋白2和低氧诱导因子1α蛋白表达高于对照组与空白组(P<0.05);移植8周后,实验组缺损区周边观察到明显的成骨反应,与对照组及空白组相比,实验组空骨陷窝率低(P<0.05),骨小梁面积百分比高(P<0.05);墨汁灌注实验显示:实验组出现血管化现象,并出现骨陷窝结构,血管间有清晰的连通脉络相互间构成网络,股骨头下血管分布及血管网的形成基本正常,实验组墨汁灌注血管面积为114.22±3.78,高于对照组(63.78±2.61)和空白组(21.39±
BACKGROUND： Bone morphogenetic protein 2（BMP-2） is the unique growth factor that independently induces osteogenesisbut its promoting angiogenesis is weakand cannot produce sufficient capillaries in the newly-born bonesthereafterother auxilliary factors are necessary. Hypoxia-inducible factor 1α（HIF-1 α） plays an important role in promoting angiogenesis. OBJECTIVE： To study the repair capacity of the adenovirus mediated BMP-2 combined with mutant HIF1α（Ad-BMP-2-IRES-HIF-1α^（mu）） transfecting bone marrow mesenchymal stem cells（BMSCs） in steroid-induced avascular necrosis of the femoral head. METHODS： Ad-BMP-2-IRES-HIF-1α^（mu） was transfected to BMSCs,the alkaline phosphatase activity and calcium nodules alizarin red staining were conducted to detect the osteogenic ability. The rabbit model of steroid-induced avascular necrosis of the femoral head was establishedand then divided into three groups： The cells were divided into three groups： Ad-BMP-2-IRES-HIF-1α^（mu） transfected BMSCs were transplanted into group Aand untransfected BMSCs were transplanted into group Band only the cell culture medium was added in the group C. At 8 weeks after transplantationthe expression levels of BMP-2 and HIF-1α were detected by western blot assay; the repair of avascular necrosis of the femoral head was observed by hematoxylin-eosin staining; the angiogenesis of the necrotic region was observed by toner perfusion transplantation vascular morphology. RESULTS AND CONCLUSION： The alkaline phosphatase activity in the transfected BMSCs and number of calcium nodules were higher than those in the untransfected BMSC,sindicating that Ad-BMP-2-IRES-HIF-1α^（mu） could promote the osteogenic differentiation of BMSCs. The expression levels of BMP-2 and HIF-1α in the group A were significantly higher than those in the groups B and C（P〈0.05）. Compared with groups B and C,obvious osteogenesis was observed in group A,the percentage of empty lacunae was significantly decreased,and trabecular b
作者
胡亮
王军海
王志烈
谢金元
陈登
丁凡
Hu Liang;Wang Jun-hai;Wang Zhi-lie;Xie Jin-yuan;Chen Deng;Ding Fan(Department of Joint Surgery;Department of Traumatic Hand and Foot Surgery,Jingmen First People＇s Hospital,Jingmen 448000,Hubei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第28期4440-4446,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金项目资助(81401827)~~
