冷冻保存的雪旺细胞对损伤后坐骨神经再生影响的实验研究

目的比较原代培养雪旺细胞(Schwanncells,SCs)和冷冻保存的 SCs移植对损伤后坐骨神经再生的作用.方法原代培养和液氮保存的SCs分别移植到桥接缺损坐骨神经的硅胶管内.在移植后不同时间,硅胶管远端神经干内注射HRP,逆行追踪背根神经节和脊髓前角的标记神经元数量;测量再生神经纤维的复合动作电位传导速度;电镜观察再生神经纤维的髓鞘形成.结果原代培养和冷冻保存SCs在移植后不同时间其背根神经节(术后 6周 3967.41± 305.91与 3890.55±315.63, t=0.55, P > 0.05;术后 8周 4029.33± 313.80与 4184.90± 305.75, t=1.11, P> 0.05)和脊髓前角神经元 HRP标记细胞数量(术后 6周 404.87± 40.05与429.77± 43.40, t=1.33,P > 0.05; 术后 8周 474.49± 42.74与 470.99±38.82,t=0.19,P > 0.05)、再生神经纤维的复合动作电位传导速度 (m/s)基本一致(术后6周 32.28± 1.96与 32.05± 2.31,t=0.24,P > 0.05; 术后 8周 43.60± 1.88与 43.84±2.19,t=0.26,P > 0.05; 术后 12周 43.78± 1.71与 43.83± 1.75,t=0.06,P > 0.05),再生神经纤维髓鞘的形成未见明显差别.结论冷冻保存的SCs仍具有促进损伤后周围神经再生的能力.

Effect of cryopreserved Schwann cells on injured sciatic nerve regeneration

AIM:To compare the effect of primary cultured Schwann cells(SCs) and cryopreserved SCs transplantation on injured sciatic nerve regeneration. METHODS:Primary cultured and liquid nitrogen kept SCs were transplanted into silica tube that bridged a sciatic nerve deficit. At different time points after the transplantation (end of 6th and 8th week after the operation), HRP was injected into the distal nerve stump in the tube. Then, the numbers of labeled neurons in dorsal root ganglion and anterior horn were traced by reversed follow-up. MCV of newly regenerated nerve fiber was measured, and myelinization of nerve fiber was studied under electron microscope. RESULTS:The number of dorsal root ganglion with primary culture and freeze kept SCs grafts at different time periods was (6 weeks after operation:3967.41± 305.91 and 3890.55± 315.63, t=0.55, P > 0.05; 8 weeks after operation: 4029.33± 313.80 and 4184.90± 305.75, t=1.11,P > 0.05) and HRP labeled neurons(6 weeks after operation: 404.87± 40.05 and 429.77± 43.40, t=1.33,P > 0.05; 8 weeks after operation: 474.49± 42.74 and 470.99± 38.82,t=0.19,P > 0.05); Both groups showed a similar MCV(6 weeks after operation: 32.28± 1.96 and 32.05± 2.31,t=0.24,P > 0.05; 8 weeks after operation: 43.60± 1.88 and 43.84± 2.19,t=0.26,P > 0.05; 12 weeks after operation: 43.78± 1.71 and 43.83± 1.75,t=0.06,P > 0.05). No apparent difference in nerve fiber myelinization was observed. CONCLUSION:Cryopreserved SCs still keep the same ability of promoting injured peripheral nerve regeneration.