CHAPS联合十二烷基硫酸钠的除垢剂动脉脱细胞支架材料制备  

Decellularization of arteries as scaffold by CHAPS and sodium dodecyl sulfate detergents

作　　者：梁远锋 文章 刘尚敏 黄淑佩 郝俊海 李东风[3] 林展翼[3] LIANG Yuan-feng;WEN Zhang;LIU Shang-min;HUANG Shu-pei;HAO Jun-hai;LI Dong-feng;LIN Zhan-yi(Guangdong Cardiovascular Institute,Guangdong General Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510100,China;South China University of Technology,Guangzhou 510641,China;Guangdong General Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510100,China;Southern Medical University,Guangzhou 510515,China)

机构地区：[1]广东省心血管病研究所广东省人民医院(广东省医学科学院),广州510100 [2]华南理工大学,广州510641 [3]广东省人民医院(广东省医学科学院),广州510100 [4]南方医科大学,广州510515

出　　处：《岭南心血管病杂志》2018年第4期450-455,共6页South China Journal of Cardiovascular Diseases

基　　金：2016广东省科技发展专项资金(项目编号:2016B070701007)

摘　　要：目的采用3-[(3-胆酰胺基丙基)二甲基铵基]-1-丙磺酸盐(CHAPS)和十二烷基硫酸钠(sodium do-decyl sulfate,SDS)的联合除垢剂方案对脐动脉进行脱细胞,评价脱细胞的有效性和支架材料的力学性能改变情况。方法分离新鲜脐带获取人脐动脉,通过对脐动脉进行CHAPS联合SDS脱细胞处理,获得脱细胞血管支架,对其进行苏木素-伊红、Masson、Verhoeff′s Van Gieson(EVG)等组织染色和扫描电镜分析,并进行DNA及胶原定量分析;于支架上接种成纤维细胞,通过细胞相容性的程度评估除垢剂残留情况;对血管支架材料进行爆破压力及应力应变曲线分析,评价支架材料力学性能改变。结果组织染色结果显示脱细胞支架材料血管壁细胞消失,与未脱细胞的脐动脉相比较,其胶原纤维和弹力纤维结构无明显差别,电镜下可见血管细胞消失,支架材料成多孔隙结构;DNA定量显示DNA浓度极低,胶原蛋白浓度无明显改变;人成纤维细胞可以很好附着于支架材料上并生长;脱细胞支架材料抗爆破压力及应力应变曲线与脐动脉比较无明显变化。结论 CHAPS联合SDS的脱细胞方法能彻底去除脐动脉血管壁细胞成分,且较为完整地保留了血管的组织结构。脱细胞后的血管支架材料具有良好的生物相容性和力学性能。Objectives To set up an ideal method for decellularization of umbilical arteries by CHAPS and sodium dodecyl sulfate(SDS)detergents and determine the mechanical properties of the acellularized vessels as the scaffolds for tissue engineering blood vessels.Methods The umbilical arteries isolated from the fresh umbilical cord were treated with CHAPS and SDS in sequence.The final decellularized scaffolds were examined with multiply histological staining[hematoxylin-eosin,Masson,Verhoeff′s Van Gieson(EVG)]and scanning electron microscope(SEM);The cellular DNA remains and collagen retention were quantified by PicoGreen and L-hydroxyproline assay,respectively.Human dermal fibroblasts(HDF)were seeded onto the acellular scaffolds for evaluation of detergent residual cell toxicity by the extent of cellular adherence and proliferation.Eventually,the burst pressure and stress-strain curve were measured by pressure transducer and INSTRON universal testing system.Results Diverse histological staining methods demonstrated that vascular cells and cellular fragments were removed completely,but the collagen and elastic fibers changed slightly compared with the fresh arteries.Micro porous construction of the acellular scaffolds was observed by SEM.Very little DNA and most collagen content remained after decellularization.HDF attached and grew well on the scaffolds.The burst pressure and stress-strain curve were not significantly varied after decellularization.Conclusions The acellular scaffolds via CHAPS and SDS retains normal microstructure and mechanical properties and shows good cell compatibility,which could be optimal scaffolds for tissue engineering blood vessels.

关 键 词：除垢剂 脱细胞 脐动脉 血管支架 

分 类 号：R331[医药卫生—人体生理学]