Characterization of a simplified ice-free cryopreservation method for heart valves
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Characterization of a simplified ice-free cryopreservation method for heart valves Agnes J. T. Huber • Timo Aberle • Martina Schleicher • Hans-Peter Wendel Kelvin G. M. Brockbank
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Received: 13 April 2012 / Accepted: 21 May 2012 Springer Science+Business Media B.V. 2012
Abstract The aim of the present study was to characterize the hemocompatibility of ice-free cryopreserved heart valves in anticipation of future human trials. Porcine pulmonary heart valves were infiltrated with either an 83 % cryoprotectant solution followed by rapid cooling and storage at -80 C or with 10 % DMSO and control rate freezing to -80 C and storage in vapor phase nitrogen as conventional frozen controls. Cryopreserved leaflets were compared with fresh, decellularized and glutaraldehyde-fixed control valve leaflets using a battery of coagulation protein assays after exposure to human blood. Von Willebrand Factor staining indicated that most of the endothelium was lost during valve processing prior to cryopreservation. Hemocompatibility, employing thrombin/ A. J. T. Huber T. Aberle M. Schleicher H.-P. Wendel Department of Thoracic, Cardiac and Vascular Surgery, University Hospital, Tu¨bingen, Germany e-mail: [email protected] K. G. M. Brockbank (&) Cell & Tissue Systems, Inc., 2231 Technical Parkway, Suite A, North Charleston, SC 29406, USA e-mail: [email protected] K. G. M. Brockbank Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USA K. G. M. Brockbank Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC, USA
antithrombin-III-complex, polymorphonuclear neutrophil-elastase, beta-thromboglobulin and terminal complement complex SC5b-9, was preserved compared with both fresh and frozen leaflets. Hemocompatibility differences were observed for cryopreserved leaflets versus both decellularized and glutaraldehyde fixed controls. In conclusion, the hemocompatibility results support the use of ice-free cryopreservation as a simplified preservation method because no statistically significant differences in hemocompatibility were observed between the two cryopreservation methods and fresh untreated controls. Keywords Cryopreservation Heart valve Hemocompatibility Endothelium Allograft Storage
Introduction Conventional frozen cryopreservation (CFC) is the method of choice for preserving allograft heart valves in tissue banks. Donor-derived human CFC valves are clinically employed because of their low incidence of thromboembolic events, resistance towards inflammatory processes such as endocarditis, and good hemodynamics. Generally, allografts, also known as homografts, have a better long term durability than porcine bioprostheses, however, allograft deterioration leading to valve dysfunction has been observed in long term follow up studies (Hopkins 2004;
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Takkenberg et al. 2002). Currently the gold standard for preserving allografts is controlled rate freezing with dimethylsul
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