基于心脏全器官脱细胞基质的组织工程支架材料制备与生物学评价  

PREPARATION AND BIO-EVALUATION OF TISSUE ENGINEERED SCAFFOLD BASED ON DECELLULARIZED WHOLE HEART EXTRACELLULAR MATRIX

作　　者：陈威震[1,2] 段翠密[2] 李霞[2] 孙红玉[2] 牟永潮[2] 舒瑶[2] 陈成[2] 冯兰兰[2] 周瑾[2] 王常勇[2] 

机构地区：[1]解放军总医院&解放军医学院,北京100853 [2]军事医学科学院基础医学研究所前沿交叉学科研究室

出　　处：《中国修复重建外科杂志》2013年第8期945-949,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划(863)资助项目(2012AA020506);国家自然科学基金青年基金资助项目(31100697)~~

摘　　要：目的探讨一种心脏全器官脱细胞方法,并对制备的心脏脱细胞基质支架进行检测与评价。方法通过联合使用胰蛋白酶、Triton X-100等洗涤剂,对成年SD大鼠离体心脏进行主动脉逆行灌流脱细胞处理。对制备的心脏脱细胞基质支架进行大体观察、甲苯胺蓝灌流染色、HE染色、扫描电镜观察、阿利新蓝染色、细胞外基质组分免疫组织化学染色等观察。结果成功制备心脏全器官脱细胞基质支架材料,其保留了原心脏的大体形态;HE染色、甲苯胺蓝染色及扫描电镜观察显示其保留了较好的脉管结构与超微空间构象;阿利新蓝染色及免疫组织化学染色结果显示,其主要的细胞外基质成分,包括Ⅰ型胶原、糖氨聚糖、纤连蛋白、层粘连蛋白保留良好。结论制备的大鼠心脏脱细胞基质支架保留了心脏原有的结构与细胞外基质成分,有望作为全器官心脏体外再造的良好组织工程支架材料。Objective To investigate a method for preparing decellularized rat heart scaffold,and to detect and evaluate the decellularized scaffold. Methods The decellularized rat heart scaffold was prepared by retrograde perfusion with a combination of enzymatic and Triton X-100 detergent methods to remove the populations of resident cells,and then the decellularized scaffold was observed by gross,toluidine blue staining,HE staining,scanning electron microcope（SEM）,Alcian blue staining,and immunohistochemisty staining to evaluate the structure and essential component of extracellular maxtix（ECM） in the scaffold. Results Tissue engineered scaffold based on decellularized whole heart ECM was successfully prepared,which maintained not only the gross morphology of the heart,but also the intact vascular structure and ultrastructural conformation that certified by toluidine blue staining,HE staining,and SEM analyses.Alcian blue staining and immunohistochemisty staining showed that the essential components of ECM,such as collagen type I,glycosaminoglycan,fibronectin,and Laminin were remained in decellularized whole heart matrix. Conclusion The decellularized whole heart ECM prepared by method mentioned can maintain the intact structure of rat heart and basic compositions of extracellular matrices,so it could be suitable for further studies of tissue engineered scaffolds for whole heart reconstruction.

关 键 词：心肌组织工程 心脏脱细胞基质支架 生物学特征 大鼠 

分 类 号：R318.08[医药卫生—生物医学工程]