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.     

The optimal pressure for initial flush with UW solution in heart procurement.

OBJECTIVE: University of Wisconsin (UW) solution is widely used in organ preservation. Some investigators have reported that high pressure during initial flush with UW solution may induce vasoconstriction and endothelial damage, because of its high potassium content and high viscosity. However, using lower pressure during the initial flush may lead to irregular distribution of the solution and incomplete flushing of blood components from coronary vascular beds. This experimental study evaluated the effects of a range of initial flush pressures during heart procurement, followed by orthotopic transplantation of the graft after 12 hours of preservation. MATERIALS AND METHODS: Twelve pairs of adult mongrel dogs, weighing 9 to 14 kg, formed the recipient-donor combinations. After determining hemodynamic status by measuring cardiac output, left ventricular pressure (LVP), and maximum positive and negative change in LVP (+/-LVdP/dt), donor hearts were excised. Coronary vascular beds were flushed with 4 degrees C UW solution at a pressure of 60 mm Hg in the low-pressure group (n = 6) and at 120 mm Hg in the high-pressure group (n = 6). After 12 hours of cold preservation, orthotopic transplantation was performed using cardiopulmonary bypass (CPB). The hemodynamics of the transplanted graft were assessed by comparing recovery rates (%) from donor hearts 2 hours after weaning from CPB. Endothelin-1 (ET-1) levels were measured in the blood obtained from the coronary sinus 30 minutes after reperfusion. The transplanted grafts were then harvested for histologic study and measurement of adenosine triphosphate (ATP) content. RESULTS: Cardiac output, LVP, LVdP/dt and myocardial tissue ATP content were significantly better (p < 0.05) in the high-pressure group than in the low-pressure group. We found no significant differences in ET-1 levels between the groups. Transmission electron microscopic findings revealed that degeneration of the mitochondria was less extensive in the high-pressure group than in the low-pressure group. We observed no obvious ultrastructural damage to the endothelial cells in either group. CONCLUSION: When using UW solution in heart procurement, high pressure is better to completely wash out the blood components and distribute the solution.     

The effect of short-term coronary perfusion using a perfusion apparatus on canine heart transplantation from non-heart-beating donors.

BACKGROUND: We investigated the effects of briefly perfusing hearts from non-heart-beating donors (NHBDs) with a Celsior solution before cardiac transplantation. METHODS: Donor hearts were left in situ for 20 minutes after cardiac arrest was induced by rapid exsanguination. Twelve donor-recipient pairs of mongrel dogs were divided into 2 groups, the simple immersion (SI, n = 6) group and the coronary perfusion (CP, n = 6) group. Both groups underwent coronary flushing with Celsior, after which hearts from the SI group were stored using simple immersion for 4 hours and hearts from the CP group underwent 1 hour of further perfusion followed by storage for 3 hours. Orthotopic transplantation was then performed. We measured cardiac output, end-systolic maximal elastance (E(max)), left ventricular pressure, and rate pressure product 1 and 2 hours after weaning from cardiopulmonary bypass (CPB). Two hours after weaning from CPB, the hearts were harvested for histopathologic study and to determine the percentage of water content. RESULTS: The cardiac output (CO) recovery rate was significantly higher in the CP group than in the SI Group 1 hour after weaning from CPB (p < 0.05). The CO recovery rate, E(max), and rate pressure product were significantly higher and the percentage of water content was significantly lower in the CP group than in the SI Group 2 hours after weaning from CPB (p < 0.05). Histopathologic damage was more severe in the SI group. CONCLUSIONS: The results of this study suggest that short-term coronary perfusion with a Celsior solution may be useful for heart transplantation from NHBDs.     

Effects of adding P38 mitogen-activated protein-kinase inhibitor to celsior solution in canine heart transplantation from non-heart-beating donors

