Mu Xiaodan, Liu Huawei, Yang Shuhui, Li Yongfeng, Xiang Lei, Hu Min, Wang Xiumei
Department of Stomotology, The First Medical Centre, Chinese PLA General Hospital, Beijing 100853, China.
Department of Materials Science and Engineering, State Key Laboratory of New Ceramics and Fine Processing, Tsinghua University, Beijing 100084, China.
ACS Omega. 2022 May 26;7(22):18509-18520. doi: 10.1021/acsomega.2c01176. eCollection 2022 Jun 7.
Facial nerve injury is a common clinical condition that leads to disfigurement and emotional distress in the affected individuals, and the recovery presents clinical challenges. Tissue engineering is the standard method to repair nerve defects. However, nerve regeneration is still not satisfactory because of poor neovascularization after implantation, especially for the long-segment nerve defects. In the current study, we aimed to investigate the potential of chitosan tubes inoculated with stem cell factor (SCF) and dental pulp stem cells (DPSCs) in facial nerve-vascularized regeneration. In the in vitro experiment, DPSCs were isolated, cultured, and then identified. The optimal concentration of SCF was screened by CCK8. Cytoskeleton and living-cell staining, migration, CCK8 test, and neural differentiation assays were performed, revealing that SCF promoted the biological activity of DPSCs. Surprisingly, SCF increased the neural differentiation of DPSCs. The migration and angiogenesis experiments were carried out to show that SCF promoted the angiogenesis and migration of human umbilical vein endothelial cells (HUVECs). In the facial nerve, 7 mm defects of New Zealand white rabbits, hematoxylin-eosin (HE), immunohistochemistry, toluidine blue staining, and transmission electron microscopy observation were performed at 12 weeks postsurgery to show more nerve fibers and better myelin sheath in the SCF + DPSC group. In addition, the whisker movements, Masson's staining, and western blot assays were performed, demonstrating functional repair and that the expression level of CD31 protein in the group SCF + DPSCs was relatively close to that in the group Autograft. In summary, chitosan tubes inoculated with SCF and DPSCs increased neurovascularization and provided an effective method for repairing facial nerve defects, indicating great promise for clinical application.
面神经损伤是一种常见的临床病症，会导致患者面部毁容并产生情绪困扰，其恢复过程面临临床挑战。组织工程是修复神经缺损的标准方法。然而，由于植入后新生血管形成不佳，神经再生效果仍不尽人意，尤其是对于长段神经缺损。在本研究中，我们旨在探究接种干细胞因子（SCF）和牙髓干细胞（DPSC）的壳聚糖管在面神经血管化再生中的潜力。在体外实验中，分离、培养并鉴定了DPSC。通过CCK8筛选出SCF的最佳浓度。进行了细胞骨架和活细胞染色、迁移、CCK8检测以及神经分化测定，结果表明SCF促进了DPSC的生物学活性。令人惊讶的是，SCF增加了DPSC的神经分化。进行迁移和血管生成实验以表明SCF促进了人脐静脉内皮细胞（HUVEC）的血管生成和迁移。在新西兰白兔的面神经7毫米缺损模型中，术后12周进行苏木精-伊红（HE）染色、免疫组织化学、甲苯胺蓝染色和透射电子显微镜观察，结果显示SCF + DPSC组有更多神经纤维和更好的髓鞘。此外，进行了触须运动、Masson染色和蛋白质印迹分析，证明了功能修复，并且SCF + DPSC组中CD31蛋白的表达水平与自体移植组相对接近。总之，接种SCF和DPSC的壳聚糖管增加了神经血管化，为修复面神经缺损提供了一种有效的方法，显示出巨大的临床应用前景。
