Altered Immunogenicity of Donor Lungs via Removal of Passenger Leukocytes Using Ex Vivo Lung Perfusion.

Passenger leukocyte transfer from the donor lung to the recipient is intrinsically involved in acute rejection. Direct presentation of alloantigen expressed on donor leukocytes is recognized by recipient T cells, promoting acute cellular rejection. We utilized ex vivo lung perfusion (EVLP) to study passenger leukocyte migration from donor lungs into the recipient and to evaluate the effects of donor leukocyte depletion prior to transplantation. For this purpose, female pigs received male left lungs either following 3 h of EVLP or retrieved using standard protocols. Recipients were monitored for 24 h and sequential samples were collected. EVLP-reduced donor leukocyte transfer into the recipient and migration to recipient lymph nodes was markedly reduced. Recipient T cell infiltration of the donor lung was significantly diminished via EVLP. Donor leukocyte removal during EVLP reduces direct allorecognition and T cell priming, diminishing recipient T cell infiltration, the hallmark of acute rejection.

Circulation stabilizing therapy and pulmonary high-resolution computed tomography in a porcine brain-dead model.

BACKGROUND: Currently 80% of donor lungs are not accepted for transplantation, often due to fluid overload. Our aim was to investigate if forced fluid infusion may be replaced by a new pharmacological therapy to stabilize circulation after brain death in an animal model, and to assess therapy effects on lung function and morphology trough blood gas parameters and state-of-the-art High-resolution CT (HRCT). METHODS: Brain death was caused by surgical decapitation. To maintain mean aortic pressure > 60 mmHg, pigs were treated with forced electrolyte solution infusion (GI; n = 6) or the pharmacological therapy (GII; n = 11). GIII (n = 11) were non-decapitated controls. Lung function was investigated with blood gases and lung morphology with HRCT. RESULTS: GI pigs became circulatory instable 4-6 h after brain death in spite of forced fluid infusion, five pigs showed moderate to severe pulmonary edema on HRCT and median final PaO2 /FiO2 was 29 kPa (Q1; Q3; range 26; 40; 17-76). GII and GIII were circulatory stable (mean aortic pressure > 80 mmHg) and median final PaO2 /FiO2 after 24 h was 72 kPa (Q1; Q3; range 64; 76; 53-91) (GII) and 66 kPa (55; 78; 43-90) (GIII). On HRCT, only two pigs in GII had mild pulmonary edema and none in GIII. More than 50% of HRCT exams revealed unexpected lung disease even in spite of PaO2 /FiO2 > 40 kPa. CONCLUSION: Pharmacological therapy but not forced fluid infusion prevented circulatory collapse and extensive HRCT verified pulmonary edema after acute brain death. HRCT was useful to evaluate lung morphology and revealed substantial occult parenchymal changes justifying efforts toward a more intense use of HRCT in the pre-transplant evaluation.

Pharmacological normalization of circulation after acute brain death.

BACKGROUND: Circulatory instability is a serious problem after brain death in organ donors. The hypotension is often counteracted with infusion of large amounts of crystalloid solutions, which may impair lung function leading to rejection of the lungs as donor organs. The aim was to show that the circulation can be normalized pharmacologically for 24 h in pigs after total removal of the brain and brainstem by decapitation (between C2 and C3). METHODS: Twenty-four 40-kg pigs (n = 8 × 3) were included: non-decapitated, decapitated, and decapitated with pharmacological treatment. All animals got the same basal fluid supply and ventilation. The pharmacological treatment consisted of the neuronal monoamine reuptake blocker cocaine and low doses of noradrenaline and adrenaline. Desmopressin, triiodothyroxine, thyroxine and cortisol were also given. RESULTS: After decapitation, a catecholamine storm occurred, with an increase of noradrenaline and adrenaline by a factor of 79 and 298, respectively. Thirty minutes later, the pigs were hypotensive. The median time to the aortic pressure that was less than 40 mmHg was 9:09 h (range 5:50 to 22:01). After 6 h, the concentration of thyroid hormones and cortisol was significantly reduced. With pharmacological treatment of decapitated animals, the aortic pressure, renal blood flow, creatinine, urine production, liver function and blood gases did not differ significantly from the non-decapitated control animals. CONCLUSION: Pharmacological substitution of pituitary gland function, blockade of peripheral catecholamine neuronal reuptake and low doses of catecholamines normalize circulation in decapitated pigs throughout a 24-h observation period, whereas untreated decapitated pigs all develop severe circulatory collapse within 12 h.

