负载外泌体FGF10的取向水凝胶调控eNSCs定向迁移及凋亡促进脊髓损伤修复机制研究

Spinal cord injury is a frequently-occurring disease in spinal surgery. At present, there is a lack of effective treatment methods. Tissue engineering technology provides the possibility for the repair of spinal cord injury. In the early stage, the research group prepared dextran-gelatin hydrogel scaffolds for the repair of spinal cord injury, which can guide the directional growth of mesenchymal stem cells. As a new hot spot, exosomes provide a new direction for the repair of spinal cord injury. Compared with cell transplantation, cell-free therapy has no risk of exogenous cell differentiation, mutation, tumorigenicity and so on. Neural stem cell exosomes FGF10 has neuroprotective effect and can activate Akt pathway. Therefore, a cell-free spinal cord regeneration strategy was proposed in this study, and a functionalized oriented hydrogel loaded with exosomes FGF10 was constructed to repair spinal cord injury. On the basis of previous work, this project intends to: (1) construct directional hydrogel scaffolds and explore the feasibility of their application in the repair of spinal cord injury; (2) load NSCs-Exos-FGF10 into hydrogel materials, to verify the possibility of repairing spinal cord injury, (3) to clarify the effect of directional hydrogel material and NSCs-Exos-FGF10 on optimizing the microenvironment of injured spinal cord, and the effect and mechanism of inducing directional migration of endogenous neural stem cells and inhibiting apoptosis, It provides a new research direction and treatment strategy for the repair of spinal cord injury.

脊髓损伤是脊柱外科多发病，目前尚缺乏有效治疗手段，组织工程技术为脊髓损伤修复提供了可能。课题组制备了用于修复脊髓损伤的葡聚糖-明胶水凝胶支架，可引导间充质干细胞取向性生长。作为新兴热点，外泌体为脊髓损伤的修复提供了新的方向，与细胞移植相比，无细胞疗法不存在外源性细胞分化、突变、致瘤性等风险。神经干细胞外泌体FGF10具有神经保护作用，同时可激活Akt通路，因此，本研究提出无细胞脊髓再生策略，拟构建负载外泌体FGF10的功能化取向水凝胶用于修复脊髓损伤。本项目拟在前期工作基础上：①构建取向性水凝胶支架材料，探讨其应用于脊髓损伤修复的可行性；②将NSCs-Exos-FGF10负载于水凝胶材料中，验证其修复脊髓损伤修复的可能性；③明确取向性水凝胶材料及NSCs-Exos-FGF10优化损伤脊髓微环境的效果，及诱导内源性神经干细胞定向迁移、抑制凋亡的效果及机制，为脊髓损伤修复提供新的研究方向。

构建取向性水凝胶支架材料，完成相关理化性能及生物学性能检测，探讨其应用于脊髓损伤修复的可行性； 采用新型的取向水凝胶生物活性脊髓作为载体，将 NSCs-Exos 负载于水凝胶材料中，验证其修复脊髓损伤修复的可能性；在专业学术刊物上发表 SCI 收录学术论文3篇；我们成功的构建了脊髓损伤特异性水凝胶支架，并构建了稳定的SD大鼠脊髓损伤模型，仿生水凝胶的设计为我们的临床转化奠定了扎实的数据基础；同时，我们通过分子模拟机制，为miRNA模拟物和抗miRNA的临床和实验设计提供了指导，并为进一步工程AGO蛋白作为基因调控领域的新工具开辟了道路，病探究了Ca2+/CaM与死亡相关蛋白激酶1 (DAPK1)结合，调节细胞内信号通路相关机制。