化学萃取法制备人类同种异体神经支架的实验研究

Experimental studies of acellular nerve autograft from human by chemical extraction

目的发展化学萃取方法去除人类周围神经中的细胞和髓鞘,获得粗大和长段的去细胞神经支架。方法取三段长10cm,直径8mm的坐骨神经,以Triton-100和脱氧胆酸钠溶液按一定浓度和程序进行萃取,萃取神经及新鲜神经于中段取材,行HE染色、纤维素染色、砂罗铬花青染色、S-100免疫组化染色,在扫描电镜及放射电镜下观察超微结构。结果萃取两次后,去细胞神经具有良好的粘性和弹性,细胞和髓鞘被彻底清除,神经纤维支架被保留,成为一种没有细胞髓鞘及其碎片的空的基底膜管架结构。结论应用Triton-100及脱氧胆酸钠萃取两次,可去除人类长段、粗大周围神经中的细胞和髓鞘而保留神经的基底膜管和纤维支架结构。

Objective To develop the procedure of chemical extraction to remove the cells and myethin in peripheral nerve of human, and to obtain a thick and long acellular nerve allograft. Methods Three segments of sciatic nerve,10cm in length and 8mm in diameter, were extracted with 4%Triton X-100 and 4% sodium deoxycholate in different times. HE, fibrin, myethin and S- 100 immunohistochemical stain were performed and checked by light microscope and electron microscope.Results The acellular nerve autograft was of fine clasticity and ductility; the cells, axons and myelin sheath were removed and basal membrane was preserved after two extraction procedures and become empty basal lamina tubes in the endoneurium.Conclusion The cells in long and thick human peripheral nerve can be removed by two chemical extractions with Triton X-100 and sodium deoxycholate, while the basal lamina tubes and fiber scaffold can be kept intact.