BACKGROUND： Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is...BACKGROUND： Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is needed. HTK is a widely used preservation solution for static cold storage in liver transplantation. The present study was to investigate the beneficial effect of warm HTK donor pretreatment on liver preservation.展开更多
BACKGROUND Traditional limitations of cold static storage(CSS)on ice at 4℃during lung transplantation have necessitated limiting cold ischemic time(CIT)to 4-6 hours.Ex vivo lung perfusion(EVLP)can extend this preserv...BACKGROUND Traditional limitations of cold static storage(CSS)on ice at 4℃during lung transplantation have necessitated limiting cold ischemic time(CIT)to 4-6 hours.Ex vivo lung perfusion(EVLP)can extend this preservation time through the suspension of CIT and normothermic perfusion.As we continue to further expand the donor pool in all aspects of lung transplantation,teams are frequently traveling further distances to procure organs.AIM To determine the effect of CSS or EVLP on donors with extended travel distance[>750 nautical miles(NM)]to recipient.METHODS Lung transplants,whose donor traveled greater than 750 NM,were identified from the United Network for Organ Sharing Database.Recipients were stratified into either:CSS or EVLP,based on preservation method.Groups were assessed with comparative statistics and survival was assessed by Kaplan-Meier methods.A 3:1 propensity match was then created,and same analysis was repeated.RESULTS Prior to matching,those in the EVLP group had significantly increased postoperative morbidity to include dialysis,ventilator use,acute rejection,and treated rejection in the first year(P<0.05 for all).However,there were no significant differences in midterm survival(P=0.18).Following matching,those in the EVLP group again had significantly increased post-operative morbidity to include dialysis,extracorporeal membrane oxygenation use,ventilator use,and treated rejection in the first year(P<0.05 for all).As before,there were no significant differences in midterm survival following matching(P=0.08).CONCLUSION While there was no significant difference in survival,EVLP patients had increased peri-operative morbidity.With the advent of changes in CSS with 10℃storage further analysis is necessary to evaluate the best methods for utilizing organs from increased distances.展开更多
Recent advances in organ transplantation,regenerative medicine,and drug discovery have emphasized the critical importance of effective preservation techniques for organs.Despite these advances,current preservation tec...Recent advances in organ transplantation,regenerative medicine,and drug discovery have emphasized the critical importance of effective preservation techniques for organs.Despite these advances,current preservation techniques have significant limitations in maintaining the viability and functional efficacy of organs over the long term.As a result,there is a pressing need to develop reliable and efficient preservation strategies for organs.Currently,the clinical standard for organ preservation involves the use of static cold storage and organ machine perfusion,but these methods can only preserve organs for a couple of days or even a few hours.Notably,the development of cryobiology has yielded promising alternatives.In this review,we aim to provide a comprehensive overview of the progression of organ preservation methods,while emphasizing the limitations of traditional approaches.Moreover,we evaluate advanced preservation techniques for organs,including kidneys,livers,hearts,lungs,and intestines.Furthermore,we share a progress perspective on the future of organ preservation,with the ultimate goal of achieving viable long-term preservation to address the pressing issue of organ shortage.展开更多
The growing demand for donor organs requires measures to expand donor pool.Those include extended criteria donors, such as elderly people, steatotic livers,donation after cardiac death, etc. Static cold storage to red...The growing demand for donor organs requires measures to expand donor pool.Those include extended criteria donors, such as elderly people, steatotic livers,donation after cardiac death, etc. Static cold storage to reduce metabolic requirements developed by Collins in late 1960 s is the mainstay and the golden standard for donated organ protection. Hypothermic machine perfusion provides dynamic organ preservation at 4°C with protracted infusion of metabolic substrates to the graft during the ex vivo period. It has been used instead of static cold storage or after it as short perfusion in transplant center. Normothermic machine perfusion(NMP) delivers oxygen, and nutrition at physiological temperature mimicking regular environment in order to support cellular function. This would minimize effects of ischemia/reperfusion injury.Potentially, NMP may help to estimate graft functionality before implantation into a recipient. Clinical studies demonstrated at least its non-inferiority or better outcomes vs static cold storage. Regular grafts donated after brain death could be safely preserved with convenient static cold storage. Except for prolonged ischemia time where hypothermic machine perfusion started in transplant center could be estimated to provide possible positive reconditioning effect. Use of hypothermic machine perfusion in regular donation instead of static cold storage or in extended criteria donors requires further investigation. Multicenter randomized clinical trial supposed to be completed in December 2021. Extended criteria donors need additional measures for graft storage and assessment until its implantation. NMP is actively evaluating promising method for this purpose.Future studies are necessary for precise estimation and confirmation to issue clinical practice recommendations.展开更多
