Cold Preservation Research Articles

Mannitol infusion within 15 min of cross-clamp improves living donor kidney preservation.

BackgroundOptical coherence tomography (OCT) revealed that cells lining proximal convoluted tubules of living donor kidneys (LDKs) procured by laparoscopic procedures were very swollen in response to the brief period of ischemia experienced between the time of arterial vessel clamping and flushing the excised kidney with cold preservation solution. Damage to the tubules as a result of this cell swelling resulted in varying degrees of acute tubular necrosis (ATN) that slowed the recovery of the donor kidneys during the first 2 weeks after their transplantation.MethodsTo prevent this cell damage during LDK procurement, we changed the protocol for intravenous administration of mannitol (i.e., 12.5 or 25 g) to the donor. Specifically, we reduced the time of mannitol administration from 30 to 15 min or less before clamping the renal artery.ResultOCT revealed that this change in the timing of mannitol administration protected the human donor proximal tubules from normothermic-induced cell swelling. An evaluation of posttransplant recovery of renal function showed that patients treated with this modified protocol returned to normal renal function significantly faster than those treated with mannitol 30 min or more before clamping the renal artery.ConclusionBecause slow graft recovery in the first weeks after transplantation represents a risk factor for long-term graft function and survival, we believe that this change in pretreatment protocol will improve renal transplants in patients receiving LDK.

Effects of Cold Preservation on Quality and Sarafloxacin and Difloxacin Residues in Beef

During cold preservation, the pH and moisture of beef decreased slowly. Changes in ash and protein content had no obvious rules, but the data analysis results showed that the effect of drugs on these indexes was significant. Ca, Fe content decreased, Zn content increased, TBARS values were significantly higher than those in the blank , and violent fluctuation. SAR and DIF had obvious bacteriostasis effect. Cold preservation had no palpable effect on fluoroquinolones residues.

Hypothermic Storage of Human Hepatocytes for Transplantation

Transplantation of human hepatocytes is gaining recognition as a bridge or an alternative to orthotopic liver transplantation for patients with acute liver failure and genetic defects. Since most patients require multiple cell infusions over an extended period of time, we investigated hepatic functions in cells maintained in University of Wisconsin solution at 4°C up to 72 h. Eleven different assessments of hepatic viability and function were investigated both pre- and posthypothermic storage, including plating efficiency, caspase-3/7 activity, ammonia metabolism, and drug-metabolizing capacity of isolated hepatocytes. Long-term function, basal, and induced cytochrome P450 activities were measured after exposure to prototypical inducing agents. Cells from 47 different human liver specimens were analyzed. Viability significantly decreased in cells cold stored in UW solution, while apoptosis level and plating efficiency were not significantly different from fresh cells. Luminescent and fluorescent methods assessed phases I and II activities both pre- and post-24-72 h of cold preservation. A robust induction (up to 200-fold) of phase I enzymes was observed in cultured cells. Phase II and ammonia metabolism remained stable during hypothermic storage, although the inductive effect of culture on each metabolic activity was eventually lost. Using techniques that characterize 11 measurements of hepatic viability and function from plating efficiency, to ammonia metabolism, to phases I and II drug metabolism, it was determined that while viability decreased, the remaining viable cells in cold-stored suspensions retained critical hepatic functions for up to 48 h at levels not significantly different from those observed in freshly isolated cells.

Analysis of the curative effect of ABO-incompatible liver transplantation in the treatment in patients with acute severe liver disease

