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Zakaria ER, Yousufzai W, Obaid O, Asmar S, Hsu CH, Joseph B. Cellular Cytosolic Energy Replenishment Increases Vascularized Composite Tissue Tolerance to Extended Cold Ischemia Time. Mil Med 2023; 188:2960-2968. [PMID: 36308325 DOI: 10.1093/milmed/usac331] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2022] [Accepted: 10/10/2022] [Indexed: 08/31/2023] Open
Abstract
BACKGROUND Vascularized composite allotransplantation (VCA) is a restorative surgical procedure to treat whole or partially disfiguring craniofacial or limb injuries. The routine clinical use of this VCA surgery is limited using compromised allografts from deceased donors and by the failure of the current hypothermic preservation protocols to extend the allograft's cold ischemia time beyond 4 h. We hypothesized that the active replenishment of the cellular cytosolic adenosine-5`-triphosphate (ATP) stores by means of energy delivery vehicles (ATPv) encapsulating high-energy ATP is a better strategy to improve allograft's tolerance to extended cold ischemia times. MATERIALS AND METHODS We utilized established rat model of isolated bilateral in-situ non-cycled perfusions of both hind limbs. Ipsilateral and contralateral limbs in the anesthetized animal were randomized for simultaneous perfusions with either the University of Wisconsin (UW) solution, with/without O2 supplementation (control), or with the UW solution supplemented with the ATPv, with/without O2 supplementation (experimental). Following perfusion, the hind limbs were surgically removed and stored at 4°C for 12, 16, or 24 hours as extended cold ischemia times. At the end of each respective storage time, samples of skin, and soleus, extensor digitalis longus, and tibialis anterior muscles were recovered for assessment using tissue histology and tissue lysate studies. RESULTS Control muscle sections showed remarkable microvascular and muscle damage associated with loss of myocyte transverse striation and marked decrease in myocyte nucleus density. A total of 1,496 nuclei were counted in 179 sections of UW-perfused control muscles in contrast to 1,783 counted in 130 sections of paired experimental muscles perfused with the ATPv-enhanced perfusate. This yielded 8 and 13 nuclei/field for the control and experimental muscles, respectively (P < .004). Oxygenation of the perfusion solutions before use did not improve the nucleus density of either the control or experimental muscles (n = 7 animals, P > .05). Total protein isolated from the muscle lysates was similar in magnitude regardless of muscle type, perfusion protocol, or duration of cold ischemia time. Prolonged static cold preservation of the hind limbs completely degraded the composite tissue's Ribonucleic acid (RNA). This supplementary result confirms the notion that that reverse transcription-Polymerase Chain Reaction, enzyme-linked immunosorbent assay, or the respiratory complex II enzyme activity techniques should not be used as indices of graft quality after prolonged static cold storage. CONCLUSIONS In conclusion, this study demonstrates that active cellular cytosolic ATP replenishment increases hind limb composite tissue tolerance to extended cold ischemia times. Quality indicators and clinically relevant biomarkers that define composite tissue viability and function during static cold storage are warranted.
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Affiliation(s)
- El Rasheid Zakaria
- Department of Surgery, Division of Trauma, Surgical Critical Care, Burns, and Acute Care Surgery, College of Medicine, The University of Arizona, Tucson, AZ 85724, USA
| | - Wali Yousufzai
- Department of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA
| | - Omar Obaid
- Department of Surgery, Division of Trauma, Surgical Critical Care, Burns, and Acute Care Surgery, College of Medicine, The University of Arizona, Tucson, AZ 85724, USA
| | - Samer Asmar
- Department of Surgery, Staten Island University Hospital, Staten Island, NY 10305, USA
| | - Chiu-Hsieh Hsu
- Department of Surgery, Division of Trauma, Surgical Critical Care, Burns, and Acute Care Surgery, College of Medicine, The University of Arizona, Tucson, AZ 85724, USA
- The Mel and Enid Zuckerman College of Public, The University of Arizona, Tucson, AZ 85724, USA
| | - Bellal Joseph
- Department of Surgery, Division of Trauma, Surgical Critical Care, Burns, and Acute Care Surgery, College of Medicine, The University of Arizona, Tucson, AZ 85724, USA
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de Vries RJ, Cronin SEJ, Romfh P, Pendexter CA, Jain R, Wilks BT, Raigani S, van Gulik TM, Chen P, Yeh H, Uygun K, Tessier SN. Non-invasive quantification of the mitochondrial redox state in livers during machine perfusion. PLoS One 2021; 16:e0258833. [PMID: 34705828 PMCID: PMC8550443 DOI: 10.1371/journal.pone.0258833] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/01/2021] [Accepted: 10/06/2021] [Indexed: 11/19/2022] Open
Abstract
Ischemia reperfusion injury (IRI) is a critical problem in liver transplantation that can lead to life-threatening complications and substantially limit the utilization of livers for transplantation. However, because there are no early diagnostics available, fulminant injury may only become evident post-transplant. Mitochondria play a central role in IRI and are an ideal diagnostic target. During ischemia, changes in the mitochondrial redox state form the first link in the chain of events that lead to IRI. In this study we used resonance Raman spectroscopy to provide a rapid, non-invasive, and label-free diagnostic for quantification of the hepatic mitochondrial redox status. We show this diagnostic can be used to significantly distinguish transplantable versus non-transplantable ischemically injured rat livers during oxygenated machine perfusion and demonstrate spatial differences in the response of mitochondrial redox to ischemia reperfusion. This novel diagnostic may be used in the future to predict the viability of human livers for transplantation and as a tool to better understand the mechanisms of hepatic IRI.
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Affiliation(s)
- Reinier J. de Vries
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
- Department of Surgery, Amsterdam University Medical Centers–Location AMC, University of Amsterdam, Amsterdam, the Netherlands
| | - Stephanie E. J. Cronin
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
| | - Padraic Romfh
- Pendar Technologies, Cambridge, MA, United States of America
| | - Casie A. Pendexter
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
| | - Rohil Jain
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
| | - Benjamin T. Wilks
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
| | - Siavash Raigani
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
- Division of Transplantation, Department of Surgery, Massachusetts General Hospital, Boston, MA, United States of America
| | - Thomas M. van Gulik
- Department of Surgery, Amsterdam University Medical Centers–Location AMC, University of Amsterdam, Amsterdam, the Netherlands
| | - Peili Chen
- Pendar Technologies, Cambridge, MA, United States of America
| | - Heidi Yeh
- Division of Transplantation, Department of Surgery, Massachusetts General Hospital, Boston, MA, United States of America
| | - Korkut Uygun
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
| | - Shannon N. Tessier
- Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, Boston, MA, United States of America
- Shriners Hospitals for Children—Boston, Boston, MA, United States of America
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3
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Xu J, Hassan-Ally M, Casas-Ferreira AM, Suvitaival T, Ma Y, Vilca-Melendez H, Rela M, Heaton N, Jassem W, Legido-Quigley C. Deregulation of the Purine Pathway in Pre-Transplant Liver Biopsies Is Associated with Graft Function and Survival after Transplantation. J Clin Med 2020; 9:jcm9030711. [PMID: 32151072 PMCID: PMC7141328 DOI: 10.3390/jcm9030711] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/20/2020] [Revised: 02/28/2020] [Accepted: 03/02/2020] [Indexed: 12/21/2022] Open
Abstract
The current shortage of livers for transplantation has increased the use of marginal organs sourced from donation after circulatory death (DCD). However, these organs have a higher incidence of graft failure, and pre-transplant biomarkers which predict graft function and survival remain limited. Here, we aimed to find biomarkers of liver function before transplantation to allow better clinical evaluation. Matched pre- and post-transplant liver biopsies from DCD (n = 24) and donation after brain death (DBD, n = 70) were collected. Liver biopsies were analysed using mass spectroscopy molecular phenotyping. Discrimination analysis was used to parse metabolites differentiated between the two groups. Five metabolites in the purine pathway were investigated. Of these, the ratios of the levels of four metabolites to those of urate differed between DBD and DCD biopsies at the pre-transplantation stage (q < 0.05). The ratios of Adenosine monophosphate (AMP) and adenine levels to those of urate also differed in biopsies from recipients experiencing early graft function (EGF) (q < 0.05) compared to those of recipients experiencing early allograft dysfunction (EAD). Using random forest, a panel consisting of alanine aminotransferase (ALT) and the ratios of AMP, adenine, and hypoxanthine levels to urate levels predicted EGF with area under the curve (AUC) of 0.84 (95% CI (0.71, 0.97)). Survival analysis revealed that the metabolite classifier could stratify six-year survival outcomes (p = 0.0073). At the pre-transplantation stage, a panel composed of purine metabolites and ALT could improve the prediction of EGF and survival.