Both cold stress and ischemia-reperfusion injury significantly contribute to poor prognosis after liver transplantation(LT).However,limited animal models incorporating both stimuli hinder the advancement of transplant...Both cold stress and ischemia-reperfusion injury significantly contribute to poor prognosis after liver transplantation(LT).However,limited animal models incorporating both stimuli hinder the advancement of transplant-related research.Here,a simplified and reproducible isolated perfused liver model is established to simulate the stresses experienced by livers maximally during transplantation.We provide a detailed protocol for a straightforward technique that requires 20e30 min for harvesting,24e48 h for static cold storage(SCS),and 2 h for normothermic machine perfusion(NMP)to induce LT-like stresses in the liver.Hepatic injury from SCS and NMP(LT-like stresses)is evaluated using three types of parameters.The pH values and hepatic enzyme levels of cold preservation solutions and perfusate serve as dynamic indicators of hepatic injury.Bile production and portal venous resistance directly reflect liver function,whereas pathological analysis visually illustrates the location and extent of injury.This animal model eliminates the influence of hemodynamic and immune factors,yielding highly reproducible results,and is strongly recommended as a standardized animal model for inducing LT-like stresses.展开更多
Static cold storage(SCS)remains the gold standard for preserving donor hearts for heart transplantation.The currently used University of Wisconsin(UW)solution has limitations in inducing rapid cardiac arrest and prese...Static cold storage(SCS)remains the gold standard for preserving donor hearts for heart transplantation.The currently used University of Wisconsin(UW)solution has limitations in inducing rapid cardiac arrest and preservation of the heart,especially when a long cold ischemic time is required.Here,we propose a novel donor heart procurement and preservation method—crystalloid cardioplegia(CC)+UW involving an initial cardioplegic flush with crystalloid cardioplegia followed by preservation in UW solution.Our results demonstrated that compared with using UW solution alone,the CC+UW method could effectively reduce metabolite consumption and suppress the activation of the NFκB signaling pathway in a pig model.NR4A3 was found to be upregulated in CC+UW,which could suppress the expression of DUSP2and thereby decrease p65 phosphorylation.In addition,the CC+UW method increased intraventricular pressure and improved electrical conduction and rhythm in Langendorff-perfused rat hearts.Mediumterm survival data for heart transplant patients also confirmed the benefits of this method.In conclusion,our study identifies the CC+UW method as an effective strategy for donor heart procurement and preservation,showing that it can reduce energy consumption and decrease inflammation;therefore,this method is a promising technique for expanding the criteria for heart transplantation and alleviating the shortage of donor hearts.展开更多
Background and Aims:End-stage liver disease is associated with disruptions in gut microbiota composition and function,which may facilitate gut-to-liver bacterial translocation,impacting liver graft integrity and clini...Background and Aims:End-stage liver disease is associated with disruptions in gut microbiota composition and function,which may facilitate gut-to-liver bacterial translocation,impacting liver graft integrity and clinical outcomes following liver transplantation.This study aimed to assess the impact of two liver graft preservation methods on fecal microbiota and changes in fecal and breath organic acids following liver transplantation.Methods:This single-center,non-randomized prospective pilot study enrolled liver transplant patients whose grafts were preserved using either static cold storage or ex situ normothermic machine perfusion(NMP).Fresh stool and breath samples were collected immediately before surgery and at postoperative months 3,6,and 12.Stool microbiota was profiled via 16S rRNA gene sequencing,stool short-chain fatty acids were measured using gas chromatography/-mass spectrometry,and breath volatile organic compounds(VOCs)were analyzed with selected-ion flow-tube mass spectrometry.Results:Both cohorts experienced a loss of microbiota diversity and dominance by single taxa.The NMP cohort demonstrated enrichment of several beneficial gut taxa,while the static cold storage cohort showed depletion of such taxa.Various gut bacteria were found to correlate with stool short-chain fatty acids(e.g.,lactic acid,butyric acid)and several VOCs.Conclusions:Fecal microbiota alterations associated with end-stage liver disease do not fully normalize to a healthy control profile following liver transplantation.However,notable differences in microbiota composition and function were observed between liver graft preservation methods.Future research with larger randomized cohorts is needed to explore whether the NMP-associated shift in gut microbiota impacts clinical outcomes and if breath VOCs could serve as biomarkers of the clinical trajectory in liver transplant patients.展开更多
