CAJ | 학술논문

目的对目前文献报道使用的5种不同方法制备的人源性同种异体去细胞外周神经材料进行综合分析比较其优劣,以确定可供临床使用的标准的制备流程.方法将人源性神经按5种不同方法进行化学萃取,制备的外周神经支架材料分别行苏木精-伊红(HE)染色、免疫组化(S-100、Col I)和透射电镜、氮含量测定等检测,观察5种方法去除许旺细胞、髓鞘和轴突等抗原成分以及基底膜保存完好的情况.结果分别萃取2次持续24 h组神经的HE染色显示去除细胞和轴突彻底,纵切片上未见任何细胞,红染的神经内膜呈波浪状纵形排列,轴突、髓鞘结构消失而形成管柱状空隙;S-100染色呈阴性;Col I染色结果可以看出其结构呈松散不规则的棕黄色结构,而其它处理组则结构相对整齐的纵向带状结构.透射电镜显示该组与各处理之间髓鞘去除差异无统计学意义.氮含量测定显示该组蛋白含量比值最低.结论使用Triton X-100处理24 h再分别脱氧胆酸钠24 h持续萃取2次,可作为供临床使用的制备人源性去细胞同种异体神经支架材料的标准流程.
목적 대목전문헌보도사용적5충불동방법제비적인원성동충이체거세포외주신경재료진행종합분석비교기우렬,이학정가공림상사용적표준적제비류정.방법 장인원성신경안5충불동방법진행화학췌취,제비적외주신경지가재료분별행소목정-이홍(HE)염색、면역조화(S-100、Col I)화투사전경、담함량측정등검측,관찰5충방법거제허왕세포、수초화축돌등항원성분이급기저막보존완호적정황.결과 분별췌취2차지속24 h조신경적HE염색현시거제세포화축돌철저,종절편상미견임하세포,홍염적신경내막정파랑상종형배렬,축돌、수초결구소실이형성관주상공극;S-100염색정음성;Col I염색결과가이간출기결구정송산불규칙적종황색결구,이기타처리조칙결구상대정제적종향대상결구.투사전경현시해조여각처리지간수초거제차이무통계학의의.담함량측정현시해조단백함량비치최저.결론 사용Triton X-100처리24 h재분별탈양담산납24 h지속췌취2차,가작위공림상사용적제비인원성거세포동충이체신경지가재료적표준류정.
Objective To analyze five kinds of allogenic acellular peripheral nerve by different meth-ods and try to establish a standard method for preparing nerve materials. Methods Five kinds of nerve ma-terial prepared by different chemical extractions according to nowaday articles were examined by HE staining. Irmnunohistochemistry and protein ratio was studied by allogenic nerves by virtue of Kjeldahl method in order to know the efficiency of these methods in removal of SCs axons and integrality of the basilar membrane. Results Myelin sheath and cytoblast in group 2 that nerves were extracted by Triton X-100 and Sodium de-oxycholate consecutively twice were completely removed, which was well demonstrated in HE staining. Per-ineurium in red staining were arranged wave-like longitudinally, axons and myelin sheath were replaced by column-like spacing. Col I staining were positive in all groups, structure of basilar membrane became loose slightly in the first and second group, and the structure of the other groups were relatively regular. Group 1 and 2 were negative in S-100 staining. There was no difference between group 2 and group 1,3,4 and 5 in sheath removal demonstrated by TEM. Protein ratio in group 2 was the lowest in the measurement with Kjel-dahl method. Conclusion The method used in group 2 that nerves were extracted by Triton X-100 and Sodium deoxycholate consecutively twice was the best in allogenic acellular peripheral nerve preparations.