组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2009, Vol. 5 ›› Issue (2): 65-69.doi: 10.3969/j.issn.1673-0364.2009.04.002

• 论著 • 上一篇    下一篇

转染人血管内皮生长因子基因的组织工程骨修复眼眶壁骨缺损的实验研究

肖彩雯，范先群，周慧芳，毕晓萍，沈琴，王业飞   

  1. 上海交通大学医学院附属第九人民医院眼科;
  • 发布日期:2020-07-23

VEGF Gene Transfected Human BMSCs Acting as Seeding Cell for Tissue Engineering Bond in Orbital Wall Bone Defects

XIAO Caiwen,FAN Xianqun,ZHOU Huifang,BI Xiaoping,SHEN Qin,WANG Yefei   

  1. Department of Ophthalmology;Shanghai Ninth People s Hospital;Shanghai Jiaotong University School of Medicine;Shanghai 200011;China;
  • Published:2020-07-23
  • Contact: 上海市重点学科建设项目(S30205) 上海交通大学博士创新基金(BXJ0826) 上海市自然科学基金(08zr1412900)

摘要： 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...

关键词: 血管内皮生长因子, 骨髓基质干细胞, 组织工程骨, 血管化, 眶壁骨缺损

Abstract: 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...

Key words: Vascular endothelial growth factor, Bone marrow stromal cells, Tissue engineered bone, Vascularity, Orbital wall bone defect

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