COMPARISON OF PULMONARY NODULE DETECTION, READING TIMES AND PATIENT DOSES OF ULTRA-LOW DOSE CT, STANDARD DOSE CT AND DIGITAL RADIOGRAPHY.

The purpose of the present work was to evaluate performance in pulmonary nodule detection, reading times and patient doses for ultra-low dose computed tomography (ULD-CT), standard dose chest CT (SD-CT), and digital radiography (DR). Pulmonary nodules were simulated in an anthropomorphic lung phantom. Thirty cases, 18 with lesions (45 total lesions of 3-12 mm) and 12 without lesions were acquired for each imaging modality. Three radiologists interpreted the cases in a free-response study. Performance was assessed using the JAFROC figure-of-merit (FOM). Performance was not significantly different between ULD-CT and SD-CT (FOMs: 0.787 vs 0.814; ΔFOM: 0.03), but both CT techniques were superior to DR (FOM: 0.541; ΔFOM: 0.31 and 0.28). Overall, the CT modalities took longer time to interpret than DR. ULD chest CT may serve as an alternative to both SD-CT and conventional radiography, considerably reducing dose in the first case and improving diagnostic accuracy in the second.

Platelet adhesion in polytetrafluoroethylene (PTFE) vascular grafts in vivo and the influence of increased intramuscular pressure--an experimental model.

Thrombogenicity of graft material is involved in early graft failure in small diameter grafts. The frequently seen postoperative swelling of the leg after distal revascularization may cause an increased intramuscular pressure and early graft failure. Pairs of 4 mm polytetrafluoroethylene (PTFE) grafts were implanted. Autologous platelets were labeled with 111In-oxine. Platelet adhesiveness onto the grafts were analyzed from gamma camera images. Intramuscular pressures were measured with wick technique. Blood flow was measured. One graft served as control the other as test graft. Ninety minutes after declamping the i.m. pressure was increased in the test-leg to 30 mmHg, and later to 60 mmHg. In the control-graft platelet uptake increased to a maximum 60 min after declamping. Blood flow and i.m. pressure remained uneffected. The test-grafts were initially similar but when i.m. pressure was increased to 30 mmHg activity in the grafts increased significantly. Blood flow decreased with 12% of initial flow. When i.m. pressure was raised to 60 mmHg platelet uptake continued to increase. An increased intramuscular pressure of 30 mmHg or more significantly increase the amount of platelets adhering onto PTFE grafts, emphasizing the need for measuring intramuscular pressures after lower limb vascular revascularizations.

Veno-right ventricular bypass as total extracorporeal lung assistance. An experimental study.

Efficacy of veno-right ventricular bypass as a total extracorporeal lung assistance was studied for a period of 24 hours in six healthy pigs with a mean weight of 60 kg. A covalently bonded heparin-coated extracorporeal membrane oxygenation system and a roller pump were used for the bypass. No local or systemic heparin was administered. The bypass was established with an open chest with two 28F venous cannulas and one 24F arterial cannula. The arterial cannula was placed in the right ventricle across the tricuspid valve. With the lung function totally disabled, this extracorporeal lung assistance maintained normal systemic arterial and mixed venous blood gases during the entire 24-hour period in all the animals. No significant tricuspid insufficiency was observed, and the animals maintained normal central hemodynamics. There was no hemolysis, and the platelet counts remained essentially unaltered. Multiple foci of clot formation were observed in all the oxygenators, but no macroscopic thrombosis or embolization was seen either in the heart or in the lungs. A veno-right ventricular bypass offers total extracorporeal lung assistance in 60 kg juvenile pigs for a period of 24 hours. Tricuspid valve competence is an important prerequisite for the success of this procedure.

Vascular function in the cadaver up to six hours after cardiac arrest.