Purpose:Ischemia and hypoxia are the main factors limiting limb replantation and transplantation.Static cold storage(SCS),a common preservation method for tissues and organs,can only prolong limb ischemia time to 4-6 ...Purpose:Ischemia and hypoxia are the main factors limiting limb replantation and transplantation.Static cold storage(SCS),a common preservation method for tissues and organs,can only prolong limb ischemia time to 4-6 h.The normothermic machine perfusion(NMP)is a promising method for the preservation of tissues and organs,which can extend the preservation time in vitro by providing continuous oxygen and nutrients.This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods.Methods:The 6 forelimbs from beagle dogs were divided into 2 groups.In the SCS group(n=3),the limbs were preserved in a sterile refrigerator at 4°C for 24 h,and in the NMP group(n=3),the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological temperature for 24 h,and the solution was changed every 6 h.The effects of limb storage were evaluated by weight gain,perfusate biochemical analysis,enzyme-linked immunosorbent assay,and histological analysis.All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance.The p value of less than 0.05 was considered to indicate statistical significance.Results:In the NMP group,the weight gained percentage was 11.72%±4.06%;the hypoxia-inducible factor-1αcontents showed no significant changes;the shape of muscle fibers was normal;the gap between muscle fibers slightly increased,showing the intercellular distance of(30.19±2.83)μm;and the vascularα-smooth muscle actin(α-SMA)contents were lower than those in the normal blood vessels.The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion,decreased after each perfusate change,and remained stable at the end of perfusion showing a peak level of 4097.6 U/L.The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L.In the SCS group,the percentage of weight gain was 0.18%±0.10%,and the contents of hypoxia-inducible factor-1αincreased gradually and reached the maximum level of(164.85±20.75)pg/mL at the end of the experiment.The muscle fibers lost their normal shape and the gap between muscle fibers increased,showing an intercellular distance of(41.66±5.38)μm.The contents of vascularα-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions:NMP caused lesser muscle damage and contained more vascularα-SMA as compared to SCS.This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.展开更多
基金supported by a grant of"Else-Kroner Fresenius Stiftung"(p49/07//A68/07)
文摘BACKGROUND： Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is needed. HTK is a widely used preservation solution for static cold storage in liver transplantation. The present study was to investigate the beneficial effect of warm HTK donor pretreatment on liver preservation.
基金Supported by The Jewel and Frank Benson Family Endowmentand The Jewel and Frank Benson Research Professorship.
文摘BACKGROUND Traditional limitations of cold static storage(CSS)on ice at 4℃during lung transplantation have necessitated limiting cold ischemic time(CIT)to 4-6 hours.Ex vivo lung perfusion(EVLP)can extend this preservation time through the suspension of CIT and normothermic perfusion.As we continue to further expand the donor pool in all aspects of lung transplantation,teams are frequently traveling further distances to procure organs.AIM To determine the effect of CSS or EVLP on donors with extended travel distance[>750 nautical miles(NM)]to recipient.METHODS Lung transplants,whose donor traveled greater than 750 NM,were identified from the United Network for Organ Sharing Database.Recipients were stratified into either:CSS or EVLP,based on preservation method.Groups were assessed with comparative statistics and survival was assessed by Kaplan-Meier methods.A 3:1 propensity match was then created,and same analysis was repeated.RESULTS Prior to matching,those in the EVLP group had significantly increased postoperative morbidity to include dialysis,ventilator use,acute rejection,and treated rejection in the first year(P<0.05 for all).However,there were no significant differences in midterm survival(P=0.18).Following matching,those in the EVLP group again had significantly increased post-operative morbidity to include dialysis,extracorporeal membrane oxygenation use,ventilator use,and treated rejection in the first year(P<0.05 for all).As before,there were no significant differences in midterm survival following matching(P=0.08).CONCLUSION While there was no significant difference in survival,EVLP patients had increased peri-operative morbidity.With the advent of changes in CSS with 10℃storage further analysis is necessary to evaluate the best methods for utilizing organs from increased distances.
