目的探讨以转染人血管内皮生长因子(Vascular endothelial growth factor,VEGF)基因的骨髓基质干细胞(Bone marrow stromal cells,BMSCs)构建的组织工程骨在犬眼眶壁骨缺损中的修复效果。方法体外分离扩增犬自体BMSCs至第2代,用腺病毒转染VEGF基因。采用Real-time PCR和Western Blot检测目的基因和蛋白表达情况。细胞接种在珊瑚支架材料上构建组织工程骨,Elisa检测转基因细胞在支架材料上的蛋白持续表达情况。成年比格犬24只,双侧眼眶内侧壁制造直径12mm圆形骨缺损模型,随机分为4组:A组植入转染VEGF的组织工程骨,B组植入未转染基因的组织工程骨,C组植入单纯珊瑚材料,D组为旷置组。分别在手术后4周、12周、24周取材,行大体观察、Micro-CT分析、免疫组化方法检测血管形成情况,以及组织学观察和组织形态学检测比较骨缺损修复效果。结果VEGF基因修饰的BMSCs能够高表达目的基因和蛋白,构建组织工程骨后能够在体外持续分泌VEGF蛋白22d。C组和D组均未修复眶壁骨缺损。A组在骨缺损修复过程中,4周时的新生血管形成量和新生骨体...

Objective To investigate the effect VEGF-gene transfected BMSCs to construct tissue engineered bone in the treatment of orbital wall bone defects.Methods In vitro isolated and expanded BMSCs of passage 2 were transfected with adenoviruses containing human VEGF gene,and the target gene expression was evaluated by Real-time PCR and Western Blot.BMSCs were harvested and seeded into the coral scaffolds to construct tissue engineered bone.VEGF protein secretion from the cells was detected using Elisa method.Bila...