Simionescu A, Simionescu D, Deac R
Cardiovascular Surgery Research Department, Academy of Medical Sciences, Tirgu-Mures, Romania.
J Biomed Mater Res. 1991 Dec;25(12):1495-505. doi: 10.1002/jbm.820251207.
Crosslinking of collagenous biomaterials currently employs the use of glutaraldehyde. The putative enhancement of glutaraldehyde crosslinking by lysine was investigated in three model systems: bovine pericardium, collagen membranes, and bovine serum albumin. Repetitive sequential treatment of bovine pericardium with glutaraldehyde and lysine and finally with formaldehyde produced a matrix which, by the two criteria used (shrinkage temperature and urea/SDS soluble collagen), was shown to be more highly crosslinked than pericardium fixed in glutaraldehyde alone. Essentially the same results were obtained when membranes prepared from pepsin-soluble pericardial collagen were subjected to sequential glutaraldehyde and lysine treatments, reaching shrinkage temperatures of more than 90 degrees C. Heart valves prepared from lysine-enhanced glutaraldehyde crosslinked bovine pericardium were tested in vitro in an accelerated fatigue tester and have been shown to behave satisfactorily after 300 million cycles. These additional crosslinks proved to be stable in saline at 37 degrees C. Studies on bovine serum albumin attempted to get an insight into the mechanisms of lysine enhancement of glutaraldehyde crosslinking by treating sequentially albumin with glutaraldehyde and lysine and analysis of the products by gel filtration and SDS-PAGE. These studies suggest that free amino groups exposed by proteins are initially reacted with glutaraldehyde and then bridged by the diamino compound (lysine) producing more extensive intermolecular crosslinking than glutaraldehyde alone.
目前，胶原生物材料的交联采用戊二醛。在三个模型系统中研究了赖氨酸对戊二醛交联的假定增强作用：牛心包、胶原膜和牛血清白蛋白。用戊二醛和赖氨酸对牛心包进行重复顺序处理，最后用甲醛处理，产生了一种基质，根据所使用的两个标准（收缩温度和尿素/十二烷基硫酸钠可溶性胶原），该基质显示出比仅用戊二醛固定的心包具有更高的交联度。当用胃蛋白酶可溶性心包胶原制备的膜进行戊二醛和赖氨酸顺序处理时，得到了基本相同的结果，收缩温度超过90℃。用赖氨酸增强戊二醛交联的牛心包制备的心脏瓣膜在体外加速疲劳试验机中进行了测试，结果表明在3亿次循环后表现令人满意。这些额外的交联在37℃的盐水中被证明是稳定的。对牛血清白蛋白的研究试图通过依次用戊二醛和赖氨酸处理白蛋白，并通过凝胶过滤和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析产物，来深入了解赖氨酸增强戊二醛交联的机制。这些研究表明，蛋白质暴露的游离氨基首先与戊二醛反应，然后由二氨基化合物（赖氨酸）桥接，产生比单独使用戊二醛更广泛的分子间交联。
