Comparison of intermittent injection of nondepolarizing solution with a single flush of UW solution for donor heart preservation

Isolated canine hearts were preserved for 6 h at 5°C followed by normothermic reperfusion for 2 h. The dogs were divided into two groups of nine hearts each; group 1 received a nondepolarizing preservation solution in multidose, and group 2 received a single flush of University of Wisconsin (UW) solution. Serum MB-CK and mitochondrial aspartate aminotransferase (m-AAT) concentrations and calcium overload during reperfusion were lower in group 1 than in group 2. At the end of reperfusion, myocardial ATP and total adenine nucleotide concentrations were higher and mitochondrial morphology appeared more intact in group 1 than in group 2. Left ventricular diatolic function was preserved better in group 1 than in group 2. These results suggest that in 6-h heart preservation, a nondepolarizing solution applied in multidose fashion protects the myocardium from the deleterious effects of hypothermia and cardioplegia better than a single flush of UW solution.     

超极化心脏保存液对离体大鼠心脏低温保存的保护作用

目的　探讨含K 通道开放剂的心脏保存液(HCS液)对大鼠离体心脏的保存作用。方法　将24只SD大鼠随机平均分为 3 组,分别用 HCS液、UW液、K H液对心脏进行保存。采用Langendorff心脏灌注和工作模型装置进行心脏功能测定,比较 24h后心功能恢复率以及细胞酶学、心肌含水量、pH值变化和能量变化,观察心肌超微结构改变。结果　心脏保存24 h后, K H组左心室功能损伤严重,UW组及HCS组左心室功能损伤较轻。HCS组冠状动脉流量(CF)恢复率较 K H组、UW组好;HCS组冠脉回流液中乳酸脱氢酶(LDH)、磷酸肌酸激酶(CPK)含量及心肌含水量与UW组比较,差异无统计学意义;HCS组 pH值变化最小, ATP含量最高;HCS组心脏复跳时间与UW组基本相同; HCS组心肌保存良好。结论　HCS保存液对大鼠心脏的保存在 24 h内有明显的保护作用,心脏功能恢复率能达到UW液的水平,在能量保护、改善酸中毒方面则优于UW液。     

Greater Hemodynamic Instability With Histidine-Tryptophan-Ketoglutarate Solution Than University of Wisconsin Solution During the Reperfusion Period in Living Donor Liver Transplantation

Objective

University of Wisconsin (UW) and histidine-tryptophan-ketoglutarate (HTK) solutions are the 2 most commonly used liver preservation solutions. The aim of this study was to compare cardiovascular stability, acid-base status, and potassium concentrations between patients who received grafts preserved in either UW or HTK solution in orthotopic liver transplantation (OLT).

Patients and Methods

In this retrospective study, 87 patients who underwent living donor OLT were divided into 2 groups: UW (n = 28) and HTK (n = 59). Group HTK was subdivided into group NF-HTK (n = 31; nonflushed before reperfusion) and group F-HTK (n = 28; flushed before reperfusion). We determined mean arterial pressure (MAP) and heart rate every minute for 5 minutes after reperfusion and the maximum change in these values and incidence of postreperfusion syndrome (PRS). Body temperature, cardiovascular and acid-base parameters, as well as potassium concentrations were compared at 5 minutes before and 5 and 30 minutes after reperfusion.

Results

The maximum decreases in MAP within 5 minutes after reperfusion were significantly greater in both the NF-HTK and the F-HTK groups. The rate of PRS was significantly greater in the NF-HTK compared with the UW group. Flushing with HTK solution decreased the rate of PRS; there was no significant difference between the F-HTK and UW groups. All serial changes in body temperature, cardiovascular and acid-base parameters, as well as potassium concentrations were similar among the 3 groups.

Conclusions

The incidence of PRS was greater using HTK compared with UW solution during the reperfusion period. Therefore, careful hemodynamic management is advised when using HTK solution.     