To analyze and evaluate the clinical effect of ABO-incompatible liver transplantation in the treatment of acute severe liver disease. A retrospective clinical study was conducted. The clinical data of 4 136 patients undergoing orthotopic liver transplantation in Organ Transplantation Center of Tianjin First Center Hospital from September 1999 to December 2013 were analyzed. The criteria of patients enrolled were as following: model for end-stage liver disease (MELD) score ≥ 20, the donor's and recipient's blood types were different, age 18-70 years, and undergone primary non-bypass orthotopic liver transplantation. According to the rate of compliance with the principles of blood transfusion, the cases were divided into two groups: ABO-compatible group (ABO-C group, n=41), ABO-incompatible group (ABO-I group, n=22). The patients in ABO-I group received basiliximab + methylprednisolone for immune induction therapy during operation, basiliximab + tacrolimus + mycophenolate + cortisol as quadruple immunosuppressive regimen after operation. They also received subcutaneous injection of low molecular heparin for anticoagulant therapy after operation, and oral warfarin or aspirin and clopidogrel bisulfate instead after 7 days. They also received routine alprostadil after operation. The remaining treatment was the same as that of ABO-C group. The clinical data, postoperative complications, rejection and survival rates of two groups were statistically analyzed. There were no significant differences in gender, age, MELD score, complicated with tumor, quality of donor liver, length of cold preservation of donor liver, duration of operation, and blood loss during operation between ABO-C and ABO-I groups. Number of splenectomy during operation was significantly higher in ABO-I group than that in ABO-C group (5 cases vs. 1 case, χ² = 4.687, P=0.030). The 3-month, 6-month, 1-year, 3-year and 5-year survival rates of ABO-C group were 89.5%, 78.3%, 72.5%, 69.1% and 61.8%, respectively, while those of ABO-I group were 78.9%, 72.9%, 65.6%, 56.2% and 46.8%, respectively. There was no significant difference in the cumulative survival rate between two groups (Log Rank, χ² = 0.647, df=1, P=0.421). The postoperative infection rate in ABO-I group was significantly higher than that of ABO-C group [63.6% (14/22) vs. 31.7% (13/41), χ² = 5.960, P=0.015]. There were no significant difference in postoperative complications of biliary tract [22.7% (5/22) vs. 12.2% (5/41), χ² = 0.531, P=0.466], vascular complications [31.8% (7/22) vs. 12.2% (5/41), χ² = 2.416, P=0.120], or rejection as diagnosed by pathology [22.7% (5/22) vs. 9.8% (4/41), χ² = 1.051, P=0.305] between ABO-I and ABO-C groups. Although ABO-incompatible liver transplantation was followed by higher postoperative infection rate and perioperative mortality, ABO-incompatible liver transplantation can still be used to save the patient with acute severe liver disease as there is a shortage of compatible donor at present.

Vernalization of Oriental hybrid lily 'Sorbonne': changes in physiology metabolic activity and molecular mechanism.

Oriental hybrid lily 'Sorbonne' was used to investigate molecular changes during the storage at 4 °C for dormancy-release besides physiology metabolic activity observations. In physiological mechanism, endogenous abscisic acid (ABA) concentration level of lily bulbs decreased as the cold preservation time increased, and it kept at a stable level after being preserved for 35 days. The level of soluble sugars concentrations also changed during the cold preservation time, and it increased as the cold preservation time raised to 49 days then decreased afterward. On molecule level, a new transcriptome providing comprehensive sequence profiling data of variation during dormancy-release in lily was constructed. 34,367 unigenes expressed differentially between the control and the treatment was analyzed. 14 genes including 8 MADS-box family genes, 4 genes related to plant hormone, and 2 DNA methylation genes were selected to identify the levels of their expression by qRT-PCR. Our results show that the decrease of ABA level during cold storage, as well as changes in plant hormone genes was correlated with dormancy-release; MADS-box family genes VRN2, FLC, FT, SOC1 a, as well as LFY, MIKC and ARF, MYB transcription factor were included in lily floral induction and DNA methylation was correlated to lily vernalization under low temperature. According to the results of the present studies, we predicted that plant hormone pathway, energy metabolic pathway, vernalization pathway, and DNA methylation played important roles during vernalization; these data provided the foundation for future studies of vernalization to induce flowering of lily.

Superior Donor Heart Preservation Using Hypothermic Oxygenated Perfusion.

