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29 September 2017 : Original article  

Machine Perfusion at 20°C Prevents Ischemic Injury and Reduces Hypoxia-Inducible Factor-1α Expression During Rat Liver Preservation

Clarissa Berardo1BCE*, Laura Giuseppina Di Pasqua1BCE, Veronica Siciliano1B, Vittoria Rizzo2BC, Plinio Richelmi1G, Andrea Ferrigno1ABCDEF, Mariapia Vairetti1ADEFG

DOI: 10.12659/AOT.904631

Ann Transplant 2017; 22:581-589

Abstract

BACKGROUND: Ischemic cholangitis is the main cause of liver failure after transplantation and subnormothermic machine perfusion may represent a better strategy than conventional cold storage, minimizing preservation injury. We compared livers preserved by machine perfusion at 20°C (MP20) or by cold storage at 4°C (CS4) with regard to hypoxia-inducible factor (HIF)-1α mRNA expression and protein stabilization in hypoxic conditions.

MATERIAL AND METHODS: Livers from male Wistar rats were stored on ice at 4°C in UW solution (CS4) or perfused with oxygenated Krebs-Henseleit buffer at 20°C (MP20) for six hours. After preservation, the livers were reperfused for two hours with oxygenated Krebs-Henseleit buffer at 37°C to simulate reimplantation. We collected bile, perfusate, and tissue samples. Transaminases, lactate dehydrogenase, bilirubin, and lactic acid were assayed in the perfusate and bile. ATP/ADP, glycogen, HIF-1α mRNA, and protein expression were measured in the tissue homogenates.

RESULTS: At the end of preservation, as well as after reperfusion, HIF-1α mRNA expression was significantly higher in the ischemic CS4 livers. Although the hypoxic conditions found in CS4 preservation stabilized HIF-1α protein was significantly higher in the CS4 livers at the end of preservation, no difference was observed after reperfusion, likely because of the oxygen in the reperfusion medium. After reperfusion, the MP20 livers released less transaminases and LDH. The MP20 livers had higher ATP/ADP, glycogen, and biliary bilirubin after both preservation and reperfusion when compared with the CS4 livers.

CONCLUSIONS: The data demonstrated that MP20 was associated with a lower HIF-1α expression and organ injury with respect to CS4, suggesting that oxygen provided by this preservation setting might approximate the organ request, thus avoiding the ischemic injury usually observed during organ preservation by cold storage.

Keywords: cold ischemia, Cryopreservation, Liver Transplantation

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Annals of Transplantation eISSN: 2329-0358
Annals of Transplantation eISSN: 2329-0358