The effect of sodium concentration on myocardial viability in donor heart preservation using a nondepolarizing solution

This study examined the effect of different sodium concentrations in a nondepolarizing solution on myocardial viability and functional recovery of the canine donor heart. Isolated canine hearts were preserved for 6 h at 5°C, followed by normothermic reperfusion for 2 h. Dogs were divided into two groups of nine dogs each: group 1 received a nondepolarizing solution with 70mm Na⁺ and group 2 with 30mm Na⁺. The myocardial Ca²⁺ concentration at the end of preservation was significantly higher in group 1 than in group 2 and increased after reperfusion in both groups without any intergroup difference. Myocardial concentrations of ATP, ADP, and total adenine nucleotide at the end of reperfusion were significantly higher in group 1 than in group 2. Myocardial cyclic adenosine monophosphate concentration was significantly higher in group 1 than in group 2 at the end of both preservation and reperfusion. The myocardial cyclic guanosine monophosphate concentration in group 1 increased and was higher than in group 2 at the end of preservation, but had returned to normal levels by the end of reperfusion. However, it remained unchanged through preservation and reperfusion in group 2. The left ventricular systolic and diastolic function, assessed by pressurevolume relationship, was better in group 1 than in group 2. Mitochondrial ultrastructural changes were similar. These results suggest that a nondepolarizing solution containing 70mm Na⁺ provides better myocardial protection than a solution containing 30mm Na⁺.     

The effect of betamethasone on myocardial viability during cold cardioplegia in canine donor heart preservation

This study examined the effect of betamethasone on myocardial viability of reperfused isolated canine hearts following a 6-h hypothermic cardioplegia. The dogs were divided into two groups: group I (n = 9) received nondepolarizing cardioplegia containing betamethasone 250 mg/l while group II (n = 7) was administered cardioplegia without betamethasone. The myocardial concentrations of calcium, ATP, ADP, total adenine nucleotide, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were identical in the two groups throughout the experiment. The coronary sinus plasma concentrations of MB fraction off creatine kinase (MB-CK), cAMP and cGMP after 2 h of reperfusion were significantly lower in group I than in group II. The myocardial mitochondrial ultrastructure, as assessed by semiquantitative morphometry, was found to be significantly better preserved in group I than in group II at the end of both preservation and reperfusion. In addition, the left ventricular end-systolic pressure volume relation (ESPVR) showed a higher slope and lower intercept in group I than in group II. These results suggest that the addition of betamethasone to nondepolarizing cold cardioplegia enhances myocardial protection via membrane stabilization without affecting the adenine nucleotide metabolism.     

Comparison of HTK- and UW-solution for liver preservation tested in an orthotopic liver transplantation model in the pig

The aim of this experimental study was to compare the preservation potency of University of Wisconsin (UW) and HTK (Bretschneider) solutions in an orthotopic liver transplantation (OLT) model in pigs. Livers were harvested using an in situ perfusion technique, where organs were flushed with the solution being tested, stored on ice — cold storage (CS) — for 2 or 24 h and then transplanted. Parameters monitored were liver enzymes in serum, hepatic water content, high energy phosphates, nuclear magnetic resonance (NMR) relaxation time T2, light microscopy and bile production. CS for 24 h is an extreme in pig liver preservation and is not compatible with animal survival. Biopsies showed drastic morphological changes and grafts did not produce bile in either group. (Bile production 2 h CS: HTK, 5.6 ± 1.8 ml/h; UW, 4.7 ± 2.3 ml/h) Enzyme release after reperfusion (ASGOT, ?LDH) was higher in long-term preservation. Hepatic tissue water content significantly decreased during CS in UW preserved livers. Edema alter reperfusion (?H₂0: HTK 24 h = + 5.6%, UW 24 h= + 4.8%) and regeneration capacity after reperfusion (UW 2 h = 63%, HTK 2 h = 55%, UW 24 h = 30%, HTK 24 h = 30%) were not significantly different. However, we did not observe major differences in preservation potency between the solutions tested. Differences were correlated, rather, with length 9 time of CS, than with the solution used. Therefore, HTK solution seemed to be a low potassium containing alternative to UW solution.     

