新方法可获得更佳的组织工程小直径血管细胞外基质支架  

Acquistion of a suitable extracellular matrix scaffold for tissue-engineered small-diameter blood vessel using a novel decellularization protocol

作　　者：蒲磊[1,2] 潘兴纳 张静[3] 吴剑[1,2] 李亚雄[1,2] 杨应南[1,2] Pu Lei;Pan Xingna;Zhang Jing;Wu Jian;Li Yaxiong;Yang Yingnan(Department of Cardiovascular Surgery,Yan’an Hospital Affiliated to Kunming Medical University, Kunming 650051, Yunnan Province, China;Yunnan ProvincialInstitute of Cardiovascular Surgery, Yan’an Hospital Affiliated to Kunming Medical University, Kunming 650051, Yunnan Province, China;Department of Anesthesiology, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, Yunnan Province, China)

机构地区：[1]昆明医科大学附属延安医院心脏大血管外科,云南省昆明市650051 [2]昆明医科大学附属延安医院云南省心血管外科研究所,云南省昆明市650051 [3]昆明医科大学第二附属医院麻醉科,云南省昆明市650101

出　　处：《中国组织工程研究》2019年第6期855-862,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:获得适宜的支架材料是构建组织工程小直径血管的基础,前期研究发现,非离子型去垢剂的脱细胞效果差,联合应用离子型去垢剂能更有效去除细胞成分。目的:比较不同脱细胞方法制备猪颈动脉脱细胞支架的有效性,探索更适宜的细胞外基质支架制备方法。方法:取普通猪颈动脉30根,随机分为5组(n=6),其中A组不进行脱细胞处理;B组采用1%十二烷基硫酸钠+1%脱氧胆酸钠进行脱细胞处理;C组采用0.5%十二烷基硫酸钠+0.5%脱氧胆酸钠进行脱细胞处理;D组采用1%十二烷基硫酸钠+1%TritonX-100进行脱细胞处理;E组采用0.5%十二烷基硫酸钠+0.5%Triton X-100进行脱细胞处理。对各组样本分别行苏木精-伊红染色、五色套染、扫描电镜、免疫组织化学及生物机械力学检测。结果与结论:(1)苏木精-伊红染色、五色套染显示,B、C组脱细胞完全,D、E组于局部可见细胞残留;(2)扫描电镜显示,脱细胞各组胶原纤维束和弹性蛋白保存完整,B组孔隙结构的形态、尺寸优于C、D、E组;(3)免疫组织化学评估显示,脱细胞各组均有异种抗原α-1,3-Gal表达,B组α-1,3-Gal抗原表达低于C、D、E组;(4)生物机械力学检测显示,脱细胞各组爆破压力、缝线保持力和缝合保留强度较A组显著下降(P <0.05),脱细胞各组间比较无差异;脱细胞各组顺应性、长轴断裂伸长率、横轴最大应力、横轴断裂伸长率、横轴杨氏模量与对照组比较无差异(P> 0.05);脱细胞各组长轴最大应力和长轴杨氏模量较A组显著下降(P <0.05),其中B、C组长轴最大应力和长轴杨氏模量下降较D、E组明显(P<0.05);(5)结果表明,基于1%十二烷基硫酸钠+1%脱氧胆酸钠的方法可获得更佳的猪颈动脉细胞外基质支架,适用于构建组织工程小直径血管。BACKGROUND:A proper scaffold is the basis for construction of tissue-engineered small diameter blood vessels.Preliminary study has shown non-ionic detergents with limited effects of decellularization,and combined application of ionic detergents exhibits more effective decellularization.OBJECTIVE:To compare the effectiveness of different preferable methods for preparing porcine carotid artery derived extracellular matrix scaffold,and to explore the optimal preparation method.METHODS:Porcine carotid arteries(n=30)were obtained and randomized into five groups(n=6per group).Then porcine carotid arteries were decellularized,followed by treatment with1%sodium dodecyl sulfate and1%sodium deoxycholate(group B),0.5%sodium dodecyl sulfate and0.5%sodium deoxycholate(group C),1%sodium dodecyl sulfate and1%Triton X-100(group D),0.5%sodium dodecyl sulfate and0.5%Triton X-100(group E).Native procine carotid aitery served as control(group A).Tissue samples of each group underwent hematoxylin-eosin staining,and Movat’s pentachrome staining,scanning electron microscope,immunohistochemistry and biomechanical test.RESULTS AND CONCLUSION:Hematoxylin-eosin staining and Movat’s staining revealed that the cellular components were completely removed in the groups B and C;however,cellular residues were visualized in the groups D and E.Under the scanning electron microscope collagen and elastin fiber bundles were well preserved in the decellularization groups;however,the morphology and size of porous structure in the group B were superior to those in the groups C,D and E.Immunohistochemistry staining showed that the xenogeneic antigen,α-1,3-Gal,expressed in the decellularization groups;however,the expression ofα-1,3-Gal was lower in the group B than the other groups.Biomechanical test indicated that burst pressure,suture-holding capacity and suture-holding strength were significantly decreased in the decellularization groups compared with group A(P<0.05).However,there was no significant difference in the compliance,longitudinal elong

关 键 词：脱细胞 细胞外基质 组织工程小直径血管 细胞外基质支架 人工血管 生物力学 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]