Nutrient-related issues affecting successful experimental orthotopic small bowel transplantation

BACKGROUND: This study tested the effectiveness of a nutrient-rich preservation solution in a small animal model of orthotopic whole small bowel transplantation. METHODS: Lewis rats received syngeneic total orthotopic small bowel graft after cold storage for 6 h. Donor small bowel was flushed vascularly with University of Wisconsin (UW) solution and flushed luminally with UW solution or an amino acid-rich (AA) solution as follows: Group 1, no luminal flush; Group 2, UW solution; Group 3, AA solution. Biopsies were taken over 14 days posttransplant; energetics, oxidative stress, neutrophil recruitment and histologic injury were assessed. RESULTS: All animals in Groups 1 and 2 failed to survive 12 h posttransplant due to hemorrhagic shock and fluid loss. In contrast, all animals in Group 3 survived the operation; survival after 14 days was 80% (4/5). In Group 3, full recovery of tissue adenylates (ATP and energy charge) to freshly isolated tissue values occurred within 3 days. Oxidative stress as assessed by malondialdehyde (MDA) levels was low in Group 3 throughout 14 d; Groups 1 and 2 exhibited high oxidative stress over the initial 35 min reperfusion (P<0.05). Neutrophil recruitment (myeloperoxidase activity) was significantly reduced in Group 3 tissues, as was histologic injury (P<0.05 compared to Groups 1 and 2). By day 14, Group 3 exhibited complete mucosal restoration. CONCLUSIONS: The data presented in this communication supports the use of an intraluminal preservation solution that is tailored to the metabolic requirements of the small bowel.     

Ameliorating Small Bowel Injury Using a Cavitary Two-Layer Preservation Method with Perfluorocarbon and a Nutrient-Rich Solution

The aim of this study was to improve small bowel (SB) quality during cold storage by combining two proven preservation strategies involving perfluorocarbon (PFC) and a novel luminal amino acid-rich solution. Rodent SB was flushed vascularly with UW solution and flushed luminally as follows: Group 1 (control)--no luminal flush, stored in UW; Group 2--luminal UW solution, stored in PFC; Group 3--luminal amino-acid (AA) solution, stored in PFC; and Group 4--luminal AA solution, stored in AA solution. Energetics, histology and mucosal function/electrophysiology were assessed over 24 h at 4 degrees C. ATP was consistently greater in Groups 2-4 than in the Control Group. Groups 3 and 4 exhibited significantly greater ATP, ATP/ADP ratios and energy charge levels after 12-h storage than in the other Groups. Histologic injury was generally lower in the AA-treated tissues (Groups 3 and 4); after 24 h, only minor epithelial clefting (Park's median grade 2) was present in Group 4; and consistent transmural infarction (grade 8) was evident in Groups 1 and 2. Combined treatment with luminal amino acid solution and oxygenated storage solution (PFC or AA solution) significantly improves energetics and mucosal function. This strategy may have implications for successful SB preservation in the clinic.     

Influence of Pituitary Adenylate Cyclase-Activating Polypeptide on the Activation of Mitogen Activated Protein Kinases Following Small Bowel Cold Preservation

Cold preservation prior to small bowel transplantation can moderate tissue oxidative injury. This stress triggers several intracellular pathways via mitogen activated protein (MAP) kinases. MAP kinases include the extracellular signal related kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAP kinase. Pituitary adenylate cyclase-activating polypeptide (PACAP) plays a central role in intestinal physiology. We sought to investigate the effect of PACAP on the activation of MAP kinases during cold preservation of the small bowel. Total orthotopic intestinal autotransplantation was performed on 40 Wistar rats. Perfused grafts were stored in University of Wisconsin (UW) solution for 1 (GI), 2 (GII), 3 (GIII), or 6 hours (GIV) without or with 30 PACAP, namely 1 (GV), 2 (GVI), 3 (GVII), or 6 hours (GVIII). After 3 hours of reperfusion in all groups, the activation of MAP kinases were measured using immunocytochemistry of small bowel tissue. Among the UW preserved grafts (GI-GIV), phosphorylated ERK1/2 level were decreased, while phosphorylated JNK1/2 and p38 MAP kinase activation were elevated compared with control levels. In GV-GVIII PACAP we observed enhanced phospho-ERK1/2 appearance with decreased JNK and p38 MAP kinase activity at the end of the reperfusion periods. We concluded that cold preservation decreased phosphorylated ERK1/2 levels and increased JNK1/2 and p38 MAP kinase activities, which meant that cold storage triggered apoptotic cell death. In contrast, PACAP treatment induced signalling pathways protective against oxidative injury by MAP kinases in bowel tissue.     

