OBJECTIVETo study the feasibility of sustaining the viable status of a liver graft in at least 96 h by extracorporeal perfusion using autologous blood.

METHODSEight extracorporeal porcine liver perfusions using autologous blood were performed, each for 96 h with hepatectomy, cold preservation, cannulation of vessels, and initiation of perfusion with normothermic oxygenated porcine blood. The graft viability was assessed by metabolic, synthetic, hemodynamic, and histologic parameters.

RESULTSAfter 96 h of normothermic, extracorporeal perfusion using autologous blood, the isolated livers maintained normal physiological levels of pH and electrolytes with sustained hepatic protein synthesis (complement and factor V) throughout the perfusion. Hemodynamic parameters maintained normal physiological ranges. Histological inspection demonstrated good preservation of the liver with a good architectural integrity.

CONCLUSIONIt is possible to sustain the viable status of a liver graft within 96 h by extracorporeal perfusion using autologous blood.

Animals ; Extracorporeal Circulation ; Liver ; Liver Transplantation ; Organ Preservation ; Organ Preservation Solutions ; Swine

OBJECTIVETo investigate the protective effect of luteolin on isolated rat heart in hypothermic preservation.

METHODSForty male SD rats were randomly divided into 4 groups (n = 10): control group, luteolin low-dose group (7.5 micromol/L), middle-dose group (15 micromol/L) and high dose group (30 micromol/L). Langendorff model of isolated rat heart was used. After 30 min basal perfusion, the hearts were stored in University of Wisconsin solution (UW solution) at 4 degrees C with luteolin (7.5, 15 and 30 micromol/L) or without luteolin for 12 h and followed by 60 min reperfusion. The recovery of cardiac contractile and diastolic function, coronary flow (CF), creatine kinase (CK) leakage in the coronary effluent, myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULTSThe results revealed that luteolin improved the recovery of left ventricular peak systolic pressure and +/- dp/dtmax dose-dependently and increased coronary flow. The leakage of creatine kinase in the coronary effluent was significantly reduced in luteolin-added hearts. Impairment of myocardial ultrastructure after 12 h hypothermic preservation was obviously alleviated in hearts luteolin-added group compared with that in control group. There were no differences between the groups in myocardial water contents.

CONCLUSIONLuteolin as a supplementation in cardiac preservation solution can significantly improve the hypothermic preservation effects on rat heart and have myocardial protection effect, especially in luteolin-added with 30 micromol/L.

Animals ; Cryopreservation ; In Vitro Techniques ; Luteolin ; pharmacology ; Male ; Myocardium ; Organ Preservation ; methods ; Organ Preservation Solutions ; Rats

BACKGROUNDOrgan preservation keeps the quality of the organs under prolonged ischemia. Continuous machine perfusions are gaining an important position in clinical research and practice. The aim of this study was to evaluate the protective effect of continuous hypothermic machine perfusion transport system (AirdriveTM) on cold ischemic injury of canine kidney.

METHODSTen kidneys of five healthy preserving canines were taken out after general anesthesia. Five kidneys were stored using common cold preservation (CCP group) by immersing it in the organ preservation solution, mixed with water and ice, and kept in a cold room at 4°C. The other five kidneys were stored using continuous machine perfusion preservation (CMP group) and were placed into the Airdrive(TM) continuous machine perfusion device at room temperature. The renal tissues were examined by histopathology, electron microscopy, and mitochondrial activity check at different time points.

RESULTSHistologic sections showed that the structures of the ten renal tissues were similar during the first 24 hours. After 48 hours, the CCP group showed more pronounced changes, as the renal tubular epithelial cells were more obvious than those in the glomeruli. Oxygen consumption rate of state III and IV respiration in the CCP group decreased after 12-48 hours and increased at 48 hours, respectively, when compared to the CMP group (P < 0.05). Cortex respiratory control ratio and phosphorus oxygen ratio were significantly higher in the CMP group at 48 hours.

CONCLUSIONWith prolonged storage time, the effect of continuous hypothermic machine perfusion transport system is better than that of common cold preservation on canine kidney.

