两种方法制备兔小肠黏膜下层的对比研究  

A comparative study of two ways of preparing small intestinal submucosa

作　　者：张龙芳[1] 陈文弦[2] 赵大庆[3] 刘志[2] 崔鹏程[2] 

机构地区：[1]兰州军区乌鲁木齐总医院耳鼻咽喉科,新疆乌鲁木齐830000 [2]第四军医大学唐都医院耳鼻咽喉科,陕西西安710038 [3]第四军医大学病理教研室,陕西西安710032

出　　处：《医学研究生学报》2007年第11期1147-1150,I0002,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助项目(批准号:30171007)

摘　　要：目的:比较两种制备方法所获兔小肠黏膜下层(SIS)的异同,为SIS作为组织工程细胞载体选择最佳制备方案。方法:分别采用消毒法(实验Ⅰ组)和顺序化学浸泡脱细胞法(实验Ⅱ组)处理机械剥离获取的兔小肠黏膜下层,进行组织学、扫描电镜检查及四甲基偶氮唑蓝(MTT)细胞毒性对比实验。结果:组织学及电镜扫描显示,与消毒法相比,经顺序化学浸泡脱细胞处理的SIS纯度高,孔隙多,安全性高,胶原纤维未受损;浸提液MTT检测各实验组相对增殖率均>1,100%SIS浸提液各实验组与对照组吸光值均有显著性差异(P<0.05),相同浓度不同SIS浸提液组间无显著性差异,表明多种化学试剂的SIS对软骨细胞不增加毒性,有一定促生长作用。结论:化学脱细胞法较消毒法制备的SIS安全性好,易保存,可操作性强,所获SIS是一种较理想的软骨细胞载体。Objective ： To compare the histological difference of the small intestinal submucosa （SIS） prepared in two ways, and to find a better means of making SIS a scaffold for tissue engineering. Methods ： SIS derived from the rabbit jejunum was treated by sterilization and steps chemical de-cellular method respectively. Then comparative examinations were performed by histological observation, scanning electron micrography and MTT cytotoxicity assay. Results： Histological observation and scanning electron micrography indicated that, compared with sterilization, steps chemical de-cellular method made SIS purer, with more ventages in it and no damage collagen fibers. MTT cytotoxicity assay showed that the relative growth rate（RGR） of each experimental group overtopped 1. The extinction value of the 100% SIS leaching liquor groups was significantly higher than that of the control （ P 〈 0.05 ）, but no significant difference was found between different SIS groups of the same concentration （ P 〉 0.05 ）, which demonstrated that SIS treated by steps chemical de-cellular method added no cytotoxicity to cartilage cells. Conclusion：Safe, convenient and easy to be preserved, steps chemical de-cellular method is a better way to make SIS a scaffold for tissue engineering.

关 键 词：小肠黏膜下层 细胞外基质 四甲基偶氮唑蓝比色 

分 类 号：R656.7[医药卫生—外科学]