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.     

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.     

Tolerance limits of liver grafts with 30 minutes of warm ischemia to cold preservation in swine

AIM: The aim of this study was to investigate the safe time limits of cold preservation in University of Wisconsin (UW) solution of liver grafts subjected to warm ischemia (WI) for 30 minutes from non-heart-beating donors (NHBDs). METHODS: The safe time limits were studied in a simple porcine orthotopic liver transplantation (LTx) model. In donors, livers were subjected to 30 minutes of WI and subsequent 6-hour (Group 1, n = 5), 10-hour (Group 2, n = 5), and 14-hour (Group 3, n = 3) cold preservation in UW solution. RESULTS: All 5 animals in Group 1 survived up to 7 days, the survey endpoint. In Group 2, only 2 animals survived to the same survey endpoint. All animals in Group 3 died within 12 hours. The 1-week survival rate of Group 1 was significantly higher than those of the other 2 groups. Group 1 showed a lower level of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) after LTx, less pathological damage, higher concentration of adenosine triphosphate (ATP), and higher microcirculation blood flux in the grafted liver tissue at 1 hour after reperfusion than the other 2 groups. CONCLUSIONS: It is concluded that about 6 hours is the safe time limit of cold preservation in UW solution for liver grafts from NHBDs subjected to WI for 30 minutes.     

A comparative study of Celsior and University of Wisconsin solutions based on 12-hr preservation followed by transplantation in canine models

BACKGROUND: Celsior is a recently developed extracellular-type preservation solution that is effective in organ preservation. This experimental study was designed to compare the effects of Celsior and University of Wisconsin (UW) solutions in myocardial protection, using 12-hour preservation followed by orthotopic transplantation. METHODS: Fourteen pairs of adult mongrel dogs were divided into 2 groups. In the UW group (n = 7), UW solution at 4 degrees C was used for coronary vascular washout and storage following cardiac arrest with glucose-insulin-potassium (GIK) solution. In the Celsior group (n = 7), Celsior solution was used to produce cardiac arrest, for coronary vascular washout, and for storage. After 12-hour cold preservation, orthotopic transplantation was performed under cardiopulmonary bypass (CPB). The rate of recovery (%) of cardiac function of donor hearts was compared 1 and 2 hours after weaning from CPB, and then the transplanted hearts were harvested for histological study. RESULTS: Hemodynamic parameters including cardiac output, left ventricular pressure (LVP), and the maximum rates of positive and negative increase of LVP after transplantation were significantly (p < 0.05) higher in the Celsior group than in the UW group 2 hours after weaning from CPB. The transmission electron microscopic study found that degeneration of the mitochondria in the Celsior group was less extensive than in the UW group. CONCLUSION: Celsior solution enhanced the cardiac function of hearts preserved for 12 hours prior to transplantation compared to UW solution. Our results indicate that Celsior solution is equivalent or superior to UW solution for cardiac preservation.     

Effect of fructose-1,6-bisphosphate in the cold storage solution after 12 and 36 hours of rat liver preservation

Fructose-1,6-bisphosphate (FBP) has been reported to have a protective effect on liver injury following ischemic/reperfusion periods because it maintains ATP levels during cold preservation. In the present study, we evaluated the effects of addition of FBP to storage solutions for cold liver preservation during 12 or 36 hours. Adult male Wistar rats were randomly divided into three experimental groups. The hepatic perfusion and preservation were performed with these solutions: UW; UW plus 10 mmol/L FBP; and FBP 10 mmol/L (FBPS) alone. The biochemical measurements of AST and ALT were performed on samples of the cold storage solution after 12- or 36-hour preservation. UW and FBPS solutions showed similar preservation grades at 12 hours. Addition of 10 mmol/L of FBP to UW solution induced liver injury and a poor preservation grade during 12 or 36 hours. UW solution was better than FBPS after 36 hours preservation. UW solution continues to offer a superior performance for liver preservation during long times; however, FBPS may be an alternative for short cold preservation times.     