BACKGROUND: The activation of p38 mitogen-activated protein kinase (MAPK) plays an important role in ischemia-reperfusion injury. This study evaluated the effects of p38 MAPK inhibition using FR167653, a novel p38 MAPK inhibitor, as an additive to Celsior solution in canine heart transplantation from non-heart-beating donors (NHBDs). METHODS: Donor hearts were left in situ for 20 minutes after cardiac arrest, which was induced by rapid exsanguination. Twelve donor-recipient pairs of mongrel dogs were divided into two groups: the control and FR167653 (FR) groups (n=6 each). In both groups, the grafts were subjected to coronary flushing and immersed in Celsior solution for 4 hours with or without FR167653. Orthotopic heart transplantation was then performed. Cardiac output (CO), left ventricular pressure (LVP), and end-systolic maximal elastance (Emax) were measured 2 hours after weaning from cardiopulmonary bypass (CPB), and the hearts were then harvested for histopathologic study. The activation of p38 MAPK was evaluated in another 20 mongrel dogs. RESULTS: In the FR group, CO, LVP recovery rate, and Emax were significantly (P<0.05) higher 2 hours after weaning from CPB, histopathologic damage was attenuated, and the activation of p38 MAPK was significantly (P<0.05) inhibited 10 minutes after reperfusion compared with the control group. CONCLUSIONS: The addition of FR167653 to Celsior solution improved heart-graft viability, probably by way of the inhibition of p38 MAPK activation, which may attenuate ischemia-reperfusion injury in heart transplantation from NHBDs.     

Cardiac transplantation following a 24-h preservation using a perfusion apparatus

BACKGROUND: We developed a new apparatus for heart preservation and have already reported successful transplantation following 12 h of preservation using this apparatus. The efficacy of coronary perfusion with an oxygenated Celsior solution was investigated through transplantation following 24 h of preservation using the apparatus. MATERIALS AND METHODS: After being harvested, grafts were preserved with a combination of immersion in a 4 degrees C Celsior solution and perfusion with an oxygenated Celsior solution using the apparatus in the coronary perfusion (CP) group and simply immersed in a 4 degrees C Celsior solution in the simple immersion(SI) group. beta-Adenosine triphosphate (beta-ATP), phosphocreatine (Pcr), and inorganic phosphate (P(i)) levels and myocardial pH (pH(i)) were measured immediately after the heart was excised and at 12 and 24 h after preservation. Following preservation, orthotopic transplantation was performed. Cardiac function was measured 2 h after weaning from cardiopulmonary bypass (CPB). RESULTS: beta-ATP/P(i), Pcr/P(i), and pH(i) levels were significantly higher in the CP group than in the SI group at 12 and 24 h after preservation. Four of six animals in the CP group and two of six in the SI group were successfully weaned from CPB. The recovery rates of cardiac function were better in the CP group than in the SI group. CONCLUSION: Twenty-four hours of heart preservation may be possible with a combination of immersion in a 4 degrees C Celsior solution and perfusion with an oxygenated Celsior solution using the perfusion apparatus.     

Effects of Celsior and University of Wisconsin preservation solutions on hemodynamics and endothelial function after cardiac transplantation in humans: a single‐center,prospective, randomized trial

Abstract Optimal preservation of the myocardium remains a major concern in clinical and experimental heart transplantation. The present study compared the efficacy of University of Wisconsin (UW) and Celsior preservation solution with respect to myocardial performance, epicardial and microvascular endothelial vasomotor function and myocardial expression of endothelin and nitric oxide synthases in humans. Forty‐one cardiac transplant recipients received either UW (n = 20) or Celsior (n = 21) preserved hearts. Catecholamine and vasodilator requirements were assessed within the first 5 postoperative days. Left ventricular performance and endothelial function was assessed 1 month after transplantation. Endothelin and nitric oxide synthase gene expression were detected in myocardial biopsy samples. Celsior preserved hearts required significantly more catecholamines and vasodilators within the first 5 postoperative days. Myocardial performance and endothelial function were comparable 1 month after transplantation. Total ischemic time correlated with impaired endothelial function in the Celsior but not in the UW group. Endothelin and inducible nitric oxide synthase gene expression were significantly higher in the Celsior group. The results of the study show that both solutions provide myocardial protection with regard to left ventricular performance and endothelial function 1 month after cardiac transplantation. The necessity for higher vasodilator and catecholamine therapy in Celsior preserved hearts suggests post‐ischemic myocardial stunning within the first 5 postoperative days. The positive correlation between impaired endothelial function and total ischemic time in the Celsior group requires longitudinal investigation in particular with regard to the development of allograft vasculopathy.     