BACKGROUND: The aim of the study was to evaluate how well vascular function is retained in a cadaver kept in a room with a temperature of 21 degrees C. METHODS: The aorta and pulmonary artery of rats were investigated in organ baths as fresh controls and after 1, 2, 3, or 6 hours' storage in the cadaver. Six-hour-old cadaver aortas were transplanted and investigated after 24 hours and 60 days. RESULTS: After 3 hours' storage there was no significant decrease in smooth muscle contractile function in either aorta or pulmonary artery. After 6 hours' storage both the aorta and the pulmonary artery demonstrated a significant decrease in smooth muscle contractile function, 30% (p < 0.05) and 44% (p < 0.001), respectively, compared to fresh controls. Storing the aorta for 2 hours and the pulmonary artery for 6 hours caused no significant decrease in endothelium-dependent relaxing function. In aorta segments investigated after 3 and 6 hours there was a significant decrease in endothelium-dependent relaxation, 12% (p < 0.05) and 29% (p < 0.001), respectively. Six-hour-old cadaver aortas transplanted and investigated after 24 hours or 60 days demonstrated no significant changes in endothelium-dependent relaxation and smooth muscle function compared to fresh controls. CONCLUSION: The pulmonary artery can tolerate 3 hours of warm ischemia in the nonheart-beating cadaver without loss of endothelium-dependent relaxation and smooth muscle function. The dysfunction seen in 6-hour-old cadaver aortas was normalized after transplantation and 24 hours of reperfusion.

Prostanoid release after lung transplantation.

BACKGROUND: Increased pulmonary vascular resistance is frequently seen after lung transplantation. Thromboxane A2 is a potent vasoconstrictor of pulmonary arteries. Thromboxane-elicited vasoconstriction can ben counteracted by prostacyclin. The effects of lung transplantation on the biosynthesis of these substances were investigated. METHODS: Pulmonary artery flush perfusion with a low-potassium dextran glucose solution was performed in six donor pigs. After a 24-hour storage period, the left lung was transplanted into a recipient, followed by right pneumonectomy, making the recipient's survival entirely dependent on the transplanted lung. A sham operation (bilateral thoracotomy, right pneumonectomy) ws done in six pigs. the urine contents of the stable thromboxane A2 metabolite 2,3-dinor-thromboxane B2 and the stable prostacyclin metabolite 2,3-dinor-6-keto-protaglandin F1 alpha were measured with a gas chromatography-mass spectrometry method. RESULTS: One to four hours after reperfusion, thromboxane A2 production reached its maximum in both groups: it ws fivefold the basal value in the transplanted group, but only twofold in the sham-operated group, the difference being significant (p < 0.005). Twenty to twenty-four hours after reperfusion, thromboxane A2 production had stabilized at about twofold the basal value in both the transplanted and in the sham-operated group. Four to eight hours after reperfusion, prostacyclin production reached 15 times the basal value in the transplanted group and twofold in the sham-operated group, the difference being significant (p < 0.05). Twenty to twenty-four hours after reperfusion, prostacyclin production was 18-fold the basal value in the transplanted group and sevenfold in the sham-operated group. No correlation was found between the thromboxane or prostacyclin production and the pulmonary vascular resistance or the mean pulmonary arterial pressure. CONCLUSIONS: The thromboxane A2 production increased fivefold after lung transplantation, with a concomitant 15-fold increase in prostacyclin synthesis, which might have counteracted the vasoconstrictor effect of thromboxane.

Bronchial healing, lung parenchymal histology, and blood gases one month after transplantation of lungs topically cooled for 2 hours in the non-heart-beating cadaver.

