Standard Preservation Solution Research Articles

Multi-Walled Carbon Nanotubes Improve the Development of Chrysanthemum × morifolium (Ramat.) Hemsl. ‘Jinba’ Inflorescences

The application of cut flower preservation technology can significantly enhance both the ornamental and economic value of fresh-cut flowers. Research on vase solutions has become a concentrated area in current studies on cut flower preservation. Multi-walled carbon nanotubes (MWCNTs), as a type of carbon nanomaterial with bactericidal and membrane-penetrating properties, can be used as a component in vase solutions. This supplementation of energy substances aims to improve antioxidant enzyme activity, thereby enhancing the postharvest quality of cut chrysanthemums. In this study, deionized water and a standard preservative solution were employed as control groups to compare the effects of MWCNTs applied at different concentrations, combined with common preservatives such as sugar and 8-hydroxyquinoline, on the postharvest flowering and preservation of Chrysanthemum × morifolium ‘Jinba’. By observing the distribution of MWCNTs in the tissues surrounding the cut and changes in water content, carbon sources, osmoregulatory substance levels, and the expression of relevant key genes, a formulation with excellent postharvest treatment effects was identified. Preliminary investigations into its action and mechanism were also conducted. The results indicated that the combined treatment with 5 mg L−1 MWCNTs, 30 g L−1 sucrose, and 0.2 g L−1 8-hydroxyquinoline effectively promoted water and sugar uptake in chrysanthemum flowers, accelerating bud opening, maintaining larger inflorescence diameter, and extending the vase life. Ultimately, this enhanced the ornamental value of cut chrysanthemums. These research findings provide theoretical and experimental foundations for the application of multi-walled carbon nanotubes as auxiliary additives to improve the ornamental quality of cut flowers.

Effect of PERLA®, a new cold-storage solution, on oxidative stress injury and early graft function in rat kidney transplantation model

BackgroundThe composition of organ preservation solutions is crucial for maintaining graft integrity and early graft function after transplantation. The aim of this study is to compare new organ preservation solution PERLA® with the gold standard preservation solution University of Wisconsin (UW) regarding oxidative stress and early graft injury.MethodsIn order to assess oxidative stress after cold storage, kidney grafts have been preserved for 18 h at 4° C in either UW solution or PERLA® solution and then assessed for oxidative stress injury (protocol 1). To assess kidney injuries and oxidative stress after reperfusion, rat kidneys were harvested, stored in cold UW or in PERLA® solutions for 18 h at 4 °C and then transplanted heterotopically for 6 h (protocol 2). PERLA® is a high Na+/low K+ solution including PEG-35 (1 g/L), trimetazidine (1 µM), carvedilol (10 µM) and tacrolimus (5 µM).ResultsOur results showed that preservation of kidneys in PERLA® solution significantly attenuates oxidative stress parameters after cold storage and reperfusion. We found a significant decrease in oxidative damage indicators (MDA, CD and CP) and a significant increase in antioxidant indicators (GPx, GSH, CAT, SOD and PSH). Moreover, PERLA® solution decreased kidney injury after reperfusion (creatinine, LDH and uric acid).ConclusionPERLA® solution was more effective than UW storage solution in preserving rat’s kidney grafts.

Lactated Ringer as Preservation Solution in Living Donor Renal Transplantation

Optimal organ preservation remains a critical hallmark event in renal transplantation as it is the supply line. Previous studies have shown that the choice of preservation solution may affect transplant outcomes. In this study, we aimed to present the early follow-up results of the graft and patients, using lactated Ringer to preserve kidney allografts with living donors. The results of 97 living donor transplant operations performed in Sanko University Hospital were evaluated retrospectively. The patient's evaluation included demographics, dialysis time duration, renal replacement method, primary disease, comorbidity, surgical and clinical complications in the acute period, graft functions, blood levels of calcineurin inhibitor drugs, anastomotic renal artery, warm ischemia, and cold ischemia times. Donor (49 men, 50.5%) and recipient (58 men, 59.7%) demographics, HLA compatibility (mismatch), hospitalization days, and length of warm and cold ischemic time are summarized in Table 1. Primary nonfunction was not defined in any patients, but delayed graft function was observed during the follow-up of 3 patients (3.09%), who were all hypotensive in the post-transplantation period, and positive inotropic infusion was needed for hemodynamic stability. Lactated Ringer demonstrated efficacy in terms of patient and graft survival, and its lower cost represents a financial advantage, so it can be used in living donor kidney transplantation because it is safe, effective, and inexpensive. Standard preservation solutions may still be recommended in cases with long cold ischemia times, such as paired exchange transplants and cadaveric transplants. Thus, randomized controlled studies are needed for further investigation.