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Affiliation(s)
- Jin Xu
- Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King’s College London, London SE1 9NH, UK; (J.X.); (M.H.-A.); (A.M.C.-F.)
| | - Mohammad Hassan-Ally
- Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King’s College London, London SE1 9NH, UK; (J.X.); (M.H.-A.); (A.M.C.-F.)
| | - Ana María Casas-Ferreira
- Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King’s College London, London SE1 9NH, UK; (J.X.); (M.H.-A.); (A.M.C.-F.)
- Department of Analytical Chemistry, Nutrition and Food Science, University of Salamanca, 37008 Salamanca, Spain
| | | | - Yun Ma
- Institute of Liver Studies, King’s College Hospital, King’s College London, London SE5 9RS, UK; (Y.M.); (H.V.-M.); (M.R.); (N.H.)
| | - Hector Vilca-Melendez
- Institute of Liver Studies, King’s College Hospital, King’s College London, London SE5 9RS, UK; (Y.M.); (H.V.-M.); (M.R.); (N.H.)
| | - Mohamed Rela
- Institute of Liver Studies, King’s College Hospital, King’s College London, London SE5 9RS, UK; (Y.M.); (H.V.-M.); (M.R.); (N.H.)
| | - Nigel Heaton
- Institute of Liver Studies, King’s College Hospital, King’s College London, London SE5 9RS, UK; (Y.M.); (H.V.-M.); (M.R.); (N.H.)
| | - Wayel Jassem
- Institute of Liver Studies, King’s College Hospital, King’s College London, London SE5 9RS, UK; (Y.M.); (H.V.-M.); (M.R.); (N.H.)
- Correspondence: (W.J.); (C.L.-Q.)
| | - Cristina Legido-Quigley
- Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King’s College London, London SE1 9NH, UK; (J.X.); (M.H.-A.); (A.M.C.-F.)
- Steno Diabetes Center Copenhagen, DK-2800 Gentofte, Denmark;
- Correspondence: (W.J.); (C.L.-Q.)
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de Vries RJ, Tessier SN, Banik PD, Nagpal S, Cronin SEJ, Ozer S, Hafiz EOA, van Gulik TM, Yarmush ML, Markmann JF, Toner M, Yeh H, Uygun K. Supercooling extends preservation time of human livers. Nat Biotechnol 2019; 37:1131-1136. [PMID: 31501557 PMCID: PMC6776681 DOI: 10.1038/s41587-019-0223-y] [Citation(s) in RCA: 115] [Impact Index Per Article: 19.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2018] [Accepted: 07/12/2019] [Indexed: 12/25/2022]
Abstract
The inability to preserve vascular organs beyond several hours contributes to the scarcity of organs for transplantation1,2. Standard hypothermic preservation at +4 °C (refs. 1,3) limits liver preservation to less than 12 h. Our group previously showed that supercooled ice-free storage at -6 °C can extend viable preservation of rat livers4,5 However, scaling supercooling preservation to human organs is intrinsically limited because of volume-dependent stochastic ice formation. Here, we describe an improved supercooling protocol that averts freezing of human livers by minimizing favorable sites of ice nucleation and homogeneous preconditioning with protective agents during machine perfusion. We show that human livers can be stored at -4 °C with supercooling followed by subnormothermic machine perfusion, effectively extending the ex vivo life of the organ by 27 h. We show that viability of livers before and after supercooling is unchanged, and that after supercooling livers can withstand the stress of simulated transplantation by ex vivo normothermic reperfusion with blood.
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Affiliation(s)
- Reinier J de Vries
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Department of Surgery, University of Amsterdam, Amsterdam, the Netherlands
- Shriners Hospital for Children, Boston, MA, USA
| | - Shannon N Tessier
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Peony D Banik
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Sonal Nagpal
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Stephanie E J Cronin
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Sinan Ozer
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Ehab O A Hafiz
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
- Department of Electron Microscopy Research, Theodor Bilharz Research Institute, Giza, Egypt
| | - Thomas M van Gulik
- Department of Surgery, University of Amsterdam, Amsterdam, the Netherlands
| | - Martin L Yarmush
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - James F Markmann
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Center for Transplant Sciences, Massachusetts General Hospital, Boston, MA, USA
| | - Mehmet Toner
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Shriners Hospital for Children, Boston, MA, USA
| | - Heidi Yeh
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA
- Center for Transplant Sciences, Massachusetts General Hospital, Boston, MA, USA
| | - Korkut Uygun
- Center for Engineering in Medicine, Harvard Medical School & Massachusetts General Hospital, Boston, MA, USA.
- Shriners Hospital for Children, Boston, MA, USA.
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5
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Bellini MI, Yiu J, Nozdrin M, Papalois V. The Effect of Preservation Temperature on Liver, Kidney, and Pancreas Tissue ATP in Animal and Preclinical Human Models. J Clin Med 2019; 8:1421. [PMID: 31505880 PMCID: PMC6780500 DOI: 10.3390/jcm8091421] [Citation(s) in RCA: 30] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/13/2019] [Revised: 08/30/2019] [Accepted: 09/02/2019] [Indexed: 02/06/2023] Open
Abstract
The recent advances in machine perfusion (MP) technology involve settings ranging between hypothermic, subnormothermic, and normothermic temperatures. Tissue level adenosine triphosphate (ATP) is a long-established marker of viability and functionality and is universal for all organs. In the midst of a growing number of complex clinical parameters for the quality assessment of graft prior to transplantation, a revisit of ATP may shed light on the underlying reconditioning mechanisms of different perfusion temperatures in the form of restoration of metabolic and energy status. This article aims to review and critically analyse animal and preclinical human studies (discarded grafts) during MP of three abdominal organs (liver, kidney, and pancreas) in which ATP was a primary endpoint. A selective review of recent novel reconditioning approaches relevant to mitigation of graft ischaemia-reperfusion injury via MP and for different perfusion temperatures was also conducted. With a current reiterated interest for oxygenation during MP, a re-introduction of tissue ATP levels may be valuable for graft viability assessment prior to transplantation. Further studies may help delineate the benefits of selective perfusion temperatures on organs viability.