Extended preservation of non-heart-beating donor livers with normothermic machine perfusion

BACKGROUND: Non-heart-beating donor (NHBD) livers represent an important organ pool, but are seldom utilized clinically and require rapid retrieval and implantation. Experimental work with oxygenated perfusion during preservation has shown promising results by recovering function in these livers. This study compared sanguinous perfusion with cold storage for extended preservation of the NHBD liver in a porcine model. METHODS: Porcine livers were subjected to 60 min of in vivo total warm ischaemia before flushing, after which they were preserved by one of two methods: group 1 (n = 4), University of Wisconsin (UW) solution by standard cold storage for 24 h; group 2 (n = 4), oxygenated autologous blood perfusion on an extracorporeal circuit for 24 h. All livers were subsequently tested on the circuit during a 24-h reperfusion phase. RESULTS: Livers in group 1 showed no evidence of viability during the reperfusion phase with no bile production or glucose utilization; they also displayed massive necrosis. Livers in group 2 demonstrated recovery of function by synthetic function, substrate utilization and perfusion haemodynamics; these livers displayed less cellular injury by hepatocellular enzymes. All differences in parameters between the two groups were statistically significant (P < 0.05). These findings were supported by histological examination. CONCLUSION: Warm ischaemia for 1 h and simple cold storage (UW solution) for 24 h renders the liver non-viable. Oxygenated, sanguinous perfusion as a method of preservation recovers liver function to a viable level after 24 h of preservation.     

Celsior solution compared with University of Wisconsin solution (UW) and histidine–tryptophan–ketoglutarate solution (HTK) in the protection of human hepatocytes against ischemia–reperfusion injury

Celsior, a new preservation solution in thoracic organ transplantation was evaluated for efficacy in cold preservation of human hepatocytes and compared with University of Wisconsin solution (UW) and histidine-tryptophan-ketoglutarate solution (HTK, Custodiol). Human hepatocyte cultures were preserved at 4 degrees C in Celsior, UW and HTK for 2, 6, 12, 24 and 48 h with 6 h of reperfusion. Levels of lactate dehydrogenase (LDH; cell necrosis), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; mitochondrial function), and adenosine 5'-triphosphate (ATP; loss of intracellular energy) were measured. Cell necrosis, mitochondrial dysfunction, and loss of ATP were significantly ( P<0.001, P<0.001, P<0.002, respectively) lower in Celsior than in HTK. The amount of cell necrosis and mitochondrial dysfunction in Celsior solution (CS) and UW was equal ( P=n.s.) up to 24 h and significantly lower in UW after 48 h ( P<0.001). Additionally, the intracellular level of ATP was significantly higher after ischemia ( P<0.001) and reperfusion from long-term ischemia (24, 48 h) ( P<0.002). We can conclude that Celsior was superior to HTK and equal to UW in the protection of human hepatocytes against cold preservation injury from ischemia and reperfusion. Furthermore, Celsior was effective in long-term preservation of human hepatocytes.     

Prolonged 24-hour subzero preservation of heterotopically transplanted rat hearts using antifreeze proteins derived from arctic fish

