不同低温冷冻预处理对异体神经移植再生影响的实验研究

Study of allogeneic neural transplantation after deep freeze pretreatment

目的观察不同冷冻温度下保存的异体神经对周围神经缺损修复的效果。方法取96只同种属雄性大鼠,其中24只为取材组,72只随机分为对照组(A、B组)和实验组(C、D、E、F组),每组12只。A组为新鲜自体神经移植组,B组为新鲜异体神经移植组,C、D、E和F组分别为经4℃、-50℃、-80℃和-196℃甘油保存3周的神经移植组。术后2周,进行免疫组织化学观察;术后9、16周进行大体观察、光镜检察、电生理测定。结果术后2周,B组移植段见明显T淋巴细胞浸润,C、D、E组次之,A组及F组未见T淋巴细胞。术后9周及16周,从神经再生效果评价看:A组神经再生效果最好,F组次之,C、D、E组再次,B组最差。结论在各冷冻组中,以-196℃组神经再生效果最好,接近新鲜自体神经移植组。

Objective To investigate the regeneration of peripheral nerve allografts stored in the glycerol of different temperature. Methods A total of 96 male Wistar rats,were randomly divided into 7 groups. Group A and B were the control groups,each of which included 12 rats;all underwent fresh femoral nerve autograft and a11ograft transplantation respectively. Twenty-four rats in group G were sacrificed to obtain 48 femoral nerves, which were randomly divided into 4 groups, 12 nerves in each group, and were stored in glycerol for 3 weeks at 4℃, -50℃, - 80℃ and -196℃ respectively. After storage for 3 weeks,4 - 12 nerves in each temperature group were randomly transplanted into 4 - 12 rats in target rats group C,D, E and F respectively. Two weeks after transplantation, two grafts of transplanted nerves in group A - F were harvested randomly and examined with immunohistochemistry to observe the infiltration of lymphocyte to the grafts and regeneration of sciatic nerve and fiber. Nine and 16 weeks after transplantation ,the regeneration of sciatic nerve was examined with histology, electro-physiology and with electron microscope. Results Wallerian degeneration was observed in the proximal and distal ends of all grafts of nerves harvested at 2 weeks post-engraftment; infiltration of T lymphocytes at the grafts in group B was significant,less T lymphocytes were seen in group C, D and E,while no T lymphocytes were seen in group A and F. Collagenoblast proliferation was observed in group B, Schwann cell proliferation in group A and F, while both collagenoblast and Schwann cell proliferation were observed in group C, D and E. Nine and 16 weeks post-engraftment,grafts in group B were severely adhered to the adjacent tissue, adherence in group A and F were slight, while that of groups C, D and E came between. Nerve regeneration of group A was the best,group F came second,the next were groups C, D and E,while group 13 was the worst. Conclusion The best storage temperature is -196℃.