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Affiliation(s)
| | - Janice Yiu
- School of Medicine, University College London, London WC1E 6BT, UK
| | - Mikhail Nozdrin
- School of Medicine, Imperial College London, London SW72AZ, UK
| | - Vassilios Papalois
- Renal and Transplant Directorate, Imperial College Healthcare NHS Trust, London W120HS, UK
- Department of Surgery and Cancer, Imperial College London, London SW72AZ, UK
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[Modern concepts for the dynamic preservation of the liver and kidneys in the context of transplantation]. DER PATHOLOGE 2019; 40:292-298. [PMID: 30976824 DOI: 10.1007/s00292-019-0595-2] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
The increasing demand on donor grafts has forced experimental research on transplantation medicine to develop more efficient organ preservation strategies. Simple cold storage of grafts rarely offers optimal conditions for extended criteria donor organs. Hypothermic, oxygenated machine perfusion (HMP) is a classical method of dynamic organ preservation, which enables the provision of oxygen and nutrients to the tissue and provides a metabolic recovery of the graft prior to implantation. A more modern approach is normothermic machine perfusion (NMP), which instead simulates physiological conditions and enables an ex vivo evaluation and treatment of organ grafts. However, studies have found that a preceding period of cold storage significantly mitigates the functional advantage of NMP. A strategy to circumvent this phenomenon is controlled oxygenated rewarming (COR). The cold-stored graft is slowly and gradually rewarmed to subnormothermic or normothermic temperatures, providing a gentle adaption of energy metabolism and counteracting events of rewarming injury.
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Burlage LC, Tessier SN, Etra JW, Uygun K, Brandacher G. Advances in machine perfusion, organ preservation, and cryobiology: potential impact on vascularized composite allotransplantation. Curr Opin Organ Transplant 2018; 23:561-567. [PMID: 30080697 PMCID: PMC6449688 DOI: 10.1097/mot.0000000000000567] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
PURPOSE OF REVIEW In this review, we discuss novel strategies that allow for extended preservation of vascularized composite allografts and their potential future clinical implications for the field of vascularized composite allotransplantation (VCA). RECENT FINDINGS The current gold standard in tissue preservation - static cold preservation on ice - is insufficient to preserve VCA grafts for more than a few hours. Advancements in the field of VCA regarding matching and allocation, desensitization, and potential tolerance induction are all within reasonable reach to achieve; these are, however, constrained by limited preservation time of VCA grafts. Although machine perfusion holds many advantages over static cold preservation, it currently does not elongate the preservation time. More extreme preservation techniques, such as cryopreservation approaches, are, however, specifically difficult to apply to composite tissues as the susceptibility to ischemia and cryoprotectant agents varies greatly by tissue type. SUMMARY In the current scope of extended preservation protocols, high subzero approaches of VCA grafts will be particularly critical enabling technologies for the implementation of tolerance protocols clinically. Ultimately, advances in both preservation techniques and tolerance induction have the potential to transform the field of VCA and eventually lead to broad applications in reconstructive transplantation.
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Affiliation(s)
- Laura C. Burlage
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA
- Department of Surgery, Section Hepato-Pancreato-Biliary Surgery and Liver Transplantation, University Medical Center Groningen, Groningen, The Netherlands
| | - Shannon N. Tessier
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA
| | - Joanna W. Etra
- Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
| | - Korkut Uygun
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA
| | - Gerald Brandacher
- Department of Plastic and Reconstructive Surgery, Vascularized Composite Allotransplantation (VCA) Laboratory, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
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Alva N, Panisello-Roselló A, Flores M, Roselló-Catafau J, Carbonell T. Ubiquitin-proteasome system and oxidative stress in liver transplantation. World J Gastroenterol 2018; 24:3521-3530. [PMID: 30131658 PMCID: PMC6102496 DOI: 10.3748/wjg.v24.i31.3521] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/05/2018] [Revised: 06/28/2018] [Accepted: 06/30/2018] [Indexed: 02/06/2023] Open
Abstract
A major issue in organ transplantation is the development of a protocol that can preserve organs under optimal conditions. Damage to organs is commonly a consequence of flow deprivation and oxygen starvation following the restoration of blood flow and reoxygenation. This is known as ischemia-reperfusion injury (IRI): a complex multifactorial process that causes cell damage. While the oxygen deprivation due to ischemia depletes cell energy, subsequent tissue oxygenation due to reperfusion induces many cascades, from reactive oxygen species production to apoptosis initiation. Autophagy has also been identified in the pathogenesis of IRI, although such alterations and their subsequent functional significance are controversial. Moreover, proteasome activation may be a relevant pathophysiological mechanism. Different strategies have been adopted to limit IRI damage, including the supplementation of commercial preservation media with pharmacological agents or additives. In this review, we focus on novel strategies related to the ubiquitin proteasome system and oxidative stress inhibition, which have been used to minimize damage in liver transplantation.
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Affiliation(s)
- Norma Alva
- Department of Cell Biology, Physiology and Immunology, University of Barcelona, Barcelona 08028, Spain
| | - Arnau Panisello-Roselló
- Experimental Pathology Department, Institute of Biomedical Research of Barcelona, Barcelona 08036, Spain
| | - Marta Flores
- Department of Cell Biology, Physiology and Immunology, University of Barcelona, Barcelona 08028, Spain
| | - Joan Roselló-Catafau
- Experimental Pathology Department, Institute of Biomedical Research of Barcelona, Barcelona 08036, Spain
| | - Teresa Carbonell
- Department of Cell Biology, Physiology and Immunology, University of Barcelona, Barcelona 08028, Spain
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Schlegel A, Muller X, Dutkowski P. Hypothermic Machine Preservation of the Liver: State of the Art. CURRENT TRANSPLANTATION REPORTS 2018; 5:93-102. [PMID: 29564206 PMCID: PMC5843682 DOI: 10.1007/s40472-018-0183-z] [Citation(s) in RCA: 63] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
PURPOSE OF REVIEW In this review, we highlight which livers may benefit from additional treatment before implantation and describe the concept of hypothermic machine liver perfusion. Furthermore, we explain why cold oxygenated perfusion concepts could potentially lead to a breakthrough in this challenging field of transplantation. Accordingly, we summarize recent clinical applications of different hypothermic perfusion approaches. RECENT FINDINGS The impact of end-ischemic, hypothermic liver perfusion in liver transplantation is currently assessed by two multicenter, randomized controlled trials. Recently, new applications of hypothermic perfusion showed promising results and recipients were protected from severe intrahepatic biliary complications, despite the use of very extended criteria grafts including donation after circulatory death livers. SUMMARY Hypothermic machine liver perfusion is beneficial for high-risk livers and protects recipients from most feared complications. Importantly, such easy approach is currently implemented in several European centers and new markers obtained from perfusate may improve the prediction of liver function in the future.
