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/L Triton 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。

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.

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）