比较两种兔颈总动脉脱细胞方法  

The comparision of two methods of decellularizing the common carotid artery in rabbits

作　　者：张安东 刘华蔚[2] 李永锋[2] 徐娟[2] 胡敏[2] ZHANG An-dong;LIU Hua-wei;LI Yong-feng;XU Juan;HU Min(Medical School of Chinese PLA,Beijing 100853,China;Department of Stomatology,the First Medical Center,Chinese PLA General Hospital,Beijing 100081,China)

机构地区：[1]解放军医学院,解放军总医院第一医学中心口腔科,北京100081 [2]解放军总医院第一医学中心口腔科,北京100081

出　　处：《口腔颌面修复学杂志》2020年第2期72-76,共5页Chinese Journal of Prosthodontics

基　　金：国家自然科学基金(项目编号:81371116);国家自然科学青年科学基金项目(项目编号:81901023)。

摘　　要：目的:摸索出更简单、高效、实用的制备脱细胞血管基质的方法,并与化学复合酶的脱细胞方法进行比较。方法:应用两种不同的脱细胞方法①0.125%胰蛋白酶+1%Triton+SDS+核酸酶;②超高压+1%糜蛋白酶+核酸酶脱除兔颈动脉细胞成分,对脱细胞后的血管进行DNA含量测定及组织学切片染色(HE、天狼星红),判断脱细胞效果;通过EVG染色、羟脯氨酸含量测定和最终抗张强度测试比较两组血管的机械性能。结果:两组的脱细胞血管基质的细胞成分被完全清除;0.125%胰蛋白酶+1%Triton+SDS+核酸酶组的胶原纤维呈波浪形且有缝隙,而超高压+1%糜蛋白酶+核酸酶组胶原纤维排列致密,组织损失量小;超高压+1%糜蛋白酶+核酸酶组的最大荷重、抗拉强度、弹性纤维及胶原纤维的量均高于对照组(P<0.05)。结论:超高压+1%糜蛋白酶+核酸酶是一种更简单、高效、实用的制备脱细胞血管基质的方法。Objective:To explore a simpler,more efficient and practical method for preparing acellular vascular matrix and compare with the decellularization method with chemical complex enzyme.Methods:Two different decellularization methods were applied:①0.125%trypsin+1%Triton+%SDS+nuclease;②ultra-high pressure+1%chymotrypsin+nuclease to remove rabbit carotid artery components,and the decellularized blood vessels were used.DNA content determination and histological section staining(HE,Sirius Red)were used to determine the decellularization effect;the mechanical properties of blood vessels in the two groups were compared by EVG staining,hydroxyproline content determination and final tensile strength test.Results:The cellular components of the acellular vascular matrix were completely eliminated in the two groups.The collagen fibers in the 0.125%trypsin+1%Triton+%SDS+nuclease group were wavy and gapy,The ultra-high pressure+1%chymotrypsin+nuclease group exibited a dense arrangement of collagen fibers and a small amount of tissue loss;Also its maximum load,tensile strength,elastic fiber and collagen fiber were higher than the control group(P<0.05).Conclusion:ultra-high pressure+1%chymotrypsin+nuclease is a simpler,more efficient and practical method for preparing acellular vascular matrix.

关 键 词：脱细胞血管基质 超高压 糜蛋白酶 

分 类 号：R780.2[医药卫生—口腔医学]