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Affiliation(s)
- Andrea Schlegel
- The Liver Unit, Queen Elizabeth University Hospital Birmingham, Birmingham, UK
- NIHR Liver Biomedical Research Unit, University Hospitals Birmingham, Birmingham, UK
| | - Xavier Muller
- Department of Surgery & Transplantation, Swiss HPB and Transplant Center, University Hospital Zurich, Raemistrasse 100, CH-8091 Zurich, Switzerland
| | - Philipp Dutkowski
- Department of Surgery & Transplantation, Swiss HPB and Transplant Center, University Hospital Zurich, Raemistrasse 100, CH-8091 Zurich, Switzerland
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10
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Abstract
PURPOSE OF REVIEW Herein, we review the field of subzero organ preservation with a focus on recent developments in hepatic supercooling. RECENT FINDINGS Organ preservation is making a rapid shift from the decade old standard of storage on ice toward techniques that improve organ availability as well as preservation time. Long-term organ preservation would have tremendous benefits to the organ transplantation field, including better organ allocation, donor-recipient matching, as well as reduced preservation injury, and subsequent improvement of donor organ use. The formation of ice has proven an important limiting factor and novel techniques attempt to control or prevent freezing using cryoprotective agents, and highly controlled cooling regimens. Various techniques have been employed over the previous decades, including true organ freezing, vitrification, and subzero nonfreezing or supercooling. For most techniques, successful transplantation following long-term subzero preservation has remained elusive. Supercooling, however, recently delivered the first promising results, yielding survival after up to 4 days of supercooled preservation at -6°C. SUMMARY As the field of organ preservation undergoes significant development, the field of subzero preservation also receives renewed interest. Although many obstacles remain to be overcome to make subzero preservation feasible, novel techniques are beginning to show their potential in achieving long-term preservation.
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11
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Bruinsma BG, Avruch JH, Sridharan GV, Weeder PD, Jacobs ML, Crisalli K, Amundsen B, Porte RJ, Markmann JF, Uygun K, Yeh H. Peritransplant Energy Changes and Their Correlation to Outcome After Human Liver Transplantation. Transplantation 2017; 101:1637-1644. [PMID: 28230641 PMCID: PMC5481470 DOI: 10.1097/tp.0000000000001699] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Abstract
BACKGROUND The ongoing shortage of donor livers for transplantation and the increased use of marginal livers necessitate the development of accurate pretransplant tests of viability. Considering the importance energy status during transplantation, we aimed to correlate peritransplant energy cofactors to posttransplant outcome and subsequently model this in an ex vivo setting. METHODS Sequential biopsies were taken from 19 donor livers postpreservation, as well as 30 minutes after portal venous reperfusion and hepatic arterial reperfusion and analyzed by liquid chromatography-mass spectrometry for energetic cofactors (adenosine triphosphate [ATP]/adenosine diphosphate [ADP]/adenosine monophosphate [AMP], nicotinamide adenine dinucleotide /NAD, nicotinamide adenine dinucleotide phosphate / nicotinamide adenine dinucleotide phosphate , flavin adenine dinucleotide , glutathione disulfide/glutathione). Energy status was correlated to posttransplant outcome. In addition, 4 discarded human donation after circulatory death livers were subjected to ex vivo reperfusion, modeling reperfusion injury and were similarly analyzed for energetic cofactors. RESULTS A rapid shift toward higher energy adenine nucleotides was observed following clinical reperfusion, with a 2.45-, 3.17- and 2.12-fold increase in ATP:ADP, ATP:AMP and energy charge after portal venous reperfusion, respectively. Seven of the 19 grafts developed early allograft dysfunction. Correlation with peritransplant cofactors revealed a significant difference in EC between early allograft dysfunction and normal functioning grafts (0.09 vs 0.31, P < 0.05). In the simulated reperfusion model, a similar trend in adenine nucleotide changes was observed. CONCLUSIONS A preserved energy status appears critical in the peritransplant period. Levels of adenine nucleotides change rapidly after reperfusion and ratios of ATP/ADP/AMP after reperfusion are significantly correlated to graft function. Using these markers as a viability test in combination with ex vivo reperfusion may provide a useful predictor of outcome that incorporates donor, preservation, and reperfusion factors.
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Affiliation(s)
- Bote G. Bruinsma
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - James H. Avruch
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Gautham V. Sridharan
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Pepijn D. Weeder
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
- Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands
| | - Marie Louise Jacobs
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Kerry Crisalli
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Beth Amundsen
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Robert J. Porte
- Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands
| | - James F. Markmann
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Korkut Uygun
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
| | - Heidi Yeh
- Transplant Center, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA
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12
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Sutherland AI, Oniscu GC. Challenges and advances in optimizing liver allografts from donation after circulatory death donors. J Nat Sci Biol Med 2016; 7:10-5. [PMID: 27003962 PMCID: PMC4780154 DOI: 10.4103/0976-9668.175017] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
In recent years, there has been a shift in the donor demographics with an increase in donation after circulatory death (DCD). Livers obtained from DCD donors are known to have poorer outcomes when compared to donors after brainstem death and currently only a small proportion of DCD livers are used. This review outlines the recent technological developments in liver DCD donation, including clinical studies using normothermic regional perfusion and extracorporal machine perfusion of livers from DCD donors.
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Affiliation(s)
| | - Gabriel C Oniscu
- Scottish Liver Transplant Unit, Royal Infirmary of Edinburgh, Edinburgh, UK
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13
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Metabolic profiling during ex vivo machine perfusion of the human liver. Sci Rep 2016; 6:22415. [PMID: 26935866 PMCID: PMC4776101 DOI: 10.1038/srep22415] [Citation(s) in RCA: 79] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/03/2015] [Accepted: 02/15/2016] [Indexed: 01/12/2023] Open
Abstract
As donor organ shortages persist, functional machine perfusion is under investigation to improve preservation of the donor liver. The transplantation of donation after circulatory death (DCD) livers is limited by poor outcomes, but its application may be expanded by ex vivo repair and assessment of the organ before transplantation. Here we employed subnormothermic (21 °C) machine perfusion of discarded human livers combined with metabolomics to gain insight into metabolic recovery during machine perfusion. Improvements in energetic cofactors and redox shifts were observed, as well as reversal of ischemia-induced alterations in selected pathways, including lactate metabolism and increased TCA cycle intermediates. We next evaluated whether DCD livers with steatotic and severe ischemic injury could be discriminated from ‘transplantable’ DCD livers. Metabolomic profiling was able to cluster livers with similar metabolic patterns based on the degree of injury. Moreover, perfusion parameters combined with differences in metabolic factors suggest variable mechanisms that result in poor energy recovery in injured livers. We conclude that machine perfusion combined with metabolomics has significant potential as a clinical instrument for the assessment of preserved livers.
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14
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Ob/ob mouse livers show decreased oxidative phosphorylation efficiencies and anaerobic capacities after cold ischemia. PLoS One 2014; 9:e100609. [PMID: 24956382 PMCID: PMC4067359 DOI: 10.1371/journal.pone.0100609] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/14/2013] [Accepted: 05/28/2014] [Indexed: 01/26/2023] Open
Abstract
Background Hepatic 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. Methods Livers 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. Conclusions Steatotic 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.