Luminal preservation of rat small intestine with University of Wisconsin or Celsior solution

AIMS: Luminal administration of a preservation solution that prevents mucosal injury may decrease posttransplant complications. However, luminal administration of University of Wisconsin solution (UW) is controversial. In this study, we examined the potential of Celsior as a luminal small bowel preservation solution in comparison to UW or UW enriched with glutamine. METHODS: Small bowels of six normal WagRij rats were excised and divided into six equal segments. Each segment was luminally flushed with 10 mL ice-cold UW, UW with glutamine (20 g/L) or Celsior, and stored for 0, 2.5, and 24 hours at 4 degrees C. LDH, glucose, and lactate concentrations were determined in the preservation solutions. Histologic changes were determined using the Park score. RESULTS: Lactate dehydrogenase (LDH) was increased in all solutions after 2.5- and after 24-hour preservation. However, LDH was lower in Celsior than UW and UW with glutamine. Furthermore, higher glucose and lactate levels were found after 2.5- and 24-hour preservation in UW and UW with glutamine compared to Celsior. Histologically, jejunal segments were more susceptible to preservation than ileal segments, irrespective of the preservation solution used. Mucosal injury was evident after 2.5 hours (Park Scale 0-3) and increased significantly after 24 hours (park scale 3-6). CONCLUSIONS: Based on the lower glucose, lactate, and LDH levels in small intestines stored in Celsior, this study suggests that Celsior is a better luminal preservation solution than UW. Unfortunately, histological evaluations still show severe mucosal injury, indicating that there is a need for better luminal preservation solutions or for concomittant intravascular delivery of a preservation solution.     

FK409 ameliorates ischemia-reperfusion injury in heart transplantation following 12-hour cold preservation.

OBJECTIVE: FK409 is the first spontaneous nitric oxide donor to increase plasma guanosine 3':5'-cyclic monophosphate. We designed this study to investigate whether the administration of FK409 during reperfusion ameliorated ischemia-reperfusion injury and enhanced post-transplant graft function in orthotopic heart transplantation following 12-hour cold preservation in a canine model. METHODS: We used 10 pairs of adult mongrel dogs, weighing 9.5 to 13.5 kg. Following cardiac arrest using cardioplegia, we washed out the coronary vascular beds with cold University of Wisconsin solution followed by 12-hour preservation. After preservation, we performed orthotopic transplantation. The experimental animals were divided into 2 groups. In the FK group (n = 5), FK409 (5 microg/kg/min) was administered intravenously, beginning 15 minutes before the onset of reperfusion and continuing for 45 minutes after reperfusion. In the control group (n = 5), saline vehicle was administered in the same manner. Two hours after transplantation, we assessed cardiac function, including cardiac output, left ventricular systolic pressure (LVP), and the maximum rates of positive and negative increase of LVP (+/-LV dP/dt) by comparing the recovery rate (%) of the cardiac function of the donor animal. We measured endothelin-1 levels in blood obtained from a catheter inserted into the coronary sinus 30, 60, and 120 minutes after reperfusion. RESULTS: Cardiac output was higher in the FK group than in the control group, but the difference was not significant (p = 0.08). Left ventricular systolic pressure and +/-LV dP/dt were significantly (p < 0.05) higher in the FK group than in the control group. Endothelin-1 levels were significantly (p < 0.05) lower in the FK group than in the control group 30 minutes after reperfusion. Transmission electron microscopy showed that the basal lamina of capillary vessels, glycogen granules, and mitochondrial structure were well-preserved in the FK group. CONCLUSIONS: In orthotopic transplantation models, FK409 is effective in ameliorating ischemia-reperfusion injury following preservation and in enhancing post-transplant cardiac function.     

Two-layer method (UW solution/perfluorochemical plus O2) for lung preservation in rat lung transplantation

