Pan Yao, Tang Li, Dong Shuxian, Xu Mengjie, Li Qiong, Zhu Guochen
Department of Otorhinolaryngology-Head and Neck Surgery, Nanjing Medical University Affiliated Wuxi No. 2 People's Hospital, Wuxi, Jiangsu, China.
Department of Otorhinolaryngology-Head and Neck Surgery, Nantong University Affiliated Wuxi Clinical College, Wuxi, Jiangsu, China.
Stem Cells Dev. 2023 Jan;32(1-2):1-11. doi: 10.1089/scd.2022.0245. Epub 2022 Dec 27.
Previous studies showed that acellular nerve allografts (ANAs) have been successfully utilized in repairing peripheral nerve defects, and exosomes produced by stem cells are useful in supporting axon regrowth after peripheral nerve injury. In this study, exosomes from hair follicle epidermal neural crest stem cells (EPI-NCSCs-Exos) combined with ANAs were used to bridge facial nerve defects. EPI-NCSCs-Exos were isolated by ultracentrifuge, and were identified. After coculture, EPI-NCSCs-Exos were internalized into dorsal root ganglions (DRGs) and schwann cells (SCs) in vitro, respectively. EPI-NCSCs-Exos elongate the length of axons and dendrites of DRGs, and accelerated the proliferation and migration of SCs, and increased neurotrophic factor expression of SCs as well. The next step was to assign 24 Sprague Dawley male rats randomly and equally into three groups: the autograft group, the ANA group, and the ANA + EPI-NCSCs-Exos group. Each rat manufactured a 5-mm gap of facial nerve defect and immediately bridged by the corresponding transplants, respectively. After surgery, behavioral changes and electrophysiological testing of each rat were observed and assessed. At 90 days postoperatively, the retrogradely fluorescent tracer-labeled neurons were successfully observed on the injured side in the three groups. Morphological changes of facial nerve regeneration were evaluated by transmission electron microscopy and semithin toluidine blue staining. The results showed that nerve fiber density, nerve fiber diameter, and myelin sheath thickness in the ANA group were significantly worse than those in the other two groups ( < 0.05). No significant difference in nerve fiber density and myelin sheath thickness was observed between the autograft group and the ANA + EPI-NCSCs-Exos group ( = 0.14; = 0.23). Our data indicated that EPI-NCSCs-Exos facilitate ANAs to bridge facial nerve defects and have the potential to replace autograft therapy in clinic.
先前的研究表明，脱细胞神经同种异体移植物（ANA）已成功用于修复周围神经缺损，并且干细胞产生的外泌体有助于支持周围神经损伤后轴突的再生。在本研究中，来自毛囊表皮神经嵴干细胞的外泌体（EPI-NCSCs-Exos）与ANA联合用于桥接面神经缺损。通过超速离心分离EPI-NCSCs-Exos，并进行鉴定。共培养后，EPI-NCSCs-Exos分别在体外被背根神经节（DRG）和雪旺细胞（SC）内化。EPI-NCSCs-Exos延长了DRG轴突和树突的长度，加速了SC的增殖和迁移，并且也增加了SC的神经营养因子表达。下一步是将24只Sprague Dawley雄性大鼠随机均分为三组：自体移植组、ANA组和ANA + EPI-NCSCs-Exos组。每只大鼠制造一个5毫米的面神经缺损间隙，并立即分别用相应的移植物进行桥接。手术后，观察并评估每只大鼠的行为变化和电生理测试。术后90天，在三组的损伤侧成功观察到逆行荧光示踪剂标记的神经元。通过透射电子显微镜和半薄甲苯胺蓝染色评估面神经再生的形态学变化。结果显示，ANA组的神经纤维密度、神经纤维直径和髓鞘厚度明显低于其他两组（<0.05）。自体移植组和ANA + EPI-NCSCs-Exos组之间在神经纤维密度和髓鞘厚度上未观察到显著差异（=0.14；=0.23）。我们的数据表明，EPI-NCSCs-Exos有助于ANA桥接面神经缺损，并且有潜力在临床上替代自体移植治疗。