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.     

供肝热缺血后对冷保存的耐受时限初步探讨

目的探讨供肝热缺血后耐受冷保存的安全时限。方法利用本组所建立的小型猪肝移植模型,设定供肝热缺血时间为20min,根据在UW液中的冷保存时间不同分为3组,分别冷保存12、16、20h,于肝移植术中及术后检测肝功能、肝脏病理、肝组织ATP含量、移植肝脏再灌注后微循环血流量及动物术后1周存活率。结果UW液冷保存12h组肝移植后小型猪1周内全部存活,而冷保存16、20h组存活率分别为20%、0%;随着冷保存时间的从12h延长到20h,ALT、AST逐渐上升,肝脏ATP含量、肝脏微循环血流量逐渐下降,形态学结果显示肝组织细胞变性、坏死及超微结构损害的程度逐渐加重。冷保存12h组与后两组上述指标存在显著性差异,生化及肝脏微循环指标的改变与病理结果及动物生存率相符合。结论在本实验条件下,热缺血时间为20min的供肝耐受冷保存的安全时限约为12h。     

Human small bowel storage: the role for luminal preservation solutions

BACKGROUND: Graft injury incurred during periods of cold storage remains a factor affecting the success of small bowel (SB) transplantation. No one preservation solution, including the gold standard University of Wisconsin (UW) solution, has been able to maintain graft integrity for storage periods paralleling that of other commonly transplanted intra-abdominal organs. We investigated the role for the luminal administration of preservation solutions in a small animal model, documenting significantly improved graft quality. The current study addresses direct clinical applicability using human SB. METHODS: Human SB was obtained at the time of standard multiviscera procurement. After a common intra-arterial UW flush, the SB was immediately removed from the abdomen, randomly divided into three segments, and treated as follows (n=6-9): group 1, no luminal flush; group 2, luminal flush with UW solution; and group 3, luminal flush with an amino acid- enriched solution. Analysis of cellular energetics, permeability, and histologic injury was performed throughout 24 hr of cold storage. RESULTS: Mucosal barrier function, measured by mannitol permeability, was significantly better overall in groups 2 and 3, with 24-hr values measuring 31 and 34 nmol/cm2/hr versus 57 nmol/cm2/hr, respectively (both P<0.05). Significantly less morphologic injury was also noted in the luminally treated specimens (groups 2 and 3) compared with the clinical standard (vascular flush with UW solution). Damage in group 1 reached gross villus denudation with an obvious elevated risk of villus tissue loss, whereas groups 2 and 3 only exhibited epithelial clefting to varying degrees. CONCLUSION: This study supports luminal administration of preservation solutions for improvement of human SB graft quality during clinically relevant periods of cold storage.     

Luminal contact with University of Wisconsin solution improves human small bowel preservation

AIM: Under clinical conditions small bowel mucosa is stored without any contact between the mucosa and the preservation solution. We evaluated the impact of luminal contact with University of Wisconsin solution (UW) on the structural quality of small bowel preservation. METHODS: Segments of ileum harvested from stable multi-organ donors were flushed with UW. For each donor, ileal segments were placed in UW without any contact between the mucosa and the preservation solution (group A), as is practiced in clinical conditions. Adjacent segments were cut on their antimesenteric side and placed in UW so that their mucosa was widely in contact with the solution (group B). The grafts preserved in ice were removed from the preservation fluid at different times (0, 3, 6, or 12 hours). Tissues were studied by optical microscopy after H&E staining of formalin-fixed, paraffin-embedded specimens. A median histologic score was calculated after examination of three random slides for each ileal segment per time point. Comparisons were performed between tissues of groups A and B from the same donor. RESULTS: Control (0h) specimens showed normal histology. As early as 3 hours of preservation, group A tissues showed detachment of the villar epithelium. At 6 and 12 hours of preservation in this group further tissue alteration were obvious with complete epithelial detachment from the basal membrane of the villi. The histologic score of the segments preserved by luminal contact with UW was always significantly higher than the controls from the same donor. CONCLUSION: Luminal contact between the mucosa of intestinal grafts and UW improves the quality of human small bowel preservation.     

