豚鼠神经干细胞修复兔面神经缺损观察

Research of guinea pig neural stem cells promoting facial nerve regeneration in rabbits

目的:观察豚鼠海马神经干细胞(NSCs)修复兔面神经缺损的效果,探讨其作为组织工程的种子细胞修复周围神经缺损的可行性。方法:健康日系大耳白兔20只,随机分为2组,每组10只,均建立一侧面神经颊支缺损10mm模型,用内置胶原蛋白海绵的硅胶管桥接面神经缺损,治疗组在上述神经导管内植入新生豚鼠海马NSCs,对照组注入生理盐水。NSCs移植前48h用5′溴尿嘧啶(BrdU)标记。术后12周,进行大体观察、神经电生理检查、BrdU和S100免疫组织化学染色和组织学观察。结果:①治疗组神经肌肉动作电位的潜伏期明显短于对照组(P<0.01),而波幅明显高于对照组(P<0.01)。②治疗组有大量BrdU阳性细胞,且部分阳性细胞同时呈现S100双标阳性。对照组未见BrdU阳性细胞。③治疗组再生有髓神经纤维数量、直径和髓鞘厚度与对照组相比明显增加(P<0.01)。结论:NSCs能明显提高面神经损伤后修复效果,可作为种子细胞应用于外周神经组织工程。

Aim ： To evaluate the feasibility of using guinea pig neuronal stem cells（NSCs） as seed cells in peripheral nerve tissue engineering. Methods： Twenty Japanese white rabbits were randomly divided into 2 groups. A 10 mm nerve defect was set up in the unilateral facial nerve of rabbit and was bridged with a 15 mm-long silicone tube filled with collagen protein sponge. NSCs were transplanted into the silicone tube in the treatment group, while normal saline for the control group. The transplanted NSCs were labeled in advance with 5-bromodeoxyuridine （BrdU）. Gross observation, electrophysiological examination, BrdU and S100 immunocytochemical staining and histological examination were carried out at 12 postoperative weeks. Results： The latency of nerve muscle action potential was shorter, while the amplitudes was higher （P 〈 0. O1 ） in the treatment group compared with those in control group; in the treatment group, there were many BrdU positive cells, moreover, some cells were S100 positive on double staining. While in control group,there were no BrdU positive cells; the number and diameter of regenerated myelinated fibers, the thickness of myelin sheath in the treatment group were significantly increased compared with control group（P 〈0. 01 ）. Conclusion ： NSCs can promote facial nerve regeneration and can be potentially used as seed cells for peripheral nerve tissue engineering.