Purpose Hypothermic machine perfusion of donor hearts enables continuous aerobic metabolism and washout of toxic metabolic byproducts. We evaluated the effect of machine perfusion on cardiac myocyte integrity in hearts preserved for 4 hours in a novel device that provides pulsatile oxygenated hypothermic perfusion (Paragonix Sherpa PerfusionTM Cardiac Transport System). Methods All pig hearts were perfused with 1L of cold Celsior® solution after aortic cross-clamping, and then harvested. They were stored conventionally in Celsior® solution on ice for 4 hours (Celsior® storage [CS], n=6) or placed in the Sherpa device for 4 hours (pulsatile perfusion [PP], n=6). After cold preservation, hearts were evaluated hemodynamically using a Langendorff system. Systolic function was measured by +dP/dt; and diastolic function was measured by -dP/dt and EDP. Three control swine hearts were also re-perfused and evaluated immediately after harvest on the Langendorff system. Biopsies were taken from the LV apex at the time of harvest, after storage (when relevant), and after reperfusion for ultra-structural analysis using electron microscopy. Results CS, PP, and control hearts did not show any significant differences in systolic or diastolic function (+dP/dt, -dP/dt, EDP). CS hearts had more arrhythmias (supraventricular and ventricular) compared to PP and control hearts. Electron microscopy revealed endothelial cell rupture and damaged muscle fibers in the CS group after reperfusion, while the cell structures were preserved in the PP group and the control group. After preservation, PP hearts showed mild edema, which was not present in CS hearts, but after reperfusion, both groups had a similar grade of interstitial edema. Conclusions Hypothermic pulsatile perfusion of donor hearts during the storage interval is a simple technique that leads to better preservation of cell structure, as compared to a conventional cold storage method. This may lead to less risk of primary graft failure after orthotopic heart transplantation. Donor hearts preserved with pulsatile perfusion also appear less prone to arrhythmias during the reperfusion period. DISCLOSURES:Anderson, L.: Other, Paragonix Technologies, President. Madsen, J.: Grant/Research Support, Paragonix Technologies.

Pretreatment of the Donor with AICAR Significantly Protects Rat Heart Graft from Prolonged Cold Ischemia/Reperfusion Injury in an Isolated Model.

Purpose: To investigate whether pretreat donor rat with AMP-activated protein kinase (AMPK) activator AICAR can reduce heart graft injury after static, prolonged cold preservation and reperfusion in an isolated model. Methods: Eighteen Sprague-Dawley(SD) heart donor rats were equally divided into two groups: AICAR pretreatment group(AICAR group, n=9) and normal saline pretreatment group(Vehicle group, n=9). After the hearts were arrested with Histidine-Tryptophan-Ketoglutarate(HTK) solution, they were excised and preserved in HTK solution for 8 hours at 4 <sup style=“font-family: Arial, sans-serif;”>o[/sup]C. After cold preservation, hearts were reperfused for 1hr in an Langendorff model, concerned parameters were examined during and after reperfusion. Result: AICAR pretreated heart showed significantly higher left ventricular developing pressure(LVDP), heart rate(HR), coronary flow(CF) than vehicle pretreated group. Caspase 3 activity and percentage of TUNEL-positive cells were significantly reduced in AICAR pretreated group. Oxidative parameter 8-hydroxy-2′-deoxyguanosine(8-OHdG) and malondialdehyde (MDA) were also significantly decreased in AICAR pretreated group. Moreover, AICAR pretreatment significantly diminished cystolic cytochrome C release. Conclusion: Pretreatment of the donor with AICAR significantly protects heart grafts from prolonged cold ischemia/reperfusion injury. This protocol may be useful and feasible in clinical heart transplantation.

CHBP in Preservation Solution and Autologous Blood Perfusate Protects Isolated Ischemic Porcine Kidneys.

