Patency and structural changes in cryopreserved arterial grafts used as vessel substitutes in the rat

Abstract

To evaluate the patency and structural changes that occur in the short- and mid-term when cryopreserved syngenic arterial grafts are implanted in an experimental animal model. Segments of iliac artery from the Spraque-Dawley rat were cryopreserved in a biological freezer according a controlled, computerized freezing protocol whereby the specimens are cooled at a rate of 1 degrees C/min. After storage at -145 degrees C in liquid N2 vapor for 30 days, the cryografts were slowly thawed. These vessels were grafted to the common iliac artery in syngenic animals. The following study groups were established: group I (GI), non-implanted cryografts; group II (GII), autografts; and group III (GIII), cryoisografts. The control group (CG) was comprised of fresh iliac arteries. The animals were sacrificed 14, 30, or 90 days post-surgery. At each of these follow-up times, graft specimens were morphologically evaluated by light and scanning and transmission electron microscopy and immunolabeling of endothelial cells (vWf). Cell damage attributed to the cryopreservation or grafting process was also determined. At the time of sacrifice, graft patency was 100% for the autografts, while 26.6% of the cryoisografts showed fully occlusive thrombosis. Among other complications, two pseudoaneurysms were detected. After cryopreservation, the grafts (GI) showed patches of endothelial denudation and good cellularity of the medial layer. The intimal hyperplasia observed in autografts implanted for 14 days (GII) was significantly delayed until day 30 when the graft was cryopreserved (GIII). Cryoisografts showed general thinning of the arterial wall and degeneration accompanied by medial layer cell loss. These grafts showed most cell damage at 90 days post-implant. Expression of the vWf in all specimens showing intimal hyperplasia was confined to the outermost graft layer. Cryopreservation modified the reparative response of the grafts. Owing to faster degeneration of the medial layer and a delay in the appearance of intimal hyperplasia, arterial wall thickness was reduced relative to that of the non-cryopreserved autografts. This thinning, at least in the short-term (90 days), does not seem to give rise to aneurysms owing to the generation of a neointima that stabilizes the vessel wall.