Characterization of a simplified ice-free cryopreservation method for heart valves
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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/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.
KeywordsCryopreservation Heart valve Hemocompatibility Endothelium Allograft Storage
The authors would like to thank Ilka Degenkolbe, Michaela Braun und Doris Armbruster for their excellent technical assistance, and the Slaughterhouse Gärtringen and Mrs. Quindt for providing tissues. This work was supported by the Deutsche Forschungsgemeinschaft (STO359/7-1).
Conflict of interest
Dr. Brockbank is an owner and employee of Cell and Tissue Systems, Inc. No competing financial interests exist for any other authors.
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