Introduction. There is an imperative need to develop new approaches to prevent ischemia reperfusion injury (IRI) during organ cold storage (CS) and reperfusion. A novel cyclic helix B peptide (CHBP) derived from erythropoietin was newly synthesized and its renoprotective roles were evaluated in an ex vivo renal model. Methods. Porcine kidneys subjected to 20 min warm ischemia were retrieved and flushed with 500 ml hyperosmolar citrate. The kidneys, as well as autologous blood, were cold preserved with or without 10.56 nmol/L CHBP for 18 hr. The kidneys were then reperfused with oxygenated autologous blood for 3 hr on an isolated organ perfusion system to assess renal function and structure, caspase-3 expression, apoptosis and inflammation. Results. CHBP significantly increased renal blood flow during 2 to 3 h reperfusion, with increased oxygen consumption and urine output, but decreased serum potassium. 32 kD precursor, as well as 12 and 17 kD active subunits of caspase-3 were all down-regulated by CHBP in after 3 h reperfusion, while only the 12 kD subunit was decreased post CS. In addition, apoptotic cells were significantly decreased in tubular areas, but increased in lumens and interstitial areas in post CS and reperfusion kidneys. Heat shock protein 70 was up-regulated; myeloperoxidase+ cells was reduced and renal tissue damage was also ameliorated by CHBP. Conclusions. The administration of CHBP during cold preservation of kidneys as well as autologous blood ameliorated IRI after hemoreperfusion with better renal blood flow, oxygenation, tubular function and tissue structure, which might be due to decreased caspase-3 protein, apoptosis and inflammation.Figure: No Caption available.

Histologic Evaluation of Post-implantation Immediate C4d Deposition in 13 Intestinal Grafts: Correlation With Cell-based Crossmatching, Cold Ischemia Time, and Preservation Injury

C4d deposits are predictive of humoral rejection in kidney and heart transplantation. The aim of this study was to identify C4d deposit patterns in intestinal mucosa of the grafts on biopsy specimens obtained immediately after implantation and to detect if it could be a valuable tool to predict humoral or acute rejection. A second objective was to search for a statistically significant relationship between positive C4d deposition and other collected variables. Thirteen immediately post-transplantation mucosal graft biopsy specimens, formalin fixed, underwent immunohistochemical stain for C4d deposits. Diffuse intense staining of capillary endothelium was considered positive and absent, focal or weak stains as negative. Preservation injury grade and cold ischemia times were registered for each case. Donor-specific preformed antibodies were detected by complement dependent cytotoxicity serologic technique (crossmatching). Another 19 endoscopic follow-up biopsy specimens from days 2 to 6 were also evaluated. Statistical studies were made using the index of correlation ρ (Spearman's test). Diffuse intense C4d deposits were observed in 2 grafts, focal and weak in 5, and completely negative in 6. The mean cold ischemia time was 327 ± 101 minutes. Two cases showed diffuse positive deposits, 1 had a positive crossmatch and the cold ischemia time was 360 minutes whereas the other had not preformed antibodies and its cold ischemia time was 475 minutes. Humoral or acute rejection was not observed in follow-up mucosal biopsy specimens. There was no statistically significant relationship between the C4d deposition, cold ischemia time, crossmatching results, and preservation injury degree. In conclusion, C4d deposition was not a helpful tool for diagnosis of humoral rejection and prediction of acute rejection during the early post-transplantation period.

Activation of Transient Receptor Potential Vanilloid Type 1 Channels by N-octanoyl Dopamine Improves Renal Function after Warm Ischemia but Not after Prolonged Cold Preservation.

Aim: N-octanoyl dopamine (NOD) improves renal function, when applied shortly before induction of acute kidney injury (AKI). It remains to be assessed how NOD convey its renoprotective properties, if NOD also protects after AKI induction, and if renal allograft recipients also benefit from NOD treatment. Methods: AKI was induced by clamping the left renal artery (45min) in unilateral nephrectomized Lewis or Spaque Dawley (SD), wild type (WT) and TRPV1-/- rats. Transplantations were performed in the Fisher to Lewis model using a standardized cold ischemia time of 20 hrs. Treatment was installed directly after restoration of organ perfusion. Renal function, histology and perfusion were assessed, by serum creatinine, microscopy and magnetic resonance imaging (MRI) using arterial spin labeling (ASL). Results: NOD significantly improved renal function in AKI in WT Lewis and SD rats, but not in TRPV1-/- SD rats. Improved renal function was paralleled by reduced renal inflammation, yet no differences were found in the expression of inflammatory mediators (adhesion molecules and cytokines). Although MRI-ASL showed a significant lower cortical perfusion in ischemic as compared to non-ischemic kidneys, no influence of NOD was observed. Even though prolonged cold storage did not abrogate TRPV1 activation by NOD, treatment of renal allograft recipients did not show a salutary effect. Conclusions: While NOD treatment improves renal function after warm ischemia induced AKI, it does not so after prolonged cold ischemia. Since the renoprotective effect of NOD depends on TRPV1 activation, it remains to be assessed why its salutary effect is lost after prolonged cold ischemia.

