类许旺细胞种植入去细胞神经桥接物后体内外标记示踪及神经功能恢复检测

目的 将类许旺细胞种值入去细胞神经桥接物内后观察体外培养下细胞存活情况及体内桥接坐骨抻终缺损后细胞存活情况及对神经功能恢复的影响。方法 通过Dezawa改良法诱导的类许旺细胞经Hochest 33342标记以10^7／ml显微注入经化学萃取的神经桥接物内，分别体外培养0、5、10、15、20、30d，苏木精-伊红染色及荧光硅微镜观察细胞在侨接物内的存活情况。同时将该神经桥接物桥接2cm长的坐骨神经缺损模型，分别于5、10、15、20、30、40d取材，观察细胞在桥接物内的存活情况、再生轴突的生长速度及再生轴突有髓纤维密度；于第30、50、70d进行神经电生理检测；于第40、60、80d进行坐骨神经功能指数测定，同时将无细胞的单纯神经桥接物桥接坐骨神经缺损及自体坐骨神经切断后原位缝合分别作为对照。结果 类许旺细胞在体内外条件下均能在化学萃取后的去细胞神经桥接物中存活。体内环境可能更有利于类许旺细胞的增殖。种植类许旺细胞的实验组各测定指标明显优于无细胞桥接对照组（P〈0．05或P〈0．01），实验组与自体神经对照组各指标近似（P〉0．05）。结论 类许旺细胞种植入去细胞神经桥接物后在体内外环境均能存话；种入类许旺细胞的去细胞神经桥接物桥接神经缺损可达到近似自体神经移植桥接的效果。

Experimental study on marking and tracing Schwann-liked cells after explantaion and implantation in acellural nerve grafts and measuring neural functional recovery after implantation

Objective To observe the Schwann-liked cells(SLCs) survival after explantation and implantation in acellural nerve grafts( ANG) and measuring the effect of the graft for promoting neural functional recovery after implantation. Methods SLCs which were achived from BMSCs by modified Dezawa method and marked with Hochest 33342 were micro-injected into ANG with 107/ml in density, and then the compound grafts were culture in vitro and SLCs' survival and migration were observed in 0,5,10,20,30 days with HE dying and fluorescence microscopy. Simultaneously, 2cm long sciatic nerve gap were bridged with the compound grafts and were observed with fluorescence microscopy in 5,10,15,20,30,40 days after implantation. Furthermore, the velocity of the axonal regeneration and the density of the myelinated fibres were measured in 5,10,20,30,40days respectively. Neural functional recovery parameters were assessed with measuring electro-physiological value in 30,50,70days and with sciatic nerve functional index in 40,60,80days. The only ANG without SLCs and autogenic sciatic nerve cut in situ bridged 2cm long sciatic nerve gaps were observed as a-bove as control groups respectively (n = 10). Results SLCs were testified the capacity to survive both in vitro and in vivo. SLCs may be more capalble to reproduce in vivo. Most of parameters from experiment group with SLCs were better than that from only ANG group without SLCs ( P <0. 05 or <0. 01) , while the parameters from experiment group with SLCs were similar to that from autogenic nerve graft group ( P > 0. 05). Conclusion SLCs could survive in ANG after explantation and implantation. To bridge the sciatic nerve gaps, the effect of ANG with SLCs is as close as that of the autogenic nerve graft.