眼部脱细胞修复材料的制备与鉴定  

Changing duramater and sclera into acellular tissue matrix

作　　者：聂丹瑶[1] 陈家祺[2] 

机构地区：[1]中国广东省深圳市眼科医院角膜及眼表疾病科,518040 [2]中山大学中山眼科中心,中国广东省广州市510060

出　　处：《国际眼科杂志》2012年第4期612-614,共3页International Eye Science

基　　金：中国卫生部临床学科重点资助项目(No.2004-468);中国国家自然科学基金资助项目(No.30271388)~~

摘　　要：目的:以不同方法制备脱细胞硬脑膜和巩膜,比较不同方法、浓度、作用时间对硬脑膜和巩膜脱细胞效果的差异,找到适合硬脑膜和巩膜的脱细胞方法。方法:将取自8只新西兰大白兔的硬脑膜和巩膜组织剪成小块,每种组织块均分为8组,每组含有硬脑膜和巩膜两种组织,4组用以5,10,20,50mL/L Triton X-100为主,辅以DNase、RNase的方法处理,另外4组用2.5g/L胰蛋白酶和1g/L SDS分别处理12h+12h,12h+24h,24h+12h,24h+24h。完成后分别从大体观察8组的外观,HE染色比较脱细胞后的细胞残留数,透射电镜观察胶原纤维的超微结构。结果:以Triton X-100处理后的组织外观与处理前相似,2.5g/L胰蛋白酶处理后的组织肿胀增厚。20,50mL/LTriton X-100两组和2.5g/L胰蛋白酶+1g/L SDS处理时间≥36h的三组组织内的细胞残留平均数都小于2个/高倍镜视野。电镜结果证实,硬脑膜和巩膜经过脱细胞处理后,胶原纤维结构同处理前相同,未受到破坏。结论:以Triton X-100处理的硬脑膜和巩膜结构紧凑,胶原纤维保存完好,其中20mL/L Triton X-100浓度处理过的组织残留细胞少,处理前后外观改变少,是适合硬脑膜和巩膜进行脱细胞处理的方法。AIM：To find best methods to prepare acellular dura and sclera,and compare the effects of different methods including processes,concentrations and time.METHODS：The duramater and sclera which came from 8 New Zealand rabbits were divided into 8 groups.Each group had dura and sclera.5,10,20,and 50mL/L Triton X-100 were used in 4 groups.Other 4 groups were treated with 2.5g/L trypsin and 1g/L SDS,and the treated time was 12h＋12h,12h＋24h,24h＋12h and 24h＋24h.After these procedures,the appearances of those tissues were observed,residual cell number was counted after HE staining,and electron microscope was used to observe the ultrastructures of dura and sclera.RESULTS：The appearances of acellular tissues treated by Triton X-100 had no obvious differences with tissues that hadn＇t been treated.Two groups treated by 20mL/L and 50mL/L Triton X-100 and three groups treated by 2.5g/L trypsin and 1g/L SDS with treated time≥36 hours had the least residual cells.After acellular procedures,dura and sclera had the same ultrastructure as pretreatment in electron microscope.CONCLUSION：The collagen fibres of dura and sclera treated by Triton X-100 were well conserved.The residual cells are less in tissues treated by 20mL/L Triton X-100.20mL/L Triton X-100 is fit for treating dura and sclera.

关 键 词：脱细胞组织基质 脱细胞巩膜 脱细胞硬脑膜 

分 类 号：R779.1[医药卫生—眼科]