十二烷基肌氨酸钠法制备带瓣管道支架及其生物学性能分析  

New preparation of acellular aortic valve conduit scaffold by using sodium n-lauroyl sarcosine and function analysis

作　　者：方宁涛[1] 孙洪亮[1] 谢尚喆[1] 吴春根[1] 王松梅[1] 高红阳[1] 潘銮凤[1] 

机构地区：[1]复旦大学上海医学院细胞与分子生物学实验室,200032

出　　处：《中华胸心血管外科杂志》2007年第4期260-262,共3页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：上海市科委重点项目基金资助(04JC14012)

摘　　要：目的尝试用十二烷基肌氨酸钠作为脱细胞试剂,制备脱细胞组织工程支架材料,并分析其生物学性能。方法用0.25%十二烷基肌氨酸钠溶液及核酸酶对猪的带瓣膜管道进行脱细胞处理,并对其进行 DNA、可溶性蛋白含量测定,HE、Movat 染色和电镜观察及生物力学测试;大鼠皮下包埋实验分析生物相容性。结果经脱细胞处理后,带瓣膜管道的瓣叶和管壁中 DNA 含量仅为正常的1.35%和2.81%,可溶性蛋白含量仅为正常的8.46%和12.65%,细胞成分几乎完全去除。HE 和 Movat 染色及电镜观察显示组织结构完整、细胞结构完全消失,胶原、弹性纤维、蛋白聚糖等主要基质成分充分保留;瓣膜生物力学性能与正常瓣膜差异无统计学意义;支架生物相容性良好,并具有低免疫原性和可降解性。结论十二烷基肌氨酸钠是一种理想的新型脱细胞试剂,以此可以建立一套低毒、高效、对组织结构无损伤的脱细胞方法,并制备出生物学性能优异的脱细胞支架材料。Objective Using sodium N-lauroyl sarcosine as new decellularization reagent to prepare decellularized matrix for tissue engineering purpose. Methods Fresh porcine aortic valve conduit were decellularized by using 0.25% sodium N-lauroyl satcosine （NKS）, RNase and DNase. The decellularized valve was evaluated by quantitative DNA/pmtein analysis, HE/Movate-pentachrome staining, electron microscopy observation, mechanical testing, and rat subdennal implantation. Results After treated by NKS solution, conformation of the conductive excellent. Histological and DNA/protein analysis showed that the treatment converted both leaflets and wails of native aortic valve conduits into cell free scaffolds, and about 98% DNA and 90% soluble protein were removed. Furthermore, the typical structures were optimally retained without apparent disruption of the leaflet histoarchitecture. Movat-pentachrome staining showed the extracellular matrix structures and main components such as collagen, elastin and glycosaminoglycans were well preserved. These facts were further verified by transmission electron microscopy. The decellularized valves have similar biomechanical characteristics to native valves, with well biocompatibility and biodegraltation. Conclusionn Sodium N-lauroyl sarcosine is an ideal decellularization reagent, by which an effective decellularization method to prepare heart valve scaffolds with low cytotoxicity and excellent biological property has been developed.

关 键 词：心脏瓣膜 人工 组织工程 细胞去除术 十二烷基肌氨酸钠 支架 

分 类 号：R318.08[医药卫生—生物医学工程]