A new preservation method using perfluorochemicals (PFC) with oxygen administered continuously was developed for lung preservation and compared with traditional cold preservation methods for rat lung transplantation. Male Sprague-Dawley rats underwent orthotopic left lung transplantations of grafts preserved in lactiated Ringers solution (LR), University of Wisconsin solution (UW), Celsior solution, or a two-layer (PFC plus O2) solution for 6 hours. One hour after reperfusion, the right pulmonary artery and bronchus were clamped and 5 minutes later we recorded peak airway pressure and PaO2 level. The isograft was excised for measurement of myeloperoxidase activity, wet-to-dry ratio, and histologic examination to evaluate isograft function. The mean peak airway pressure was 29.80+/-6.72 mm H2O in the LR group, 28.80+/-5.76 mm H2O in the UW group, 33.60+/-5.17 mm H2O in the Celsior group, and 32.40+/-2.60 in the two-layer group. The mean PaO2 level was 99.78+/-76.09 mm Hg in the LR group, 87.84+/-33.58 mm Hg in the UW group, 104.50+/-72.93 mm Hg in the Celsior group, and 62.08+/-31.34 mm Hg in PFC and UW solution plus O2 group (two layers). The mean net myeloperoxidase activity OD level was 0.110+/-0.104 in the LR group, 0.392+/-0.328 in the UW group, 0.351+/-0.620 in the Celsior group, and 0.532+/-0.616 in the two-layer group. The mean wet-to-dry ratio was 7.47+/-1.60 in the LR group, 6.56+/-0.62 in the UW group, 7.54+/-2.19 in the Celsior group, and 5.32+/-2.20 in the two-layer group. The differences between groups in these parameters were not significant. Upon histologic examination, more inflammatory cell aggregates were seen in the two-layer group, less in the LR and the Celsior groups. The function of the lung graft after 6 hours of storage was not better using this two-layer method for preservation than traditional preservation methods in rat lung transplantation. Histologic examination revealed more inflammatory cell aggregates in the lung graft preserved using a two-layer method.     

Celsior solution provides superior post-ischemic right ventricular function as compared with UW solution in a porcine heart transplantation model.

BACKGROUND: Use of the new cardioprotective Celsior solution has been suggested for organ preservation in cardiac transplantation, but selective data for right ventricular function, of special interest in the clinical setting, have not been evaluated. METHODS: Celsior solution was compared with the clinical standard University of Wisconsin solution (UW) in a porcine allogenic heart transplantation model with accurate isovolumic measurement of right ventricular (RV) function. RESULTS: Maximum RV developed pressures were significantly different between Celsior and UW groups (51.1 +/- 9.6 mm Hg vs 42.2 +/- 15.4 mm Hg after 1 hour, respectively, and 55.6 +/- 7.8 mm Hg vs 45.1 +/- 16.2 mm Hg after 2 hours, respectively; p = 0.02, 2-way analysis of variance). CONCLUSIONS: Celsior significantly improves post-ischemic right ventricular function when compared with UW solution in an experimental heart transplantation model.     

Improved preservation of the rat liver for orthotopic liver transplantation: use of University of Wisconsin-lactobionate solution and retrograde reflushing

This study investigated cold preservation and reflushing before orthotopic liver transplantation by examining (1) new University of Wisconsin solution (UW) versus Euro-Collins solution (EC), (2) retrograde reflushing (RR) versus antegrade reflushing (AR), and (3) the addition of a platelet-activating inhibitor (PAF), superoxide disumatase (SOD), or SOD + catalase to UW. Syngeneic, male Lewis rats (200 to 400 gm) were used. Preservation for 9, 12, 18, or 24 hours in UW or EC with RR (through the inferior vena cava) was used. The 9- and 12-hour groups experienced a significant decrease in the weight of the grafts preserved in UW. The 3-week survival rate after 9 hours of preservation (n = 6) in UW was 66%, and the survival rate with EC was 0% (p less than 0.025). After 12 hours of preservation, recipient survival rate was 70% (n = 10) with UW versus 0% (n = 4) with EC (p less than 0.025). RR of the graft with cold lactated Ringer's solution immediately before reimplantation significantly improved 3-week survival in the 12-hour group to the level of the control group (no preservation time, 69%). Preservation for 12 hours in UW followed by AR yielded a 3-week survival of 14%; 3-week survival for the RR group was 70% (p less than 0.025). Furthermore, RR allowed a 3-week survival of 33% and 20% after 18 and 24 hours of UW preservation, respectively. In the 24-hour RR/UW group, donor pretreatment with SRI 63-441 (20 mg/kg, intravenously) and recipient treatment with SOD (15 mg/kg, intravenously) or SOD + catalase (15 mg/kg and 5000 units/kg, intravenously) produced a 3-week survival comparable to preservation in UW followed by RR alone. These studies show that UW is a profound improvement over EC for cold preservation of liver and that the new application of RR to rat orthotopic liver transplantation improves survival. However, the addition of free-radical scavengers or PAF does not improve organ function or recipient survival in this model.     

