脐带Wharton胶来源的仿生化支架/干细胞复合构建组织工程神经及其体内外生物学评价

Although various kinds of methods including autologous nerve grafting were available for the peripheral nerve lesions repair. Unfortunately, the result of nerve function recovery was still undesirable. The tissue engineering nerve technique has provided a promising alternative for nerve repair. However, there are many problems which should be resolved as soon as possibly: a new type scaffold biomaterial should be researched and developed; the never scaffold was designed with biomimetic chemical composition and microstructure; the stem cell was induced effectively to neural cell and induction result should be detected exactly. To aim directly at above-mentioned difficulties and according to related researches of microfluidi、decellularization and fabrication of oriented scaffold，our research team plan to make a exploratory study as follows: the umbilical cord mesenchymal stem cells (UCMSCs) and Wharton's jelly were got out from umbilical cord; the optimal induction concentration of UCMSCs being induced into Schwann cell by bFGF and PDGF was detected by Microfluidic Technology; the ECM origined from decellularized Wharton's jelly was fabricated into tissue engineering nerve conduit scaffold with biomimetic orientation microchannel structure. The microstructure and property of nerve conduit was assessed biologically in vivo and in vitro; The tissue engineering nerve was constructed by conduit scaffold with implanted induced UCMSCs.The cell affinity, orientation and function in tissue engineering nerve was evaluated. We investigated the feasibility that the UCMSCs or combined structure (oriented Wharton's jelly ECM conduit scaffold implanted induced UCMSCs) could be applied as tissue engineering nerve. The experimental results provided theoretical and technological base for experiment of animal nerve repair and clinical application.

包括自体神经移植在内的各种方法应用于周围神经损伤修复，但神经功能恢复效果仍然不理想。组织工程神经技术为解决该难题提供了可能，但目前仍然存在急需解决的难题：研发新型支架材料、支架成份及结构的仿生化设计、干细胞向神经细胞的有效诱导及准确检测等。课题组拟针对以上难题,结合前期微流控芯片、脱细胞及仿生化定向支架制备技术，进行以下探索性研究：脐带中提取脐带间充质干细胞（UCMSCs）及脐带Wharton胶；通过新型微流控芯片技术，筛选bFGF联合PDGF诱导UCMSCs向雪旺细胞转化的最佳诱导浓度；Wharton胶脱细胞获得细胞外基质材料，并利用其制备仿生化定向结构的神经导管组织工程支架，对其结构及性质进行体内外生物学评价；将诱导转化的UCMSCs种植于支架，评价细胞亲和性、排列及功能情况。探讨其在神经组织工程应用的可行性。为进一步神经修复动物实验及临床应用提供理论与技术基础。

周围神经损伤修复后，神经功能恢复效果不理想。组织工程神经技术为解决该难题提供了可能。目前仍然存在急需解决的难题：包括研发新型支架材料、支架成份及结构仿生化设计、干细胞向神经细胞诱导分化等。针对以上难题,结合前期微流控芯片、脱细胞及仿生化定向支架制备技术工作。课题组设立了脐带Wharton胶来源的仿生化支架/干细胞复合构建组织工程神经及其评价的可行性研究方案。课题组按计划完成了研究内容，包括：脐带Wharton胶中优化分离培养并鉴定脐带间充质干细胞（UCMSCs），研制微流控芯片并筛选UCMSCs向雪旺细胞分化的诱导浓度，利用脱细胞脐带Wharton胶制备仿生化取向神经导管支架，进行支架的表征及评价。研究结果显示：脐带Wharton胶中既分离UCMSCs又获得脱细胞Wharton胶ECM材料；UCMSCs具备干细胞特性，微流控芯片优化了UCMSCs向雪旺细胞分化的诱导浓度；制备的神经导管支架具有仿生化取向结构及良好生物学性质。本项基金支持下发表SCI论文4篇。本课题较全面的回答了课题立项时提出的问题，按照课题任务要求完成了各项指标。