Characterization of a simplified ice-free cryopreservation method for heart valves.
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.
Department of Thoracic, Cardiac and Vascular Surgery, University Hospital, Tübingen, Germany. agnes.huber@med.uni-tuebingen.de
Agnes J T
AJ
Huber
Timo
T
Aberle
Martina
M
Schleicher
Hans-Peter
HP
Wendel
Kelvin G M
KG
Brockbank
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism / Q000473:pathology; D006351:Heart Valves / Q000187:drug effects* / Q000473:pathology / Q000637:transplantation*; D007053:Ice / Q000009:adverse effects; D023421:Models, Animal; D013552:Swine; D014841:von Willebrand Factor / Q000378:metabolism
D003451:Cryoprotective Agents; D007053:Ice; D014841:von Willebrand Factor; D004121:Dimethyl Sulfoxide