Long-term preservation using a new apparatus combined with suppression of pro-inflammatory cytokines improves donor heart function after transplantation in a canine model.

OBJECTIVE: We developed a new apparatus for long-term heart preservation that combines simple immersion with coronary perfusion. In a previous study, we reported that suppression of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), improved results after transplantation. In this study, we evaluated whether long-term preservation using our apparatus for continuous coronary perfusion, combined with suppression of pro-inflammatory cytokines, improves donor heart function after transplantation in a canine model. METHODS: We used adult mongrel dogs in this study. Coronary vascular beds were washed with University of Wisconsin (UW) solution after arresting hearts with glucose-insulin-potassium solution. The heart was then excised and preserved for 12 hours with a combination of immersion and coronary perfusion using a preservation apparatus. Adult mongrel dogs were divided into 2 groups: the coronary perfusion (CP) group (n = 7) and the FR167653 (FR-CP) group (n = 6). In the CP group, we used a 4 degrees C UW solution for immersion and coronary perfusion. In the FR-CP group, we used a 4 degrees C UW solution supplemented with 20 mg/liter of the anti-inflammatory agent FR167653 for immersion and coronary perfusion. At 2 and at 3 hours after orthotopic transplantation, we compared hemodynamic parameters with pre-operative values in donor animals, with right atrial pressure at 10 mm Hg and with 5 microg/kg/min dopamine infusion. We compared serum concentrations of TNF-alpha from the coronary sinus and compared electron microscopic studies between the 2 groups. RESULTS: Three hours after transplantation, cardiac output (CO), left ventricular pressure (LVP), and -LVdp/dt were significantly greater (p < 0.05) in the FR-CP group than in the CP group (CO, 178% +/- 65% vs 93% +/- 40%; LVP, 115% +/- 22% vs 73% +/-26%; -LVdp/dt, 168% +/- 13% vs 61% +/- 17%, respectively). Electron microscopic studies showed that glycogen was well preserved in the FR-CP group compared with the CP group. Serum concentrations of TNF-alpha were decreased significantly in the FR-CP group compared with the CP group at 3 hours after reperfusion (161 +/- 54 pg/dl vs 642 +/- 636 pg/dl, respectively). CONCLUSION: Hemodynamics after transplantation were significantly better in the FR-CP group than in the CP group. The combined preservation method of continuous perfusion and immersion using our apparatus in conjunction with suppression of pro-inflammatory cytokines improves donor heart function after transplantation.     

The effect of short-term coronary perfusion using oxygenated diluted blood following cold storage for long-term heart preservation

BACKGROUND: The aim of this study was to compare the results obtained from the use of both University of Wisconsin (UW) solution and diluted blood in short-term coronary perfusion following 12-hour cold storage. METHODS: Following coronary vascular washout of adult mongrel dogs with the UW solution, the heart was excised and immersed in a cold (4 degrees C) UW solution for 12 hours followed by 1-hour of coronary perfusion. Two different solutions were used for the coronary perfusion; a 4 degrees C oxygenated UW solution (Group U, n=7) and 15 degrees C oxygenated diluted blood (Group B, n=7). Myocardial high energy phosphate (HEP) levels, tissue water content (TWC), interstitial tissue space (ITS) rates and histological findings were evaluated at 0- and 12-hour cold storage and also following coronary perfusion. The preserved graft was then evaluated through orthotopic transplantation. The control group in this experiment consisted of seven hearts transplanted after 12-hour cold storage without coronary perfusion. RESULTS: Myocardial HEP levels significantly decreased after 12-hour cold storage. The recovery rate of myocardial HEP levels after coronary perfusion was significantly (p<0.05) higher in Group B than in Group U. The increase of myocardial TWC during coronary perfusion was significantly (p<0.01) higher in Group B than in Group U. After 1-hour coronary perfusion, the subendocardial ITS rate was significantly (p<0.01) higher compared with the value at 0-hour cold storage in Group U, whereas it demonstrated no significant change in Group B. PAS stain revealed the glycogen content of the subendocardial tissues was higher in Group B than in Group U. The recovery rate of hemodynamic parameters 2 hours after heart transplantation was higher in Group U and significantly (p<0.05) higher in Group B than in the control. CONCLUSIONS: Myocardial HEP levels recovered significantly after additional coronary perfusion. Though the UW solution prevented myocardial cellular edema, subendocardial perfusion was incomplete and the recovery rate of myocardial HEP levels was lower, suggesting that diluted blood may become the solution of choice as a perfusate.     

