OBJECTIVES AND DESIGN: At the present time there are two waiting list for patients with vascular prosthetic infection indicated for arterial transplantation in the Czech Republic. The inclusion of each patient for cold-stored or cryopreserved arterial transplantation is the preference of indicating surgeon. In this experimental work we studied the immunogenicity of rat aortal allografts treated by our new clinical cryopreservation/slow thawing protocol. MATERIAL AND METHODS: Brown-Norway (BN) (N = 6, 203-217 g) or Lewis (LEW) (N = 6, 248-254 g) abdominal aortal grafts treated in accordance with our new clinical cryopreservation/slow thawing protocol were orthotopically transplanted to Lewis recipients (N = 12, 191-245 g). Aortal wall histology and infiltration by recipient immune cells, as well as donor specific anti MHC class I and II antibodies in recipient serum were studied in both isografts and allografts on day 30 postransplant. Core data of cryopreserved allografts were compared to our previous data of cold-stored aortal allografts treated in accordance with our clinical cold-storage protocol. RESULTS: Cryopreserved allografts showed regular morphology of aortal wall with clear differentiation of all three basic anatomical layers on day 30 postransplant. Intimal layer showed no hyperplasia, luminal surface was covered by endothelial cells. No statistical difference was observed in tunica media thickness between isografts and allografts. The medial layer showed no necrosis, shrinkage or immunoglobuline G deposition in any experimental group. The adventitial infiltration by immune cells was significantly higher (P<0.05) in allografts. Cryopreserved allografts showed significant lower activation of both cell- and antibody mediated immunity compared to historical data of cold-stored allografts. CONCLUSION: Aortal wall histology of rat allografts treated by our new standardized clinical cryopreservation/slow thawing protocol was comparable to that of the cryopreserved isografts on day 30 posttranspant. The immunogenicity of cryopreserved aortal allografts was significantly lower compared to that of cold-stored aortal allografts.

2nd Department of Surgery Department of Cardiovascular Surgery 1st Faculty of Medicine Charles University Prague and General University Hospital Prague Czech Republic

Department of Cardio Vascular Surgery Nord Hospital and Karl Landsteiner Institute for Cardio Vascular Research Vienna Austria

Department of Surgery Campus Charité Mitte and Campus Virchow Klinikum Charité Universitätsmedizin Berlin Berlin Germany

Institute of Physiology 1st Faculty of Medicine Charles University Prague

Teaching Center Medizinische Universität Wien Vienna Austria

Technology Centre of the Academy of Sciences of the Czech Republic Prague Czech Republic

Tissue Bank Faculty Hospital Hradec Kralove Charles University Faculty of Medicine in Hradec Kralove Hradec Kralove Czech Republic

Translational Centre for Regenerative Medicine University of Leipzig Leipzig Germany

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Berger P, Vaartjes I, Moll FL, De Borst GJ, Blankensteijn JD, Bots ML. Cumulative incidence of graft infection after primary prosthetic aortic reconstruction in the endovascular era. Eur J Vasc Endovasc Surg. 2015;49: 581–5. 10.1016/j.ejvs.2015.01.001 PubMed DOI

Antonopoulos CN, Papakonstantinou NA, Hardy D, Lyden SP. Editor’s Choice—Cryopreserved Allografts for Arterial Reconstruction after Aorto-Iliac Infection: A Systematic Review and Meta-Analysis. Eur J Vasc Endovasc Surg. 2019;58: 120–128. 10.1016/j.ejvs.2019.03.003 PubMed DOI

Erb S, Sidler JA, Elzi L, Gurke L, Battegay M, Widmer AF, et al. Surgical and antimicrobial treatment of prosthetic vascular graft infections at different surgical sites: A retrospective study of treatment outcomes. PLoS One. 2014;9. PubMed PMC

Kieffer E, Gomes D, Chiche L, Fléron M-H, Koskas F, Bahnini A. Allograft replacement for infrarenal aortic graft infection: early and late results in 179 patients. J Vasc Surg. 2004;39: 1009–1017. 10.1016/j.jvs.2003.12.040 PubMed DOI

Touma J, Cochennec F, Parisot J, Fialaire Legendre A, Becquemin J-P, Desgranges P. In Situ Reconstruction in Native and Prosthetic Aortic Infections Using Cryopreserved Arterial Allografts. Eur J Vasc Endovasc Surg. 2014;48: 292–299. 10.1016/j.ejvs.2014.04.023 PubMed DOI

Heo S-H, Kim Y-W, Woo S-Y, Park Y-J, Kim D-K, Chung D-R. Recent Results of In Situ Abdominal Aortic Reconstruction with Cryopreserved Arterial Allograft. Eur J Vasc Endovasc Surg. 2017;53: 158–167. 10.1016/j.ejvs.2016.07.090 PubMed DOI

