基于内皮祖细胞及VEGF165基因促血管生成作用促进组织工程神经修复犬大段周围神经缺损

Clinical medicine has urgent needs for large tissue engineering nerve defects regeneration; however, the vascularization problem of current tissue engineered nerve has not solved, thus limiting the progress of large nerve defect repairing. Previously, we successfully constructed tissue engineered nerve. In this project, we think the early vascularization play an important role in surviving of tissue engineered nerve in vivo, and propose a new method for large nerve defect reparing by vascularized tissue engineered peripheral nerve. We use recombinant adenovirus carrying VEGF165 gene to transfect autologous endothelial progenitor cells, and obtain Schwann-like cells induced by autologous adipose-derived stem cells; the two kinds of cells were used as the seed cells together. The allogeneic acellular nerve scaffolds were prepared used improved method by our research group. Then, the vascularized tissue engineered peripheral nerves were constructed. The dog large sciatic nerve defect was repaired by them. General observation, nerve electrophysiology and histological observation were used to evaluate the repairing effect. Investigate the reparing effect on vascularized tissue engineered peripheral nerve based on the angiogenesis effect of endothelial progenitor cell and VEGF165. The project would provide new methods and thinking for large peripheral nerve repair and reconstruction.

临床医学对大段周围神经缺损的修复有迫切需求，然而目前构建的神经移植物的血管化问题是困扰大段周围神经缺损修复的重要瓶颈之一。本项目在前期构建组织工程化周围神经的基础上，认为移植物早期血管化对组织工程化周围神经在体内存活起重要作用，提出促血管化组织工程化周围神经促进大段周围神经缺损修复的新方法，即利用携带血管内皮生长因子165基因（VEGF165）的重组腺病毒转染自体内皮祖细胞（EPCs），并与自体脂肪干细胞（ADSCs）诱导后的类许旺细胞共同作为种子细胞，利用本课题组改良制备的同种异体脱细胞神经支架材料作为载体，构建促血管化组织工程化周围神经，并修复犬坐骨神经大段缺损（6cm），通过大体观察、神经电生理检测、组织学观察等方法评估神经修复情况。探讨基于VEGF165基因及EPCs促血管生成作用构建的神经移植物对动物体内大段神经缺损修复的影响。本研究将为大段周围神经缺损的修复提供新的方法和思路。

临床医学对大段周围神经缺损的修复有迫切需求，然而目前构建的神经移植物的血管化问题是困扰大段周围神经缺损修复的重要瓶颈之一。本项目在前期构建组织工程化周围神经的基础上，认为移植物早期血管化对组织工程化周围神经在体内存活起重要作用，提出促血管化组织工程化周围神经促进大段周围神经缺损修复的新方法。本课题利用携带血管内皮生长因子165基因（VEGF165）的重组腺病毒转染自体内皮祖细胞（EPCs），并与自体脂肪干细胞（ADSCs）诱导后的类许旺细胞共同作为种子细胞，利用本课题组改良制备的同种异体脱细胞神经支架材料作为载体，成功构建了促血管化组织工程化周围神经，并成功修复了大鼠坐骨神经缺损（2cm）。分别以自体神经移植及普通组织工程化周围神经为对照组，通过对各组大体观察、SFI坐骨神经功能评价试验、超声下观察、神经电生理检测、腓肠肌湿重比、组织学观察及分子生物学检测等来比较各组移植物血管化程度及神经修复情况。结果表明基于VEGF165基因及EPCs促血管生成作用构建的血管化组织工程化周围神经能明显提高动物体内大段神经缺损修复的效果。本研究将为大段周围神经缺损的修复提供新的方法和思路。

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