Short Oxygenated Warm Perfusion With Prostaglandin E1 Administration Before Cold Preservation Facilitates Resuscitation of Liver Grafts From Non-Heart-Beating Donor Rats.

Short Oxygenated Warm Perfusion With Prostaglandin E1 Administration Before Cold Preservation Facilitates Resuscitation of Liver Grafts From Non-Heart-Beating Donor Rats.

The First Clinical Case of Intermediate Ex Vivo Normothermic Perfusion in Renal Transplantation

A short period of ex vivo normothermic perfusion (EVNP) immediately before transplantation can revive the kidney and reduce the effects of cold ischemic (CI) injury. Herein, we report a clinical case of EVNP carried out at an intermediate period of the preservation interval. The kidney was retrieved from a 63-year-old extended criteria donor. After 10 h 29 min of CI the kidney underwent EVNP with 1 unit of compatible packed red blood cells mixed with a priming solution at 35.0°C while the recipient was being prepared for surgery. The mean renal blood flow was 93.6 mL/min/100 g and the kidney produced 60 mL of urine. Shortly after the start of surgery the first intended recipient became unfit for transplantation. After 60 min EVNP the kidney was flushed with cold preservation solution and re-packed in ice. The second period of CI was 5 h and 21 min. The kidney was transplanted without any complications into a 54-year-old predialysis patient. The recipient had immediate graft function with serum creatinine levels falling from 315 to 105 µmol/L by day 7. This is the first report of an intermediate period of EVNP in clinical renal transplantation. This case demonstrates the feasibility and safety of the technique.

N-octanoyl Dopamine Improves Vascular Barrier Function Independent of Its Cytoprotective Properties in the Setting of Static Cold Preservation.

N-octanoyl Dopamine Improves Vascular Barrier Function Independent of Its Cytoprotective Properties in the Setting of Static Cold Preservation.

Porcine Cytokine Profiles Associated With Ischemia Reperfusion Injury in Liver Transplantation Using Donation After Cardiac Death.

Introduction: Optimizing the use of marginal liver graft has been a problem due to organ shortage and high mortality rate during transplant waiting list. A close homology to humans including similar organ size, liver enzymes or immune system has given porcine models significantly increasing the use in scientific preclinical research. However, in the models it has been difficult to analyze simultaneously multiple cytokines associated with ischemia reperfusion injury and to investigate the mechanisms. Method: Porcine liver transplantation was performed by using donation one hour after cardiac death and 5-hour cold preservation. Serum levels of 9 cytokines including IFN-α, γ, IL-4, 6, 8, 10, IL-12p40, IL-1β and TNF-α were measured simultaneously by Multiplex immunobead assay technology at 30 minutes, 1, 2, 3 and 6hour after reperfusion of the liver graft. Results: Four transplantation were performed, and all of them had to be sacrificed within a day. Soon after reperfusion of the marginal graft, IL-6 and IL-12p40 were considerably expressed, one hour after reperfusion. There were no correlation between the cytokines and liver enzyme or time survived in this model. Conclusion: Multiplex immunobead assay successfully measured multiple cytokines simultaneously and identify specific ones in mini pig models. This latest method support enormous potentialities to investigate ischemia reperfusion injury with DCD liver transplantation

Kidneys From Uncontrolled Donors After Cardiac Death Provide the Same Results as Kidneys Form Donors With Extended Criteria.

