【摘要】 目的:探讨戊二醛交联时间对人脱细胞真皮基质(ADM)生物学性质的影响。方法:用低浓度戊二醛交联高渗盐-NaOH消蚀法制备的人ADM,根据交联时间的不同分为交联10、15、20、30min组,大体观察各组的物理性状;免疫组织化学检测各组Ⅰ型、Ⅲ型胶原蛋白的表达;MTT细胞毒性实验评价4组ADM的细胞毒性反应;将NIH3T3细胞接种于各组ADM上,比较24h细胞贴壁数并观察细胞生长情况;将人成纤维细胞与交联10、15min组ADM体外复合培养,观察细胞生长和增殖情况。结果:随着交联时间延长,ADM的韧性逐渐增加;各组ADMⅠ型、Ⅲ型胶原蛋白的表达阳性;ADM细胞毒性反应交联10和15min组0～1级、20min组1～2级、30min组1～3级;交联10、15min组分别与20min组24h3T3细胞贴壁数比较有统计学意义;而交联10、15min两组间无统计学意义;3T3细胞能够在交联10、15min组ADM表面贴附、增殖;免疫荧光法显示鼠抗人波形蛋白抗体呈阳性,有较多的人成纤维细胞黏附于交联10和15min组ADM上,生长和增殖良好。结论:戊二醛化学交联10～15min的人ADM蕴涵丰富的生物信息,细胞毒性小,生物相容性好,是一种很好的生物支架材料。更多还原

【Abstract】 Objective:This study was performed to compare the biological characteristics of the four different acellular dermal matrix(ADM)dealed by glutaraldehyde.Methods:Human skin was treated with hypertonic saline followed by NaOH maceration.The obtained ADM were divided into four groups according to different glutaraldehyde cross-linking time,including 10,15,20 and 30 minutes and then were observed and characterized by immunohistochemistry.The cytotoxicity of them was evaluated by methyl thiazolyl tetrazolium(MTT)colorimetry and then cell compatibility was analysized by cell culture.NIH3T3 cells were cultured on different ADM and the adhering number was counted.Human fibroblast cells were cultured on the optimized ADM and observed.Results:The longer the cross linkage was,the more tenacious was.Collagen type Ⅰand Ⅲ were observed in four groups.The cytotoxicity scores of group 10 minutes and 15 minutes were grade 0-1,while those of group 20 minutes and group 30 minutes were grade 1-2 and grade 1-3 respectively.There was significant difference in the adhering numbers between group 10 minutes and 15 minutes,compared with group 20 minutes.There was no significant difference in the adhering numbers between group 10 minutes and 15 minutes.NIH3T3 grew well in group 10 minutes and 15 minutes.Human fibroblast cells proliferated well in the ADM cross-linked for 10 minutes and 15 minutes.Conclusion:ADM dealed by glutaraldehyde for 10 minutes and 15 minutes was good tissue engineering biomaterial with little cytotoxicity and plenty of biological characterization.更多还原