Luminal administration of biliverdin ameliorates ischemia-reperfusion injury following intestinal transplant in rats

BackgroundIntestinal grafts are susceptible to ischemia-reperfusion injury, resulting in the loss of mucosal barrier function and graft failure. Biliverdin is known to exert a variety of cytoprotective functions against oxidative tissue injury. Because the mucosal layer is the primary site of ischemia-reperfusion injury, mucosa-targeting strategies by luminal delivery of reagents might be beneficial. We tested whether intraluminal administration of biliverdin as an adjuvant to standard preservation solutions protected against ischemia-reperfusion injury. MethodsOrthotopic syngeneic intestinal transplants were performed on Lewis rats after 6 hours of cold preservation. Saline containing biliverdin (10 μM) or without biliverdin was introduced into the lumen of the intestinal grafts immediately before cold preservation. ResultsDamage to the intestinal mucosa caused by ischemia-reperfusion injury resulted in severe morphological changes, including blunting of the villi and erosion, and led to significant loss of gut barrier function 3 hours after reperfusion. These changes to the mucosa were notably ameliorated by intraluminal administration of biliverdin. Biliverdin also effectively inhibited upregulation of messenger RNAs for interleukin-6, inducible nitric oxide synthase, and C-C motif chemokine 2. Additionally, biliverdin treatment prevented the loss of expression of claudin-1, a transmembrane, tight-junction barrier protein. The 14-day survival of recipients of biliverdin-treated grafts was significantly improved as compared with the recipients of saline-treated control grafts (83.3% vs 38.9%, P = .030). ConclusionThis study demonstrated that luminally delivered biliverdin provides beneficial effects during the transplant of rat small intestinal grafts and could be an attractive therapeutic option in organ transplantation.

Beneficial effects of Aucubin on restoration of rabbits with cartilage defect.

Osteochondral grafts are suitable materials for repair of articular cartilage defect and plastic and reconstructive surgery. In our study, osteochondral allografts from rabbits were preserved in vitro for 28days, and chondrocyte death, degradation of collagen and proteoglycan, morphological alterations, and inflammatory reaction were observed in the grafts. Supplementing of Aucubin with 10 or 20μM in the preservation solution inhibited chondrocyte death, matrix degradation, pathological alterations and inflammation in allografts preserved in vitro, compared with that preserved in standard preservation solution. In addition, after transplantation of 20μM Aucubin-treated allografts, the osteochondral repair and regeneration of rabbits with knee joint defect were accelerated. In conclusion, Aucubin was beneficial for maintaining chondrocyte viability and normal morphology, and inhibiting inflammatory occurrence in rabbit osteochondral grafts preserved in vitro, and facilitated osteochondral repair and regeneration of rabbits with knee defect. These findings might provide novel insights for preservation of grafts for clinical articular cartilage repair and plastic surgery.

Development of a Novel Perfusable Solution for ex vivo Preservation: Towards Photosynthetic Oxygenation for Organ Transplantation.

Oxygen is the key molecule for aerobic metabolism, but no animal cells can produce it, creating an extreme dependency on external supply. In contrast, microalgae are photosynthetic microorganisms, therefore, they are able to produce oxygen as plant cells do. As hypoxia is one of the main issues in organ transplantation, especially during preservation, the main goal of this work was to develop the first generation of perfusable photosynthetic solutions, exploring its feasibility for ex vivo organ preservation. Here, the microalgae Chlamydomonas reinhardtii was incorporated in a standard preservation solution, and key aspects such as alterations in cell size, oxygen production and survival were studied. Osmolarity and rheological features of the photosynthetic solution were comparable to human blood. In terms of functionality, the photosynthetic solution proved to be not harmful and to provide sufficient oxygen to support the metabolic requirement of zebrafish larvae and rat kidney slices. Thereafter, isolated porcine kidneys were perfused, and microalgae reached all renal vasculature, without inducing damage. After perfusion and flushing, no signs of tissue damage were detected, and recovered microalgae survived the process. Altogether, this work proposes the use of photosynthetic microorganisms as vascular oxygen factories to generate and deliver oxygen in isolated organs, representing a novel and promising strategy for organ preservation.