Animals ; Dogs ; Kidney ; Kidney Transplantation ; Male ; Organ Preservation ; methods ; Organ Preservation Solutions

Although lung transplantation (LTx) has been established as a therapeutic approach for end-stage respiratory failure, several problems remain to be solved. In addition to the serious problem of donor shortage, primary graft failure, which is mostly caused by ischemia-reperfusion injury, a serious problem, and represents one of the most frequent causes of early mortality. The development of a highly reliable organ preservation solution that reduces ischemia-reperfusion injury will improve the functioning of transplanted organs and alleviate the donor shortage. We first evaluated the importance of saccharides and electrolytes in the lung preservation solution. We proved the superiority of trehalose, a non-reducing disaccharide, and the efficiency of the extracellular-type (low potassium) ion composition, and we also developed an extracellular-type trehalose containing Kyoto (ET-Kyoto) solution. Furthermore, several agents for vascular endothelial protection were evaluated, and finally, a more effective solution named "new ET-Kyoto solution" was developed, by adding N-acetylcysteine, dibutyryl adenosine 3', 5'-cyclic monophosphate, and nitroglycerin to the "conventional" ET-Kyoto solution. The new ET-Kyoto solution enabled canine LTx to last up to 30 hours. ET-Kyoto solution has so far been used and produced good results in five clinical LTx throughout Japan and South Korea. Although it was initially developed for lung preservation, its effectiveness in the preservation of various organs/ tissues, such as the trachea, kidney, skin/muscle flap, amputated digits, liver, and pancreas, has also been experimentally and clinically shown. In this paper, clinical and experimental findings with ET-Kyoto solution have been accumulated to further analyze its effect, safety, and chemical stability. We hope to provide ET-Kyoto solution as the standard organ/tissue preserving solution throughout the world.
Animals ; Humans ; Japan ; *Organ Preservation Solutions ; *Technology, Pharmaceutical ; *Universities

OBJECTIVETo investigate whether the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) opener diazoxide as an additive to cardioplegia solution could enhance myocardial protection during hypothermic preservation of the rat heart.

METHODSThe Langendorff model of isolated rat heart was used. After equilibrium, the hearts were stored in Celsior cardioplegia solution at 4 degree with or without supplement of diazoxide for 3 or 8 h followed by 60 minutes reperfusion. The recovery of cardiac contractile function, myocardial enzyme leakage in the coronary effluent, and myocardial water content were determined. The myocardial ultrastructure was also observed.

RESULT(1) Treatment of diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes, lactate dehydrogenase (LDH) and creatine kinase (CK), at the 2nd and 4th minute of reperfusion of rat heart after hypothermic preservation for 3 h. (2) After hypothermic preservation for 8 h, diazoxide improved the recovery of left ventricular developed pressure and decreased the leakage of myocardial enzymes (LDH, CK and glutamic oxalic transaminase) during reperfusion. Moreover, left ventricular end-diastolic pressure was significantly lower in diazoxide-treated hearts than that of hearts in Celsior solution. (3) Diazoxide significantly decreased the water content of myocardium and increased coronary flow of the hearts compared with those in control after hypothermic preservation for 8 h. (4) Impairment of myocardial ultrastructure after 8 h hypothermic preservation was alleviated in hearts treated with 30 mol/L diazoxide. (5) The cardiac effects of 30 mol/L diazoxide were attenuated by a mitoK(ATP) blocker 5-hydroxydecanoate (100 micromol/L).

CONCLUSIONDiazoxide as a supplementation in cardioplegia solution could enhance myocardial protection during hypothermic heart preservation via opening of mitochondrial K(ATP) channel.