A new model for human intestinal preservation: comparison of University of Wisconsin and Celsior preservation solutions

AIM: We compared University of Wisconsin (UW) and Celsior preservation solutions using a new model of human intestinal preservation that mimics the clinical conditions of small bowel procurement. METHODS: Intestinal grafts were harvested from four multiorgan donors. After classic warm dissection for organ procurement, an ileal segment of 50 cm was immediately flushed with Celsior. After the perfusion of the abdominal organs with UW, a second segment of adjacent ileum was harvested. The two intestinal grafts were then divided into segments by stapling, before immersion into the corresponding preservation solution (Celsior or UW) for 0-, 6-, 12-, or 24-hour incubation at 4 degrees C. A histological score was graded after blinded examination of three random specimens within each ileal graft for each duration of preservation. RESULTS: Control specimens showed normal histology. After 6 hours of preservation, most villi showed complete epithelial detachment although the crypts appeared intact. After 12 hours of preservation, a larger proportion of the villi showed extensive epithelial sloughing. After 24 hours, the damage involved the entire mucosa with the crypt epithelium largely detached from the basal membrane. No statistical difference in histological score was observed between the two preservation solutions. CONCLUSION: This study showed severe histological alterations of graft mucosa after short periods of preservation by UW or Celsior solutions. This model may be useful to evaluate improvements in the quality of preservation of human intestinal transplants.     

Advantages of Celsior solution in graft preservation from non-heart-beating donors in a canine liver transplantation model.

BACKGROUND: The optimal method for preserving livers from non-heart-beating donors (NHBD) is still unknown. We compared the Celsior solution, a new extracellular-type, low-potassium, low-viscosity preservation solution, with the University of Wisconsin (UW) solution in a canine orthotopic liver transplantation from NHBD. MATERIALS AND METHODS: Fourteen adult mongrel dogs, weighing 9 to 17 kg, were divided into two groups: the Celsior or the UW group (n = 7 each). The thoracic descending aorta and supradiaphragmatic inferior vena cava were cross-clamped for 20 min to induce warm ischemia as a NHBD model. The liver was flushed with the respective cold preservation solution and then stored at 4 degrees C for 4 h. The grafts were transplanted using the piggy-back technique under portal decompression by leaving the native right lobe as a temporary shunt. RESULTS: The duration of liver flushing out (min) was shorter (P < 0.05), and the serum glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactate dehydrogenase, lactate, and alpha-glutathione S-transferase concentrations 2 and 6 h after reperfusion of the graft (RPF) were lower (P < 0.05) in the Celsior group than in the UW group. Hepatic tissue blood flow (HTBF) did not deteriorate as much (P < 0.05) in the Celsior group. The serum endothelin-1 level was lower (P < 0.05) in the Celsior group 2 h after RPF. Histopathology of liver specimens revealed portal congestion and hepatocyte necrosis with neutrophil infiltration in the UW group, while these findings were mild in the Celsior group. CONCLUSIONS: The Celsior solution improves vascular endothelial injury in livers from NHBDs and may have advantages in graft flush and preservation of grafts from NHBDs.     

Ischemic preconditioning with opening of mitochondrial adenosine triphosphate-sensitive potassium channels or Na/H exchange inhibition: which is the best protective strategy for heart transplants?