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15
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Bruinsma B, Yeh H, Özer S, Martins P, Farmer A, Wu W, Saeidi N, op den Dries S, Berendsen T, Smith R, Markmann J, Porte R, Yarmush M, Uygun K, Izamis M. Subnormothermic machine perfusion for ex vivo preservation and recovery of the human liver for transplantation. Am J Transplant 2014; 14:1400-9. [PMID: 24758155 PMCID: PMC4470578 DOI: 10.1111/ajt.12727] [Citation(s) in RCA: 157] [Impact Index Per Article: 14.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2013] [Revised: 02/25/2014] [Accepted: 02/26/2014] [Indexed: 01/25/2023]
Abstract
To reduce widespread shortages, attempts are made to use more marginal livers for transplantation. Many of these grafts are discarded for fear of inferior survival rates or biliary complications. Recent advances in organ preservation have shown that ex vivo subnormothermic machine perfusion has the potential to improve preservation and recover marginal livers pretransplantation. To determine the feasibility in human livers, we assessed the effect of 3 h of oxygenated subnormothermic machine perfusion (21°C) on seven livers discarded for transplantation. Biochemical and microscopic assessment revealed minimal injury sustained during perfusion. Improved oxygen uptake (1.30 [1.11-1.94] to 6.74 [4.15-8.16] mL O2 /min kg liver), lactate levels (4.04 [3.70-5.99] to 2.29 [1.20-3.43] mmol/L) and adenosine triphosphate content (45.0 [70.6-87.5] pmol/mg preperfusion to 167.5 [151.5-237.2] pmol/mg after perfusion) were observed. Liver function, reflected by urea, albumin and bile production, was seen during perfusion. Bile production increased and the composition of bile (bile salts/phospholipid ratio, pH and bicarbonate concentration) became more favorable. In conclusion, ex vivo subnormothermic machine perfusion effectively maintains liver function with minimal injury and sustains or improves various hepatobiliary parameters postischemia.
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Affiliation(s)
- B.G. Bruinsma
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA,Department of Surgery (Surgical Laboratory), Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
| | - H. Yeh
- Transplant Center, Massachusetts General Hospital, Boston, MA, USA
| | - S Özer
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA
| | - P.N. Martins
- Organ Transplant Surgery, UMass Memorial Medical Center, Boston, MA, USA
| | - A. Farmer
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA
| | - W. Wu
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA
| | - N. Saeidi
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA
| | - S. op den Dries
- Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands
| | - T.A. Berendsen
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA
| | - R.N. Smith
- Department of Pathology, Massachusetts General Hospital, Boston, MA, USA
| | - J.F. Markmann
- Transplant Center, Massachusetts General Hospital, Boston, MA, USA
| | - R. Porte
- Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands
| | - M.L. Yarmush
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA,Department of Biomedical Engineering, Rutgers University, Piscataway, NJ, USA
| | - K. Uygun
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA,Corresponding authors Korkut Uygun, PhD , Maria-Louisa Izamis, PhD
| | - M.L. Izamis
- Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Boston, MA, USA,Corresponding authors Korkut Uygun, PhD , Maria-Louisa Izamis, PhD
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16
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García-Gil FA, Albendea CD, López-Pingarrón L, Royo-Dachary P, Martínez-Guillén J, Piedrafita E, Martínez-Díez M, Soria J, García JJ. Altered cellular membrane fluidity levels and lipid peroxidation during experimental pancreas transplantation. J Bioenerg Biomembr 2012; 44:571-7. [PMID: 22986734 DOI: 10.1007/s10863-012-9459-7] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2012] [Accepted: 05/30/2012] [Indexed: 12/13/2022]
Abstract
Although the pathogenesis of ischemia reperfusion (IR) injury is based on complex mechanisms, free radicals play a central role. We evaluated membrane fluidity and lipid peroxidation during pancreas transplantation (PT) performed in 12 pigs (six donors and six recipients). Fluidity was measured by fluorescence spectroscopy, and malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA) concentrations were used as an index of lipid oxidation. Pancreatic tissues were collected as follows: (A) donor, immediately before vascular clamping; (B) graft, following perfusion lavage with University of Wisconsin preservation fluid; (C) graft, after 16 h of cold ischemia; and (D) recipient, 30 min vascular postreperfusion. Fluidity and MDA and 4-HDA concentrations were similar in cases A, B, and C. However, there was significant membrane rigidity and increased lipid peroxidation after reperfusion (D). These findings suggest that reperfusion exaggerates oxidative damage and may account for the rigidity in the membranes of allografts during PT.
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Affiliation(s)
- F A García-Gil
- Department of Surgery, Gynaecology and Obstetrics, University of Zaragoza, Zaragoza, Spain
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17
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Mendes-Braz M, Elias-Miró M, Jiménez-Castro MB, Casillas-Ramírez A, Ramalho FS, Peralta C. The current state of knowledge of hepatic ischemia-reperfusion injury based on its study in experimental models. J Biomed Biotechnol 2012; 2012:298657. [PMID: 22649277 PMCID: PMC3357607 DOI: 10.1155/2012/298657] [Citation(s) in RCA: 112] [Impact Index Per Article: 8.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/01/2011] [Accepted: 02/23/2012] [Indexed: 12/11/2022] Open
Abstract
The present review focuses on the numerous experimental models used to study the complexity of hepatic ischemia/reperfusion (I/R) injury. Although experimental models of hepatic I/R injury represent a compromise between the clinical reality and experimental simplification, the clinical transfer of experimental results is problematic because of anatomical and physiological differences and the inevitable simplification of experimental work. In this review, the strengths and limitations of the various models of hepatic I/R are discussed. Several strategies to protect the liver from I/R injury have been developed in animal models and, some of these, might find their way into clinical practice. We also attempt to highlight the fact that the mechanisms responsible for hepatic I/R injury depend on the experimental model used, and therefore the therapeutic strategies also differ according to the model used. Thus, the choice of model must therefore be adapted to the clinical question being answered.
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Affiliation(s)
- M. Mendes-Braz
- Departamento de Patologia e Medicina Legal, Faculdade de Medicina, Universidade de Sao Paulo, 14040-900 Ribeirão Preto, SP, Brazil
| | - M. Elias-Miró
- Institut d'lnvestigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
| | - M. B. Jiménez-Castro
- Institut d'lnvestigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
| | - A. Casillas-Ramírez
- Institut d'lnvestigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
| | - F. S. Ramalho
- Departamento de Patologia e Medicina Legal, Faculdade de Medicina, Universidade de Sao Paulo, 14040-900 Ribeirão Preto, SP, Brazil
| | - C. Peralta
- Institut d'lnvestigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 08036 Barcelona, Spain
- Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, 08036 Barcelona, Spain
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18
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Giannone FA, Treré D, Domenicali M, Grattagliano I, Baracca A, Sgarbi G, Maggioli C, Longobardi P, Solaini G, Derenzini M, Bernardi M, Caraceni P. An innovative hyperbaric hypothermic machine perfusion protects the liver from experimental preservation injury. ScientificWorldJournal 2012; 2012:573410. [PMID: 22593698 PMCID: PMC3345935 DOI: 10.1100/2012/573410] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/01/2011] [Accepted: 12/18/2011] [Indexed: 02/07/2023] Open
Abstract
Purpose. Hypothermic machine perfusion systems seem more effective than the current static storage to prevent cold ischemic liver injury. Thus, we test an innovative hyperbaric hypothermic machine perfusion (HHMP), which combines hyperbaric oxygenation of the preservation solution and continuous perfusion of the graft. Methods. Rat livers were preserved with Celsior solution according to 4 different modalities: normobaric static preservation; hyperbaric static preservation at 2 atmosphere absolute (ATA); normobaric dynamic preservation, with continuous perfusion; hyperbaric dynamic preservation, with continuous perfusion at 2 ATA. After 24 h cold preservation, we assessed different parameters. Results. Compared to baseline, livers preserved with the current static storage showed severe ultrastructural damage, glycogen depletion and an increased oxidative stress. Normobaric perfused livers showed improved hepatocyte ultrastructure and ameliorated glycogen stores, but they still suffered a significant oxidative damage. The addition of hyperbaric oxygen produces an extra benefit by improving oxidative injury and by inducing endothelial NO synthase (eNOS) gene expression. Conclusions. Preservation by means of the present innovative HHMP reduced the liver injury occurring after the current static cold storage by lowering glycogen depletion and oxidative damage. Interestingly, only the use of hyperbaric oxygen was associated to a blunted oxidative stress and an increased eNOS gene expression.
