INTRODUCTIONAlthough liver transplantation for irreversible liverdiseases is increasingly prevalent worldwide,patientdie while waiting for donors because of organshortages.One important problem commonlyencountered is ...INTRODUCTIONAlthough liver transplantation for irreversible liverdiseases is increasingly prevalent worldwide,patientdie while waiting for donors because of organshortages.One important problem commonlyencountered is that fatty livers often affect theoutcome of liver transplantation.It is reported thatthe incidence of abnormal fatty livers in autopsiesafter accidental death ranged from 15% to 24%.Since fatty livers may result in a primarynonfunction (PNF)liver graft,which contributes展开更多
AIM To develop a simplified bioartificial liver(BAL) device prototype, suitable to use freshly and preserved liver Microorgans(LMOs) as biological component. METHODS The system consists of 140 capillary fibers through...AIM To develop a simplified bioartificial liver(BAL) device prototype, suitable to use freshly and preserved liver Microorgans(LMOs) as biological component. METHODS The system consists of 140 capillary fibers through which goat blood is pumped. The evolution of hema-tocrit, plasma and extra-fiber fluid osmolality was evaluated without any biological component, to characterize the prototype. LMOs were cut and cold stored 48 h in BG35 and Via Span~? solutions. Fresh LMOs were used as controls. After preservation, LMOs were loaded into the BAL and an ammonia overload was added. To assess LMOs viability and functionality, samples were taken to determine lactate dehydrogenase(LDH) release and ammonia detoxification capacity. RESULTS The concentrations of ammonia and glucose, and the fluids osmolalities were matched after the first hour of perfusion, showing a proper exchange between blood and the biological compartment in the minibioreactor. After 120 min of perfusion, LMOs cold preserved in BG35 and Via Span~? were able to detoxify 52.9% ± 6.5% and 53.6% ± 6.0%, respectively, of the initial ammonia overload. No significant differences were found with Controls(49.3% ± 8.8%, P < 0.05). LDH release was 6.0% ± 2.3% for control LMOs, and 6.2% ± 1.7% and 14.3% ± 1.1% for BG35 and Via Span~? cold preserved LMOs, respectively(n = 6, P < 0.05). CONCLUSION This prototype relied on a simple design and excellent performance. It’s a practical tool to evaluate the detoxification ability of LMOs subjected to different preservation protocols.展开更多
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
Mitochondria were isolated from normal and cold adapted rat livers.The respiratory func-tion of mitochondria in rat livers,including ADP:O ratio(P/O)and the respiratory control ratio(RCR),was determined by oxygen elec...Mitochondria were isolated from normal and cold adapted rat livers.The respiratory func-tion of mitochondria in rat livers,including ADP:O ratio(P/O)and the respiratory control ratio(RCR),was determined by oxygen electrode method,The protein synthesis in mitochondria wasstudied by observing the incorporation of[~3H]-Leucine into mitochondria.Polyacrylamide gelelectrophoresis was carried out to detect the changes of the inner membrane proteins.It was shownthat the P/O and RCR decreased in cold adapted rats in the 2nd and 4th weeks and returned tothe control level in the 6th week,the protein synthesis of mitochondria decreased significantly incold adapted rats in 1,2 and 4 weeks;the electrophoretic pattern of the inner membrane proteinsin mitochondria from cold adapted rat livers revealed some new bands.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
AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin col...AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin coldstorage solution(UW) or histidine-tryptophan-ketoglutarate solution(HTK) as the perfusate. Each group was divided into two subgroups: static cold storage(SCS) and HMP(n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate(0-4?℃) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase(AST), alanine transaminase(ALT) and lactate dehydrogenase(LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight(D/W) ratio, and malondialdehyde(MDA) and adenosine-triphosphate(ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period(SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level(saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline(vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently(vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best(vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.展开更多