Animals ; Cardioplegic Solutions ; Cryopreservation ; Diazoxide ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Potassium Channels ; drug effects ; Rats ; Rats, Sprague-Dawley

Generally, mammalian cells and living tissues can be cryopreserved in a frozen state at very low temperatures over a long storage term. The survival rate of cell suspensions is often acceptable however, living tissues suffer a variety of injuries. In this paper, it was demonstrated that the addition of polyphenols extracted from green tea to conventional cell culture medium and tissue compatible liquid, can control cell proliferation and also preserve tissues for several months at ordinary room temperature, including such tissues as blood vessels, cartilage, islet cells and corneas. This protocol allows a non-frozen living tissue bank to be established using the preservation fluid described.
Animals ; Flavonoids/*pharmacology ; Humans ; Organ Preservation Solutions/*pharmacology ; Phenols/*pharmacology ; Tea/*chemistry ; *Tissue Banks ; *Tissue Preservation ; *Tissue Transplantation ; Transplantation, Homologous

OBJECTIVETo investigate the effective method of preserving composite facial allograft so as to attenuate ischemic injury.

METHODSThe composite facial allografts were harvested from dog, perfused and preserved with 4 degrees C physiological sodium chloride and UW solution respectively. Immediately after the removal of the flap, after 12, 24, 48 h of preservation, MTT assay was used to determine the viability of several kinds of tissue, including skin, mucosa, muscle, bleed vessel, nerve and gland. The results of the two groups were compared in term of viability percentage. The pathology of several tissues were observed after 24 and 48 h of storage.

RESULTSThe viability percentage of every tissue conserved in UW solution for 48 hours was more than 75%. There was significant difference between physiological sodium chloride group and UW group (P < 0.05). Some changes, including Porous arrangement of fibers in connective tissue of skin and mucosa, hyalinization of tissue around the hair follicle and edema of cell in hair follicle, enlargement of space between muscle bundles and unclearness of boundary of acinus could be seen in physiological sodium chloride group while no significant change in UW group.

CONCLUSIONSUW solution could be considered as preservation solution for composite facial allograft.

Adenosine ; Allopurinol ; Animals ; Dogs ; Face ; Female ; Glutathione ; Insulin ; Male ; Organ Preservation Solutions ; Raffinose ; Tissue Preservation ; methods ; Transplantation, Homologous

No abstract available.
Adenosine ; Allopurinol ; Glutathione ; Humans ; Insulin ; Organ Preservation Solutions ; Periodontal Ligament ; Raffinose

Drugs, Chinese Herbal ; pharmacology ; Heart ; Humans ; Liver ; Organ Preservation Solutions ; Panax ; Pancreas ; Salvia miltiorrhiza

Organ preservation solutions currently in use have been quite successful clinically, but it is true that improvements can still be made in preservation time and postpreservation function of the graft. The aim of this study is to compare the effectiveness of two domestic organ preservation solutions, SNU-I and SNU-II, with that of UW solution in a swine autotransplantation model. SNU-I is a domestic reproduction of UW solution and SNU-II is an theoretically improved regimen with the addition of pyruvate and aspartate. Unilateral nephrectomy was done in 14 female pigs and the grafts were cold-preserved with UW (n=3), SNU-I (n=6) and SNU-II (n=5) solution for 24 hours before being autotransplanted back to the same animal. The groups were compared in posttransplant serum BUN/creatinine level and in the degree of tissue injury in graft specimens taken 14 days after the transplantation. The peak values for the BUN/creatinine level were the highest in the SNU-II group, reaching 80 mg/dl for BUN and 6.43 mg/dl for creatinine. Those for the UW and SNU-I groups were lower, but the differences were not statistically significant. The tissues were embedded in paraffin, stained with H/E and then were subjected to light microscopic examination (X200). When the degree of tissue injury was scored on the items of tubular necrosis, intratubular cell detachment, and brush border integrity, the SNU-II group displayed higher scores than the other two. The SNU-II group was also given higher scores in the degree and aggressiveness of the inflammatory process, but the difference between the three groups were not statistically significant in all items. In conclusion, UW and SNU-I solutions appear to have the same potency as organ preservation solutions. Further investigations are needed for the efficacy of SNU-II solution, including longer term preservation or warm ischemia models.
Animals ; Aspartic Acid ; Autografts* ; Creatinine ; Female ; Humans ; Kidney Transplantation ; Microvilli ; Necrosis ; Nephrectomy ; Organ Preservation Solutions* ; Organ Preservation* ; Paraffin ; Pyruvic Acid ; Reproduction ; Swine ; Transplants ; Warm Ischemia