Faggioli G L, Gargiulo M, Giardino R, Pasquinelli G, Preda P, Fini M, Corbascio M, Stella A, D'Addato M, Ricotta J J
Department of Surgery, State University of New York at Buffalo.
Cardiovasc Surg. 1994 Apr;2(2):259-65.
This study was undertaken to determine the effect of long-term cryopreservation on graft ultrastructure and endothelial cell viability in an animal model. The jugular veins from 12 New Zealand White rabbits were excised with a 'no-touch' technique and divided into four groups: control group (fresh veins); group 1, veins cryopreserved for 1 month; group 2, veins cryopreserved for 2 months; and group 3, veins cryopreserved for 3 months. Cryopreservation was accomplished by rapid freezing (-5 degrees C s-1 to -196 degrees C) in a solution of 17.5% dimethylsulphoxide and 20% fetal bovine serum and by storage in liquid nitrogen. Veins were then implanted as a carotid autograft (three grafts/group). At the time of graft implantation a segment of the paired matched vein was perfusion-fixed and evaluated by scanning and transmission electron microscopy, whereas the remainder were subjected to endothelial cell culture techniques to determine cell viability. Autografts were removed 1 month after implantation and subjected to similar evaluations. Histological changes seen in cryopreserved veins were dependent on preservation time and included focal endothelial cell blebbing, cytoplasmic vacuolization and disruption of cell-to-cell contacts. Smooth muscle cells showed mitochondrial swelling. Patency was identical in all groups (66.6%). Explants at 1 month were similar in histological appearance to fresh veins with a smooth endothelial cell lining arranged longitudinally and intact cell junctions. Endothelial cells could be cultured from fresh veins and 1-month-old explants but not from the cryopreserved graft surface before implantation. the present technique of cryopreservation leads to some damage of graft architecture and loss of endothelial cell viability.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究旨在确定在动物模型中，长期冷冻保存对移植物超微结构和内皮细胞活力的影响。采用“无接触”技术切除12只新西兰白兔的颈静脉，并将其分为四组：对照组（新鲜静脉）；第1组，冷冻保存1个月的静脉；第2组，冷冻保存2个月的静脉；第3组，冷冻保存3个月的静脉。通过在含有17.5%二甲基亚砜和20%胎牛血清的溶液中快速冷冻（-5℃/秒至-196℃）并储存在液氮中来实现冷冻保存。然后将静脉作为颈动脉自体移植物植入（每组3个移植物）。在移植物植入时，对配对匹配静脉的一段进行灌注固定，并通过扫描和透射电子显微镜进行评估，而其余部分则采用内皮细胞培养技术来确定细胞活力。自体移植物在植入1个月后取出并进行类似评估。冷冻保存静脉中观察到的组织学变化取决于保存时间，包括局灶性内皮细胞起泡、细胞质空泡化和细胞间接触的破坏。平滑肌细胞显示线粒体肿胀。所有组的通畅率相同（66.6%）。1个月时的移植物在组织学外观上与新鲜静脉相似，内皮细胞衬里光滑，纵向排列且细胞连接完整。内皮细胞可以从新鲜静脉和1个月大的移植物中培养出来，但不能从植入前冷冻保存的移植物表面培养出来。目前的冷冻保存技术会导致移植物结构的一些损伤和内皮细胞活力的丧失。（摘要截断于250字）