BACKGROUND: The aim of this study was to investigate, in an experimental survival model, the functional and morphologic results of lung transplantation using lungs from non-heart-beating donors. METHODS: Left lungs, topically cooled to 25 degrees C for 2 hours in situ after 5 minutes of circulatory arrest followed by 26 minutes of unsuccessful cardiopulmonary resuscitation, were transplanted into syngeneic rats. Five weeks after the transplantation, right pneumonectomy was performed and blood gases measured after 60 minutes. In a control group, fresh donor lungs were used for transplantation and comparison was made with the cadaver group and a group of normal rats after right pneumonectomy. Morphologic changes were evaluated by semiquantitative scoring of 13 different parameters to obtain a total histologic index for each rat. RESULTS: Computerized tomography scans of the chest made during the third post-operative week showed normal lung parenchyma in both groups, and at 5 weeks there were no significant differences in blood gases. The bronchial anastomoses showed normal healing in all cases. The histologic changes in the lung parenchyma were generally mild and focal, primarily consisting of interstitial and perivascular mononuclear inflammation, bronchial inflammation and athelectasis. Surprisingly, the transplanted controls demonstrated the most pronounced changes, although only the difference in total histologic index between groups was significant. CONCLUSIONS: Lungs from non-heart-beating donors, topically cooled in the cadaver for two hours after failed resuscitation, showed normal bronchial healing and favorable parenchymal histology compared to transplanted control lungs 5 weeks after transplantation.

Importance of calcium in long-term preservation of the vasculature.

BACKGROUND: The aim was to investigate the effect of calcium in organ preservation solutions with respect to 36-hour preservation of vascular smooth muscle function and endothelium-dependent relaxation. METHODS: The infrarenal aortas of 60 Sprague-Dawley rats were studied in organ baths as fresh controls and after 36 hours of cold (4 degrees C) storage in different preservation solutions with and without calcium. The thromboxane A2 analogue U-46619 was used to study contractility. Endothelium-dependent relaxation was tested by the cumulative addition of acetylcholine. Papaverine hydrochloride was used to elicit endothelium-independent relaxation. RESULTS: Krebs solution was the only solution able to fully preserve contractility. Krebs solution without calcium gave poor preservation. After the addition of 1.5 mmol/L of calcium to University of Wisconsin solution and to Perfadex, both these solutions became fully able to preserve contractility. None of the solutions (with or without calcium) were fully able to preserve endothelium-dependent relaxation, although University of Wisconsin solution gave good preservation and Perfadex, fair preservation. Euro-Collins solution and K+ (124 mmol/L)-enriched Krebs solution were not able to preserve smooth muscle function or endothelium-dependent relaxation. CONCLUSIONS: Calcium is essential for long-term preservation of vascular smooth muscle function but not for long-term preservation of endothelium-dependent relaxation.

Endothelium-mediated relaxation in transplanted aorta.

This study compares the vasodilating effect of endothelium-derived relaxing factors (EDRFs) in free arterial grafts with that in their normal control vessels. The infrarenal aorta of Sprague-Dawley rats was transplanted into the same position in other inbred recipient rats. A Krebs buffer solution (4 degrees C) served as the preservation solution. The ischemic time for the grafts (n = 8) was 42 +/- 1 minutes. Two grafts were studied after 3 days and six grafts, after 60 days. Ring segments were cut from all vessels, and isometric contractions were recorded in organ baths. The vessel segments were constricted with noradrenaline, a thromboxane A2 mimic (U-46619), or prostaglandin F2 alpha. Concentration-response curves with acetylcholine, which was used as the endothelium-stimulating substance causing release of EDRFs, were obtained. The patency of the grafts was 100%. Acetylcholine induced relaxation in all vessel segments with intact intima, whereas no relaxation was seen when the endothelium was manually removed. No significant differences were found between the grafts and the normal control vessels. Histology of the 60-day grafts showed elastomuscular arteries without intimal thickening and a media consisting of eight to ten muscle layers interrupted by five to six elastic lamellae. Scanning electron microscopy showed no major differences between normal endothelium and the endothelium of 3-day or 60-day grafts. This study indicates that free elastomuscular arterial grafts, in which the morphology of the intima is preserved, will retain their full ability to release EDRFs.

Endothelium-dependent relaxation after short-term preservation of vascular grafts.

