title: Does prolonged post-mortem cold ischemic harvesting time influence cryopreserved pulmonary homograft tissue integrity?
abstract: This study investigated cryopreserved pulmonary homograft (CPA) structural integrity after prolonged cold ischemic harvesting times in a juvenile sheep model. Three groups with different post-mortem cold ischemic harvesting times were studied, i.e. Group 1 (24 h, n = 10); group 2 (48 h, n = 10); group 3 (72 h, n = 10). In each group, 5 CPAs were studied in vitro after cryopreservation and thawing. The other 5 CPAs were implanted in juvenile sheep for a minimum of 180 days. Serology samples were obtained and echocardiography was performed before euthanasia. Hematoxylin and eosin (H&E), scanning electron microscopy (SEM), von Kossa, Picrosirius red, α-actin, immunohistochemistry [von Willebrand factor (vWF), CD4, CD31 and CD34] and calcium content analyses were performed on explanted CPAs. The in vitro and in vivo studies failed to demonstrate any change in tensile strength, Young's Modulus and thermal denaturation (Td) results between the groups. SEM demonstrated a reduction in endothelial cells (50 % at 24 h, 60.9 % at 48 h and 40.9 % at 72 h), but H&E could not demonstrate autolysis in any CPA in vitro. All cultures were negative. In the explanted groups, IgE, IgM and IgG results were inconclusive. Echocardiography demonstrated normal valve function in all groups. H&E and Picrosirius red staining confirmed tissue integrity. vWF, CD31 and CD34 staining confirmed a monolayer of endothelial cells in all explanted valves. Calcium content of explanted CPA leaflets was similar. This experimental study supports the concept of prolonging the cold ischemic harvesting time of cryopreserved homografts to reduce homograft shortage.
authors: Department Cardiothoracic Surgery, Faculty of Health Sciences, University of the Free State (UFS), Bloemfontein, South Africa.
authors: Francis Edwin
authors: FE
authors: Smit
authors: Department Cardiothoracic Surgery, Faculty of Health Sciences, University of the Free State (UFS), Bloemfontein, South Africa.
authors: Dreyer
authors: D
authors: Bester
authors: Department Cardiothoracic Surgery, Faculty of Health Sciences, University of the Free State (UFS), Bloemfontein, South Africa.
authors: Johannes Jacobus
authors: JJ
authors: van den Heever
authors: Department of Cardiac Surgery, Heart Centre Leipzig, University of Leipzig, Leipzig, Saxony, Germany.
authors: Franziska
authors: F
authors: Schlegel
authors: Department of Health Sciences, Central University of Technology (CUT), Free State, Private Bag X20539, Bloemfontein, South Africa. lbotes@cut.ac.za.
authors: Lezelle
authors: L
authors: Botes
authors: Department Cardiothoracic Surgery, Faculty of Health Sciences, University of the Free State (UFS), Bloemfontein, South Africa.|Department of Cardiac Surgery, Heart Centre Leipzig, University of Leipzig, Leipzig, Saxony, Germany.
authors: Pascal Maria
authors: PM
authors: Dohmen
mesh_terms: D064591:Allografts; D000818:Animals; D050377:Cold Ischemia / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D055119:Elastic Modulus; D006085:Graft Survival / Q000502:physiology*; D008297:Male; D011180:Postmortem Changes; D011664:Pulmonary Valve / Q000166:cytology / Q000502:physiology* / Q000637:transplantation*; D012756:Sheep; D013718:Tensile Strength
keywords: Cryopreserved homografts; Juvenile sheep model; Right ventricular outflow tract reconstruction; Warm ischemic time