The urgent need to expand the donor pool in order to attend to the growing demand for liver transplantation has obliged physicians to consider the use of suboptimal liver grafts and also to redefine the preservation s...The urgent need to expand the donor pool in order to attend to the growing demand for liver transplantation has obliged physicians to consider the use of suboptimal liver grafts and also to redefine the preservation strategies. This review examines the different methods of liver graft preservation, focusing on the latest advances in both static cold storage and machine perfusion(MP). The new strategies for static cold storage are mainly designed to increase the fatty liver graft preservation via the supplementation of commercial organ preservation solutions with additives. In this paper we stress the importance of carrying out effective graft washout after static cold preservation, and present a detailed discussion of the future perspectives for dynamic graft preservation using MP at different temperatures(hypothermia at 4 ℃, normothermia a t 3 7 ℃ and subnormothermia at 20 ℃- 2 5 ℃). Finally, we highlight some emerging applications of regenerative medicine in liver graft preservation. In conclusion, this review discusses the "state of the art" and future perspectives in static and dynamic liver graft preservation in order to improve graft viability.展开更多
Background/aim:Currently, the liver is cold-preserved at 0~4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was be...Background/aim:Currently, the liver is cold-preserved at 0~4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion. Methods:In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A:45 min and Group B:24 h) or by slow cooling HTK solution (from 13 °C to 3 °C) during the initial 45 min of preservation (Group C:24 h). In the second set of experiments, additional groups of livers were evaluated:Group BB—preservation according to Group B and Group CC—preservation according to Group C. Further, some livers were preserved at 13 °C for 24 h. Livers were then reperfused using a blood-free perfusion model. Results:Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2~3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 °C for 24 h exhibited severe ischemic injury. Conclusion:These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.展开更多
AIM: To compare, in a pig liver transplantation model, the protective effect of UW with that of IGL-1, a high-sodium preservation solution containing polyethylene glycol (PEG) as an oncotic supply. METHODS: All livers...AIM: To compare, in a pig liver transplantation model, the protective effect of UW with that of IGL-1, a high-sodium preservation solution containing polyethylene glycol (PEG) as an oncotic supply. METHODS: All livers were harvested and grafted orthotopically according to standard techniques. The livers were washed out and preserved for 7 h in IGL-1 (n = 6) or in UW solution (n = 7) at 4℃. In a sham group (n = 4), the livers underwent a 60-min warm ischemia at 37℃. The hepatocellular injury was assessed in organ preservation solution washed out from the graft at the end of ischemic storage (before revascularization), and in serum 2 h after reperfusion and daily for up to 6 d. RESULTS: Livers preserved in IGL-1 solution released markedly less AST than that preserved in the UW solution before and after revascularization (P < 0.05). Besides, the activity of creatine kinase-BB, a marker of sinusoidal lining cells injury, was higher in the UW group than in the IGL-1 group (P < 0.05). Histological results showed less necrotic regions in livers preserved in IGL-1 solution; however, no difference was observed for in? ammation. CONCLUSION: IGL-1 liquid effectively protects parenchymal and non-parenchymal cells against prese-rvation-reperfusion injuries.展开更多