Matia I, Adamec M, Varga M, Janousek L, Lipar K, Viklicky O. Aortoiliac reconstruction with allograft and kidney transplantation as a one-stage procedure: long term results. Eur J Vasc Endovasc Surg. 2008;35: 353–7. 10.1016/j.ejvs.2007.09.022 PubMed DOI

Špaček M, Měřička P, Janoušek L, Štádler P, Adamec M, Vlachovský R, et al. Current vascular allograft procurement, cryopreservation and transplantation techniques in the Czech Republic. Adv Clin Exp Med. 2019;28: 529–534. 10.17219/acem/90037 PubMed DOI

Špaček M, Měřička P, Janoušek L, Štádler P, Adamec M, Vlachovský R, et al. Organization model for allotransplantations of cryopreserved vascular grafts in Czech Republic. Cell Tissue Bank. 2018;19: 437–445. 10.1007/s10561-018-9691-4 PubMed DOI

Rodríguez M, Pascual G, Pérez-Köhler B, Cifuentes A, Garcia-Honduvilla N, Bellón JM, et al. Immune response to the long-term grafting of cryopreserved small-diameter arterial allografts. Histol Histopathol. 2012;27: 873–84. 10.14670/HH-27.873 PubMed DOI

Konrad H, Wahle A, Altermann W, Schlaf G. Strong Humoral Anti-HLA Immune Response Upon Arbitrarily Chosen Allogeneic Arterial Vessel Grafts. J Clin Cell Immunol. 2017;8: 1–10.

Szilagyi DE, Rodriguez FJ, Smith RF, Elliott JP. Late fate of arterial allografts. Observations 6 to 15 years after implantation. Arch Surg. 1970;101: 721–33. 10.1001/archsurg.1970.01340300077014 PubMed DOI

Matia I, Lodererova A, Adamec M. Delayed administration of FK 506 is sufficient to suppress acute rejection changes after aortal transplantation in rats. Transpl Int. 2007;20: 371–80. 10.1111/j.1432-2277.2006.00446.x PubMed DOI

Spunda R, Hruby J, Mericka P, Mlcek M, Pecha O, Splith K, et al. Immunosuppressive protocols with tacrolimus after cryopreserved aortal allotransplantation in rats. Stepkowski S, editor. PLoS One. 2018;13: PubMed PMC

Matia I, Fellmer P, Splith K, Varga M, Adamec M, Kämmerer I, et al. Immunosuppressive protocol with delayed use of low-dose tacrolimus after aortic transplantation suppresses donor-specific anti-MHC class I and class II antibody production in rats. Ann Transplant. 2014;19: 225–32. 10.12659/AOT.889870 PubMed DOI

Pascual G, Jurado F, Rodríguez M, Corrales C, López-Hervás P, Bellón JM, et al. The use of ischaemic vessels as prostheses or tissue engineering scaffolds after cryopreservation. Eur J Vasc Endovasc Surg. 2002;24: 23–30. 10.1053/ejvs.2002.1663 PubMed DOI

Knight RJ, Liu H, Fishman E, Reis ED. Cold ischemic injury, aortic allograft vasculopathy, and pro-inflammatory cytokine expression. J Surg Res. 2003;113: 201–7. 10.1016/s0022-4804(03)00199-9 PubMed DOI

Buján J, Pascual G, López R, Corrales C, Rodríguez M, Turégano F, et al. Gradual thawing improves the preservation of cryopreserved arteries. Cryobiology. 2001;42: 256–65. 10.1006/cryo.2001.2329 PubMed DOI

Novotny R, Slizova D, Hlubocky J, Krs O, Spatenka J, Burkert J, et al. Cryopreserved human aortic root allografts arterial wall: Structural changes occurring during thawing. PLoS One. 2017;12: e0175007 10.1371/journal.pone.0175007 PubMed DOI PMC

Thaunat O, Louedec L, Dai J, Bellier F, Groyer E, Delignat S, et al. Direct and Indirect Effects of Alloantibodies Link Neointimal and Medial Remodeling in Graft Arteriosclerosis. Arterioscler Thromb Vasc Biol. 2006;26: 2359–2365. 10.1161/01.ATV.0000241980.09816.ac PubMed DOI

Lou H, Kodama T, Zhao YJ, Maurice P, Wang YN, Katz N, et al. Inhibition of Transplant Coronary Arteriosclerosis in Rabbits by Chronic Estradiol Treatment Is Associated With Abolition of MHC Class II Antigen Expression. Circulation. 1996;94: 3355–3361. 10.1161/01.cir.94.12.3355 PubMed DOI

Viability of Human Arterial Grafts Monitored by Comet Assay

Cell Viability Assessment Using Fluorescence Vital Dyes and Confocal Microscopy in Evaluating Freezing and Thawing Protocols Used in Cryopreservation of Allogeneic Venous Grafts