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Decellularized aortic conduits: could their cryopreservation affect post-implantation outcomes? A morpho-functional study on porcine homografts

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Abstract

Decellularized porcine aortic valve conduits (AVCs) implanted in a Vietnamese Pig (VP) experimental animal model were matched against decellularized and then cryopreserved AVCs to assess the effect of cryopreservation on graft hemodynamic performance and propensity to in vivo repopulation by host’s cells. VPs (n = 12) underwent right ventricular outflow tract substitution using AVC allografts and were studied for 15-month follow-up. VPs were randomized into two groups, receiving AVCs treated with decellularization alone (D; n = 6) or decellularization/cryopreservation (DC; n = 6), respectively. Serial echocardiography was carried out to follow up hemodynamic function. All explanted AVCs were processed for light and electron microscopy. No signs of dilatation, progressive stenosis, regurgitation, and macroscopic calcification were echocardiographically observed in both D and DC groups. Explanted D grafts exhibited near-normal features, whereas the presence of calcification, inflammatory infiltrates, and disarray of elastic lamellae occurred in some DC grafts. In the unaltered regions of AVCs from both groups, almost complete re-endothelialization was observed for both valve cusps and aorta walls. In addition, side-by-side repopulation by recipient’s fibroblasts, myofibroblasts, and smooth muscle cells was paralleled by ongoing tissue remodeling, as revealed by the ultrastructural identification of typical canals of collagen fibrillogenesis and elastogenesis-related features. Incipient neo-vascularization and re-innervation of medial and adventitial tunicae of grafted aortic walls were also detected for both D and DC groups. Cryopreservation did not affect post-implantation AVC hemodynamic behavior and was topically propensive to cell repopulation and tissue renewal, although graft deterioration including calcification was present in several areas. Thus, these preliminary data provide essential information on feasibility of decellularization and cryopreservation coupling in the perspective of treatment optimization and subsequent clinical trials using similarly treated human allografts as innovative heart valve substitutes.

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    Authors and Affiliations

    1. Division of Cardiac Surgery, Department of Cardiac, Thoracic and Vascular Sciences, University of Padua, Via Giustiniani 2, 35128, Padua, Italy

      Michele Gallo, Filippo Naso, Tomaso Bottio, Roberto Bianco, Laura Iop & Gino Gerosa

    2. Department of Experimental and Clinical Medicine, University of Udine, Udine, Italy

      Antonella Bonetti, Maurizio Marchini & Fulvia Ortolani

    3. Department of Animal Medicine, Productions and Health, University of Padua, Legnaro, Italy

      Helen Poser, Paolo Franci & Roberto Busetto

    4. Department of Biomedical Sciences, University of Padua, Padua, Italy

      Michele Spina

    5. Tissue Bank of Veneto Region, Treviso Regional Hospital, Treviso, Italy

      Adolfo Paolin

    6. Department of Cardiac, Thoracic and Vascular Sciences, Biostatistics, Epidemiology and Public Health Unit, University of Padua, Padua, Italy

      Anna Chiara Frigo

    7. Department of Cardiothoracic Surgery, Royal Children’s Hospital, Murdoch Children’s Research Institute, University of Melbourne, Melbourne, Australia

      Edward Buratto

    8. Cardiovascular Regenerative Medicine, Venetian Institute of Molecular Medicine (VIMM), Biomedical Campus Pietro d’Abano, Padua, Italy

      Laura Iop & Gino Gerosa

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    1. Michele Gallo
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    Correspondence to Michele Gallo.

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    Funding

    This study was funded by the RSF 286/08 Regione Veneto Grant and Azione Biotech III.

    Conflict of interest

    The authors do not have any conflicts to disclose. No competing financial interests exist.

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    All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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    M. Gallo and A. Bonetti contributed equally to this work.

    F. Ortolani, L. Iop and G. Gerosa are equally responsible for the concept and design of the study.

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    Gallo, M., Bonetti, A., Poser, H. et al. Decellularized aortic conduits: could their cryopreservation affect post-implantation outcomes? A morpho-functional study on porcine homografts. Heart Vessels 31, 1862–1873 (2016). https://doi.org/10.1007/s00380-016-0839-5

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