BACKGROUND:Ischemia-related biliary tract complications remain high after orthotopic liver transplantation.Severe ischemic biliary complications often involve the hepatic duct bifurcation and left hepatic duct,resulti...BACKGROUND:Ischemia-related biliary tract complications remain high after orthotopic liver transplantation.Severe ischemic biliary complications often involve the hepatic duct bifurcation and left hepatic duct,resulting finally in obstructive jaundice.Prevention and management of such complications remain a challenge for transplant surgeons.METHODS:All 160 patients were followed up for at least 180 days after transplantation.One-way analysis of variance (ANOVA) and comparative univariate analysis were made using 3 groups (no complications;mild complications;severe complications),to analyze risk factors associated with biliary complications.Multiple logistic regression and linear regression analysis were used to analyze independent risk factors for severe ischemic biliary complications,after excluding other confounding factors.RESULTS:By ANOVA and comparative univariate analysis,the risk factors associated with biliary complications were preoperative bilirubin level (P=0.007) and T-tube stenting of the anastomosis (P=0.016).Multiple logistic regression analysis showed that the use of T-tube and preoperative serum bilirubin were not independent risk factors for severe ischemic biliary complications after orthotopic liver transplantation.Chi-square analysis indicated that in the incidence of severe ischemic biliary lesions,bile duct second warm ischemic time longer than 60 minutes was a significant risk factor.Linear regression demonstrated a negative correlation between cold preservation time and warm ischemia time.CONCLUSIONS:Preoperative serum bilirubin level and the use of T-tube stenting of the anastomosis were independent risk factors for biliary complications after liver transplantation,but not for severe ischemic biliary complications.The second warm ischemia time of bile duct longer than 60minutes and prolonged bile duct second warm ischemia time combined with cold preservation time were significant risk factors for severe ischemic biliary complications after liver transplantation with grafts from non-heart-beating donors.展开更多
BACKGROUND: To evaluate feasibility and safety of venovenous bypass prior to mobilization of the liver during orthotopic liver transplantation (OLT). METHODS: Fifty-four patients were classified into two groups. Group...BACKGROUND: To evaluate feasibility and safety of venovenous bypass prior to mobilization of the liver during orthotopic liver transplantation (OLT). METHODS: Fifty-four patients were classified into two groups. Group A consisted of 23 patients receiving OLT with classical venovenous bypass. Group B consisted of 31 patients who received a modified-procedure: venovenous bypass ahead of the mobilization of the liver during ULT. The blood loss, duration of venovenous bypass, cold ischemia time, anhepatic phase, and transfusion during operation in the two groups were compared. Complications after the operation were also compared between the two groups. RESULTS: The duration of venovenous bypass and cold ischemia time in group A were longer than those in group B [(99.78±21.36 min) vs (96.32±22.25 min) and (484.78±134.01 min) vs (443.15± 85.27 min)]. The anhepatic phase lasted for about 100 min averagely in the two groups. The volumes of blood loss and transfusion during the operation were larger in group A than in group B [(5096±4243 ml) vs (1726±1125 ml) and (3676±2938.74 ml) vs (1217.69±829.72 ml)]. Postoperative complications occurred in 26 patients of group A and in 19 patients of group B. CONCLUSION: This modified-procedure or venovenous bypass ahead of mobilization of the liver in OLT can reduce the blood loss during OLT and the incidence of postoperative complications without prolongation of the anhepatic phase and duration of venovenous bypass.展开更多
AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex...AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.展开更多
