Effects of cryopreservation on the immunogenicity of porcine arterial allografts in early stages of transplant vasculopathy

Abstract

Background The number of revascularization procedures including coronary and lower extremity bypass, have increased greatly in the last decade. It suggests a growing need for vascular grafts. Cryopreserved allografts could represent a viable alternative but their immunologic reactivity remains controversial. Methods 71 pigs (40 recipients and 31 donors) were used. Two femoral grafts per recipient animal were implanted for 3, 7, and 30 days. Types of grafts: fresh autograft as a control graft ( n = 19), fresh allograft ( n = 31) and cryopreserved allograft ( n = 30). Histological and immunohistochemical studies were performed. Results Fresh allografts compared to autografts showed intimal inflammatory infiltration at 3 days (328 vs. 0 macrophages/mm 2; P < 0.05) and 7 days (962 vs. 139 T lymphocytes/mm 2; P < 0.05) post-transplantation. At 30 days, there was a loss of endothelial cells, presence of luminal thrombus and aneurismal lesions (total area = 15.8 vs. 8.4 mm 2; P < 0.05). Cryopreservation did not reduce these lesions nor modify endothelial nitric oxide synthase (eNOS) expression nor modify the number of animals that developed anti-SLA antibodies. Moreover, at 7 days, cryopreserved allografts compared to fresh allografts showed a higher expression of P-selectin (5 out of 5 vs. 1 out of 5; P < 0.05) and, at 30 days, a greater inflammatory reactivity (2692 vs. 1107 T lymphocytes/mm 2 in media; P < 0.05) with a trend towards a higher presence of multinucleated giant cells than in the fresh ones. Conclusions The cryopreservation method used maintained immunogenicity of allografts and increased the inflammatory reactivity found in fresh allografts up to 30 days of vascular transplantation.