小肠黏膜下层的制备及其在心肌组织工程中的应用  

Small intestinal submucosa:Preparation and application in cardiomyocyte tissue engineering

作　　者：郭勇[1] 张西正[1] 李瑞欣[1] 孙鹃[2] 魏严[1] 郭春[1] 闫玉仙[2] 

机构地区：[1]军事医学科学院卫生装备研究所四室,天津300161 [2]武警医学院训练部实验管理中心

出　　处：《解放军医学杂志》2008年第11期1304-1306,共3页Medical Journal of Chinese People's Liberation Army

基　　金：天津市自然科学基金重点项目(06YFJZJC02000);全军医药卫生科研基金面上项目(06MA344);国家自然科学基金面上项目(30700156)

摘　　要：目的制备小肠黏膜下层生物材料,并以其为支架材料体外构建工程化心肌组织。方法将猪小肠黏膜下层以0.25%胰酶和0.5%SDS各处理24h,制备出脱细胞小肠黏膜下层。用材料试验机沿平行或垂直于管腔的方向进行单轴拉伸,检测其力学性能,同时检测其细胞毒性和组织相容性,然后将3日龄新生大鼠心肌细胞接种于小肠黏膜下层支架上,体外构建工程化心肌组织薄片,倒置显微镜下观察。经Masson组织化学染色、肌节型肌动蛋白免疫组化染色和透射电镜观察对获得的心肌薄片进行评价。结果小肠黏膜下层的力学性能良好,已经完全脱去细胞,无细胞毒性,组织相容性良好;接种12h后,心肌细胞开始在小肠黏膜下层上搏动,2d后心肌细胞-支架复合物(工程化心肌组织薄片)开始自发搏动,可搏动14d。构建的工程化心肌组织薄片由多层心肌细胞构成并有大量胞外胶原。结论成功制备出力学性能和生物相容性良好的小肠黏膜下层生物支架材料,并以其为支架成功构建出工程化心肌组织薄片。Objective To prepare the small intestinal submucosa （SIS） to be used as a scaffold in constructing engineered cardiac tis sue. Methods Small intestinal submucosa of pig was treated with 0.25% trypsin and then 0.5% SDS for 24 hours each to obtain decellu- larized SIS. The obtained SIS was tested for its mechanical strength by stretching both horizontally and longitudinally with a material test machine. The cytotoxicity and histocompatibility of the material was also examined. Then the cardiomyocytes harvested from 3-day old neonatal rats were seeded on the SIS to construct engineered cardiac tissue sheets. These acquired engineered cardiac tissue sheets were immunohistochemically stained and observed with inversion microscope and transmission electromicroscope to evaluate their characteristics. Results SIS was decellularized completely. The mechanical capability of the decellularized SIS was satisfactory. Under 15% stretching, its strain and stress was nearly linear. SIS showed no cytotoxicity and inflammatory response. After 12 hours, the cardiomyocytes seeded in the SIS scaffold began to beat spontaneously. Two days later, the cardiomyocytes-SIS scaffold composite （engineered cardiac tissue sheet） began to beat spontaneously, and could beat spontaneously for 14 days. The constructed engineered cardiac tissue sheets consisted of layers of cardiomyocytes and with abundant extracellular collagen in the sheet. Conclusions SIS with good mechanical capability and biocompati- bility has been prepared successfully, and then the engineered cardiac tissue has been constructed successfully based on the SIS scaffold.

关 键 词：小肠黏膜下层 支架 肌细胞 心脏 组织工程 

分 类 号：R318.11[医药卫生—生物医学工程]