OBJECTIVE: This study was designed to compare ischemic preconditioning with opening of mitochondrial adenosine triphosphate-sensitive potassium channels and Na(+)/H(+) exchange inhibition in an isolated heart model of cold storage, simulating the situation of cardiac allografts. METHODS: Sixty-seven isolated isovolumic buffer-perfused rat hearts were arrested with and stored in Celsior solution (Imtix-Sangstat) at 4 degrees C for 4 hours before a 2-hour reperfusion. Group I hearts served as controls and were arrested with and stored in Celsior solution. In group II, hearts were preconditioned by two 5-minute episodes of global ischemia, each separated by 5 minutes of reperfusion before arrest with Celsior solution. Group III hearts were arrested with and stored in Celsior solution supplemented with 100 micromol/L of the mitochondrial adenosine triphosphate-sensitive potassium channel opener diazoxide. In group IV, hearts received an infusion of diazoxide (30 micromol/L) during the first 15 minutes of reperfusion. Group V hearts underwent a protocol combining both interventions used in groups III and IV. In group VI, hearts were arrested with and stored in Celsior solution supplemented with 1 micromol/L of the Na(+)/H(+) exchange inhibitor cariporide. Group VII hearts received an infusion of cariporide (1 micromol/L) during the first 15 minutes of reperfusion. In group VIII, hearts underwent a protocol combining both interventions used in groups VI and VII. Group IX hearts were ischemically preconditioned as in group II, and sustained Na(+)/H(+) exchange inhibition during both storage and early reperfusion was used as in group VIII. RESULTS: On the basis of comparisons of postischemic left ventricular contractility and diastolic function, coronary flow, total creatine kinase leakage, and myocardial water content, values indicative of improved protection were obtained by combining ischemic preconditioning with Na(+)/H(+) exchange inhibition by cariporide given during storage and initial reperfusion. The endothelium-dependent vasodilatory postischemic responses to 5-hydroxytryptamine or acetylcholine and endothelium-independent responses to papaverine were not affected by these interventions. CONCLUSIONS: These data suggest that cardioprotection conferred by the Na(+)/H(+) exchange inhibitor cariporide is additive to that of ischemic preconditioning and might effectively contribute to improve donor heart preservation during cardiac transplantation.     

Long-term heart preservation using a new portable hypothermic perfusion apparatus.

OBJECTIVE: Perfusion storage is not often used clinically compared with simple immersion because of complicated circuits and demanding management. We developed a new apparatus for preservation combined with simple immersion and continuous coronary perfusion. METHODS: The main characteristics of this apparatus are as follows: (1) hypothermic storage, (2) does not require any energy source, (3) variable perfusion pressure, and (4) portability. The perfusion apparatus is composed of a storage chamber, a cooling chamber, and metal bars from which a perfusate bag is suspended. Adult mongrel dogs were divided into two groups: the coronary perfusion group (CP, n = 6) and the simple immersion group (SI, n = 6). Coronary vascular beds of the dog were washed out with a University of Wisconsin (UW) solution following cardiac arrest obtained using a GIK solution. The hearts were then excised. In the CP group, the heart graft, which was immersed in a 4 degrees C UW solution, was perfused with the same solution at a flow rate of 35 approximately 50 ml/hr. In the SI group, the heart graft was immersed in a 4 degrees C UW solution only. The heart graft was preserved for 12 hours in both groups. Beta-adenosine triphosphate (beta-ATP), phosphocreatine (Pcr), and inorganic phosphate (Pi) levels were measured immediately after excision of the heart, and at 3, 6, and 12 hours after preservation. Beta-ATP, Pcr, and Pi values were expressed as a percentage of control values, which had been obtained immediately after excision of the heart. Water content of the myocardium was measured prior to and after 12-hour preservation. The preserved graft was then evaluated through orthotopic transplantation. RESULTS: Beta-ATP/Pi levels at 6 and 12 hours after preservation were significantly higher in the CP group than in the SI group (62 +/- 5 versus 39 +/- 7%, 48 +/- 5 versus 22 +/- 8%, respectively, p < 0.05). Pcr/Pi levels at 6 and 12 hours after preservation were 30 +/- 9% and 22 +/- 8%, respectively in the CP group, while Pcr/Pi levels in the SI group were detected in only one case. There was no significant difference in water content either prior to or after 12-hour preservation between the two groups. Histopathologically, irregular expansion and/or contraction of myocardial fibers were more severe in the SI group than in the CP group. The recovery rate of hemodynamic parameters 2 hours after heart transplantation was significantly (p < 0.05) higher in the CP group than in the SI group. CONCLUSION: Stable and safe long-term canine heart preservation with continuous coronary perfusion associated with immersion is possible using this new apparatus, and may have broad clinical application.     