BACKGROUND: Arctic fish survive subzero temperatures by producing a family of antifreeze proteins (AFPs) that noncolligatively lower the freezing temperature of their body fluids. We report 24-hour storage of mammalian hearts for transplantation at subzero temperatures using AFPs derived from arctic fish. METHODS: Forty-two heterotopic transplantations were performed in isoimmune Sprague-Dawley rats. Harvested hearts were retrogradely infused with cold 4 degrees C University of Wisconsin (UW) solution and were preserved in a specialized cooling bath at two target temperatures, 4 degrees C and -1.3 degrees C for 12,18, and 24 hours (6 experiments/group). Preservation solutions were UW alone for the 4 degrees C group, and UW with 15 mg/mL AFP III for the -1.3 degrees C group. After hypothermic storage the hearts were heterotopically transplanted into isoimmune rats. Viability was assessed and graded on a scale of 0 to 6 (0 = no contractions to 6 = excellent contractions). Transplanted hearts were then fixed in vivo and were subject to electron microscopy and histopathologic examination. RESULTS: None of the hearts preserved at -1.3 degrees C in UW/AFP III solution froze. All control hearts preserved at -1.3 degrees C without AFP protection froze and died at reperfusion. Viability of hearts preserved at -1.3 degrees C in UW/AFP III solution was significantly better after 18 hours of preservation, 30 and 60 minutes after reperfusion (median, 5 versus 3 and 6 versus 3, respectively; p < 0.05) and after 24 hours of preservation 30 and 60 minutes after reperfusion (median, 4.5 versus 1.5 and 5 versus 2, respectively; p < 0.05). Histologic and electron microscopy studies demonstrated better myocyte structure and mitochondrial integrity preservation with UW/AFP III solution. CONCLUSIONS: Antifreeze proteins prevent freezing in subzero cryopreservation of mammalian hearts for transplantation. Subzero preservation prolongs ischemic times and improves posttransplant viability.     

A non-heart-beating donor model to evaluate functional and morphologic outcomes in resuscitated pig hearts.

A non-heart-beating donor model was considered to examine whether pig hearts from the abattoir could be resuscitated by whole blood reperfusion. For preservation, machine perfusion using University of Wisconsin (UW) solution was compared with storage on ice. Nineteen hearts from abattoir pigs, harvested 25 +/- 3 min after exsanguination, were harvested and transported to the laboratory. Controls (n = 7) were immediately reperfused with homologous whole pig blood in an isolated heart model for 60 min with monitoring of left ventricular developed pressure (LVDP), contractility, and coronary flow. UW solution hearts (UW, n = 6) were perfused for 4 h with 10 degrees C cold UW solution before blood reperfusion. In the cold storage group (CS, n = 6), the organs were stored for an additional 4 h on ice before blood reperfusion. In all hearts, histology was performed after 60 min of blood reperfusion to evaluate myocardial reperfusion injury. All three groups showed significant increases in LVDP (p <.001), although this functional recovery was earliest in the control group and latest in the UW group. Significant declines were observed for both LVDP and contractility from the peak values in each group to the end of blood reperfusion. Coronary flow increased steadily over the time course for the UW group, whereas in the control and CS groups flow increased during the first 15 min of blood reperfusion and then decreased. In the UW and CS groups, there were significant positive correlations between coronary flow and LVDP (p <.001). Microscopic examination revealed no differences between the three groups. Thus, hearts from an abattoir with 25 min of warm ischemic time can be resuscitated. For storage of these organs, continuous machine perfusion with UW solution is superior to cold storage on ice.     

Comparison of Celsior and UW solution in experimental pancreas preservation

BACKGROUND: The University of Wisconsin solution (UW) is the gold standard for pancreas preservation. Celsior (CEL) was formulated specifically for heart preservation. Recently, experimental and clinical experience has been reported on the application of CEL to abdominal organs. In this animal study, pancreas preservation with CEL was compared with that in UW solution. PATIENTS AND MATERIALS: Heterotopic, allogeneic pancreaticoduodenal transplantation was performed in female G?ttingen Minipigs (n = 12 donors, n = 12 recipients). The grafts were flushed and stored for 6 h at 4 degrees C in UW or CEL. The recipients were randomized into two groups receiving either UW (n = 6)- or CEL (n = 6)-preserved grafts with a follow-up of 5 days. Blood flow (laser Doppler), partial oxygen tension, histological changes, endothelin-1 (plasma, immunohistochemistry), lipase, amylase, trypsinogen activation peptide, and C-reactive protein (CRP) were measured. RESULTS: Partial oxygen tension was lower in the CEL group (P < 0.05). However, blood flow did not differ between UW- and CEL-preserved organs. The histomorphologic analysis of the pancreatic grafts revealed significantly less edema in the UW-preserved organs. Serum levels of amylase, lipase, CRP, and TAP taken from the central venous blood were comparable in the two groups, except for higher amylase values 36 h after reperfusion in the CEL group compared to the UW group (P < 0.05). Likewise, TAP taken from the portal venous effluent of the graft was found to be higher in the CEL group than in UW (P < 0.05). Endothelin-1 serum levels rose significantly during reperfusion without differences between the two groups. ET-1 immunohistochemistry revealed increased local ET-1 during reperfusion in all grafts. However, the ET-1 immunostaining in the CEL group was more pronounced than that in the UW group (P < 0.05). CONCLUSIONS: Our results suggest that CEL solution is not as effective in preventing pancreatic ischemia/reperfusion damage as the standard UW solution in experimental pancreas transplantation. Increased ET-1 immunostaining and reduced p(ti)O(2) in the CEL group indicate increased microcirculatory damage in the CEL group.     

