骨髓间充质干细胞与血管内皮生长因子165基因转染人脐静脉内皮细胞共移植促进缺血皮瓣血管的生成  

Promoting angiogenesis by con-transplantation of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells transfected with vascular endothelial growth factor 165 gene

作　　者：先德彬 明华伟 徐荣胜 夏德林 Xian De-bin;Ming Hua-wei;Xu Rong-sheng;Xia De-lin(Department of Oral and Maxillofacial Surgery,Affiliated Stomatological Hospital of Southwest Medical University,Luzhou 646000,Sichuan Province,China)

机构地区：[1]西南医科大学附属口腔医院口腔颌面外科,四川省泸州市646000

出　　处：《中国组织工程研究》2018年第1期46-52,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:如何促进大块组织工程骨早期血管化是目前研究的热点。通过细胞共培养以及添加生物活性因子来促进血管生成均是很好地促进早期血管化的方法。目的:探讨骨髓间充质干细胞与血管内皮生长因子165(vascular endothelial growth factor,VEGF165)基因转染的人脐静脉内皮细胞共移植在体内促进血管生成的能力,为构建血管化组织工程骨修复大段骨缺损提供理论依据和实验基础。方法:50只SD大鼠完全随机分入5组,每组10只,于SD大鼠背部中线上建立一个4 cm×1.5 cm大小的缺血皮瓣模型,分别移植骨髓间充质干细胞与VEGF165基因转染的人脐静脉内皮细胞(A组)、单独转染VEGF165基因的人脐静脉内皮细胞(B组)、骨髓间充质干细胞与未转染VEGF165基因的人脐静脉内皮细胞(C组)、未转染VEGF165基因的人脐静脉内皮细胞(D组)、DMEM培养基(E组)。ELISA检测外周血VEGF水平,组织学观察其皮瓣存活状况及微血管密度。结果与结论:(1)术后A组皮瓣存活质量较其他4组高;(2)A组、B组术后第2,4,7,14天外周血中的VEGF持续高表达且逐渐升高,第7天时达到峰值,第14天时较术后第2天明显下降。不同时期A组外周血中VEGF水平明显高于其他4组,差异有显著性意义(P<0.05);(3)术后第11天,A组皮瓣存活率高于其他4组,差异有显著性意义(P<0.05);(4)术后第11天,A组微血管密度远远大于其他4组,差异有显著性意意义(P<0.05);(5)结果表明,骨髓间充质干细胞与VEGF165基因转染人脐静脉内皮细胞共移植可促进缺血皮瓣血管化,提高缺血皮瓣存活率。BACKGROUND:How to promote the early vascularization of large tissue-engineered bone has become the hotspot of current research.Cell co-culture and the addition of bioactive factors to promote angiogenesis are very good methods to promote early vascularization.OBJECTIVE:To explore the ability of angiogenesis by co-transplantation of bone marrow mesenchymal stem cells(BMSCs)and human umbilical vein endothelial cells(HUVECs)which were transfected with vascular endothelial growth factor 165(VEGF165)gene in v iv o,to move forward a single step to offer theoretical basis and experimental basis to build vascularized tissue-engineered bone which can be used to repair large segmental bone defects.METHODS:We built an ischemic skin flap with 4 cm><1.5 cm in the back of Sprague-Dawley rats,and then BMSCs+VEGF165-transfected HUVECs(group A),VEGF165-transfected HUVECs(group B),BMSCs+non-transfected HUVECs(group C),non-transfected HUVECs(group D),DMEM(group E)were respectively transplanted.ELISA method was used to detect peripheral blood VEGF level.Histologically,survival and microvessel density of the flap were observed.RESULTS AND CONCLUSION:(1)The flap survival quality of group A was better than that in the other groups.VEGF exhibited high expression continuously high expression at 2,4,7,14 days after transplantation,and reached the peak at 7 days,but the expression level at 14 days was obviously lower than that at 2 days postoperatively.The VEGF level of group always exceeded that in group B at different time points(P<0.05).The flap survival rate and microvessel density of group A was significantly higher than that in the other groups at 11 days postoperatively(both P<0.05).In summary,co-transplantation of BMSCs and VEGF165-transfected HUVECs can promote survival of an ischemic flap in v iv o through pro-angiogenic actions.

关 键 词：骨髓 间质干细胞 脐静脉 内皮细胞 血管内皮生长因子类 转染 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]