As the integrity of graft endothelium seems to be essential to successful long-term patency in coronary operations, its preservation demands the utmost care. The aim of the present study was to investigate the effects of currently used solutions on endothelium-dependent relaxation after short-term storage of vessels at room temperature or at 4 degrees C. The infrarenal rat aorta was selected for study because its use enabled standardization of the investigation, which was performed in organ baths on 672 vessel segments from 112 Sprague-Dawley rats. Stable vasoconstriction was obtained with the thromboxane analogue U-46619. Acetylcholine was used to elicit endothelium-dependent relaxation. The results obtained for vessels preserved for 2 hours were compared with those for autologous vessels studied immediately after harvesting. Vessel contractility was unaffected by the preservation solutions, except in the Ringer's acetate group, where it was reduced by 50% (p < 0.05). Endothelium-independent relaxation, tested with papaverine, was unaffected in all groups. Ringer's lactate, Krebs solution, and Perfadex (a low-potassium-dextran-glucose solution) did not significantly affect endothelium-dependent relaxation either at room temperature or at 4 degrees C, although a tendency to impaired relaxation was seen in these three groups after cold storage. Standard Ringer's solution and fresh heparinized blood each significantly reduced endothelium-dependent relaxation in vessels stored at room temperature (p < 0.05), but not in those stored at 4 degrees C. Endothelium-dependent relaxation was significantly reduced after storage in normal saline solution (p < 0.05) and in Ringer's acetate (p < 0.01), both at room temperature and at 4 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

Pulmonary vascular resistance related to endothelial function after lung transplantation.

In 8 donor pigs, flush perfusion was performed with a low-potassium-dextran solution. Ring segments were taken from a small intralobar pulmonary artery in the right lung immediately after perfusion and after 24 hours of cold storage for studies in organ baths. Stable vasoconstriction was induced with the thromboxane mimic U-46619, and acetylcholine was used to induce endothelium-dependent relaxation. The maximum relaxation was significantly reduced after flush perfusion compared with fresh nonperfused controls, and a significant additional reduction was seen after the 24-hour storage period. The left donor lung was transplanted into a recipient after 24 hours of cold storage. Contralateral pneumonectomy was then performed, making the recipient entirely dependent on the transplanted lung for survival. All 8 pigs were in good condition throughout the 24-hour observation period, with arterial oxygen tension of around 165 mm Hg (range, 80 to 275 mm Hg; inspired oxygen fraction, 0.5) and pulmonary vascular resistance of around 450 dyne.s.cm-5 (range, 260 to 730 dyne.s.cm-5). The maximum endothelium-dependent relaxation for each donor was checked for correlation to pulmonary vascular resistance and to systolic, mean, and diastolic pulmonary artery pressures as recorded at 4-hour intervals. Regression analyses showed arterial oxygen tension to be unrelated to pulmonary vascular resistance and endothelial dysfunction to be unrelated to pulmonary artery pressure but to correlate to pulmonary vascular resistance, this correlation being significant after reperfusion for 16 hours (p < 0.05) and highly significant after 24 hours (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of topical cooling in lung preservation: is a reappraisal due?

The aim of the present study was to test the efficacy of topical cooling as the only viable lung preservation method using the most challenging evaluation method, namely single-lung transplantation followed by immediate contralateral pneumonectomy. Ten domestic pigs (5 donors and 5 recipients) with a mean body weight of 57 kg (range, 53 to 59 kg) were used. After we administered systemic heparin (4 mg/kg), the lungs were harvested and placed in an atelectatic state under cold (8 degrees to 9 degrees C) low-potassium-dextran solution for 12 hours. Left lung transplantation was then done in the recipient pig followed by right pneumonectomy, thus making the recipient 100% dependent on the transplanted donor lung. No operative mortality or morbidity occurred. All animals were in excellent condition throughout the 24-hour observation period. They had normal blood gases which did not differ significantly from the preoperative blood gases obtained from the 5 recipients before transplantation (ie, when they had their own two lungs). A moderate increase (p < 0.05) in pulmonary vascular resistance was seen as compared with sham-operated animals. To conclude, topical cooling to 8 degrees C provides excellent lung preservation for 12 hours in pigs. If similar results can be obtained with other species, the currently accepted 6-hour limit for safe clinical lung preservation may be extended to 12 hours. It seems also warranted to critically reconsider which factors, apart from cooling alone, actually contribute favorably to 12-hour lung preservation.

Long-term preservation of vascular endothelium and smooth muscle.

