小口径组织工程血管脱细胞生物支架材料的研究  

Acellular Organismic Scaffolds for Small-diameter Vascular Tissue Engineering

作　　者：刘国锋[1] 杨大平[1] 郭铁芳[1] 郝晨光[1] 聂春磊[1] 何志娟[2] 

机构地区：[1]哈尔滨医科大学附属第二医院整形外科,黑龙江哈尔滨市150086 [2]哈尔滨医科大学附属第一医院妇产科,黑龙江哈尔滨市150010

出　　处：《中国康复理论与实践》2008年第3期234-236,F0003,共4页Chinese Journal of Rehabilitation Theory and Practice

基　　金：国家杰出青年科学基金(30325042);哈尔滨医科大学附属第二医院博士科研基金项目(BS2007-37)

摘　　要：目的探索利用生物酶类联合消化法制备猪颈总动脉脱细胞基质材料的方法,为小口径组织工程血管提供理想的生物支架材料。方法利用胰酶和核酸酶连续消化法脱除成年猪颈总动脉的细胞成分;通过组织学染色和定量DNA分析,检测细胞及细胞碎片的去除情况;利用扫描电镜观察脱细胞支架材料的空间结构和细胞去除情况。结果组织学染色结果显示,制备的猪颈总动脉脱细胞支架材料不含有细胞成分,细胞外基质成分没有明显的改变,保留了脱细胞前动脉的空间结构;DNA定量分析结果显示,支架材料的DNA含量小于0.1%;扫描电镜结果显示,支架材料的内表面完全去除了内皮细胞成分,显示出良好的三维空间结构。结论利用胰酶和核酸酶连续生物酶消化法制备的猪颈总动脉细胞外基质支架材料中细胞成分去除完全,细胞外基质的空间结构保持良好,制备的支架材料符合小口径组织工程血管对生物支架材料的基本要求。Objective To prepare the acellular carotid artery of adult pig using biotic enzymes for small-diameter vascular tissue-engineering scaffold.Methods Carotid artery of adult pig was removed cells by using Trypsin/EDTA,ribonuclease and desoxyribonuclease.Residual cellular composition was studied with quantitative DNA analysis and the Hematoxylin and Eosin（H＆E）staining.Extracellular matrix composition was evaluated with elastin van Gieson＇s（VG）staining and scanning electron microscopy（SEM）.Results Quantitative DNA analysis and H＆E staining confirmed that carotid arteries were completely decellularized.Elastin van Gieson＇s staining demonstrated that elastin morphology appeared unchanged.Scanning electron microscopy examination of the acellular scaffolds revealed a well-oriented porous decellularized structure that maintained natural architecture of the aorta.Conclusion Carotid artery of adult pig rendered acellular with Trypsin/EDTA,ribonuclease and desoxyribonuclease has well-preserved extracellular matrix for vascular scaffold,which can be used as a scaffold for further small-diameter vascular tissue engineering.

关 键 词：组织工程 血管移植 支架 细胞外基质 脱细胞 

分 类 号：R318.11[医药卫生—生物医学工程]