Kidney donation after uncontrolled circulatory determination of death (uCDD) is an activity limited by logistical concerns and questioning concerning the evolution of the transplants. We have started the uCDD program in our two transplant centers since May 2008. All donors (n=35) were from Maastricht I and II categories, aged of 18 to 55 and presented a no flow period less than 30 min. In May 2011, in situ cold preservation was changed for normothermic regional extracorporeal circulation (NRC). Main recipient inclusion criteria were non sensitized patients, aged from 18 to 60 and awaiting first kidney transplantation. Fifty recipients from uCDD (G1) were compared to patients having received the transplants from standard (G2, n = 74) or extended criteria (ECD, G3, n=74) dead donors. One year patient and graft survivals were not significantly different between the 3 groups. PNF was more frequently observed in G1 (6% vs 0% and 2.7% in G2 and G3). The delay to obtain a sCreatinine below 250mmol/l was longer in G1 (21,5±11 days, vs 5±5 et 10±9 in G2 and G3, p<10-4). At one and two years after KTx, renal function was similar between G1 (178±68 and 166±67) and G3 (158±60 and 163±68 mmol/l) but remained significantly inferior compared to G2 (138±48 and 140±66 mmol/l, p<0.01). Finally, CNR use was associated with a shortened delay graft function time and an improvement of renal function at one year (145±35 vs 187±77 mmol/l). One year protocol graft biopsies (G1, n=24, G2, n=41 and G3, n=37) evidenced a more severe progression of arterial fibrous intimal thickening in G1 than in G2 or G3 but scoring for other histological lesions remained similar. The results of the KTx from uCDD program get closer to those from ECD. It seems coherent to propose these transplants to the recipients having the strongest probability to receive ECD transplants. However, the best quality of transplants taken under CNR will allow proposing these kidneys to younger recipients.

Ob/ob mouse livers show decreased oxidative phosphorylation efficiencies and anaerobic capacities after cold ischemia.

BackgroundHepatic steatosis is a major risk factor for graft failure in liver transplantation. Hepatic steatosis shows a greater negative influence on graft function following prolonged cold ischaemia. As the impact of steatosis on hepatocyte metabolism during extended cold ischaemia is not well-described, we compared markers of metabolic capacity and mitochondrial function in steatotic and lean livers following clinically relevant durations of cold preservation.MethodsLivers from 10-week old leptin-deficient obese (ob/ob, n = 9) and lean C57 mice (n = 9) were preserved in ice-cold University of Wisconsin solution. Liver mitochondrial function was then assessed using high resolution respirometry after 1.5, 3, 5, 8, 12, 16 and 24 hours of storage. Metabolic marker enzymes for anaerobiosis and mitochondrial mass were also measured in conjunction with non-bicarbonate tissue pH buffering capacity.Results Ob/ob and lean mice livers showed severe (>60%) macrovesicular and mild (<30%) microvesicular steatosis on Oil Red O staining, respectively. Ob/ob livers had lower baseline enzymatic complex I activity but similar adenosine triphosphate (ATP) levels compared to lean livers. During cold storage, the respiratory control ratio and complex I-fueled phosphorylation deteriorated approximately twice as fast in ob/ob livers compared to lean livers. Ob/ob livers also demonstrated decreased ATP production capacities at all time-points analyzed compared to lean livers. Ob/ob liver baseline lactate dehydrogenase activities and intrinsic non-bicarbonate buffering capacities were depressed by 60% and 40%, respectively compared to lean livers.ConclusionsSteatotic livers have impaired baseline aerobic and anaerobic capacities compared to lean livers, and mitochondrial function indices decrease particularly from after 5 hours of cold preservation. These data provide a mechanistic basis for the clinical recommendation of shorter cold storage durations in steatotic donor livers.

Ischemia/Reperfusion during normothermic perfusion

Cold storage of organs for preservation and transplantation is reaching its limits especially with extended criteria for heart beating donors and donation after cardiac death. We will discuss recent findings and perspectives in normothermic kidney preservation. A literature review was performed from original articles and syntheses selected by the search engine PubMed. Keywords used were: cold ischemia; warm ischemia, normothermic, organ preservation, preconditioning, organ perfusion. We identified several ways to improve kidney preservation: Ischemic normothermic preconditioning; Pharmacologic normothermic preconditioning; Ex vivo normothermic reperfusion; Remote ischemic transplantation preconditioning; Ischemic postconditioning. In clinical practice, only uses of ECMO for organ preconditioning or ex vivo normothermic organ perfusion were used. Promising experimental and clinical results make challenge cold preservation. The most suitable and physiological method seems to be a normothermic perfusion and conservation with autologous oxygenated blood using Extra Corporeal Membrane Oxygenation or Regional Normothermic Circulation.