Pancreas preservation: clinical practice and future developments.

To summarize recently published studies of preservation strategies including machine perfusion in pancreas transplantation. The shortage of conventional donors is leading units to use extended criteria donors (ECDs) and donors after cardiac death (DCD). Static cold storage (SCS) is still the standard method of preservation for pancreases and University of Wisconsin remains the gold standard preservation solution. In experimental studies, oxygen delivered during preservation reduced tissue injury and improved islet cell yield and function. Hypothermic machine perfusion of discarded human pancreases has been shown to improve adenosine triphosphate levels without adversely effect histology and oedema compared with SCS. Normothermic machine perfusion of discarded human organs has so far been challenging and led to increasing injury, rather than preservation. There are currently no clinical studies in pancreas transplant with the exception of a small number of pancreases being transplanted following normothermic regional perfusion. The storm of new organ preservation methods is now being more widely studied in the pancreas, with some promising results. These new strategies have the potential to allow expansion of the donor pool and greater utilization of ECD and DCD organs.

Can we continue blaming on the static preservation solution as the fingle most important cause of DGF or PNF for liver trasplant?

Presenter: Gabriel Gondolesi | Hospital Universitario Fundación Favaloro Background: DGF and PNF have shown in different studies the impact they have on early and long term outcomes after liver transplantation. UW has been considered as the Gold Standard preservation solutions until discontinuation. HTK has been the alternative and only available option in different countries. Although controversies have been raised regarding its indications of use and the possible negative impact on graft survival; the preservation solution has been consider by us as just one piece in a very complex procedure, with other variables like DRI, MELD score, logistics, costs (among others) that also have a role as risk factor for graft failure. We aim to analyze the variables involved in a liver transplant procedure during two different eras under the same medical group, but with different preservations solutions (PS), available, in order to evaluate their impact as independent risk factor for graft loss. Methods: A retrospective database analysis of adult (>18 years old) patients receiving a cadaveric donor organ for a primary liver transplant. Diagnoses, MELD score, BMI, time on waiting list (WL), DRI, national, regional or local procurement; preservation solution used total ischemia time (TIT), warm ischemia time (WIT), cold ischemia time (CIT), delay graft function (DGF), primary non function (PNF), LFT’s, biliary complications, incidence of acute and chronic rejection, patient and graft survival, were analyzed comparing the two study periods (UW: 2009-11; HTK: 2016-19). All analyses were performed using IBM SPSS v20.0. Results: A total of 290 liver transplant were performed between 2009-11 and 2016-19; 176 (60.7%) were adult primary liver transplants (102 males (58%) and 74 females (42%)). Diagnoses were: viral infections (n=52, 30%), alcoholic liver disease (n=44, 26%), autoimmune hepatitis (n=30, 18%), metabolic diseases (n=24, 14%), cryptogenic cirrhosis (n=18, 10%) and acute liver failure (n=4, 2%). Table 1 summarizes Donor and Recipient variables. We observed a difference on LFTS`s when we compared the preservation solution used and the primary disease that lead to the liver transplant (Table 1). There were no differences in biliary or venous complications, DGF, PNF or rejection in regards to donor location. Quite the contrary, there was an increased TGO mean value 48 hs post op. and a higher rate of arterial complications (p: 0.0001), and recurrence of the primary disease (p:0.008) in the UW group. Long term patient and graft survival showed no significant difference in favor of the use of HTK (Figure 1), even when MELD score was considerably worse for this group. Conclusion: Liver transplantation is a complex multifactorial procedure. Preservation solution is only “one of the multiple players” involved in it, and its use might depend on availability; but other hospital policies and actions like logistics improvement, reduction of TIT and surgical times should be done in order to maintain expected results, since other variables like DRI and recipient MELD can not be modified.

Hearts Donated After Circulatory Death and Reconditioned Using Normothermic Regional Perfusion Can Be Successfully Transplanted Following an Extended Period of Static Storage.