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Affiliation(s)
- Ferdinando A Giannone
- Department of Clinical Medicine, Alma Mater Studiorum University of Bologna, 40138 Bologna, Italy
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19
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Iwai S, Kikuchi T, Kasahara N, Teratani T, Yokoo T, Sakonju I, Okano S, Kobayashi E. Impact of normothermic preservation with extracellular type solution containing trehalose on rat kidney grafting from a cardiac death donor. PLoS One 2012; 7:e33157. [PMID: 22457739 PMCID: PMC3310060 DOI: 10.1371/journal.pone.0033157] [Citation(s) in RCA: 27] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/23/2011] [Accepted: 02/06/2012] [Indexed: 12/30/2022] Open
Abstract
Background The aim of this study was to investigate factors that may improve the condition of a marginal kidney preserved with a normothermic solution following cardiac death (CD) in a model of rat kidney transplantation (RTx). Methods Post-euthanasia, Lewis (LEW) donor rats were left for 1 h in a 23°C room. These critical kidney grafts were preserved in University of Wisconsin (UW), lactate Ringer's (LR), or extracellular-trehalose-Kyoto (ETK) solution, followed by intracellular-trehalose-Kyoto (ITK) solution at 4, 23, or 37°C for another 1 h, and finally transplanted into bilaterally nephrectomized LEW recipient rats (n = 4–6). Grafts of rats surviving to day 14 after RTx were evaluated by histopathological examination. The energy activity of these marginal rat kidneys was measured by high-performance liquid chromatography (HPLC; n = 4 per group) and fluorescence intensity assay (n = 6 per group) after preservation with UW or ETK solutions at each temperature. Finally, the transplanted kidney was assessed by an in vivo luciferase imaging system (n = 2). Results Using the 1-h normothermic preservation of post-CD kidneys, five out of six recipients in the ETK group survived until 14 days, in contrast to zero out of six in the UW group (p<0.01). Preservation with ITK rather than ETK at 23°C tended to have an inferior effect on recipient survival (p = 0.12). Energy activities of the fresh donor kidneys decreased in a temperature-dependent manner, while those of post-CD kidneys remained at the lower level. ETK was superior to UW in protecting against edema of the post-CD kidneys at the higher temperature. Luminescence intensity of successful grafts recovered within 1 h, while the intensity of grafts of deceased recipients did not change at 1 h post-reperfusion. Conclusions Normothermic storage with extracellular-type solution containing trehalose might prevent reperfusion injury due to temperature-dependent tissue edema.
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Affiliation(s)
- Satomi Iwai
- Laboratory of Small Animal Surgery I, School of Veterinary Medicine, Kitasato University, Aomori, Japan
| | - Takeshi Kikuchi
- Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan
| | - Naoya Kasahara
- Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan
| | - Takumi Teratani
- Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan
| | - Takashi Yokoo
- Project Laboratory for Kidney Regeneration, Institute of DNA Medicine, Department Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
| | - Iwao Sakonju
- Laboratory of Small Animal Surgery I, School of Veterinary Medicine, Kitasato University, Aomori, Japan
| | - Shouzou Okano
- Laboratory of Small Animal Surgery II, School of Veterinary Medicine, Kitasato University, Aomori, Japan
| | - Eiji Kobayashi
- Center for Development of Advanced Medical Technology, Jichi Medical University, Tochigi, Japan
- * E-mail:
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20
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Sgarbi G, Giannone F, Casalena GA, Baracca A, Baldassare M, Longobardi P, Caraceni P, Derenzini M, Lenaz G, Trerè D, Solaini G. Hyperoxia fully protects mitochondria of explanted livers. J Bioenerg Biomembr 2011; 43:673-82. [PMID: 22015484 DOI: 10.1007/s10863-011-9390-3] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2011] [Accepted: 09/28/2011] [Indexed: 12/18/2022]
Abstract
Liver ischemia-reperfusion injury is still an open problem in many clinical circumstances, including surgery and transplantation. This study investigates how mitochondrial structure, mass and oxidative phosphorylation change and may be preserved during a brief period of ischemia followed by a long period of reperfusion, an experimental model that mimics the condition to which a liver is exposed during transplantation. Livers were explanted from rats and exposed for 24 h to three different oxygen availability conditions at 4 °C. Mitochondrial mass, respiration, oxidative phosphorylation (OXPHOS), and levels of OXPHOS complexes were all significantly altered in livers stored under the currently used preservation condition of normoxia. Remarkably, liver perfusion with hyperoxic solutions fully preserved mitochondrial morphology and function, suggesting that perfusion of the graft with hyperoxic solution should be considered in human transplantation.
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Affiliation(s)
- G Sgarbi
- Department of Biochemistry, University of Bologna, Via Irnerio, 48, 40126, Bologna, Italy
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21
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Abstract
Due to the critical shortage of deceased donor grafts, clinicians are continually expanding the criteria for an acceptable liver donor to meet the waiting list demands. However, the reduced ischemic tolerance of those extended criteria grafts jeopardizes organ viability during cold storage. Machine perfusion has been developed to limit ischemic liver damage but despite its proven biochemical benefit, machine liver perfusion is not yet considered clinically due to its low practicability. In this review, we summarize our understanding of the role of machine perfusion in marginal liver preservation. The goal is to highlight advantages or disadvantages of current perfusion techniques and to explain the underlying mechanisms. We provide evidence for the need of a liver perfusion performance shortly before implantation, and point out promising designs.
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Affiliation(s)
- P Dutkowski
- Swiss HPB (Hepato-Pancreato-Biliary) Center, Department of Visceral and Transplantation Surgery, University Hospital Zürich, Zurich, Switzerland
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22
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Manekeller S, Seinsche A, Stegemann J, Hirner A. Optimising post-conditioning time of marginal donor livers. Langenbecks Arch Surg 2008; 393:311-6. [PMID: 18283484 DOI: 10.1007/s00423-008-0288-4] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/11/2008] [Accepted: 01/17/2008] [Indexed: 01/14/2023]
Abstract
BACKGROUND Due to the discrepancy between organ donors and receptors, the use of marginal livers (e.g., non-heart-beating-donor grafts) for transplantation purpose increased. The potential of a short-term aerobic machine perfusion (post-conditioning) for "less than optimal" grafts after cold storage (CS) was recently demonstrated. In our study, the optimal time course of post-conditioning (PC) is to be evaluated. MATERIALS AND METHODS Livers from male Wistar rats were withdrawn 30 min after cardiac arrest and flushed with histidine tryptophan ketoglutarate (HTK) solution. Then they were stored in HTK at 4 degrees C for 18 h. After 16 h, some livers were put on PC by cold perfusion with HTK for 0.5, 1, 2 or 3 h. Afterwards, the viability of the organs was estimated by warm reperfusion (2 h) in vitro. RESULTS After 1 h of PC, a significant increase in bile production and a decrease in enzyme release could be detected in comparison to CS. The adenosine triphosphate content of the PC livers after 1 h of treatment was significant higher than in CS organs. No markers for apoptosis could be detected after 1 h PC. CONCLUSION It can be concluded that a PC of 1 h after cold storage can ameliorate the organ viability of marginal livers. The extension or abbreviation of PC time seems to have no further beneficial effects.
