具有血管化功能/干细胞的复合组织工程化神经支架修复坐骨神经缺损的实验研究

The research concerning repairment of nerve deficit by tissue engineering artificial nerves progressed rapidly.However,the clinical applications were confined due to unsufficient active Schwann cells,as well as poor vascularization.Thus, we aim at promoting the vascularization and Schwann cells proliferation of tissue engineering artificial nerves.It was verified that BDNF is capable of facilitating vascularization and Schwann cells proliferation, and VPA prossesses the characteristic of inducing Schwann cells proliferation. In our study, endothelial progenitor cells (EPCs) transfected by BDNF gene were cultured on the surface of scaffold possessing the ability of controlled-release of VPA, on which the EPCs can secrete BDNF continously and efficiently.Sequently,the Schwann cells differentiated from adipose-derived stem cells (ADSC-SCs) were cultured on the previous scaffold for additional two to four days.The ability that BDNF secreted by EPCs has was evaluated by means of MTT and subcutaneous transplantation of host, and the new gene-enhanced tissue engineering nerve scaffold was used to bridge the nerve dificit with long segment.The effects of BDNF on vascularization and Schwann cells proliferation of gene-enhanced tissue engineering nerve were evaluated, which provided technical support and theoretical direction for protential clinical application of tissue engineering nerve scaffold.

采用组织工程化神经材料修复神经缺损的实验研究进展较快，但其临床应用受限的主要原因有组织工程化神经的血管化较差及缺乏充足的活性雪旺细胞。如何加快组织工程化神经的血管生成以及如何保证组织工程化神经植入体内后活性雪旺细胞数量，仍为制约组织工程化神经临床应用的难题。研究证明BDNF可加速组织工程化神经的血管生成和雪旺细胞增殖，我们前期研究已证实VPA亦具有促雪旺细胞增殖的功能。本研究以一种可缓释VPA的可吸收复合材料作为支架，将转染BDNF基因的内皮祖细胞种植于该支架，使其在此支架上持续高效分泌BDNF；再将诱导分化而来的ADSC-SCs种植于支架表面，进行体外培养。通过各项试验评价转基因内皮祖细胞分泌BDNF的促血管化和雪旺细胞增殖能力，并将基因增强的新型组织工程化神经支架植入修复长段神经缺损，探讨该组织工程化神经支架的有效性，为临床功能性组织工程化神经的制备提供技术支持和理论指导。

采用组织工程化神经材料修复神经缺损的实验研究进展较快，但其临床应用受限的主要原因有组织工程化神经的血管化较差及缺乏充足的活性雪旺细胞。如何加快组织工程化神经的血管生成以及如何保证组织工程化神经植入体内后活性雪旺细胞数量，仍为制约组织工程化神经临床应用的难题。研究证明BDNF可加速组织工程化神经的血管生成和雪旺细胞增殖，我们前期研究已证实VPA亦具有促雪旺细胞增殖的功能。本研究以一种可缓释VPA的可吸收复合材料作为支架，将转染BDNF基因的内皮祖细胞种植于该支架，使其在此支架上持续高效分泌BDNF；再将诱导分化而来的ADSC-SCs种植于支架表面，进行体外培养。通过各项试验评价转基因内皮祖细胞分泌BDNF的促血管化和雪旺细胞增殖能力，并将基因增强的新型组织工程化神经支架植入修复长段神经缺损，探讨该组织工程化神经支架的有效性，为临床功能性组织工程化神经的制备提供技术支持和理论指导。