改良化学去细胞同种异体神经制备方法的实验研究  

Experimental study of the improved preparative method of chemically extracted acelluar nerve allograft

作　　者：王冠军[1] 孙明学[1] 卢世璧[1] 许文静[1] 赵斌[1] 彭江[1] 张莉[1] 黄靖香[1] 

机构地区：[1]解放军总医院全军骨科研究所,北京100853

出　　处：《中国矫形外科杂志》2007年第12期929-932,I0001,共5页Orthopedic Journal of China

基　　金：国家自然科学基金(NO:30571875);北京市自然科学基金重点项目资助(NO:03G502)

摘　　要：[目的]改进去细胞异体神经化学萃取制备方法,制备出细胞及髓鞘去除完全、神经机构保存完好的去细胞异体神经移植物。[方法]SD大鼠14只,取双侧坐骨神经(共28根),分3组进行化学萃取去细胞处理:Son-dell法组(10根)、改良法组(10根)和对照组(8根)。Sondell法组所用萃取剂为Triton X-100和脱氧胆酸钠;改良法组所用萃取剂为Triton X-200、SB-10和SB-16;对照组未进行化学处理。处理后神经行HE染色、快蓝染色、基底膜素免疫组化染色及环境扫描电镜观察,并从去细胞程度、髓鞘染色分级和结构完整性三方面综合评价。[结果]在去细胞异体神经物理性状方面,改良法制备的去细胞异体神经韧性弹性略好于Sondell法;HE染色表明,2种方法的去细胞效果均较好,但改良法对神经结构的破坏较小;快蓝染色、基底膜素免疫组化染色、环境扫描电镜结果均表明,改良法在对髓鞘的去除、基底膜素的保留及神经结构的保存方面均优于Sondell法;综合质量评分结果表明,2种方法在去细胞效果方面相似(P1>0.05),髓鞘去除及神经结构的保存方面改良法优于Sondell法(P2、P3<0.05),综合去细胞程度、髓鞘染色分级和结构完整性3方面,改良法制备的去细胞异体神经质量优于Sondell法(P4<0.05)。[结论]综合运用萃取剂Triton X-200、SB-10和SB-16的化学萃取方法,是一种较为理想的去细胞异体神经制备方法,能够制备出免疫物质清除完全、神经结构较好保存的去细胞异体神经移植物,为自体神经移植找到了较好的替代解决方法。[ Objective]To improve the preparative method of chemically extracted acelluar nerve allograft, and to develop a graft from which Schwann cells and myelin could be removed while the nerve structures（ basal lamina tubes and extracellular matrix） remained intact. [ Method] Twenty-eight fresh sciatic nerves were obtained form 14 adult SD rats and divided into 3 groups： Sondell method group（ 10 nerve segments） , in which the nerve segments were chemically extracted with the detergents of Triton X-100 and deoxycholate. Improved method group （ 10 nerve segments ） , with the detergents of Triton X 200, sulfobetaine-10 （SB-10） and sulfobetaine-16 （SB-16）, and control group（8 nerve segments） , in which the nerve were underwent no chemical treatment. The quality of the acellular nerve allograft was revealed by HE staining, fast-blue staining, laminin immunohistochemistry, and scanning electron microscopy. [ Result] The acelluar nerve allografts in improved method group possessed better tenacity and elasticity than that of in the Sondell method group. HE staining demonstrated that the Schwann cells were completely removed from the nerves in the two groups and there were no significant difference in the effect of cell removal between the two groups （ P1〉 0.05 ）. There were better effect of removal of myelin （ P2 〈 0.05 ） and preservation of laminin and nerve structures （P3 〈 0.05 ） in the improved method group through the fast-blue staining, laminin immunohistochemistry, and scanning electron microscopy. The comprehensive quality evaluation of the effect of removal of cells and myelin and preservation of laminin and nerve structures showed that the quality of acelluar nerve allografts in the improved group exceeded that of in the Sondell method group（ P4 〈 0.05）. [ Conclusion ] This new chemical extraction procedure with the detergents of Triton X 200 , SB - 10 and SB - 16 is a ideal paradigni to develop high quality acellular nerve allograft with the immunologic substa

关 键 词：周围神经 化学萃取 去细胞异体神经 

分 类 号：R651.3[医药卫生—外科学]