高精度DLP打印的导电神经导管联合电刺激促面神经再生及机制研究

面神经缺损导致的面瘫是临床治疗的难点，目前自体神经移植及组织工程神经导管均无法完成带分支神经缺损的修复。本人前期通过水凝胶收缩制造技术构建了10μm级高精度的导管（Nat Commun，2020），并成功使用DLP打印技术制造了内壁带微条纹的导管，定向生长的神经干细胞可高表达神经相关标志物；并发现初级纤毛可感知电信号。据此推测DLP制造的带分支PEDOT/GelMA神经导管可在结构与功能上高度仿生，细胞可通过初级纤毛感受电刺激并调控细胞内钙离子及下游信号，促神经向分化。为验证此假说，本研究拟构建带分支的导电神经导管，探讨电刺激对神经修复的作用，重点研究初级纤毛上PC1/PC2复合体如何受电信号调节，CaMKII如何整合钙离子信号并控制CREB行为，阐明初级纤毛-钙离子-CaMKII-CREB的新型信号通路促神经生长的作用，深化电信号调控细胞行为的分子机制，为新型神经导管的临床应用提供依据。

It is difficult to treat facial paralysis caused by facial nerve defect. In the present study, neither autologous nerve transplantation nor tissue-engineered nerve guidance conduits can complete facial nerve repair with branches. Before, I constructed conduits with a complexation-induced resolution enhancement manufacturing technology (precision of 10μm) and named it shrinking printing (Nat Commun, 2020). In addition, I successfully used DLP printing technology to manufacture multi-branch nerve conduits with directional microgroove on the inner wall. The neural stem cells could grow in a direction, and highly express neuro-related markers. The primary cilia may be involved in sensing electrical signals. Therefore, it is speculated that personalized PEDOT/GelMA nerve conduits can be highly biomimetic in structure and function, and cells can sense electrical signals through primary cilia, thus regulate intracellular Ca2+ and downstream signals to promote neurogenic differentiation. In order to prove this hypothesis, PEDOT/GelMA nerve conduits with great electrical conductivity are constructed to investigate the effect of electrical stimulation on nerve repair. Meanwhile, we will focus on the molecular mechanism of nerve cells sensing electrical signals, discuss how PC1/PC2 complex on primary cilia is regulated by electrical signals, how CaMKII integrates calcium signal and controls CREB behavior, and clarify the role of a new signaling pathway of primary cilia-Ca2+-CaMKII-CREB in promoting neurite growth. This study deepens the molecular mechanism of electrical signals regulating cell behavior, and provides theoretical support and experimental basis for the clinical application of new tissue-engineered nerve guidance conduits.