基金financially supported by the National Key Research and Development Program of China(2022YFC2100800)the National Natural Science Foundation of China(22478296,22078238,52373117,and U23B20121)+1 种基金the Haihe Laboratory of Sustainable Chemical Transformations(24HHWCSS00005)the Open Funding Project of the National Key Laboratory of Biochemical Engineering。
文摘Recent advances in organ transplantation,regenerative medicine,and drug discovery have emphasized the critical importance of effective preservation techniques for organs.Despite these advances,current preservation techniques have significant limitations in maintaining the viability and functional efficacy of organs over the long term.As a result,there is a pressing need to develop reliable and efficient preservation strategies for organs.Currently,the clinical standard for organ preservation involves the use of static cold storage and organ machine perfusion,but these methods can only preserve organs for a couple of days or even a few hours.Notably,the development of cryobiology has yielded promising alternatives.In this review,we aim to provide a comprehensive overview of the progression of organ preservation methods,while emphasizing the limitations of traditional approaches.Moreover,we evaluate advanced preservation techniques for organs,including kidneys,livers,hearts,lungs,and intestines.Furthermore,we share a progress perspective on the future of organ preservation,with the ultimate goal of achieving viable long-term preservation to address the pressing issue of organ shortage.
文摘The growing demand for donor organs requires measures to expand donor pool.Those include extended criteria donors, such as elderly people, steatotic livers,donation after cardiac death, etc. Static cold storage to reduce metabolic requirements developed by Collins in late 1960 s is the mainstay and the golden standard for donated organ protection. Hypothermic machine perfusion provides dynamic organ preservation at 4°C with protracted infusion of metabolic substrates to the graft during the ex vivo period. It has been used instead of static cold storage or after it as short perfusion in transplant center. Normothermic machine perfusion(NMP) delivers oxygen, and nutrition at physiological temperature mimicking regular environment in order to support cellular function. This would minimize effects of ischemia/reperfusion injury.Potentially, NMP may help to estimate graft functionality before implantation into a recipient. Clinical studies demonstrated at least its non-inferiority or better outcomes vs static cold storage. Regular grafts donated after brain death could be safely preserved with convenient static cold storage. Except for prolonged ischemia time where hypothermic machine perfusion started in transplant center could be estimated to provide possible positive reconditioning effect. Use of hypothermic machine perfusion in regular donation instead of static cold storage or in extended criteria donors requires further investigation. Multicenter randomized clinical trial supposed to be completed in December 2021. Extended criteria donors need additional measures for graft storage and assessment until its implantation. NMP is actively evaluating promising method for this purpose.Future studies are necessary for precise estimation and confirmation to issue clinical practice recommendations.
基金supported by the National Natural Science Foundation of China(No.82400773,82400745,82370646)Young Talent Support Program of Henan Association for Science and Technology(No.2025HYTP083).
文摘Both cold stress and ischemia-reperfusion injury significantly contribute to poor prognosis after liver transplantation(LT).However,limited animal models incorporating both stimuli hinder the advancement of transplant-related research.Here,a simplified and reproducible isolated perfused liver model is established to simulate the stresses experienced by livers maximally during transplantation.We provide a detailed protocol for a straightforward technique that requires 20e30 min for harvesting,24e48 h for static cold storage(SCS),and 2 h for normothermic machine perfusion(NMP)to induce LT-like stresses in the liver.Hepatic injury from SCS and NMP(LT-like stresses)is evaluated using three types of parameters.The pH values and hepatic enzyme levels of cold preservation solutions and perfusate serve as dynamic indicators of hepatic injury.Bile production and portal venous resistance directly reflect liver function,whereas pathological analysis visually illustrates the location and extent of injury.This animal model eliminates the influence of hemodynamic and immune factors,yielding highly reproducible results,and is strongly recommended as a standardized animal model for inducing LT-like stresses.
基金supported by the National Key Research and Development Program(2021YFA1101900)Key Research Development Program of Hubei Province(2020BCA061)+1 种基金the National Natural Science Foundation of China(82200410,82470423,82204659,and 82300413)Hubei Provincial Natural Science Foundation Projects(JCZRYB202500142)。
文摘Static cold storage(SCS)remains the gold standard for preserving donor hearts for heart transplantation.The currently used University of Wisconsin(UW)solution has limitations in inducing rapid cardiac arrest and preservation of the heart,especially when a long cold ischemic time is required.Here,we propose a novel donor heart procurement and preservation method—crystalloid cardioplegia(CC)+UW involving an initial cardioplegic flush with crystalloid cardioplegia followed by preservation in UW solution.Our results demonstrated that compared with using UW solution alone,the CC+UW method could effectively reduce metabolite consumption and suppress the activation of the NFκB signaling pathway in a pig model.NR4A3 was found to be upregulated in CC+UW,which could suppress the expression of DUSP2and thereby decrease p65 phosphorylation.In addition,the CC+UW method increased intraventricular pressure and improved electrical conduction and rhythm in Langendorff-perfused rat hearts.Mediumterm survival data for heart transplant patients also confirmed the benefits of this method.In conclusion,our study identifies the CC+UW method as an effective strategy for donor heart procurement and preservation,showing that it can reduce energy consumption and decrease inflammation;therefore,this method is a promising technique for expanding the criteria for heart transplantation and alleviating the shortage of donor hearts.