Preservation injury of jejunal grafts and its modulation by custodiol and university of wisconsin perfusion solutions in wistar rats

BACKGROUND: The two preservation solutions most commonly used in human transplantation surgery are University of Wisconsin (UW) and Custodiol (histidine-tryptophan-ketoglutarate; HTK). The aim of our study was to compare the protective effect of UW and HTK solutions on preservation-induced injury of jejunal grafts, as evaluated by the histological changes (semiquantitative method) and small bowel mucosal serotonin levels (as a possible new quantitative method). METHODS: Male Wistar rats (n = 50) weighing 316 +/- 52 g were divided into two main groups according to which preservation solution was used, i.e. UW (n = 25) or HTK (n = 25), and each of these groups was divided into five subgroups according to cold ischemic time (0, 1, 6, 9 and 12 h). Jejunal mucosa biopsy specimens were obtained to determine the serotonin concentration in mucosa and for standard light histology. To grade histological changes in mucosa, Park's small bowel injury grading system was used. RESULTS: Histological examination revealed that injury increased with cold ischemic time in the UW as well as in the HTK group, and there were no significant differences in injury between the two groups, except for the 6-hour cold ischemic period (p < 0.05), when HTK-preserved grafts showed a lower degree of injury (0.97 +/- 0.41) compared with UW-preserved grafts (1.25 +/- 0.39). The mucosal serotonin concentration decreased with cold ischemic time in both groups, and there were significant differences (p < 0.05) in concentrations between the groups after 9 and 12 h of cold ischemia. A significantly higher concentration was measured in grafts preserved in UW solution at these time points. CONCLUSION: The concentration of mucosal serotonin in rat small bowel grafts preserved for 9 and 12 h in UW preservation solution was significantly higher than that in HTK solution. These findings indicate a better protective effect of UW solution on small bowel injury after 9 h of cold ischemia.     

Delivery of the bioactive gas hydrogen sulfide during cold preservation of rat liver: effects on hepatic function in an ex vivo model