Liver transplantation(LT)is the best treatment for endstage hepatic failure,with an excellent survival rates over the last decade.Biliary complications after LT pose a major challenge especially with the increasing nu...Liver transplantation(LT)is the best treatment for endstage hepatic failure,with an excellent survival rates over the last decade.Biliary complications after LT pose a major challenge especially with the increasing number of procured organs after circulatory death.Ischaemic cholangiopathy(IC)is a set of disorders characterized by multiple diffuse strictures affecting the graft biliary system in the absence of hepatic artery thrombosis or stenosis.It commonly presents with cholestasis and cholangitis resulting in higher readmission rates,longer length of stay,repeated therapeutic interventions,and eventually re-transplantation with consequent effects on the patient’s quality of life and increased health care costs.The pathogenesis of IC is unclear and exhibits a higher prevalence with prolonged ischaemia time,donation after circulatory death(DCD),rejection,and cytomegalovirus infection.The majority of IC occurs within 12 mo after LT.Prolonged warm ischaemic times predispose to a profound injury with a subsequently higher prevalence of IC.Biliary complications and IC rates are between 16%and 29%in DCD grafts com-pared to between 3%and 17%in donation after brain death(DBD)grafts.The majority of ischaemic biliary lesions occur within 30 d in DCD compared to 90 d in DBD grafts following transplantation.However,there are many other risk factors for IC that should be considered.The benefits of DCD in expanding the donor pool are hindered by the higher incidence of IC with increased rates of re-transplantation.Careful donor selection and procurement might help to optimize the utilization of DCD grafts.展开更多
基金the National Natural Science Foundation of China,No.39470682
文摘INTRODUCTIONAlthough liver transplantation for irreversible liverdiseases is increasingly prevalent worldwide,patientdie while waiting for donors because of organshortages.One important problem commonlyencountered is that fatty livers often affect theoutcome of liver transplantation.It is reported thatthe incidence of abnormal fatty livers in autopsiesafter accidental death ranged from 15% to 24%.Since fatty livers may result in a primarynonfunction (PNF)liver graft,which contributes
文摘AIM To develop a simplified bioartificial liver(BAL) device prototype, suitable to use freshly and preserved liver Microorgans(LMOs) as biological component. METHODS The system consists of 140 capillary fibers through which goat blood is pumped. The evolution of hema-tocrit, plasma and extra-fiber fluid osmolality was evaluated without any biological component, to characterize the prototype. LMOs were cut and cold stored 48 h in BG35 and Via Span~? solutions. Fresh LMOs were used as controls. After preservation, LMOs were loaded into the BAL and an ammonia overload was added. To assess LMOs viability and functionality, samples were taken to determine lactate dehydrogenase(LDH) release and ammonia detoxification capacity. RESULTS The concentrations of ammonia and glucose, and the fluids osmolalities were matched after the first hour of perfusion, showing a proper exchange between blood and the biological compartment in the minibioreactor. After 120 min of perfusion, LMOs cold preserved in BG35 and Via Span~? were able to detoxify 52.9% ± 6.5% and 53.6% ± 6.0%, respectively, of the initial ammonia overload. No significant differences were found with Controls(49.3% ± 8.8%, P < 0.05). LDH release was 6.0% ± 2.3% for control LMOs, and 6.2% ± 1.7% and 14.3% ± 1.1% for BG35 and Via Span~? cold preserved LMOs, respectively(n = 6, P < 0.05). CONCLUSION This prototype relied on a simple design and excellent performance. It’s a practical tool to evaluate the detoxification ability of LMOs subjected to different preservation protocols.
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.
文摘Mitochondria were isolated from normal and cold adapted rat livers.The respiratory func-tion of mitochondria in rat livers,including ADP:O ratio(P/O)and the respiratory control ratio(RCR),was determined by oxygen electrode method,The protein synthesis in mitochondria wasstudied by observing the incorporation of[~3H]-Leucine into mitochondria.Polyacrylamide gelelectrophoresis was carried out to detect the changes of the inner membrane proteins.It was shownthat the P/O and RCR decreased in cold adapted rats in the 2nd and 4th weeks and returned tothe control level in the 6th week,the protein synthesis of mitochondria decreased significantly incold adapted rats in 1,2 and 4 weeks;the electrophoretic pattern of the inner membrane proteinsin mitochondria from cold adapted rat livers revealed some new bands.