基金supported in part by a grant from the Lerner Research Institute,Cleveland Clinic to GAMC and CQ.
文摘Background and Aims:End-stage liver disease is associated with disruptions in gut microbiota composition and function,which may facilitate gut-to-liver bacterial translocation,impacting liver graft integrity and clinical outcomes following liver transplantation.This study aimed to assess the impact of two liver graft preservation methods on fecal microbiota and changes in fecal and breath organic acids following liver transplantation.Methods:This single-center,non-randomized prospective pilot study enrolled liver transplant patients whose grafts were preserved using either static cold storage or ex situ normothermic machine perfusion(NMP).Fresh stool and breath samples were collected immediately before surgery and at postoperative months 3,6,and 12.Stool microbiota was profiled via 16S rRNA gene sequencing,stool short-chain fatty acids were measured using gas chromatography/-mass spectrometry,and breath volatile organic compounds(VOCs)were analyzed with selected-ion flow-tube mass spectrometry.Results:Both cohorts experienced a loss of microbiota diversity and dominance by single taxa.The NMP cohort demonstrated enrichment of several beneficial gut taxa,while the static cold storage cohort showed depletion of such taxa.Various gut bacteria were found to correlate with stool short-chain fatty acids(e.g.,lactic acid,butyric acid)and several VOCs.Conclusions:Fecal microbiota alterations associated with end-stage liver disease do not fully normalize to a healthy control profile following liver transplantation.However,notable differences in microbiota composition and function were observed between liver graft preservation methods.Future research with larger randomized cohorts is needed to explore whether the NMP-associated shift in gut microbiota impacts clinical outcomes and if breath VOCs could serve as biomarkers of the clinical trajectory in liver transplant patients.
基金supported by the National Key R&D Program of China(2019YFA0110704)the Military Logistic(s)Scientific Research Program of China(BHJ20J002).
文摘Purpose:Ischemia and hypoxia are the main factors limiting limb replantation and transplantation.Static cold storage(SCS),a common preservation method for tissues and organs,can only prolong limb ischemia time to 4-6 h.The normothermic machine perfusion(NMP)is a promising method for the preservation of tissues and organs,which can extend the preservation time in vitro by providing continuous oxygen and nutrients.This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods.Methods:The 6 forelimbs from beagle dogs were divided into 2 groups.In the SCS group(n=3),the limbs were preserved in a sterile refrigerator at 4°C for 24 h,and in the NMP group(n=3),the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological temperature for 24 h,and the solution was changed every 6 h.The effects of limb storage were evaluated by weight gain,perfusate biochemical analysis,enzyme-linked immunosorbent assay,and histological analysis.All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance.The p value of less than 0.05 was considered to indicate statistical significance.Results:In the NMP group,the weight gained percentage was 11.72%±4.06%;the hypoxia-inducible factor-1αcontents showed no significant changes;the shape of muscle fibers was normal;the gap between muscle fibers slightly increased,showing the intercellular distance of(30.19±2.83)μm;and the vascularα-smooth muscle actin(α-SMA)contents were lower than those in the normal blood vessels.The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion,decreased after each perfusate change,and remained stable at the end of perfusion showing a peak level of 4097.6 U/L.The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L.In the SCS group,the percentage of weight gain was 0.18%±0.10%,and the contents of hypoxia-inducible factor-1αincreased gradually and reached the maximum level of(164.85±20.75)pg/mL at the end of the experiment.The muscle fibers lost their normal shape and the gap between muscle fibers increased,showing an intercellular distance of(41.66±5.38)μm.The contents of vascularα-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions:NMP caused lesser muscle damage and contained more vascularα-SMA as compared to SCS.This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.