Effect of hyperbarically oxygenated-perfluorochemical with University of Wisconsin solution on preservation of rat small intestine using an original pressure-resistant portable apparatus

BACKGROUND: Perfluorochemicals (PFC) are chemical substances that have a higher oxygen solubility under hyperbaric oxygen (HBO) pressure. This study investigated the effect of cold HBO-PFC/University of Wisconsin (UW) solution on preservation of rat small intestinal graft. METHODS: We manufactured an air-tight, pressure-resistant tank made of stainless steel with high thermal conductivity. Rat ileal grafts were placed in a custom-made silicon-gum bag with UW solution, which was immersed in 5 atm HBO-PFC solution in the tank (Group P-5). The tank was kept at 4 degrees C. We compared the ATP concentration and mucosal permeability in Group P-5 with grafts preserved in 1 atm oxygenated-PFC/UW solution (Group P-1) and simple cold storage in UW solution (Group C). Histologic study was also performed. RESULTS: PO(2) in UW solution after 48 h preservation were 1852 +/- 37, 499 +/- 13, and 173 +/- 3 mmHg (Group P-5, P-1 and C, respectively, mean +/- SD). At 48 h of preservation, graft ATP concentration was significantly greater in Group P-5 compared to that in Group P-1 and Group C. Mucosal hyperpermeability as well as mucosal morphologic changes were also ameliorated in Group P-5. CONCLUSION: HBO-PFC can supply a greater amount of oxygen to UW solution. Indirect measures of oxygen metabolism such as ATP content and lactate production suggested improvement in maintaining graft oxygen metabolism.     

Pancreas preservation by the 2-layer cold storage method before islet isolation protects isolated islets against apoptosis through the mitochondrial pathway

BACKGROUND: Apoptosis in isolated islets has been implicated in primary nonfunction or early graft failure after islet transplantation. Recently, pancreas preservation by the 2-layer method (TLM) before islet isolation has been proved to improve the islet yield, quality, and transplant results not only in experimental models, but also in clinical settings. We examined the influence of TLM on apoptosis of isolated islets. METHOD: Rat islets freshly isolated and after pancreas preservation by TLM or conventional cold storage in University of Wisconsin solution (UW) were examined and compared. Islet apoptosis was assessed by TUNEL and annexin V assays. The apoptosis pathways involved were investigated by measurement of caspase 3, 8, and 9 activities and by immunoblotting for total and phosphorylated c-Jun NH2-terminal kinase (JNK) and p38. RESULTS: Islet apoptosis in the UW group was significantly increased compared with the fresh and TLM groups. Both caspase 3 and 9 activities in the UW group were higher than in the fresh and TLM groups with an approximate increase of 2- to 3-fold. On the other hand, there was no significant difference in caspase 8 activity among these 3 groups. JNKs were strongly activated both in the TLM and UW groups; although they were not activated in the fresh group, p38 was activated to almost the same levels in these 3 groups. CONCLUSIONS: Pancreas preservation by TLM before islet isolation protects isolated islets against apoptosis mainly through the mitochondrial pathway. Pancreas storage before islet isolation even with TLM triggers activation of JNKs in isolated islets.     

Successful 96-Hr cold-storage preservation of canine pancreas with UW solution containing the thromboxane A2 synthesis inhibitor OKY046.