There has been an increased interest in donation after circulatory death (DCD) to expand donor pool for cardiac transplantation. Normothermic regional perfusion (NRP) allows in situ assessment of DCD hearts, allowing only acceptable organs to be procured. We sought to determine if extended cold storage was possible for DCD hearts following NRP and to compare hearts stored using standard cold storage with a novel cardioprotective solution designed for room temperature storage. Donor pigs underwent hypoxic cardiac arrest (DCD) followed by 15 minutes of warm ischemia and resuscitation on NRP. They were then randomly assigned to static storage with histidine-tryptophan-ketoglutarate (HTK) at 4°C (HTK group, n=5) or SOM-TRN-001 at 21°C (SOM group, n=5). Conventional beating-heart donations were used as controls (n=4). Fourteen transplants were successfully performed. HTK hearts showed initial dysfunction following reperfusion; however, they demonstrated significant recovery up to 3 hours post-transplant. No significant differences were seen between HTK and control hearts post-transplantation (cardiac index: control 49.5±6% and HTK 48.5±5% of baseline). SOM improved myocardial preservation; hearts showed stable contractility after transplantation (cardiac index: 113.0±43% of NRP function) and improved diastolic function compared with HTK. Preservation in SOM also significantly reduced proinflammatory cytokine production and release following transplantation and partially prevented endothelial dysfunction. DCD hearts stored using a standard preservation solution demonstrated comparable post-transplantation myocardial function to standard controls. Thus, short periods of cold storage following successful NRP and documented adequate function is an acceptable strategy for DCD hearts. Preservation in SOM at room temperature is feasible and can improve cardiac recovery by minimizing endothelial dysfunction and tissue injury.

Effect of improved preservation solution with methoxy polyethylene glycol succinimidyl propionate on rat cornea.

To observe the effect of DMEM/F12 pegylated with methoxy polyethylene glycol succinimidyl propionate (mPEG-SPA) on the biophysical and immune characteristics of the rat cornea preserved in it. Corneal grafts were harvested from Wistar rat and preserved in the DMEM/F12 plus mPEG-SPA, DMEM/F12 without mPEG-SPA, and standard Optisol-GS solution at 4°C for 14days, referred as plus-PEG, minus-PEG, and Optisol grafts, respectively. The biophysical properties of those grafts, including transmittance, thickness, water content, and biomechanics were investigated. The survival of those grafts was observed in the high-risk corneal transplantation model. Transmittance and biomechanics did not show any differences among those grafts. Thickness and water content of plus-PEG grafts were slightly improved. Proliferation and activation of lymphocytes were lower while they were incubated with plus-PEG grafts, compared with minus-PEG grafts and Optisol grafts. The mean survival time was significantly prolonged in plus-PEG grafts. DMEM/F12 solution plus mPEG-SPA improved the survival of corneal grafts and maintained the comparative biophysical characteristics of them, compared with the standard preservation solution.

Prevention of ischemia-reperfusion lung injury during static cold preservation by supplementation of standard preservation solution with HEMO2life® in pig lung transplantation model

We describe the results of adding a new biological agent HEMO2life® to a standard preservation solution for hypothermic static lung preservation aiming to improve early functional parameters after lung transplantation. HEMO2life® is a natural oxygen carrier extracted from Arenicola marina with high oxygen affinity developed as an additive to standard organ preservation solutions. Standard preservation solution (Perfadex®) was compared with Perfadex® associated with HEMO2life® and with sham animals after 24 h of hypothermic preservation followed by lung transplantation. During five hours of lung reperfusion, functional parameters and biomarkers expression in serum and in bronchoalveolar lavage fluid (BALF) were measured. After five hours of reperfusion, HEMO2life® group led to significant improvement in functional parameters: reduction of graft vascular resistance (p < .05) and increase in graft oxygenation ratio (p < .05). Several ischemia-reperfusion related biomarkers showed positive trends in the HEMO2life® group: expression of HMG B1 in serum tended to be lower in comparison (2.1 ± 0.8 vs. 4.6 ± 1.5) with Perfadex® group, TNF-α and IL-8 in BALF were significantly higher in the two experimental groups compared to control (p < .05). During cold ischemia, expression of HIF1α and histology remained unchanged and similar to control. Supplementation of the Perfadex® solution by an innovative oxygen carrier HEMO2life® during hypothermic static preservation improves early graft function after prolonged cold ischemia in lung transplantation.

Evaluation of Perfluorohexyloctane/Polydimethylsiloxane for Pancreas Preservation for Clinical Islet Isolation and Transplantation.