Edaravone, a Free Radical Scavenger, Improves the Graft Viability on Liver Transplantation From Non–heart-beating Donors in Pigs

BackgroundAlthough liver transplantation from non–heart-beating donors (NHBDs) is an effective way to overcome shortage of donors, primary graft nonfunction is often noted in these grafts. We have previously reported that edaravone, a free radical scavenger, has a cytoprotective effect on warm ischemia-reperfusion injury and improves the function of liver grafts from NHBDs in a rat model of ischemia-reperfusion. The purpose of this study was to investigate the effects of edaravone on liver transplantations from NHBDs. MethodsPigs were divided into three groups: (1) a heart-beating (HB) group (n = 5), in which liver grafts were retrieved from HB donors; (2) a non-heart-beating (NHB) group (n = 4), in which liver grafts were retrieved under apnea-induced NHB conditions; and (3) an edaravone-treated (ED) group (n = 5), in which liver grafts were retrieved in the same manner as the NHB group and treated with edaravone at the time of perfusion (3 mg/L in University of Wisconsin [UW] solution), cold preservation (1 mg/L in UW solution), and after surgery (1 mg/kg/d). The grafts from all groups were transplanted after 4 hours of cold preservation. ResultsIn the ED group, the 7-day survival rate was significantly higher than that in the NHB group (80% versus 0%, P = .0042, Kaplan-Meier log-rank test). Furthermore, on histologic examination, the structure of sinusoids in the ED group was well preserved and similar to that in the HB group. ConclusionsEdaravone may improve the viability of liver grafts from NHBDs.

Impact of Machine Perfusion Preservation of Liver Grafts From Donation After Cardiac Death

Because of the critical shortage of deceased donor grafts, using a donation after cardiac death (DCD) donor is an important resource. However, the ischemic damage of those DCD grafts jeopardizes organ viability during cold storage. Maintaining organ viability after donation until transplantation is important for optimal graft function and survival. This review describes the effective preservation in transplantation for DCD livers. Concepts and development of machine perfusion for DCD liver grafts to reduce ischemia/reperfusion injury are discussed. Despite the fact that hypothermic machine perfusion might be superior to static cold preservation, DCD livers are exposed to hypothermia-induced damage. Recently, some groups introduced the beneficial effects of normothermic or subnormothermic machine perfusion in DCD liver preservation and transplantation.

Angiopoietin-2 Inhibition Prevents Transplant Ischemia-Reperfusion Injury and Chronic Rejection in Rat Cardiac Allografts

Transplant ischemia-reperfusion injury (Tx-IRI) and allograft dysfunction remain as two of the major clinical challenges after heart transplantation. We investigated the role of angiopoietin-2 (Ang2) in Tx-IRI and rejection using fully MHC-mismatched rat cardiac allografts. We report that plasma levels of Ang2 were significantly enhanced in the human and rat recipients of cardiac allografts, but not in the rat recipients of syngrafts, during IRI. Ex vivo intracoronary treatment of rat cardiac allografts with anti-Ang2 antibody before 4-h cold preservation prevented microvascular dysfunction, endothelial cell (EC) adhesion molecule expression and leukocyte infiltration, myocardial injury and the development of cardiac fibrosis and allograft vasculopathy. Recipient preoperative and postoperative treatment with anti-Ang2 antibody produced otherwise similar effects without effecting microvascular dysfunction, and in additional experiments prolonged allograft survival. Recipient preoperative treatment alone failed to produce these effects. Moreover, ex vivo intracoronary treatment of allografts with recombinant Ang2 enhanced Tx-IRI and, in an add-back experiment, abolished the beneficial effect of the antibody. We demonstrate that neutralization of Ang2 prevents EC activation, leukocyte infiltration, Tx-IRI and the development of chronic rejection in rat cardiac allografts. Our results suggest that blocking Ang2 pathway is a novel, clinically feasible, T cell-independent strategy to protect cardiac allografts.