转染血管内皮细胞生长因子骨髓间充质干细胞与牛松质骨支架复合组织工程骨的血管形成

摘要

背景:小块组织工程骨植入动物体内后,早期依靠组织液的渗透可获得营养,但大块组织工程骨的营养仅靠组织液的渗透是远远不够的,必须通过血管再生来获得.目的:观察转染血管内皮细胞生长因子表达载体骨髓间充质干细胞复合组织工程骨植入动物体内后的血管形成能力.方法:制作日本大耳白兔双侧尺骨中段骨缺损模型,左侧尺骨缺损植入转染血管内皮细胞生长因子表达载体的自体骨髓间充质干细胞复合脱钙脱脂去蛋白的牛松质骨支架组织工程骨为实验组,右侧尺骨缺损植入自体骨髓间充质干细胞复合脱钙脱脂去蛋白的牛松质骨支架组织工程骨为对照组.术后12周行X射线摄片观察、大体标本观察、苏木精-伊红染色和Masson染色组织切片观察、MiFas图像分析系统定量分析.结果与结论:实验组与对照组尺骨缺损处均有连续骨痂形成;组织切片经苏木精-伊红染色、Masson三色法染色及MiFas图像分析系统定量分析可见实验组和对照组均有大量的新生骨,但实验组新生血管明显多于对照组(P < 0.01),且血管较粗大,而且与新生骨接近.说明将转染血管内皮细胞生长因子表达载体的骨髓间充质干细胞复合在组织工程骨上植入动物体内可明显促进新生血管的形成.%BACKGROUND: After a small tissue engineering bone transplanted into animals, which can obtain enough nutrition by interstitialfluid penetration, but if a big one, it is insufficient by interstitial fluid penetration and must obtained by revascularization.OBJECTIVE: To observe the ability of bone marrow mesenchymal stem cells (BMSCs) transfected with vascular endothelialgrowth factor (VEGF) expression vector to promote vascularization on tissue engineering bone.METHODS: Bone defect model of the middle piece of both hand side ulnar bones of Japanese big ear rabbits was made.Transfected VEGF expression vector of autologous BMSCs to the protein of defatted bovine demineralized cancellous bonetissue engineering bone scaffold was implanted into left ulna defect as the experimental group. Autologous BMSCs combinedwith decalcifying, defatted and protein-free bovine cancellous bone scaffold of tissue engineering bone as the control group. At12 weeks after operation, X ray photo observation, general specimen observation, hematoxylin-eosin staining and Massonstaining histological section observation, MiFas picture analysis system quantitative analysis were performed.RESULTS AND CONCLUSION: The continuous calluses were formed in ulnar bone defect between experimental and controlgroups. Tissue section was stained with hematoxylin-eosin staining and Masson staining, and observed by MiFas pictureanalysis system quantitative analysis, which showed a large number of new bones between experimental and control groups, butnew vessels in experimental group was significantly more than that in control group (P ＜ 0.01); vessles were thick and closed tonew bones. It is indicated that BMSCs of transfected VEGF expression vector combined with tissue engineering bone andimplanted into animals can obviously promote the formation of new vessels.