周围神经缺损的治疗是尚未解决的世界性难题。自体神经移植作为临床治疗的金标准，受到供体来源不足等诸多因素的限制，亟需一种有效的替代物。因此，开发能够重建受损神经形态连续性、为神经再生提供良好的再生微环境的组织工程神经具有重要意义。本课题围绕"周围神经损伤修复"这一重大科学问题，拟构建电导活性人工神经，并将其与持续电刺激联合应用，为周围神经再生提供包括结构引导、电学微环境以及持续作用的生物学微环境为一体的多重、立体、交叉的刺激环境，以加快神经再生，并进行动物实验验证其修复有效性。同时，为阐明"电信号- - 生物学行为"的具体机制，分别以雪旺细胞和神经元为研究对象，深入展开电信号调控细胞的分子机制研究，为利于电信号缩短受损神经元休克期，重建再生微环境，加快神经再生，以及电导活性人工神经的临床应用提供理论基础和实验依据

The treatment of peripheral nerve defect is one of the unsolved clinical problems worldwide. The current gold standard for nerve defect is autologous nerve transplantation, whose application was restricted by limited donor nerve supply. Therefore, it is imperative to develop an alternative to autograft, which can restore the morphological structure of the damaged nerve, as well as provide favourable microenvironment for nerve regeneration. The present grant focused on "periphral nerve injury repair". We fabricated an electrically conductive activated nerve scaffold, which was seeded Schwann cells.Then Schwann cells was activated by electrical stimulation to build an multiple regenerative environment at the local site of nerve defect. The electrically conductive conduit seeded with electrically activated Schwann cells was used to bridge a large nerve defect to identify its efficacy in promtoing nerve regeneration in animals. The mechanism underlying the benificial effect of electrical stimulation on neurons and Schwann cells was explored to provide evidence for the application of electrically conductive nerve conduit in clinical settings

本研究从电刺激对神经细胞的调控作用入手，拟构建体内可以进行电刺激操作的电导活性人工神经，并评估其促进神经再生的有效性及机制。本研究首先发现适宜的电刺激可以通过上调BDNF等分子表达加快神经轴突的生长速度，进一步研究通过高通量miRNA芯片技术筛选了与电刺激加快神经轴突生长相关的miRNA分子群，通过对16种显著变化的miRNA进行验证及功能分析，最终发现2个重要的miRNA分子（miR-363-5p及miR-3102）在电刺激促进神经轴突生长中发挥关键作用。在此基础上，发现适宜的电刺激可促进雪旺细胞合成及分泌NGF及BDNF。钙成像结果表明：电刺激可开放细胞膜T型电压门控钙离子通道，同时活化细胞内IP3敏感性钙库及Ryanodine敏感性钙库，升高细胞内钙浓度，刺激雪旺细胞分泌NGF。而且电刺激可以调控雪旺细胞增殖、迁移、粘附等生物学活性。最后，本课题制备可降解导电材料PPy/chitosan，并将该导电材料制备成具有轴向矩阵排列微管样结构的电导活性人工神经，应用电导活性人工神经桥接大鼠15mm坐骨神经缺损，发现电导活性人工神经可显著加快神经再生，促进神经缺损后神经再生和功能恢复，为周围神经损伤修复提供了新思路。