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 National Science and Technology Major Project,No.2012ZX10002-017Natural Science Foundation of China for Innovative Research Group,No.81121002+4 种基金National Natural Science Foundation of China,No.81000137 and No.81470891The Qianjiang Talent Program of Zhejiang Province,China,No.2012R10045the Scientific Research Program for the Returned Overseas Chinese Scholars,Ministry of Health,China,No.J20112008National High Technology Research and Development Program of China for Young Scientists(863 Program),No.2015AA020923Ministry of Education,Zhejiang Province,China,No.Y201328095
文摘AIM: To optimize the perfusates used for hypothermicmachine perfusion(HMP).METHODS: Sprague-Dawley rats were assigned randomly to three groups(n = 12 per group) that received either saline, University of Wisconsin coldstorage solution(UW) or histidine-tryptophan-ketoglutarate solution(HTK) as the perfusate. Each group was divided into two subgroups: static cold storage(SCS) and HMP(n = 6 per subgroup). The liver graft was retrieved according to the method described by Kamada. For the SCS group, the graft was directly placed into cold perfusate(0-4?℃) for 6 h after liver isolation while the portal vein of the graft was connected to the perfusion machine for the HMP group. Then the perfusates were collected at different time points for analysis of aspartate aminotransferase(AST), alanine transaminase(ALT) and lactate dehydrogenase(LDH) levels. Liver tissues were obtained for evaluation of histology, dry/wet weight(D/W) ratio, and malondialdehyde(MDA) and adenosine-triphosphate(ATP) levels. The portal vein pressure and velocity were monitored in real time in all HMP subgroups.RESULTS: Comparison of HMP and SCS: Regardless of the perfusate, HMP improved the architecture of donor graft in reducing the congestion around sinusoids and central vein and maintaining sinusoid lining in morphology; HMP improved liver function in terms of ALT, AST and LDH, especially during the 3-6 h period(SCS vs HMP using saline: ALT3, 225.00 ± 105.62 vs 49.50 ± 18.50, P = 0.047; LDH3, 1362.17 ± 563.30 vs 325.75 ± 147.43, P = 0.041; UW: LDH6, 2880.14 ± 948.46 vs 2135.00 ± 174.27, P = 0.049; HTK, AST6, 307.50 ± 52.95 vs 185.20 ± 20.46, P = 0.041); HMP decreased MDA level(saline, 2.79 ± 0.30 vs 1.09 ± 0.09, P = 0.008; UW, 3.01 ± 0.77 vs 1.23 ± 0.68, P = 0.005; HTK, 3.30 ± 0.52 vs 1.56 ± 0.22, P = 0.006). Comparison among HMP subgroups: HTK showed less portal vein resistance than UW and saline(vs saline, 3.41 ± 0.49 vs 5.00 ± 0.38, P < 0.001; vs UW, 3.41 ± 0.49 vs 4.52 ± 0.63, P = 0.007); UW reduced edema most efficiently(vs saline, 0.68 ± 0.02 vs 0.79 ± 0.05, P = 0.013), while HTK maintained ATP levels best(vs saline, 622.60 ± 29.11 vs 327.43 ± 44.66, P < 0.001; vs UW, 622.60 ± 29.11 vs 301.80 ± 37.68, P < 0.001).CONCLUSION: HMP is superior to SCS in maintaining both architecture and function of liver grafts. Further, HTK was found to be the optimal perfusate for HMP.
基金Supported by Grant from Fondo de Investigaciones Sanitarias,No.FIS PI12/00519Eirini Pantazi is the recipient of a fellowship from Agència de Gestiód’Ajuts Universitaris i de Recerca,No.2012FI_B00382,Generalitat de Catalunya,Barcelona,Spain
文摘The urgent need to expand the donor pool in order to attend to the growing demand for liver transplantation has obliged physicians to consider the use of suboptimal liver grafts and also to redefine the preservation strategies. This review examines the different methods of liver graft preservation, focusing on the latest advances in both static cold storage and machine perfusion(MP). The new strategies for static cold storage are mainly designed to increase the fatty liver graft preservation via the supplementation of commercial organ preservation solutions with additives. In this paper we stress the importance of carrying out effective graft washout after static cold preservation, and present a detailed discussion of the future perspectives for dynamic graft preservation using MP at different temperatures(hypothermia at 4 ℃, normothermia a t 3 7 ℃ and subnormothermia at 20 ℃- 2 5 ℃). Finally, we highlight some emerging applications of regenerative medicine in liver graft preservation. In conclusion, this review discusses the "state of the art" and future perspectives in static and dynamic liver graft preservation in order to improve graft viability.
