Pascual G, García-Honduvilla N, Rodríguez M, Turégano F, Bujan J, Bellón J M
Department of Morphological Sciences and Surgery (Surgical Research Laboratory), Medical School, University of Alcala, Crta. Madrid-Barcelona Km 33,600, E-28871 Alcalá de Henares, Madrid, Spain.
Ann Vasc Surg. 2001 Nov;15(6):619-27. doi: 10.1007/s100160010130.
This study was designed to explore the changes that occur in cryopreserved pig arteries following different thawing procedures, before and after being placed in an in vitro flow circuit. Segments of minipig iliac artery were cryopreserved in complete minimal essential medium plus 10% dimethylsulphoxide and stored in liquid nitrogen at -196 degrees C for 30 days. Three study groups were established according to whether the arterial specimens were fresh (control, n = 20), cryopreserved and rapidly thawed (RT) at 37 degrees C (n = 22) or cryopreserved and subjected to controlled, automated slow thawing (ST) (n = 22). Half of the specimens of each group were subsequently placed in the flow circuit for 72 hr. Evaluation was made of morphological and ultrastructural changes. Cell damage was established using the TUNEL method. All cryopreserved specimens showed endothelial denudation that was most extensive in those subjected to rapid thawing. Slowly thawed specimens showed improved cell viability and organization of the vessel wall, compared to those thawed rapidly. Under conditions of flow, the damage induced by the freezing/thawing process was enhanced. These findings suggest that (a) slow thawing of cryopreserved arteries results in improved preservation of the structure and viability of vessels, and (b) the damage induced by freezing/thawing is enhanced when vessels are subjected to flow in an in vitro circuit.
本研究旨在探讨在体外血流循环装置中,不同解冻程序对冷冻保存的猪动脉解冻前后所产生的变化。小型猪髂动脉段在完全最小必需培养基加10%二甲亚砜中进行冷冻保存,并在-196℃液氮中储存30天。根据动脉标本是否新鲜(对照组,n = 20)、在37℃下冷冻保存并快速解冻(RT,n = 22)或冷冻保存并进行可控的自动缓慢解冻(ST,n = 22),设立了三个研究组。随后,每组一半的标本被置于血流循环装置中72小时。对形态学和超微结构变化进行评估。采用TUNEL法确定细胞损伤情况。所有冷冻保存的标本均出现内皮剥脱,其中快速解冻的标本内皮剥脱最为广泛。与快速解冻的标本相比,缓慢解冻的标本细胞活力和血管壁结构有所改善。在血流条件下,冻融过程所导致的损伤会加剧。这些研究结果表明:(a)冷冻保存动脉的缓慢解冻可改善血管结构和活力的保存情况;(b)当血管在体外循环装置中处于血流状态时,冻融所导致的损伤会加剧。