The objective of this paper is to investigate the possibility and efficacy of recurrent laryngeal nerve repair by transplantation of co-cultured Schwann cells and neural stem cells (NSCs) in laminin-chitosan-poly-lactic-co-glycolic acid (laminin-chitosan-PLGA) nerve conduits in rats. A laminin-chitosan-PLGA conduit was used in a rat recurrent laryngeal nerve transection model. The rat recurrent laryngeal nerve was dissected to generate a 5 mm defect. Then, a laminin-chitosan-PLGA nerve conduit with or without Schwann cells and NSCs in the lumen was transplanted into the defect. A total of 96 female rats were randomised into six groups: co-culture of NSCs and Schwann cells in the nerve conduit group (CO), Schwann cells only in the nerve conduit group (SC), neural stem cells only in the nerve conduit group (NSC-only), nerve conduit group (null), autologous nerve graft group (autograft) and sham operation group (sham). Regenerated nerves were evaluated by histological and functional assessment at 8 and 12 weeks after surgery. The diameter and area of the regenerated myelin sheath, as well as the secretion of brain-derived neurotrophic factor and glial cell-derived neurotrophic factor in laryngeal muscle or regenerated nerve tissue in the CO group, were significantly better than they were in the SC, NSC-only and null groups (all P values < 0.05). Immunofluorescence showed that the CO group had significantly more neurofilament-200 immunoreactive and S-100 immunoreactive fibres than the SC, NSC-only and null groups (all P values < 0.05). The performance of the CO groups and autograft groups was found to be similar by laryngoscopy. Arytenoid cartilage motion recovery in these two groups was significantly better than it was in the other groups (all P values < 0.05). Our results indicated that co-culture of Schwann cells and NSCs in laminin-chitosan-PLGA conduits might promote injured nerve regeneration. This method might be a promising alternative for defective nerve repair.

本文的目的是研究通过在层粘连蛋白-壳聚糖-聚乳酸-乙醇酸（层粘连蛋白-壳聚糖- PLGA) 大鼠神经导管。层粘连蛋白-壳聚糖-PLGA导管用于大鼠喉返神经横断模型。解剖大鼠喉返神经以产生 5 mm 缺损。然后，将管腔中有或没有雪旺氏细胞和 NSCs 的层粘连蛋白-壳聚糖-PLGA 神经导管移植到缺损处。共96只雌性大鼠随机分为六组：神经导管组（CO）NSCs与雪旺细胞共培养，神经导管组（SC）仅施万细胞，神经导管组仅神经干细胞(NSC-only), 神经导管组 (null), 自体神经移植组（autograft）和假手术组（sham）。在手术后 8 周和 12 周通过组织学和功能评估评估再生神经。CO组再生髓鞘的直径和面积，以及喉肌或再生神经组织中脑源性神经营养因子和神经胶质细胞源性神经营养因子的分泌均明显优于SC组，仅 NSC 和空组（所有P值 < 0.05)。免疫荧光显示，CO 组比 SC、仅 NSC 和无效组具有显着更多的神经丝 200 免疫反应和 S-100 免疫反应纤维（所有P值 < 0.05）。喉镜检查发现 CO 组和自体移植组的表现相似。这两组的杓状软骨运动恢复明显好于其他组（所有P值 < 0.05）。我们的结果表明雪旺细胞和 NSC 在层粘连蛋白-壳聚糖-PLGA 导管中的共培养可能会促进受损神经再生。这种方法可能是有缺陷的神经修复的有希望的替代方法。