基金Project supported by the Ministry of Health of the Slovak Republicunder the project of Modulation of Heat Transfer in Isolated Liver (No.2005/32-SZU-10)the VEGA Grant (No.1/1158/04),Slo-vakia
文摘Background/aim:Currently, the liver is cold-preserved at 0~4 °C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion. Methods:In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A:45 min and Group B:24 h) or by slow cooling HTK solution (from 13 °C to 3 °C) during the initial 45 min of preservation (Group C:24 h). In the second set of experiments, additional groups of livers were evaluated:Group BB—preservation according to Group B and Group CC—preservation according to Group C. Further, some livers were preserved at 13 °C for 24 h. Livers were then reperfused using a blood-free perfusion model. Results:Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2~3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 °C for 24 h exhibited severe ischemic injury. Conclusion:These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.
文摘AIM: To compare, in a pig liver transplantation model, the protective effect of UW with that of IGL-1, a high-sodium preservation solution containing polyethylene glycol (PEG) as an oncotic supply. METHODS: All livers were harvested and grafted orthotopically according to standard techniques. The livers were washed out and preserved for 7 h in IGL-1 (n = 6) or in UW solution (n = 7) at 4℃. In a sham group (n = 4), the livers underwent a 60-min warm ischemia at 37℃. The hepatocellular injury was assessed in organ preservation solution washed out from the graft at the end of ischemic storage (before revascularization), and in serum 2 h after reperfusion and daily for up to 6 d. RESULTS: Livers preserved in IGL-1 solution released markedly less AST than that preserved in the UW solution before and after revascularization (P < 0.05). Besides, the activity of creatine kinase-BB, a marker of sinusoidal lining cells injury, was higher in the UW group than in the IGL-1 group (P < 0.05). Histological results showed less necrotic regions in livers preserved in IGL-1 solution; however, no difference was observed for in? ammation. CONCLUSION: IGL-1 liquid effectively protects parenchymal and non-parenchymal cells against prese-rvation-reperfusion injuries.
文摘BACKGROUND:Ischemia-related biliary tract complications remain high after orthotopic liver transplantation.Severe ischemic biliary complications often involve the hepatic duct bifurcation and left hepatic duct,resulting finally in obstructive jaundice.Prevention and management of such complications remain a challenge for transplant surgeons.METHODS:All 160 patients were followed up for at least 180 days after transplantation.One-way analysis of variance (ANOVA) and comparative univariate analysis were made using 3 groups (no complications;mild complications;severe complications),to analyze risk factors associated with biliary complications.Multiple logistic regression and linear regression analysis were used to analyze independent risk factors for severe ischemic biliary complications,after excluding other confounding factors.RESULTS:By ANOVA and comparative univariate analysis,the risk factors associated with biliary complications were preoperative bilirubin level (P=0.007) and T-tube stenting of the anastomosis (P=0.016).Multiple logistic regression analysis showed that the use of T-tube and preoperative serum bilirubin were not independent risk factors for severe ischemic biliary complications after orthotopic liver transplantation.Chi-square analysis indicated that in the incidence of severe ischemic biliary lesions,bile duct second warm ischemic time longer than 60 minutes was a significant risk factor.Linear regression demonstrated a negative correlation between cold preservation time and warm ischemia time.CONCLUSIONS:Preoperative serum bilirubin level and the use of T-tube stenting of the anastomosis were independent risk factors for biliary complications after liver transplantation,but not for severe ischemic biliary complications.The second warm ischemia time of bile duct longer than 60minutes and prolonged bile duct second warm ischemia time combined with cold preservation time were significant risk factors for severe ischemic biliary complications after liver transplantation with grafts from non-heart-beating donors.