The insults sustained by transplanted livers (hepatectomy, hypothermic preservation, and normothermic reperfusion) could compromise hepatic function. Hydrogen sulfide (H₂S) is a physiologic gaseous signaling molecule, like nitric oxide (NO) and carbon monoxide (CO). We examined the effect of diallyl disulfide as a H₂S donor during hypothermic preservation and reperfusion on intrahepatic resistance (IVR), lactate dehydrogenase (LDH) release, bile production, oxygen consumption, bromosulfophthalein (BSP) depuration and histology in an isolated perfused rat liver model (IPRL), after 48 h of hypothermic storage (4°C) in University of Wisconsin solution (UW, Viaspan). Livers were retrieved from male Wistar rats. Three experimental groups were analyzed: Control group (CON): IPRL was performed after surgery; UW: IPRL was performed in livers preserved (48 h—4°C) in UW; and UWS: IPRL was performed in livers preserved (48 h—4°C) in UW in the presence of 3.4 mM diallyl disulfide. Hypothermic preservation injuries were manifested at reperfusion by a slight increment in IHR and LDH release compared with the control group. Also, bile production for the control group (1.32 µL/min/g of liver) seemed to be diminished after preservation by 73% in UW and 69% in UW H₂S group at the end of normothermic reperfusion. Liver samples analyzed by hematoxylin/eosin clearly showed the deleterious effect of cold storage process, partially reversed (dilated sinusoids and vacuolization attenuation) by the addition of a H₂S delivery compound to the preservation solution. Hepatic clearance (HC) of BSP was affected by cold storage of livers, but there were no noticeable differences between livers preserved with or without diallyl disulfide. Meanwhile, livers preserved in the presence of H₂S donor showed an enhanced capacity for BSP uptake (k_ACON = 0.29 min^?1; k_AUW = 0.29 min^?1; k_AUWS = 0.36 min^?1). In summary, our animal model suggests that hepatic hypothermic preservation for transplantation affects liver function and hepatic depuration of BSP, and implies that the inclusion of an H₂S donor during hypothermic preservation could improve standard methods of preparing livers for transplant.     

Endothelium-dependent relaxation of canine pulmonary artery after prolonged lung graft preservation in University of Wisconsin solution: role of L-arginine supplementation.

BACKGROUND: The University of Wisconsin (UW) solution has been demonstrated to enhance pulmonary allograft preservation. Endothelial nitric oxide (NO) production has been shown to be significantly impaired after ischemia and reperfusion (I/R) injury. The present experiments aimed to determine the protective effects of pulmonary endothelium-dependent function by using supplemental NO in University of Wisconsin (UW) solution following prolonged lung graft preservation. METHODS: Thirty-six healthy mongrel dogs underwent thoracotomy to expose the left lung. In addition to a group given UW solution (n = 4), 100 micromol/liter l-arginine, (n = 7), 100 micromol/liter N(G)-monomethyl-l-arginine (l-NMMA n = 7) and 1.0 micromol/liter 3-morpholinosydnonimine (SIN-1, n = 18 respectively, were added to UW solution, and infused from the aortic root and pulmonary artery to the pulmonary vein. The perfused lung was then allowed to inflate to its maximum volume for 24-hour oxygenated preservation in each supplemented condition of UW solution at 4 degrees C. In the SIN-1 group, the preservation period was further divided into 8 hours and 16 hours, respectively. Rings of the third-order pulmonary artery of the inflated lung were then suspended in organ chambers to measure isometric force. RESULTS: Endothelium-dependent relaxation (EDR) to acetylcholine, adenosine diphosphate and sodium fluoride of the pulmonary rings in the l-arginine group was significantly preserved compared with UW-solution-only group. The l-NMMA group showed significant EDR impairment after 24-hour preservation compared with the UW solution group. Similar to the l-arginine group, the SIN-1 group showed significant EDR protection with 8-hour preservation, but not with 24-hour preservation. In contrast, EDR to calcium ionophore A23187 showed no EDR changes after 24-hour preservation in any of the supplemented groups. CONCLUSIONS: Supplemental l-arginine in UW solution ameliorates impaired pulmonary EDR following prolonged lung preservation of up to 24 hours.     

The effect of Celsior solution on 12-hour cardiac preservation in comparison with University of Wisconsin solution

BACKGROUND: Celsior is a new extracellular-type preservation solution which has been developed to act not only as a storage medium but also as a perfusion fluid during initial donor heart arrest, poststorage graft reimplantation and early reperfusion. We designed this experimental study to evaluate the effect of the Celsior solution in comparison with the University of Wisconsin solution from the viewpoint of energy depletion. METHODS: Adult mongrel dogs weighing 9 to 13 kg were divided into two groups. In the UW group (n=7), a 4 degrees C University of Wisconsin solution was used for coronary vascular washout and storage following cardiac arrest using a glucose-insulin-potassium solution. In the Celsior group (n=7), the Celsior solution was used to obtain cardiac arrest, coronary vascular washout and storage. High energy phosphate levels and myocardial pH were measured using (31)P-nuclear magnetic resonance spectroscopy immediately after preservation and at 3, 6 and 12 hours after preservation. After 12-hour cold storage, left ventricular free wall tissues were harvested for histological examination. RESULTS: High energy phosphate levels and myocardial pH were significantly better preserved in the Celsior group than in the UW group. In the histological findings, glycogen granules were preserved well in the Celsior group. CONCLUSIONS: We conclude from our study that the Celsior solution is comparable to the University of Wisconsin solution for use in 12-hour heart preservation in canine models.     

