软骨素酶ABC处理化学去细胞异体神经后的形态学观察  

Histomorphology observation of chemically extracted acellular nerve allograft treated with chondroitinase ABC

作　　者：赵斌[1] 王玉[1] 彭江[1] 张莉[1] 赵喆[1] 许文静[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京市复兴路28号100853

出　　处：《中国矫形外科杂志》2010年第18期1547-1549,共3页Orthopedic Journal of China

基　　金：国家863计划项目(编号:2009AA03Z312);军队十一五计划项目(编号:06Z057);国家支撑计划项目(编号:2009BAI87B02)

摘　　要：[目的]观察软骨素酶ABC处理化学去细胞异体神经后,硫酸软骨素蛋白多糖(CSPG)去除效果以及对活性成分层粘蛋白(Laminin)的影响。[方法]取12根新鲜猪坐骨神经,每段长60 mm,应用7%脱氧胆酸钠与7%Triton X-100制备化学去细胞异体神经后,随机分为两组。实验组(n=6):软骨素酶ABC处理组,将6根去细胞异体神经浸泡在包软骨素酶ABC(2 U/ml)的PBS(pH7.4);对照组(n=6)为PBS液浸泡。两组处理时间均为16 h37℃水浴。分别对两组神经行HE染色以及CSPG、Laminin免疫荧光染色观察。[结果]实验组与对照组HE纵切片均未能观察到细胞,红染的神经基底膜呈波浪状纵形排列,轴突消失而形成管柱状空隙;对照中CSPG免疫荧光染色阳性,经软骨素酶ABC处理后CSPG免疫荧光染色阴性;而实验组和对照组基底膜管成分层粘蛋白免疫荧光染色显示为阳性。[结论]经软骨素酶ABC处理的去细胞异体神经特异性降解了不利于神经生长的成分CSPG,而对去细胞神经中促神经生长成分Laminin无影响,可能是修复周围神经缺损的良好的移植物。[Objective]To evaluate the histomorphological changes of chemically extracted acellular nerve treated with chondroitinase ABC(ChABC) and to observe the removal condition of chondroitin sulfate proteoglycans(CSPG) and laminin.[Method]The fresh porcine sciatic nerves were isolated free of the underlying fascia and 60-mm-long nerve segments were made.After cleaned from external debris and treated with chemical detergents,acellular nerve allografts were incubated in phosphate-buffered(PBS) containing 2 U/ml ChABC or in PBS alone for 16 h at 37℃.The former was experimental group and the latter was control group.Hematoxylin and eosin(H&E) staining and immunofluorescence histological staining for CSPG and laminin were done for observation of general morphology.[Result]After histological staining,it was found that cells were completely removed while the basal lamina tube remained intact in acellular nerve group and ChABC-treated acellular nerve groups.Acellular nerve group demonstrated abundant expression of CSPG.Acellular nerve treatment with ChABC showed no expression of CSPG.[Conclusion]The acellular nerve treatment with ChABC can completely degrade inhibitory CSPG and preserve bioactive laminin component.A decellularized and ChABC processed nerve graft represents the new generation of circumspectly optimized,immunotolerated allografts.

关 键 词：化学去细胞异体神经 软骨素酶ABC 组织学观察 

分 类 号：R651.3[医药卫生—外科学]