Prostanoids, such as prostacyclin (PGI) and thromboxane A2 (TxA), have been recently suggested to play an important role in preservation-induced injury of pancreas grafts. We have previously shown that the TxA synthesis inhibitor OKY046 prevents a decrease of both the PGI/TxA ratio and blood flow in pancreas grafts after 24-hr preservation with Euro-Collins solution. In our present study, we analyzed whether OKY046 added to University of Wisconsin (UW) solution could extend successful cold-storage preservation of segmental canine pancreas grafts, compared with UW alone. We divided 30 dogs into four preservation groups: Group 1, UW solution for 72 hr (n = 7); Group 2, UW solution for 96 hr (n = 8); Group 3, UW solution plus OKY046 (10(-4) M) for 72 hr (n = 7); and Group 4, UW solution plus OKY046 (10(-4) M) for 96 hr (n = 8). After the cold storage period, segmental pancreas auto-transplantation with immediate completion pancreatectomy was done. Preservation was deemed successful if serum glucose less than 150 mg/dl was maintained for at least 5 days. Intravenous glucose tolerance tests and biopsies were done in those dogs with functioning grafts 14 days post-transplant. Successful preservation rates were as follows: Group 1, 57.1%; Group 2, 12.5%; Group 3, 100%; and Group 4, 75%. The mean K values (+/- standard error) were: Group 1, 1.54 +/- 0.13; Group 2, 0.59; Group 3, 1.54 +/- 0.14; and Group 4, 1.59 +/- 0.24 (not statistically different).(ABSTRACT TRUNCATED AT 250 WORDS)     

A novel technique of hypothermic luminal perfusion for small bowel preservation

BACKGROUND: This study improved small bowel preservation using University of Wisconsin (UW) solution in conjunction with hypothermic luminal perfusion. METHODS: Small bowels from Sprague-Dawley rats (n=4) were flushed vascularly with modified UW solution and flushed luminally: group 1, none (clinical control); group 2, UW solution; group 3, 1-hr oxygenated perfusion then static storage with UW; and group 4, 24-hr continuous oxygenated perfusion with UW. Energetics, lipid peroxidation, and histology were assessed during 24 hr at 4 degrees C. RESULTS: After 12 hr, adenosine triphosphate ranged from 0.5 to 0.8 micromol/g in groups 1 to 3 compared with 1.5 micromol/g in group 4. Even after 24 hr, levels in group 4 were more than twofold greater than levels in groups 1 to 3. Energy charge values ([adenosine triphosphate+adenosine diphosphate/2]/total adenylates) decreased from fresh tissue values of 0.69 in all groups except group 4 throughout 24-hr perfusion. Malondialdehyde (MDA; a product of lipid peroxidation) doubled within 4 hr in group 1 and remained high throughout storage. In groups 3 and 4, MDA levels increased as the time of perfusion increased; group 2 showed no elevated MDA levels at any time. After 12 hr, histologic integrity was superior in groups 3 and 4; however after 24 hr, the best Park's grade was observed in group 3 (median grade 4) compared with groups 1 (grade 7) and 4 (grade 6). CONCLUSIONS: Our data indicate that perfusion clearly improves tissue energetics; however, mucosal integrity is superior with only a brief 1-hr period of luminal perfusion, despite limited improvements in energetics.     

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.     

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.     

Comparison of histidine-tryptophan ketoglutarate and University of Wisconsin solutions as primary preservation in renal allografts undergoing pulsatile perfusion

INTRODUCTION: University of Wisconsin (UW) solution is the standard preservation solution for organ transplantation. Histidine-tryptophan ketogluatarate (HTK) solution has been used increasingly for kidney, pancreas, and liver transplantation. This study compared HTK and UW used during kidney procurement with subsequent pulsatile perfusion. METHODS: Between January and October 2003, 91 deceased renal and simultaneous kidney pancreas transplants were performed (UW, n = 41, and HTK, n = 50). There were no differences with regard to donor and recipient demographics or cold ischemia. RESULTS: Delayed graft function occurred in 3 (7%) of UW and 4 (8%) of HTK-preserved kidneys (P = NS). There were no significant differences between patient or graft survival. There was an anticipated difference between total preservative volumes used (HTK: 4.1 +/- 1.0 vs UW: 3.0 +/- 0.5; P < .005). CONCLUSION: UW and HTK appear to have similar efficacy in kidney preservation with pulsatile perfusion. HTK preservation solution can be used safely in conjunction with pulsatile preservation for cold storage of renal allografts.