This study aimed to evaluate a 50:50 mix of perfluorohexyloctane/polydimethylsiloxane 5 (F6H8S5) preservation of pancreases in a clinical setting compared with standard solutions for 1) cold ischemia time (CIT) 10 h and 2) an extended CIT 20 h. Procured clinical-grade pancreases were shipped in either F6H8S5 or in standard preservation solutions, that is, University of Wisconsin (UW) or Custodiol. F6H5S5 was preoxygenated for at least 15 min. Included clinical-grade pancreases were procured in UW or Custodiol. Upon arrival at the islet isolation laboratory, the duodenum was removed followed by rough trimming while F6H8S5 was oxygenated for 1520 min. Trimmed pancreases were immersed into oxygenated F6H8S5 and stored at 4C overnight followed by subsequent islet isolation. Pancreas preservation using F6H8S5 proved as effective as UW and Custadiol when used within CIT up to 10 h, in terms of both isolation outcome and islet functionality. Preservation in F6H8S5 of pancreases with extended CIT gave results similar to controls with CIT 10 h for both isolated islet functionality and isolation outcome. This study of clinically obtained pancreases indicates a clear benefit of using F6H8S5 on pancreases with extended CIT as it seems to allow extended cold ischemic time without affecting islet function and islet numbers.

Liver Transplantation From Donor With Situs Inversus Totalis: Is It a Safety Procedure?

Background: The situs inversus is a rare condition of unknown cause that occurs in less than 0.1% of the population. A standard operation procedure has not been established for liver transplantation when the graft comes from a donor with situs inversus. Counting with this, there are nine cases of transplant with liver graft from deceased donors with situs inversus totalis reported in literature. Methods - Case Report: During organs harvesting of a young male, it was found that all organs of the abdominal and thoracic cavities were located in changed positions, as if it were a mirror image, diagnosing the donor with situs inversus totalis. The liver was flushed with standard preservation solution and procured using standard techniques. The recipient was a female with cirrhosis caused by Wilson’s disease. She was Child-Pugh C and MELD SCORE 32. Hepatectomy was performed using a piggyback technique with preservation of the vena cava after complete liver mobilization. The liver graft was brought onto the field and rotated 180° in a ventral caudal fashion (backwards), maintaining the largest lobe in the right upper quadrant. Twenty-eight months after transplant, the graft liver function was still normal and there were no biliary complications or rejection reactions. Conclusion: Liver transplantation using a graft from donor with a situs inversus totalis seems to be a safe surgical procedure.

High-Molecular-Weight Polyethylene Glycol Enhances Hypothermic Storage of Feline Kidney Cells

ABSTRACTPhosphate-buffered sucrose (PBSc) solution is effective for short-term hypothermic preservation of tissue during feline kidney transplantation. A high-molecular-weight polyethylene glycol (35,000 Da, PEG35) reportedly enhanced the protective effects against cold-induced tubular injuries in animal kidney transplantation models. We investigated the ability of PBSc solution containing PEG35 to preserve cultured feline kidney cells using in vitro WST-8 cell proliferation assays. PEG35 significantly improved cell viability during 24 hr of cold preservation. PBSc containing 20 g/l PEG35 achieved an effect almost equal to that of University of Wisconsin (UW) solution, the gold standard preservation solution used in human clinical kidney transplantation, for up to 24 hr of preservation. Our results suggest that PBSc containing PEG35 provides an excellent medium for graft cold storage during feline kidney transplantation.

Improvement of Functional Recovery of Donor Heart Following Cold Static Storage with Doxycycline Cardioplegia

Injury to the donor heart during cold preservation has a negative impact on graft survival before transplantation. This study aims to examine whether doxycycline, known as an MMP-2 inhibitor, has a positive effect on donor heart preservation via its antioxidant action when added to standard preservation solution. Hearts were obtained from 3-month-old male Wistar rats and randomly divided into three groups: hearts stored for 1 h at 4 °C (1) with doxycycline preservation solution (DOX cardioplegia) with low Ca2+; (2) with standard cardioplegia with low Ca2+; and (3) unstored hearts. All hearts were perfused in working mode, arrested at 37 °C, removed from the perfusion system, reattached in Langendorff perfusion system, and converted to working mode for 1 h. At the end of the storage period, hearts preserved in DOX cardioplegia had significantly less weight gain than those preserved in the standard cardioplegia. DOX cardioplegia-induced preservation resulted in significantly higher heart rates and better recovery quality during reperfusion in aortic flow compared to the standard cardioplegia group. Recovery in the left ventricular function and Lambeth Convention Arrhythmia scores during 1 h reperfusion were also significantly better in the DOX cardioplegia group. Biochemical data showed that DOX cardioplegia prevented an increase in MMP-2 activity and blocked apoptosis through increased activity of the pro-survival kinase Akt in the donor heart homogenates. DOX cardioplegia also led to a balanced oxidant/antioxidant level in the heart homogenates. This is the first study to report that cardioplegia solution containing doxycycline provides better cardioprotection via the preservation of heart function, through its role in controlling cellular redox status during static cold storage.

