Bellón J M, Gimeno M J, Pascual G, Garcia-Honduvilla N, Dominguez B, Buján J
Department of Morphological Sciences and Surgery, Faculty of Medicine, University of Alcalá de Henares, Madrid, Spain.
Eur J Vasc Endovasc Surg. 1999 Feb;17(2):136-43. doi: 10.1053/ejvs.1998.0739.
The aim of the present study was to investigate the changes which occur to the arterial wall following cryopreservation and thawing and to determine whether these changes are reversible after a week of culture in an organ bath.
Rat iliac arterial segments were cryopreserved. Once thawed, the arterial segments were cultured for a period of 0, 1, 2, 4 or 7 days. Freshly isolated rat iliac vessels cultured for 7 days served as the control group. Evaluation was made of ultrastructural changes, the expression of metalloproteinase activity (MMP-1, MMP-3 and MMP-9) and the apoptotic state of cells.
The freezing-thawing process induced damage to the arterial segments compared to fresh control vessels. After 1 week of culture, arteries showed a high degree of tissue degeneration. Only a few individual endothelial cells remained on the luminal surface. There was a gradual increase in the proportion of apoptotic cells. The sequential expression of MMP-1 during the first 2 days and subsequent expression of MMP-3 and MMP-9 were of most significance.
Cryopreservation induced damage to the vessels which could not be reversed by organ culture. The changes observed in the expression of metalloproteinases may be indicative of the degenerative process which occurs in the extracellular matrix.
本研究旨在调查冷冻保存和解冻后动脉壁发生的变化,并确定在器官浴中培养一周后这些变化是否可逆。
对大鼠髂动脉段进行冷冻保存。解冻后,将动脉段培养0、1、2、4或7天。将新鲜分离的大鼠髂血管培养7天作为对照组。对超微结构变化、金属蛋白酶活性(MMP-1、MMP-3和MMP-9)的表达以及细胞凋亡状态进行评估。
与新鲜对照血管相比,冻融过程对动脉段造成了损伤。培养1周后,动脉出现高度组织变性。管腔表面仅残留少数单个内皮细胞。凋亡细胞比例逐渐增加。MMP-1在最初2天的顺序表达以及随后MMP-3和MMP-9的表达最为显著。
冷冻保存导致血管损伤,器官培养无法使其逆转。金属蛋白酶表达的变化可能表明细胞外基质中发生的退行性过程。