Viability assessment of the liver during ex-situ machine perfusion prior to transplantation

Purpose of review:In an attempt to reduce waiting list mortality in liver transplantation, less-than-ideal quality donor livers from extended criteria donors are increasingly accepted. Predicting the outcome of these organs remains a challenge. Machine perfusion provides the unique possibility to assess donor liver viability pretransplantation and predict postreperfusion organ function. Recent findings: Assessing liver viability during hypothermic machine perfusion remains challenging, as the liver is not metabolically active. Nevertheless, the levels of flavin mononucleotide, transaminases, lactate dehydrogenase, glucose and pH in the perfusate have proven to be predictors of liver viability. During normothermic machine perfusion, the liver is metabolically active and in addition to the perfusate levels of pH, transaminases, glucose and lactate, the production of bile is a crucial criterion for hepatocyte viability. Cholangiocyte viability can be determined by analyzing bile composition. The differences between perfusate and bile levels of pH, bicarbonate and glucose are good predictors of freedom from ischemic cholangiopathy. Summary:Although consensus is lacking regarding precise cut-off values during machine perfusion, there is general consensus on the importance of evaluating both hepatocyte and cholangiocyte compartments. The challenge is to reach consensus for increased organ utilization, while at the same time pushing the boundaries by expanding the possibilities for viability testing.</p

The Emerging Role of Viability Testing During Liver Machine Perfusion

The transplant community continues to be challenged by the disparity between the need for liver transplantation and the shortage of suitable donor organs. At the same time, the number of unused donor livers continues to increase, most likely attributed to the worsening quality of these organs. To date, there is no reliable marker of liver graft viability that can predict good posttransplant outcomes. Ex situ machine perfusion offers additional data to assess the viability of donor livers before transplantation. Hence, livers initially considered unsuitable for transplantation can be assessed during machine perfusion in terms of appearance and consistency, hemodynamics, and metabolic and excretory function. In addition, postoperative complications such as primary nonfunction or posttransplant cholangiopathy may be predicted and avoided. A variety of viability criteria have been used in machine perfusion, and to date there is no widely accepted composition of criteria for clinical use. This review discusses potential viability markers for hepatobiliary function during machine perfusion, describes current limitations, and provides future recommendations for the use of viability criteria in clinical liver transplantation

The first case of ischemia-free organ transplantation in humans:A proof of concept

With great interest we have read the article by Xiaoshun He and colleagues regarding their first patient who underwent successful ischemia-free organ transplantation (IFOT).1 This group transplanted a liver donated after brain death to a 51-year-old patient with decompensated cirrhosis and hepatocellular carcinoma without any ischemic episode or interruption of the blood circulation. The graft was procured, ex-situ preserved and implanted under continuous normothermic machine perfusion using the Liver Assist device (Organ Assist, Groningen, The Netherlands). The donor liver had 85-95% macrovesicular steatosis, however there was no post-reperfusion syndrome observed after revascularization of the graft and the recipient had an uneventful recovery. This article is protected by copyright. All rights reserved

Oxygenated versus non-oxygenated flush out and storage of donor livers:An experimental study