Influence of preservation solution on human islet isolation outcome

BACKGROUND: The influence of the preservation solution used for in situ perfusion of the donor and pancreas storage on islet isolation has received little attention. METHODS: In this prospective controlled trial, we compared the outcome of human islet isolation from pancreata perfused with University of Wisconsin (UW) solution or Celsior, an alternative colloid-free extracellular solution. RESULTS: At the 1-year interim analysis, the viability and insulin secretion of islets isolated from donors perfused with UW (n=19) or Celsior (n=5) were identical. However, total islet recovery (IEQ) and isolation yield (IEQ/g) were 1.8-fold and 2.1-fold inferior in the Celsior group (P<0.05 vs. UW). Overall, 13 (68%) of islet preparations were effectively transplanted from the UW group vs. none from the Celsior group (P=0.01). The clinical study was discontinued and the causes of these differences were further explored in the pig (n=14). In contrast to UW, Celsior induced cell swelling and pancreas edema after only four hours of cold storage. These abnormalities were delayed when the donor was perfused with Solution de Conservation d'Organes et de Tissus (SCOT), an extracellular solution containing polyethylene glycol. CONCLUSIONS: Our results suggest that colloid-free preservation solutions might be suboptimal for pancreas perfusion and cold storage prior to islet isolation and transplantation. Because pancreata are now frequently recovered for islet transplantation, preliminary experimental and clinical data about islet isolation should be obtained prior to the routine implementation of new preservation solutions for abdominal perfusion during multiorgan recovery.     

The effect of graft perfusion with warm blood cardioplegia for cadaver heart transplantation

This study was designed to verify the effect of reperfusion of donor hearts in a perfusion apparatus after 60 min of global ischemia prior to heart transplantation. Thirteen dogs were exsanguinated from the femoral artery and cardiac arrest was achieved. The hearts were left in situ at room temperature (25°C)for 60 min. In group A (n=7), the hearts were excised and reperfused 60 min after cardiac arrest in the perfusion apparatus with substrate-enriched warm blood cardioplegia (WBCP) containing a hydroxyl radical scavenger, EPC, followed by 45 min of blood perfusion, Next, the hearts were preserved in cold (4°C) University of Wisconsin (UW) solution. In group B (n=6), the hearts were perfused with cold (4°C) St. Thomas' solution 60 min after cardiac arrest and preserved in cold UW solution. Thereafter, all hearts in both groups were transplanted orthotopically to recipient dogs. In group A, 6 of 7 dogs were weaned from cardiopulmonary bypass (CPB). In group B, only 2 of 6 dogs were weaned from CPB. Moreover, 3 of the 6 hearts in group B did not start beating after transplantation (stone heart). This study suggested reperfusion of the donor heart in the perfusion apparatus with WBCP to be a beneficial preconditioning method when utilizing 60-min arrested hearts for transplantation. This study was supported in part by Senju Pharmaceutical Co. Ltd., Osaka, Japan     

Pancreas preservation with University of Wisconsin and Celsior solutions: a single-center, prospective, randomized pilot study

BACKGROUND: Celsior is an extracellular-type, low-viscosity, preservation solution already used for heart, lung, liver, and kidney transplantation. We report the results of a single-center, prospective, randomized pilot study specifically designed to compare the safety profile of Celsior solution with University of Wisconsin (UW) solution in clinical pancreas transplantation. METHODS: A total of 105 consecutive procurements were randomized to graft preservation with UW (n=53) solution or Celsior (n=52) solution. The groups were comparable with regard to all donor and recipient characteristics. RESULTS: Five grafts were discarded and 100 grafts (50 UW vs. 50 Celsior) were transplanted. Mean cold and warm ischemia times were 11.0 +/- 2.1 hr and 37.2 +/- 6.0 min for UW compared with 10.8 +/- 1.8 hr and 38.1 +/- 5.9 min for Celsior (P =not significant). Delayed endocrine pancreas function was recorded in one graft preserved with UW solution. Eleven recipients (UW 12% vs. Celsior 10%, P =not significant) required a relaparotomy. The mean serum levels of glucose, amylase, and lipase remained comparable between the study arms at equivalent intervals after transplantation. One recipient died with functioning grafts in each study arm; two further grafts were lost to arterial thrombosis (Celsior) and chronic rejection (UW), respectively. Actuarial 1-year patient and graft survival rates overlapped in the two study arms (98% and 96%, respectively). CONCLUSIONS: Within the range of cold ischemia time reported in this study, UW and Celsior solutions have similar safety profiles for pancreas preservation.