raffinose and Pulmonary-Edema

Ischaemia/reperfusion (I/R) lung injury using University of Wisconsin solution (UW) as perfusate has not been well studied. Isolated rat lungs were challenged with various periods of ischaemia and/or reperfusion. Haemodynamics, lung weight gain (LWG), capillary filtration coefficient (K(fc)), tissue pathology, the concentrations of cytokines in the perfusate, and mRNAs for the various cytokines in the lung tissues were measured. I/R induced a permeability type of pulmonary oedema, as reflected by increases in LWG and K(fc). LWG and K(fc) in the I(45)R(60)(UW) group (45 min of ischaemia followed by 60 min of reperfusion with UW) were only 2% and 5% respectively of those in the I(45)R(60)(NS) group (where NS is normal saline). LWG and K(fc) in the UW group had both increased by 180 min, to values similar to those in the I(45)R(60)(NS) group. However, these findings show that UW was remarkably effective at preventing LWG after 60 min of reperfusion, and was more than 3-fold more effective than NS in delaying LWG. For longer ischaemic times only, or the same period of ischaemia followed by longer reperfusion periods, greater lung injury occurred. I/R lung injury also induced increased concentrations of tumour necrosis factor-alpha (TNF-alpha), interleukin 1 and interleukin 6 in the perfusate, and increased the mRNAs for these cytokines in lung tissue. A significant correlation was obtained between TNF-alpha concentration and LWG. TNF-alpha production in the I(45)R(60)(UW) group was only 7% of that in the I(45)R(60)(NS) group. However, TNF-alpha mRNA expression in the I(45)R(60)(UW) group was 80% of that in the I(45)R(60)(NS) group. This indicates that transcription/translation do not correlate well with cytokine production, and also suggests that one reason for the effectiveness of UW in delaying LWG may be because it delays TNF-alpha production. In summary, ischaemia or I/R caused a permeability-type pulmonary oedema that was associated with leucocyte infiltration and the up-regulation of various cytokines, regardless of the perfusion fluid. Except for pulmonary hypertension, less severe I/R lung injury and delayed cytokine production in lungs perfused with UW, the pattern of injury associated with I/R challenge was similar to that in lungs perfused with NS. We propose that more or long-acting protective agents are required as additives in order to modify UW to produce an optimal preservation solution.

Topics: Adenosine; Allopurinol; Animals; Cytokines; Glutathione; Hemodynamics; Insulin; Lung; Male; Organ Preservation Solutions; Organ Size; Polymerase Chain Reaction; Pulmonary Edema; Raffinose; Rats; Rats, Sprague-Dawley; Reperfusion Injury; RNA, Messenger; Up-Regulation

Macromolecules are present in lung preservation solutions to limit liquid filtration out of the pulmonary circulation and minimize pulmonary edema. We tested the effectiveness of these molecules by measuring interstitial edema in rat lungs perfused with macromolecular solutions (University of Wisconsin [UW] solution and Euro-Collins solution supplemented with modified pentastarch [pentafraction, PEN]) or with solutions that lacked macromolecules (UW solution with PEN and Euro-Collins solution.) The lungs were inflated with air and perfused with one of the test solutions, then rapidly frozen and prepared for histological analysis. From tissue sections, we measured cross-sectional areas of pulmonary arteries and veins, and also measured cross-sectional areas of the interstitial spaces surrounding arteries and veins. We then calculated the interstitium-to-vessel cross-sectional area ratio. In lungs perfused with macromolecular solutions these ratios were 0.09+/-0.15 and 0.53+/-0.56 (mean +/- SD) for UW solution and Euro-Collins solutions solution with PEN, respectively (P

Topics: Adenosine; Allopurinol; Animals; Glutathione; Hypertonic Solutions; Insulin; Male; Organ Preservation; Organ Preservation Solutions; Perfusion; Pulmonary Edema; Raffinose; Rats

Despite suggestions of a connection between endothelial damage and permeability alterations after ischemia and reperfusion in pulmonary tissue undergoing transplantation, no direct correlation between vascular endothelial discontinuity and parenchymal edema has yet been shown.. Forty-two rat lungs were harvested and stored for 48 or 72 hours under hypothermic and ischemic conditions. Stored pulmonary tissue was studied before transplantation and 5 minutes or 24 hours after transplantation by light microscopy and scanning electron microscopy of arterial vascular endothelium.. Stored lungs not subjected to revascularization showed moderate perivascular edema, with small intercellular gaps in endothelial monolayers. Five minutes after transplantation, pulmonary tissue appeared congested, with perivascular and alveolar edema. Examination of vascular endothelium by scanning electron microscopy showed detachment of endothelial cells. Twenty-four hours after transplantation, edema, hemorrhage, and vascular congestion were found in all specimens. Arterial vascular endothelium showed weak intercellular connections, numerous intercellular gaps, and widespread cell detachment. Bronchial epithelial cells appeared damaged after storage, with loss of cilia, blebbing of apical cytoplasm, and cellular rounding. These changes were maintained 5 minutes after transplantation but appeared totally reversed after 24 hours in specimens stored 48 hours, whereas bronchial denudation was observed in 72-hour stored lungs. Statistically significant positive correlations (Kendall p < 0.001) between revascularization time and alveolar edema and hemorrhage were found for both storage periods.. The results from this study demonstrate correlation between loss of endothelial monolayer continuity and histologic evidence of vascular permeability increases in pulmonary tissue before and after lung transplantation.