A Non-Heart-Beating Donor Model to Evaluate Functional and Morphologic Outcomes in Resuscitated Pig Hearts

A non-heart-beating donor model was considered to examine whether pig hearts from the abattoir could be resuscitated by whole blood reperfusion. For preservation, machine perfusion using University of Wisconsin (UW) solution was compared with storage on ice. Nineteen hearts from abattoir pigs, harvested 25 &#45 3 min after exsanguination, were harvested and transported to the laboratory. Controls ( n = 7) were immediately reperfused with homologous whole pig blood in an isolated heart model for 60 min with monitoring of left ventricular developed pressure (LVDP), contractility, and coronary flow. UW solution hearts (UW, n = 6) were perfused for 4 h with 10°C cold UW solution before blood reperfusion. In the cold storage group (CS, n = 6), the organs were stored for an additional 4 h on ice before blood reperfusion. In all hearts, histology was performed after 60 min of blood reperfusion to evaluate myocardial reperfusion injury. All three groups showed significant increases in LVDP ( p < .001), although this functional recovery was earliest in the control group and latest in the UW group. Significant declines were observed for both LVDP and contractility from the peak values in each group to the end of blood reperfusion. Coronary flow increased steadily over the time course for the UW group, whereas in the control and CS groups flow increased during the first 15 min of blood reperfusion and then decreased. In the UW and CS groups, there were significant positive correlations between coronary flow and LVDP ( p < .001). Microscopic examination revealed no differences between the three groups. Thus, hearts from an abattoir with 25 min of warm ischemic time can be resuscitated. For storage of these organs, continuous machine perfusion with UW solution is superior to cold storage on ice.     

Postoperative recovery of mitochondrial function of the human liver graft procured and preserved with University of Wisconsin (UW) solution

Changes in arterial blood ketone body ratio (KBR) were investigated in 47 human liver transplantations. Of the 20 grafts preserved with University of Wisconsin (UW) solution, 10 had a cold preservation period of less than 10 h (UWS group) and 10 of more than 10 h (UWL group). In 27 other cases, grafts were preserved with EuroCollins (EC) solution for less than 10 h (EC group). In the EC group, KBR increased over 0.7 within 6 h after reperfusion of the graft in 17 cases (63%) and within 24 h in 7 cases (26%). In the 3 other cases, KBR failed to recover, and these patients underwent retransplantation. In the UW group, KBR recovered within 6 h in 13 cases (65%) and within 24 h in 7 cases (35%). There were no significant differences between the UWS and UWL groups. It is shown that the mitochondrial function of liver grafts preserved with UW solution can be well maintained even after extended preservation periods of more than 10 h.     

Postoperative recovery of mitochondrial function of the human liver graft procured and preserved with University of Wisconsin (UW) solution

Abstract. Changes in arterial blood ketone body ratio (KBR) were investigated in 47 human liver transplantations. Of the 20 grafts preserved with University of Wisconsin (UW) solution, 10 had a cold preservation period of less than 10 h (UWS group) and 10 of more than 10 h (UWL group). In 27 other cases, grafts were preserved with EuroCollins (EC) solution for less than 10h (EC group). In the EC group, KBR increased over 0. 7 within 6h after reperfusion of the graft in 17 cases (63%) and within 24 h in 7 cases (26%). In the 3 other cases, KBR failed to recover, and these patients underwent retransplantation. In the UW group, KBR recovered within 6 h in 13 cases (65%) and within 24 h in 7 cases (35%). There were no significant differences between the UWS and UWL groups. It is shown that the mitochondrial function of liver grafts preserved with UW solution can be well maintained even after extended preservation periods of more than 10 h.     