Organ Biology-New Development.

Organ Biology-New Development.

P57 Hydrogen sulfide treatment reduces activation of ischemia–reperfusion injury-induced tissue inflammatory processes and resultant recipient adaptive immune response to donor tissue during allogeneic renal transplantation

Background Organ procurement is inherently associated with ischemia–reperfusion injury (IRI), resulting from loss and subsequent restoration of blood flow and is detrimental to short- and long-term graft function and survival. Treatment of donor organs with endogenously produced gasotransmitters such as hydrogen sulfide (H 2 S) is a novel method of mitigating IRI during transplantation, as previously shown in a model of murine syngeneic renal transplantation (RTx) [1] . The protective effects of H 2 S against IRI during allogeneic RTx, in which genetically dissimilar grafts are targeted by the recipient immune system causing organ rejection, have not yet been studied. Since H 2 S is known to exhibit anti-inflammatory actions during tissue IRI, we postulated that H 2 S treatment during allogeneic RTx could reduce graft inflammation and modulate the resultant recipient immune response against donor tissue. Methods Following bilateral native nephrectomy, recipient Lewis rats underwent RTx with left kidneys obtained from Brown Norway donor rats that were flushed, at the time of procurement, with either cold (4°C) University of Wisconsin standard preservation solution (UW group) or cold UW + H 2 S donor molecule (150 μ M NaHS) (H 2 S group) and stored for 6 h at 4°C in the same solution. Sham animals were also followed. Renal grafts were obtained at post-operative day 2 and RNA was isolated from renal homogenates for gene microarray analysis (Affymetrix Rat Gene 1.0 ST Microarray). Additionally, serum was collected post-operatively at day 1 to 2 and at time of sacrifice to determine the relative abundance of allo-reactive serum antibodies. TIB-120 cells (ATCC), a rat B-cell line heterozygous for Brown Norway and Lewis alleles, were incubated with serum from transplant recipients in each treatment group, stained for bound rat IgG/M antibodies and analyzed via flow cytometry. Results Upon microarray analysis both treatment groups exhibited significant enrichment in genes involved in inflammatory and innate immune responses as well as cytokine signaling and antigen processing at day 2 compared to Sham (p 2 S treated grafts had markedly reduced enrichment scores in each of these pro-inflammatory processes compared to UW. Upon allo-antibody binding analysis both UW and H 2 S treatment groups exhibited similar trends, with more cells unbound by allo-antibodies at day 1 to 2 and few unbound cells at time of sacrifice. However, while cells incubated with UW serum at day 1 to 2 exhibited markedly more allo-antibody binding (27% unbound cells) compared to cells incubated with naive Sham serum (37% unbound cells), cells incubated with H 2 S serum at day 1 to 2 exhibited similar levels of allo-antibody binding (40% cells unbound) as those incubated with Sham serum. These findings indicate that H 2 S treated animals experienced less acute graft inflammation, leading to a delayed induction of adaptive immune response to donor tissue compared to UW alone. Conclusions These findings are the first to report that H 2 S treatment may modulate the recipient immune response to donor tissue during allogeneic RTx. This phenomenon could represent an important underlying mechanism of H 2 S-mediated protection against renal injury early in the transplant curriculum, ultimately prolonging the survival and function renal allografts.

Static Cold Storage of Brain Dead Donor Hearts: Can ‘Pharmacological Conditioning’ Overcome Primary Graft Failure?

