Seoul National University Hospital, Clinical Research Institute, Xenotransplantation Research Center, Seoul, Korea.
Korean Circ J. 2012 Mar;42(3):154-63. doi: 10.4070/kcj.2012.42.3.154. Epub 2012 Mar 26.
Glutaraldehyde (GA) has been used as a representative method of tissue preservation in cardiovascular surgery. However, GA has showed limited durability including calcification, mechanical failure and toxicity. To overcome those unsolved problems, we analyzed the crosslinking differences of primary amines, GA and genipin in their mechanical and biochemical properties with a single or double crosslinking agent for clinical application.
Samples were divided into 3 groups; control, single crosslinking fixation and double crosslinking fixation after decellurarization using bovine pericardium. For analysis of the biochemical and mechanical properties of each crosslinking method, tensile strength, percentage strain, thermal stability, resistance to pronase, nynhydrin and cytotoxicity test were studied.
Combined hexamethylene diamine and suberic acid in the carbodiimide hydrochloride/N-hydroxysucinimide solution (EDC/NHS) after decellurarization, tensile strength and strain percentage were not statistically significant compared to the single crosslinking treated groups (p>0.05). Tissue crosslinking stability was weak in single treatment of diphenylphosphoryl azide, suberic acid, low concentration of EDC, hexamethylene diamine and procyanidin groups, but thermal stability and resistance to the pronase and ninhydrin were markedly increased in concentrated EDC/NHS or after combined double treatment with low concentration of GA or genipin (p<0.001).
Single or double crosslinking with low concentration of carbodiimide, diphenylphosphonyl azide, procyanidin, suberic acid and hexane diamine were not as effective in mechanical, biochemical, cytotoxic and crosslinking properties compared to GA or genipin fixation, but their mechanical and chemical properties were much improved when combined with low concentrations of GA or genipin in the double crosslinking process.
戊二醛(GA)已被用作心血管手术中组织保存的代表性方法。然而,GA 显示出有限的耐久性,包括钙化、机械故障和毒性。为了克服这些未解决的问题,我们分析了伯胺、GA 和京尼平在其机械和生化特性方面的交联差异,使用单一或双交联剂进行临床应用。
样品分为 3 组:脱细胞牛心包后使用对照组、单交联固定组和双交联固定组。为了分析每种交联方法的生化和机械性能,研究了拉伸强度、应变百分比、热稳定性、对糜蛋白酶、戊二醛和细胞毒性试验的抗性。
脱细胞后,在碳二亚胺盐酸盐/N-羟基琥珀酰亚胺溶液(EDC/NHS)中加入己二胺和琥珀酸,与单交联处理组相比,拉伸强度和应变百分比没有统计学意义(p>0.05)。在单处理二苯膦酰叠氮、琥珀酸、低浓度 EDC、己二胺和原花青素组中,组织交联稳定性较弱,但在高浓度 EDC/NHS 或在低浓度 GA 或京尼平联合双处理后,热稳定性和对糜蛋白酶和戊二醛的抗性显著增加(p<0.001)。
与 GA 或京尼平固定相比,低浓度碳二亚胺、二苯膦酰叠氮、原花青素、琥珀酸和己二胺的单一或双交联在机械、生化、细胞毒性和交联性能方面的效果并不理想,但当与低浓度 GA 或京尼平在双交联过程中结合使用时,其机械和化学性能得到了很大改善。