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Affiliation(s)
- Steffen Manekeller
- Department of Surgery, Faculty of Medicine, Rheinische Friedrich-Wilhelms-Universität, Bonn, Germany.
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23
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Manekeller S, Schuppius A, Stegemann J, Hirner A, Minor T. Role of perfusion medium, oxygen and rheology for endoplasmic reticulum stress-induced cell death after hypothermic machine preservation of the liver. Transpl Int 2007; 21:169-77. [PMID: 18005084 DOI: 10.1111/j.1432-2277.2007.00595.x] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022]
Abstract
Recently, the endoplasmic reticulum (ER) has been disclosed as subcellular target reactive to ischaemia/reperfusion and possibly influenced by hypothermic machine preservation. Here, the respective role of perfusate, perfusion itself, and the effect of continuous oxygenation to trigger ER-stress in the graft should be investigated. Livers were retrieved 30 min after cardiac arrest of male Wistar rats and preserved by cold storage (CS) in histidine-tryptophan-ketoglutarate (HTK) for 18 h at 4 degrees C. Other organs were subjected to aerobic conditions either by oxygenated machine perfusion with HTK (MP-HTK) or Belzer solution (MP-Belzer) at 4 degrees C or by venous insufflation of gaseous oxygen during cold storage (VSOP). Viability of livers was evaluated upon reperfusion in vitro according to previously validated techniques for 120 min at 37 degrees C. Oxygenation during preservation (MP-HTK, MP-Belzer or VSOP) concordantly improved functional recovery (bile flow, ammonia clearance), reduced parenchymal enzyme leakage and histological signs of necrosis and significantly attenuated mitochondrial induction of apoptosis (cleavage of caspase 9) compared to CS. However, MP with either medium produced about 500% elevated protein expression of CHOP/GADD153, suggesting pro-apoptotic ER-stress responses, paralleled by a significant elevation of caspase-12 enzyme activity compared to CS or VSOP. Although MP also promoted a slight (20%) induction of the cytoprotective ER-protein Bax inhibitor protein (BI-1), prevailing of proapoptotic reactions was seen by increased cleavage of caspase-3 and poly (ADP-Ribase)-polymerase (PARP) in both MP-groups. Endoplasmic stress activation is conjectured a specific side effect of long-term machine preservation irrespective of the medium, actually promoting cellular apoptosis via activation of caspase-12. The simple insufflation of gaseous O2 may be considered a feasible alternative, apparently indifferent to the endoplasmic reticulum.
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Boros P, Liu J, Li Y, Bromberg JS. Organ transplantation in rodents: novel applications of long-established methods. Transpl Immunol 2007; 18:44-52. [PMID: 17584602 PMCID: PMC2737136 DOI: 10.1016/j.trim.2007.03.004] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/21/2007] [Accepted: 03/13/2007] [Indexed: 01/27/2023]
Abstract
Rodent models of solid organ transplantation have been used for many decades. Standardized operative techniques resulting in highly reproducible survival rates have been developed for several organs. This allowed scientists to investigate many clinically relevant problems, test new drugs and establish novel treatment regimens. Recently, many studies used these models to explore novel issues such as graft modification by pharmaceutical, surgical or genetic engineering methods, post-transplant regeneration, leukocyte trafficking or interactions between the innate and allo-specific arms of the immune response. The results from these studies clearly facilitate a more complex and comprehensive understanding of existing problem. The long-established methods of rodent organ transplantation, combined with the newest achievements in surgical techniques, biotechnology and imaging, will remain indispensable tools of transplantation biology.
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Affiliation(s)
- Peter Boros
- Microvascular Surgery, Shared Research Facilities, Mount Sinai School of Medicine, New York, NY 10029, United States.
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25
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't Hart NA, der van Plaats A, Leuvenink HGD, van Goor H, Wiersema-Buist J, Verkerke GJ, Rakhorst G, Ploeg RJ. Determination of an adequate perfusion pressure for continuous dual vessel hypothermic machine perfusion of the rat liver. Transpl Int 2007; 20:343-52. [PMID: 17326775 DOI: 10.1111/j.1432-2277.2006.00433.x] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Hypothermic machine perfusion (HMP) provides better protection against ischemic damage of the kidney compared to cold-storage. The required perfusion pressures needed for optimal HMP of the liver are, however, unknown. Rat livers were preserved in University of Wisconsin organ preservation solution enriched with acridine orange (AO) to stain viable cells and propidium iodide (PI) to detect dead cells. Perfusion pressures of 12.5%, 25% or 50% of physiologic perfusion pressures were compared. Intravital fluorescence microscopy was used to assess liver perfusion by measuring the percentage of AO staining. After 1-h, the perfusion pressure of 12.5% revealed 72% +/- 3% perfusion of mainly the acinary zones one and two. The perfusion pressure of 25% and 50% showed complete perfusion. Furthermore, 12.5% showed 14.7 +/- 3.6, 25% showed 3.7 +/- 0.9, and 50% showed 11.2 +/- 1.4 PI positive cells. One hour was followed by another series of experiments comprising 24-h preservation. In comparison with 24-h cold-storage, HMP at 25% showed less PI positive cells and HMP at 50% showed more PI positive cells. In summary, perfusion at 25% showed complete perfusion, demonstrated by AO staining, with minimal cellular injury, shown with PI. This study indicates that fine-tuning of the perfusion pressure is crucial to balance (in)complete perfusion and endothelial injury.
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Affiliation(s)
- Nils A 't Hart
- Surgery Research Laboratory, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
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Dutkowski P, Furrer K, Tian Y, Graf R, Clavien PA. Novel short-term hypothermic oxygenated perfusion (HOPE) system prevents injury in rat liver graft from non-heart beating donor. Ann Surg 2006; 244:968-76; discussion 976-7. [PMID: 17122622 PMCID: PMC1856639 DOI: 10.1097/01.sla.0000247056.85590.6b] [Citation(s) in RCA: 123] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
OBJECTIVE To assess a machine perfusion system in rescuing liver grafts from non-heart-beating donors (NHBD). SUMMARY BACKGROUND DATA The introduction of extracorporeal liver perfusion systems in the clinical routine depends on feasibility. Conceivably, perfusion could be performed during recipient preparation. We investigated whether a novel rat liver machine perfusion applied after in situ ischemia and cold storage can rescue NHBD liver grafts. METHODS We induced cardiac arrest in male Brown Norway rats by phrenotomy and ligation of the subcardial aorta. We studied 2 experimental groups: 45 minutes of warm in situ ischemia + 5 hours cold storage versus 45 minutes of warm in situ ischemia + 5 hours cold storage followed by 1 hour hypothermic oxygenated extracorporeal perfusion (HOPE). In both groups, livers were reperfused in a closed sanguineous isolated liver perfusion device for 3 hours at 37 degrees C. To test the benefit of HOPE on survival, we performed orthotopic liver transplantation in both experimental groups. RESULTS After cold storage and reperfusion, NHBD livers showed necrosis of hepatocytes, increased release of AST, and decreased bile flow. HOPE improved NHBD livers significantly with a reduction of necrosis, less AST release, and increased bile flow. ATP was severely depleted in cold-stored NHBD livers but restored in livers treated by HOPE. After orthotopic liver transplantation, grafts treated by HOPE demonstrated a significant extension on animal survival. CONCLUSIONS We demonstrate a beneficial effect of HOPE by preventing reperfusion injury in a clinically relevant NHBD model.
