改良萃取法制备脱细胞神经支架的去髓鞘效果评价  

Demyelination Efficiency of Optimized Extractive Method in Preparation of Acellular Nerve Scaffold

作　　者：侯博[1] 李文胜[1] 秦峰[1] 王辉[1] 何海勇[1] 龚谨[1] 叶卓鹏[1] 张保豫[1] 郭英[1] 

机构地区：[1]中山大学附属第三医院神经外科,广东广州510630

出　　处：《中山大学学报（医学科学版）》2012年第3期322-325,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30672152);广东省科技计划项目(2007B031003002);广东省产学研结合项目(2011B090400098)

摘　　要：【目的】评价改良化学方法制备脱细胞神经支架的去髓鞘与脱细胞效果。【方法】12只成年SD大鼠(230～280g),取双侧坐骨神经,随机分为3组,每组8例,分别采用不同的处理方法。正常对照组:不作任何处理;Hudson组:采用Triton X-200、磺基三甲铵乙内酯-10(SB-10)、磺基三甲铵乙内酯-16(SB-16)处理;改良组:采用Triton X-200、SB-10、SB-16联合脱氧胆酸钠(SDC)及过氧乙酸(PAA)处理。处理后行HE染色、甲苯胺蓝染色及透射电镜检查,观察脱细胞、去髓鞘程度及基底膜保存情况。【结果】形态学检查显示,与对照组相比,Hudson组中的细胞核结构完全消失,甲苯胺蓝染色为散在同心圆状的髓鞘结构,透射电镜下显示髓鞘板层成分保留,但有崩解断裂,轴突依然存在;改良组细胞核完全去除,与Hudson组不同的是,甲苯胺蓝染色显示为不规则多孔状结构,髓鞘成分消失,透射电镜下显示髓鞘及轴突成分彻底去除,基底膜管壁结构清晰并完整保留。【结论】改良萃取法制备脱细胞神经支架能够有效地去除细胞和髓鞘成分,同时基底膜管结构保留完整。[Objective] To evaluate the competence of modified extractive method in removing myelin sheath, thereby seek a kind of effective extractive method to prepare acellular nerve scaffold. [ Methods ] Twelve adult SD rats weighing 230-280 g were used. The bilateral sciatic nerves were harvested and divided evenly into three groups according to preparation methods： control group were fresh nerve; Hudson group were treated by Triton X-200; sulfobetaine-10 （SB-10）, sulfobetaine-16 （SB-16） and modified group were processed by Triton X-200, SB-10, SB-16, sodium deoxycholate （SDC） and peroxyacetie acid （PAA）. HE, toluidine blue, and transmission electron microscope （TEM） were used to monitor the efficiency of extractive method by assess the degrees of deeellularization, demyelination and integrity of basal lamina tube. [ Results ] In Hudson group, HE staining showed that cell nucleis totally disappeared. Toluidine blue staining revealed concentrically ringed myelin sheath debris, stratiform structure of myelin sheath was distinct apart from a little ineompact and axons were still visible in TEM detection. Modified group presented the same HE results, while Toluidine blue staining and TEM showed sample exhibited irregularly porous structure, myelin sheath and axons were completely removed and basal lamina tubes were integrally preserved, when compared to Hudson group. [ Conclusion ] Modified extractive method exhibited a good efficiency in decellularization and demyelination, and provided protection for extracellular matrix.

关 键 词：萃取法 神经 脱细胞支架 髓鞘 基底膜管 

分 类 号：R318.08[医药卫生—生物医学工程]