This study was performed in organ baths on 400 ring segments of infrarenal aorta taken from 40 Sprague-Dawley rats that had been randomized into five groups. Contractility was tested with the thromboxane analogue U-46619. Acetylcholine was used to elicit endothelium-dependent relaxing factor (EDRF). The results obtained from vessels preserved at 4 degrees C for 6, 12, 24, and 36 hours were compared with those from autologous vessels studied immediately after harvesting. Vessels preserved in Euro-Collins solution showed a 46% (p < 0.01) decrease in contractility after 12 hours of storage; after 24 hours only weak contractions could be elicited, and after 36 hours they had lost their ability to contract. The EDRF function was slightly reduced after 12 hours and could not be investigated after 24 and 36 hours. With the University of Wisconsin solution (UW) and the low-potassium-dextran-glucose solution Perfadex no decrease in contractility was seen in the first 24 hours, but at 36 hours the vessels preserved in UW had lost 40% (p < 0.01) and those preserved in Perfadex 30% (p < 0.05) of their contractility. The EDRF function was significantly reduced by about 15% after 6, 12, and 24 hours in both the UW and the Perfadex groups. At 36 hours, vessels stored in Perfadex had lost 41% (p < 0.001) and those stored in UW 17% (p < 0.01) of their EDRF function.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhalation of nitric oxide after lung transplantation.

BACKGROUND: Pulmonary hypertension is a postoperative complication that may adversely affect the outcome of lung transplantation. The effect of nitric oxide (NO) inhalation on pulmonary hemodynamic indices after lung transplantation was studied and compared with findings in control pigs. METHODS: Varying concentrations of NO were inhaled by 5 pigs after left lung transplantation and right pneumonectomy and by 5 controls after right pneumonectomy at an inspired oxygen fraction of 0.21 and 0.5. Hemodynamic data were recorded continuously, and fast circulatory courses were analyzed. RESULTS: Inhalation of NO reduced pulmonary vascular resistance and mean pulmonary arterial pressure in all pigs, but the decrease was pronounced and dose dependent only at an inspired oxygen fraction of 0.21 in the pigs that had transplantation. These were the only pigs that became hypoxic. With the termination of NO, there was a dose-independent rebound pulmonary vasoconstriction in the controls, especially at an inspired oxygen fraction of 0.21, but not in the pigs that had transplantation. This response was transient and could be blunted with a higher inspired oxygen fraction. CONCLUSION: Inhalation of NO reduced pulmonary vascular resistance in the transplanted lung and may be useful in the treatment of pulmonary hypertension after lung transplantation. The rebound pulmonary vasoconstriction with the termination of NO inhalation stresses the need to be aware of this effect and to wean NO carefully in clinical situations. This study showed oxygen dependency, which has to be taken into consideration in dose-response studies involving NO inhalation.

Safe pulmonary preservation for 12 hours with low-potassium-dextran solution.

The function of porcine left lung allografts was studied after perfusion with (150 mL/kg) and storage for 12 hours in a 4 degrees to 6 degrees C low-potassium-dextran solution (Perfadex; Kabi Pharmacia AB, Uppsala, Sweden). After a left lung transplantation, an artificial lung in the form of venoarterial extracorporeal membrane oxygenation was established. The artificial lung has a "biological" heparin-coated surface (Carmeda AB, Stockholm, Sweden), and there is no need for systemic anticoagulation. Immediately thereafter, pneumonectomy of the normal right lung was done. All the animals were weaned from the artificial lung within 1 hour after the pneumonectomy. Six animals were followed up for 24 hours. They were in good condition throughout the 24-hour observation period with arterial oxygen tensions around 200 mm Hg (inspired oxygen fraction = 0.4) and arterial carbon dioxide tensions around 40 mm Hg. This study demonstrates a reliable method for continuous evaluation of the function of a transplanted lung immediately after transplantation and over the ensuing postoperative period. Safe 12-hour lung preservation can be obtained with the low-potassium-dextran solution Perfadex.

Perfadex is superior to Euro-Collins solution regarding 24-hour preservation of vascular function.