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Affiliation(s)
- Philipp Dutkowski
- Swiss Hepato-Pancreato-Biliary (HPB) Center, Laboratory for HPB and Transplantation Surgery, Department of Visceral and Transplantation Surgery, University Hospital Zurich, Zurich, Switzerland.
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Manekeller S, Minor T. Possibility of conditioning predamaged grafts after cold storage: influences of oxygen and nutritive stimulation. Transpl Int 2006; 19:667-74. [PMID: 16827684 DOI: 10.1111/j.1432-2277.2006.00320.x] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Abstract
The potential of short-term oxygenated perfusion after cold storage (CS) to reverse deleterious priming of nonheart beating donors grafts should be investigated, addressing the respective role of oxygenation and nutrients or metabolic charge. Livers were retrieved 30 min after cardiac arrest of male Wistar rats and preserved with histidine tryptophan ketoglutarate (HTK)-solution for 18 h by CS. After 16 h, some livers were put on an oxygenated machine-preservation-circuit for the last 2 h and conditioned by cold perfusion with either HTK (conHTK), HTK supplemented with adenosine, phosphate and glucose (conHTK+) or Williams-E solution (conWE). Upon warm reperfusion, postconditioning with any of the solutions led to a significant (three- to fivefold) reduction of parenchymal damage (ALT, GLDH-release) compared with CS. Metabolic recovery (bile production) was also significantly enhanced compared with CS, with best results found after conHTK. The beneficial effect of postconditioning with HTK was associated with a significantly mitigated cleavage of caspase 12 and 3. We conclude from these data that conditioning of predamaged livers is possible even after CS by short-term oxygenated perfusion in the cold and, under these conditions, not depending on energetic support or nutritive stimulation.
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Affiliation(s)
- Steffen Manekeller
- Surgical Research Division, University Clinic of Surgery, Bonn, Germany.
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Minor T, Manekeller S, Sioutis M, Dombrowski F. Endoplasmic and vascular surface activation during organ preservation: refining upon the benefits of machine perfusion. Am J Transplant 2006; 6:1355-66. [PMID: 16686759 DOI: 10.1111/j.1600-6143.2006.01338.x] [Citation(s) in RCA: 49] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
The endoplasmic reticulum (ER) represents a subcellular target reactive to various cytosolic impairments. The involvement of ER-stress in organ preservation was investigated, comparing machine preservation, cold storage (CS) and a novel concept of only temporary perfusion after procurement. Rat livers were retrieved 30 min after cardiac arrest and preserved for 18 h by CS, oxygenated machine perfusion for 18 h (18 h MP) or for 2 h with subsequent CS for 16 h (2 h MP + 16 h CS). Upon reperfusion, 18 h MP significantly improved enzyme leakage (ALT, LDH) and promoted a 2-fold increase of metabolic recovery compared to CS. However, vascular stress, evaluated by endothelin-release, was significantly elevated after 18 h MP. Interestingly, better viability was obtained using the short-term perfusion protocol (2 h MP + 16 h CS), which further reduced enzyme leakage, maintained energetic recovery and mitigated endothelin-release compared to 18 h MP. Caspase 12-mRNA was upregulated in the 18 h MP-group but unchanged after CS or 2 h MP + 16 h CS. Activation/cleavage of caspase 12 protein was significantly enhanced after 18 h MP and very low in the 2 h MP + 16 h CS-group. Correspondingly, electron microscopy showed ultrastructural alterations of ER after CS and especially after 18 h MP but not after 2 h MP + 16 h CS. At this time mitochondrial appearance was unaffected in all groups, suggesting the ER to be an early subcellular target of preservation injury. In our model, ER and vascular endothelium were best protected by only temporary machine perfusion, which also maintained overall graft viability.
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Affiliation(s)
- T Minor
- Surgical Research Division, University Clinic of Surgery, Bonn, Germany.
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Ijichi H, Taketomi A, Soejima Y, Yoshizumi T, Uchiyama H, Shimada M, Maehara Y. Effect of hyperbaric oxygen on cold storage of the liver in rats. Liver Int 2006; 26:248-53. [PMID: 16448464 DOI: 10.1111/j.1478-3231.2005.01218.x] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/13/2023]
Abstract
BACKGROUND The depletion of biochemical energy stores during prolonged cold storage is one of the most critical events of cold ischemia-reperfusion (CI/R) injury. The aim of this study was to evaluate the effect of hyperbaric oxygen (HBO) treatment on CI/R injury. METHODS Livers were harvested from male Wistar rats and stored for 24 h at 4 degrees C in University of Wisconsin solution (Group 1). Others were additionally treated with HBO during the preservation period (Group 2). At the end of the 24 h cold preservation, the concentrations of hepatic enzymes and lipid peroxidation (LPO) in the effluent and the hepatic adenosine triphosphate (ATP) levels were measured. After preservation, the livers were reperfused for 90 min with an oxygenated Krebs-Henseleit bicarbonate buffer. Perfusate samples were obtained serially, and portal flow rates were also recorded. RESULTS In group 2, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and LPO into the effluent at the end of preservation were decreased and the depletion of ATP was prevented (P<0.05). After reperfusion, the portal flow was significantly improved in group 2 (P<0.05). The time-dependent increase of alanine aminotransferase levels (ALT) observed in group 1 was suppressed significantly in group 2, and total bile production during 90 min of reperfusion was significantly greater in group 2 (P<0.05). The structure of the livers in group 2 was significantly well maintained, and the liver weight change ratio was significantly greater in group 1 (P<0.05). CONCLUSIONS HBO treatment during cold storage seems to prevent hepatic ischemic injury and have protective effects against CI/R injury by attenuating the depletion of energy stores.
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Affiliation(s)
- Hideki Ijichi
- The Department of Surgery, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
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't Hart NA, van der Plaats A, Leuvenink HGD, Wiersema-Buist J, Olinga P, van Luyn MJA, Verkerke GJ, Rakhorst G, Ploeg RJ. Initial blood washout during organ procurement determines liver injury and function after preservation and reperfusion. Am J Transplant 2004; 4:1836-44. [PMID: 15476484 DOI: 10.1111/j.1600-6143.2004.00580.x] [Citation(s) in RCA: 36] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/25/2023]
Abstract
Organ procurement is the first step toward effective liver preservation and comprises a thorough washout of blood components from the microvasculature. To study the efficacy of optimal blood washout of the liver, three groups were compared including low-pressure perfusion with UW-CSS (12 mmHg, group A), which is the routine method in clinical practice, high-pressure perfusion with UW-CSS (100 mmHg, group B) and low-pressure perfusion with modified UW solution (12 mmHg, group C). After procurement all livers were preserved in original UW-CSS for 0, 24 or 48 h, followed by reperfusion in oxygenated Williams Medium E for 24 h at 37 degrees C. Histology results of livers procured in group A, showed good hepatocyte viability but also remaining erythrocytes. However, injury parameters were high and ATP concentrations were low. No functional differences were found. Group B, high pressure, and group C, modified UW-CSS, both showed better results. High-pressure washout is preferable since the warm ischemia time during procurement is short. We propose to use high-pressure UW-CSS perfusion for the initial blood washout of the donor liver instead of the usually used low-pressure washout.
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Affiliation(s)
- Nils A 't Hart
- University Hospital Groningen, Surgery Research Laboratory, Hanzeplein 1, 9713 GZ Groningen, The Netherlands.
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