Špaček Miroslav, Měřička Pavel, Janoušek Libor, Dalecká Markéta, Benda Aleš, Krs Otakar, Slížová Dáša, Špunda Rudolf, Hrubý Jan, Matia Ivan, Honegrová Barbora, Lindner Jaroslav
2nd Department of Surgery-Department of Cardiovascular Surgery, 1st Medical Faculty, Charles University and General University Hospital, Prague, Czech Republic.
Tissue Bank University Hospital Hradec, Hradec Králové, Czech Republic.
Ann Vasc Surg. 2020 Apr;64:347-354. doi: 10.1016/j.avsg.2019.11.026. Epub 2019 Nov 16.
The aim of our study was to assess the impact of different thawing protocols on morphological changes arising in cryopreserved human saphenous vein grafts.
The study was performed in 12 saphenous vein grafts harvested in brain death donors. Storage in the vapor phase of liquid nitrogen for 3 or 5 years followed. Two thawing protocols were tested: slow thawing in a refrigerator at temperature +4°C for 2 hr and rapid thawing-in a water bath at +37°C. Grafts were processed for scanning electron microscopy. Comparisons of continuous parameters under study between experimental groups were performed using the t-test (age, cold ischemia time, exposure to cryoprotectant, time of storage, total thawing time, mean thawing rate, morphology scoring of thawed HSVG) and the median test (HSVG length). Categorical parameters (sex and blood group) were formally tested using the chi-square test.
All samples were evaluated according to morphological changes and scored in terms of morphologically intact endothelium, confluent endothelium with structural inhomogeneity, disruption of the intercellular contacts, separation of the endothelial cells, complete loss of the endothelium, and damage of the subendothelial layers. There is no statistically significant difference between the sample sets at the significance level of 0.05. There was no association with donors' age, sex, and time of storage.
Human cryopreserved saphenous vein grafts in our experimental work showed no difference in terms of structural deterioration of the endothelial surface and basal membrane depending on different thawing protocols used.
我们研究的目的是评估不同解冻方案对冷冻保存的人隐静脉移植物中出现的形态学变化的影响。
该研究在12例从脑死亡供体获取的隐静脉移植物中进行。随后在液氮气相中储存3年或5年。测试了两种解冻方案:在+4°C冰箱中缓慢解冻2小时和在+37°C水浴中快速解冻。对移植物进行扫描电子显微镜处理。使用t检验(年龄、冷缺血时间、接触冷冻保护剂的时间、储存时间、总解冻时间、平均解冻速率、解冻后隐静脉移植物的形态学评分)和中位数检验(隐静脉移植物长度)对实验组之间研究的连续参数进行比较。使用卡方检验对分类参数(性别和血型)进行正式检验。
根据形态学变化对所有样本进行评估,并根据内皮形态完整、内皮融合但结构不均匀、细胞间接触破坏、内皮细胞分离、内皮完全丧失以及内皮下层损伤进行评分。在0.05的显著性水平下,样本集之间没有统计学上的显著差异。与供体的年龄、性别和储存时间无关。
在我们的实验工作中,冷冻保存的人隐静脉移植物在使用不同解冻方案时,在内皮表面和基底膜的结构恶化方面没有差异。