Background: During donor organ procurement and subsequent static cold storage (SCS), hepatic adenosine triphosphate (ATP) levels are progressively depleted, which contributes to ischemia-reperfusion injury (IRI). We sought to investigate a simple approach to prevent ATP depletion and IRI using a porcine donation after circulatory death (DCD) liver reperfusion model. Methods: After 30 min warm ischemia, porcine livers were flushed via the portal vein with cold (4 degrees C) non-oxygenated University of Wisconsin (UW) preservation solution (n = 6, control group) or with oxygenated UW (n = 6, OxyFlush group). Livers were then subjected to 4 h SCS in non-oxygenated (control) or oxygenated (OxyFlush) UW, followed by 4 h normothermic reperfusion using whole blood. Hepatic ATP levels were compared, and hepatobiliary function and injury were assessed. Results: At the end of SCS, ATP was higher in the OxyFlush group compared to controls (delta ATP of +0.26 vs. -0.68 mu mol/g protein, p = 0.04). All livers produced bile and metabolized lactate, and there were no differences between the groups. Grafts in the OxyFlush group had lower blood glucose levels after reperfusion (p = 0.04). Biliary pH, glucose and bicarbonate were not different between the groups. Injury markers including liver transaminases, lactate dehydrogenase, malondialdehyde, cell-free DNA and flavin mononucleotide in the SCS solution and during reperfusion were also similar. Histological assessment of the parenchyma and bile ducts did not reveal differences between the groups. Conclusion: Oxygenated flush out and storage of DCD porcine livers prevents ATP depletion during ischemia, but this does not seem sufficient to mitigate early signs of IRI

Heterogeneity of the humoral immune response following Staphylococcus aureus bacteremia

Expanding knowledge on the humoral immune response in Staphylococcus aureus-infected patients is a mandatory step in the development of vaccines and immunotherapies. Here, we present novel insights into the antibody responses following S. aureus bacteremia. Fifteen bacteremic patients were followed extensively from diagnosis onwards (median 29 days, range 9-74). S. aureus strains (median 3, range 1-6) and serial serum samples (median 16, range 6-27) were collected. Strains were genotyped by pulsed-field gel electrophoresis (PFGE) and genes encoding 19 staphylococcal proteins were detected by polymerase chain reaction (PCR). The levels of IgG, IgA, and IgM directed to these proteins were determined using bead-based flow cytometry. All strains isolated from individual patients were PFGE-identical. The genes encoding clumping factor (Clf) A, ClfB, and iron-responsive surface-determinant (Isd) A were detected in all isolates. Antigen-specific IgG levels increased more frequently than IgA or IgM levels. In individual patients, different proteins induced an immune response and the dynamics clearly differed. Anti-ClfB, anti-IsdH, and anti-fibronectin-binding protein A IgG levels increased in 7 of 13 adult patients (p < 0.05). The anti-IsdA IgG level increased in 12 patients (initial to peak level: 1.13-10.72 fold; p < 0.01). Peak level was reached 7-37 days after diagnosis. In a bacteremic 5-day-old newborn, antistaphylococcal IgG levels declined from diagnosis onwards. In conclusion, each bacteremic patient develops a unique immune response directed to different staphylococcal proteins. Therefore, vaccines should be based on multiple components. IsdA is immunogenic and, therefore, produced in nearly all bacteremic patients.

Transplantation of Severely Steatotic Liver Grafts After Machine Perfusion Remains a Risky Challenge

Liver transplantation is the treatment of choice for patients with end-stage liver disease. However, donor shortages have increased the use of high-risk and extended criteria donor livers, including livers donated after circulatory death and those with severe steatosis. Severe donor liver steatosis is associated with poor outcomes due to high susceptibility to ischemia-reperfusion injury. Ex situ machine perfusion, combining hypothermic oxygenated perfusion and normothermic perfusion (termed the DHOPE-COR-NMP protocol), has emerged as a promising strategy to mitigate injury, assess liver viability, and improve transplant outcomes. Here, we present two patients who received very steatotic donor livers following resuscitation and viability assessment using DHOPE-COR-NMP. Although both steatotic donor liver functioned well during NMP and met all of our clinically validated viability criteria, the outcome after transplantation was complicated. One recipient suffered from pulmonary fat emboli syndrome, likely due to significant loss of fat from the donor liver. The second patient required retransplantation and histopathological examination of the donor liver revealed massive lipopeliosis in zones III of the explanted liver. With the increasing incidence of steatotic donor livers, further research to prevent steatosis-related posttransplant complications is becoming progressively important. At present, transplantation of severely steatotic liver grafts remains a risky challenge, even after ex situ machine perfusion.</p