Purpose Static cold storage of hearts from brain dead donors is still the mainstay in cardiac transplantation. The risk of ischaemia-reperfusion injury (IRI) and primary graft failure (PGF) are significant disadvantages of this method. We found that three ‘conditioning’ agents–Glyceryl trinitrate (G), Erythropoietin€ and Zoniporide (Z), when added to standard preservation solutions, attenuate IRI in rat hearts. We aimed to test their efficacy in hearts from brain dead rats in prolonged cold storage. Methods and Materials Rats were subjected to brain death (BD) by inflation of a subdural embolectomy catheter. Invasive haemodynamic changes are shown in Figure 1 . Cardiac function was assessed on an isolated working heart model (IWHM), then hearts arrested and preserved in Celsior for 1, 3 or 6 hours (n=6 each sub-group). Additional hearts from BD donors were arrested and stored for 3 or 6 hours using Celsior with G+E+Z (n=6 for 3h, 6h) and function re-assessed on an IWHM. Results BD hearts had inferior recovery of cardiac output (CO) cf with shams. Supplementing Celsior with G+E+Z significantly improved CO in BD hearts (p Conclusions G, E & Z that activate ‘conditioning’ pathways significantly improve recovery of BD rat hearts after prolonged static cold storage. This data warrants assessing these strategies clinically. [ figure 2 ]

An Experimental Porcine Model of Heterotopic Renal Autotransplantation

ObjectiveThe aim of the present study was to validate an experimental model of heterotopic renal allotransplantation. Such a model, more relevant to the human situation, has never been previously described. Materials and MethodsPietrin pigs (40 to 50 kg) were used in the study. Through a midline incision, the left kidney was removed, washed, and preserved in a standard preservation solution (Celsior, Genzyme, France) for 20 hours at 4°C. Heterotopic autotransplantation was performed into the right iliac fossa onto the external iliac vessels with an end-to-side anastomosis and a nonstented uretero-ureteral anastomosis was performed. ResultsTwenty-five renal allotransplantations were performed over a 5-month time period. Mean operating time progressively decreased and stabilized after 15 procedures (mean ± SD: 78.2 ± 19 minutes and 187.4 ± 18 minutes for left nephrectomy and transplantation, respectively) as morbidity decreased concomitantly. Suturing times for end-to-side anastomosis of the renal artery and vein onto the external iliac artery and vein were 21.9 ± 7 minutes and 34 ± 8 minutes (mean ± SD), respectively. Ten pigs died before the end of the experiment. ConcIusionsWe have developed and validated the first nonrodent animal model of heterotopic renal autotransplantation relevant to the human anatomy and physiology. The procedure was easy to learn and safe. This model could be used to teach junior surgeons renal transplantation techniques and could also be used as a model to study ischemia-reperfusion injury in renal transplantation.

Carbon Monoxide Releasing Molecules Inhibit Cell Death Resulting from Renal Transplantation Related Stress

Organ cold storage and subsequent transplantation are associated with significant ischemia-reperfusion injury, leading to cell death, graft inflammation and decreased graft function. CORM-3s reduce oxidative stress and prevent inflammation in kidneys stored at 4C and subsequently transplanted. Graft survival and function are markedly improved compared to kidneys preserved and stored in University of Wisconsin solution alone. We determined whether CORM-3 has direct antiapoptotic effects on in vitro preparations of human HUVECs exposed to anoxic conditions. We also determined whether direct administration of CORM-3 to renal grafts before and/or after cold storage would prevent renal damage during the transplantation process. CORM-3 supplementation led to a significantly increased frequency of live cells (mean ± SD 72.3% ± 1.9%, p <0.01), reduced apoptosis (14.9% ± 6.1%, p <0.01) and decreased mitochondrial transmembrane potential (40.2% ± 7.2%, p <0.05) in HUVECs exposed to 20 hours of cold storage compared to controls (11.6% ± 3.5%, 82.2% ± 2.3% and 78.2% ± 3.2%, respectively). In keeping with this antiapoptotic effect CORM-3 supplementation led to a mean 7.4 ± 2.1-fold up-regulation in Bcl-2 gene expression. CORM-3 supplementation in standard preservation solution was most beneficial at initial ischemic injury and before cold storage exposure. However, additional reflushing before vascular reperfusion showed an additive benefit to graft survival and function after transplantation. This was confirmed by decreased glomerular and tubular necrosis, and apoptosis in double flushed grafts. CORM-3 supplementation in standard University of Wisconsin solution has a significant impact on decreasing cellular and graft injury, and improving survival through its antiapoptotic effects, which are likely mediated through mitochondrial membrane stabilization.