文摘BACKGROUND: To evaluate feasibility and safety of venovenous bypass prior to mobilization of the liver during orthotopic liver transplantation (OLT). METHODS: Fifty-four patients were classified into two groups. Group A consisted of 23 patients receiving OLT with classical venovenous bypass. Group B consisted of 31 patients who received a modified-procedure: venovenous bypass ahead of the mobilization of the liver during ULT. The blood loss, duration of venovenous bypass, cold ischemia time, anhepatic phase, and transfusion during operation in the two groups were compared. Complications after the operation were also compared between the two groups. RESULTS: The duration of venovenous bypass and cold ischemia time in group A were longer than those in group B [(99.78±21.36 min) vs (96.32±22.25 min) and (484.78±134.01 min) vs (443.15± 85.27 min)]. The anhepatic phase lasted for about 100 min averagely in the two groups. The volumes of blood loss and transfusion during the operation were larger in group A than in group B [(5096±4243 ml) vs (1726±1125 ml) and (3676±2938.74 ml) vs (1217.69±829.72 ml)]. Postoperative complications occurred in 26 patients of group A and in 19 patients of group B. CONCLUSION: This modified-procedure or venovenous bypass ahead of mobilization of the liver in OLT can reduce the blood loss during OLT and the incidence of postoperative complications without prolongation of the anhepatic phase and duration of venovenous bypass.
基金Supported by the National Natural Science Foundation,No.81470896the Project of Development and Innovation Team of Ministry of Education,No.IRT_16R57
文摘AIM To compare the effect of University of Wisconsin(UW) solution with or without metformin, an AMP-activated protein kinase(AMPK) activator, for preserving standard and marginal liver grafts of young and aged rats ex vivo by hypothermic machine perfusion(HMP).METHODS Eighteen young(4 mo old) and 18 aged(17 mo old)healthy male SD rats were selected and randomly divided into three groups: control group, UW solution perfusion group(UWP), and UW solution with metformin perfusion group(MUWP). Aspartate aminotransferase(AST), alanine aminotransferase(ALT), lactate dehydrogenase(LDH), interleukin-18(IL-18), and tumor necrosis factor-alpha(TNF-α) in the perfused liquid were tested. The expression levels of AMPK and endothelial nitric oxide synthase(e NOS) in liver sinusoidal endothelial cells were also examined.Additionally, microscopic evaluation of the harvested perfused liver tissue samples was done. RESULTS AST, ALT, LDH, IL-18 and TNF-α levels in the young and aged liver-perfused liquid were, respectively,significantly lower in the MUWP group than in the UWP group(P < 0.05), but no significant differences were found between the young and aged MUWP groups.Metformin increased the expression of AMPK and e NOS protein levels, and promoted the extracellular release of nitric oxide through activation of the AMPK-e NOS mediated pathway. Histological examination revealed that in the MUWP group, the extent of liver cells and tissue damage was significantly reduced compared with the UWP group.CONCLUSION The addition of metformin to the UW preservative solution for ex vivo HMP can reduce rat liver injury during cold ischemia, with significant protective effects on livers, especially of aged rats.
文摘Liver transplantation(LT)is the best treatment for endstage hepatic failure,with an excellent survival rates over the last decade.Biliary complications after LT pose a major challenge especially with the increasing number of procured organs after circulatory death.Ischaemic cholangiopathy(IC)is a set of disorders characterized by multiple diffuse strictures affecting the graft biliary system in the absence of hepatic artery thrombosis or stenosis.It commonly presents with cholestasis and cholangitis resulting in higher readmission rates,longer length of stay,repeated therapeutic interventions,and eventually re-transplantation with consequent effects on the patient’s quality of life and increased health care costs.The pathogenesis of IC is unclear and exhibits a higher prevalence with prolonged ischaemia time,donation after circulatory death(DCD),rejection,and cytomegalovirus infection.The majority of IC occurs within 12 mo after LT.Prolonged warm ischaemic times predispose to a profound injury with a subsequently higher prevalence of IC.Biliary complications and IC rates are between 16%and 29%in DCD grafts com-pared to between 3%and 17%in donation after brain death(DBD)grafts.The majority of ischaemic biliary lesions occur within 30 d in DCD compared to 90 d in DBD grafts following transplantation.However,there are many other risk factors for IC that should be considered.The benefits of DCD in expanding the donor pool are hindered by the higher incidence of IC with increased rates of re-transplantation.Careful donor selection and procurement might help to optimize the utilization of DCD grafts.