The effect of perfusion with UW solution on the skeletal muscle and vascular endothelial exocrine function in rat hindlimbs

BACKGROUND: The effect of University of Wisconsin (UW) solution perfusion for extremity preservation is still unknown although it is widely used. The purpose of this study is to examine the effect of UW solution perfusion on skeletal muscle preservation in a rat model. MATERIALS AND METHODS: Rat hindlimbs were amputated and either preserved with UW solution perfusion (UW perfusion group) or given no perfusion (no-perfusion group) for 5 h at 25 degrees C. They were then transplanted to other isogeneic rats. ATP in the muscle and serum creatine phosphokinase were measured after 24 h of reperfusion. The vascular endothelial function of the femoral artery rings was measured before and after 24 h of reperfusion in the presence or absence of indomethacin (cyclooxygenase inhibitor) and L-NMMA (nitric oxide synthase inhibitor). TEA (calcium-activated potassium channel inhibitor) was also used to verify the vasodilator function. Reperfusion blood flow was monitored during the first 2 h of reperfusion. RESULTS: ATP in the UW perfusion group was significantly decreased after 24 h of reperfusion, while that in the no-perfusion group recovered. Reperfusion blood flow in the UW solution perfusion group was significantly lower than that in the no-perfusion group. Acetylcholine-induced relaxation in the UW perfusion group was significantly reduced before and after 24 h of reperfusion compared to that in the no-perfusion group and was mostly diminished by indomethacin and L-NMMA administration. CONCLUSIONS: Skeletal muscle injury is augmented by UW solution perfusion, probably due to deterioration of the vascular endothelial function resulting in blood supply diminution.     

Apoptosis is caused by prolonged organ preservation and blocked by apoptosis inhibitor in experimental rat pancreatic grafts

Apoptosis is a major mode of cell death after ischemia/reperfusion injury in several organs. The aims of this study were to evaluate apoptosis induction after different conditions of pancreas preservation and to investigate the impact of a caspase inhibitor on apoptosis induction in pancreatic grafts. METHODS: Inbred male Lewis rats served as donors and recipients. Apoptosis was detected using an in situ cell death detection kit and an apoptotic DNA ladder kit (Roche, Germany). An apoptotic index (AI) was defined as the number of apoptotic cells per field (400x) under a light microscope. The five groups included: group 1 (n = 5), normal pancreata; group 2 (n = 7), pancreata stored in University of Wisconsin (UW) solution (4 degrees C) for 6 hours (hr); group 3 (n = 7), pancreata preserved in UW solution (4 degrees C) for 18 hr; group 4 (n = 7), pancreata preserved in 0.9% saline (4 degrees C) for 6 hours; and group 5 (n = 5), pancreata preserved in 0.9% saline (4 degrees C) for 6 hours with Z-Asp-2,6-dichlorobenzoyloxymethylketone (caspase inhibitor) treatment. The pancreatic grafts in all experimental groups underwent a 2-hr period of reperfusion after transplantation. RESULTS: The results in this study showed that the AI was not significantly increased among pancreatic grafts in group 2 compared to group 1 (P >.05). However, AI in group 3 was significantly higher than that in group 2 (P <.05). The highest AI was observed in group 4. Interestingly, AI in group 5 was significantly lower than that in group 4 (P <.01), and not significantly different from group 1 (P >.05). Apoptotic DNA ladders were detected in the pancreatic grafts in group 2, 3, and 4, but not in group 1 and 5. CONCLUSIONS: Prolonged preservation of pancreata in cold UW solution induces a dramatic increase in apoptotic cell death among the pancreatic grafts following Tx. This observation may well be an important mechanism of graft damage. A caspase inhibitor might be useful to inhibit apoptosis induction in pancreatic grafts.