BACKGROUND: The aim of this study was to compare Perfadex with Euro-Collins solution regarding 24-hour preservation of endothelium-dependent relaxation and vascular smooth muscle function. METHODS: The infrarenal aorta of 72 isogenic rats was studied in organ baths as fresh controls, after 24 hours of cold (4 degrees C) storage, and after 24-hour storage followed by transplantation and examination after 7 or 30 days. The thromboxane A2 analogue U-46619 was used to test contractility. Acetylcholine chloride was used to elicit endothelium-dependent relaxation and papaverine hydrochloride, to elicit endothelium-independent relaxation. RESULTS: With both solutions, all grafts were patent after 7 and 30 days. Vessels preserved in Euro-Collins solution for 24 hours lost 95% (p < 0.001) of their contractility compared with fresh controls; 7 days after transplantation, they had regained 40% of initial contractility, and after 30 days, there was no significant decrease in contractility. Vessels preserved in Perfadex manifested no significant decrease in contractility at any time. Endothelium-dependent relaxation could not be evaluated in vessels stored for 24 hours in Euro-Collins solution because they had lost almost all contractility; 7 days after transplantation, endothelium-dependent relaxation was reduced by 65% (p < 0.001), but at 30 days after transplantation, there was no significant decrease in endothelium-dependent relaxation. Vessels preserved in Perfadex for 24 hours lost 17% (p < 0.05) of endothelium-dependent relaxation, but 7 and 30 days after transplantation, there was no significant decrease in endothelium-dependent relaxation. CONCLUSIONS: Perfadex, but not Euro-Collins solution, has the capacity to preserve vascular function after 24 hours of storage followed by in vivo reperfusion.

Might free arterial grafts fail due to spasm?

The rat femoral artery was used as a free graft and was studied after 2, 7, 14, 30, and 60 days. The patency of the grafts was 100% (2 days, n = 6), 78% (7 days, n = 9), 63% (14 days, n = 8), 33% (30 days, n = 12), and 18% (60 days, n = 11). Histology showed an intimal thickening after 14 days and the media, which in the controls consisted of eight to ten layers of myocytes, was reduced to six to eight cell layers. During the first 2 weeks the graft segments had an impaired contraction when exposed to Krebs solution with 124 mmol/L K+, whereas after 1 month and later the graft segments approached the controls or had even higher contractile force. The thromboxane mimic U-46619 elicited full contractile force at all times whereas the potency was significantly lower during the first 14 days. Noradrenaline was unable to induce contraction in the graft segments during the first 14 days, but at 30 and 60 days it had regained full contractile force and was significantly more potent (approximately 60 times) in the graft segments compared with the controls. This study suggests that intimal thickening and hypercontractility might be a problem in free muscular arterial grafts.

Inhaled nitric oxide reveals and attenuates endothelial dysfunction after lung transplantation.

BACKGROUND: Maintaining endothelial function within transplanted organs may be critical to successful preservation. In this study we have evaluated the relationship between the effect of inhalation of nitric oxide and the degree of endothelial dysfunction after lung transplantation. METHODS: A left lung, which had been preserved for 24 hours, was transplanted and a right pneumonectomy was performed in 5 pigs. After a 24-hour observation period the pigs inhaled 5, 20, and 80 ppm nitric oxide, and pulmonary vascular resistance was recorded continuously. From the same donors preserved pulmonary arteries from the contralateral lung were studied simultaneously in organ baths. Acetylcholine chloride was used to elicit endothelium-dependent relaxation in vessel segments contracted with the thromboxane A2 analogue U-46619. RESULTS: Maximal endothelium-dependent relaxation decreased in the preserved lungs and correlated to the pulmonary vascular resistance in the simultaneously transplanted lungs. Inhalation of nitric oxide in the pigs that had received transplants caused the pulmonary vessels to dilate in proportion to the endothelial dysfunction. CONCLUSIONS: Preservation of lung for transplantation induces an endothelial dysfunction, and the degree of the decrease in pulmonary vascular resistance caused by nitric oxide inhalation may be an indication of the degree of this endothelial damage. The vasodilation caused by inhaled nitric oxide increases as the endothelial function deteriorates.