Topics: Adenosine; Allopurinol; Animals; Bronchi; Capillary Permeability; Cold Temperature; Endothelium; Endothelium, Vascular; Female; Glutathione; Insulin; Lung; Lung Transplantation; Microscopy, Electron, Scanning; Organ Preservation; Organ Preservation Solutions; Pulmonary Edema; Raffinose; Rats; Rats, Inbred Lew; Reperfusion Injury

University of Wisconsin (UW) solution has been demonstrated to enhance and extend the preservation of the hepatic, pancreatic, renal, and cardiac allografts. Prostaglandin E1 (PGE1), which has been used to produce pulmonary vasodilatation, has improved preservation in lung transplantation. Experiments were designed to evaluate their potential role in 24-h preservation of the allografts in lung transplantation. Thirty-six dogs underwent left lung transplantation. The donor lungs (n = 6 in each group) were flushed with UW solution (group 1 and 2) or modified Euro-Collins (EC) solution (group 3) after PGE1 infusion (500 micrograms). In group 1, donor lungs were transplanted immediately. Lung allografts of group 2 and 3 were cold stored (4 degrees C) in the same preservation solution for 24 h in the inflated state and then transplanted. The right pulmonary artery and right main bronchus were ligated 1 h after completion of the transplantation, forcing the recipient dogs to survive with the transplanted left lung. The recipients were ventilated with an inspired oxygen fraction of 0.4 and end-expiratory pressure 10 cm H2O. Two dogs died prematurely in group 3, whereas all dogs in group 1 and 2 survived the experimental period. The arterial oxygen tension and saturation and dynamic lung compliance were significantly higher in dogs of group 1 and 2. Transplanted lungs of group 1 and 2 had significantly lower pulmonary vascular resistance, alveolar-arterial oxygen difference, and wet/dry lung weight ratio. Histologically, pulmonary edema, congestion, sloughing of bronchial mucosa, and peribronchial and peripulmonary arterial hemorrhage were shown in lungs of group 3, but not in those of group 1 and 2. Airway mucosa and pulmonary vascular structure were well preserved in lungs of group 1 and 2. There was no significant difference between group 1 and 2 in lung functions, hemodynamics, or morphologic features. We concluded that PGE1 and UW solution could effectively extend lung preservation up to 24 h in this in vivo canine lung allotransplantation model.

Topics: Adenosine; Allopurinol; Alprostadil; Animals; Blood Pressure; Bronchi; Cardioplegic Solutions; Dogs; Endothelium, Vascular; Glutathione; Hypertonic Solutions; Insulin; Lung; Lung Compliance; Lung Transplantation; Organ Preservation Solutions; Organ Size; Oxygen; Pulmonary Artery; Pulmonary Edema; Raffinose; Tissue Preservation; Vascular Resistance

Topics: Adenosine; Allopurinol; Animals; Dogs; Glutathione; Insulin; Lipid Peroxides; Lung; Lung Compliance; Organ Preservation; Organ Preservation Solutions; Pulmonary Circulation; Pulmonary Edema; Raffinose; Reperfusion Injury; Solutions; Time Factors; Vascular Resistance

Belzer's University of Wisconsin cold storage solution (UWCSS) has proved useful in extending the shelf life of organs in extrathoracic transplantation and more recently has also been shown to be useful in heart transplantation. I investigated the effect of 4 degrees C UWCSS on the vascular and interstitial properties of the lung to see whether it affected the pulmonary microcirculation or caused pulmonary edema. Infusion of UWCSS was associated with a slight decrease in oxygen tension, but the final oxygen tension was no different from that previously demonstrated with Euro-Collins solution. Vascular conductance was not affected by UWCSS, but average vascular closure increased slightly, indicating that increased vascular tone occurs. This effect is similar to but less than that previously observed with Euro-Collins solution. Based on comparisons of wet to dry weight ratios, estimates of interstitial compliance, transvascular fluid flux, and microvascular filtration coefficient, it does not appear that UWCSS causes pulmonary edema. Further investigation into the usefulness of UWCSS in lung transplantation is therefore warranted.

Topics: Adenosine; Allopurinol; Animals; Blood Gas Analysis; Dogs; Drug Evaluation, Preclinical; Glutathione; Hematocrit; Insulin; Lung; Microcirculation; Organ Preservation Solutions; Pulmonary Circulation; Pulmonary Edema; Raffinose; Solutions; Tissue Preservation; Vascular Resistance; Vasoconstriction