The Journal of Heart and Lung Transplantation, Vol 32, No 4S, April 2014. (AI: 1.6% vs 3.6%, iNOS: 9 vs 25). AI values and iNOS expression were.
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The Journal of Heart and Lung Transplantation, Vol 32, No 4S, April 2014
Methods: Rats were divided into control (n = 5), non-plasmin (n = 7), and plasmin (n = 7) groups. In the non-plasmin and plasmin groups, cardiac arrest was induced by withdrawal of mechanical ventilation. After 120 min of warm ischemia, the lungs were ventilated and flushed. Heart and lungs were excised en bloc and perfused and ventilated in the EVLP for 30 min, and plasmin or placebo was administered upon EVLP initiation. After the EVLP period, the lungs inflated with air were stored at 4°C for 90 min. Finally, all lungs were perfused with rat blood for 80 min. Correlations of physiological data at the end of EVLP and reperfusion were assessed and histological investigation after reperfusion was also done. Results: Throughout the reperfusion period, the plasmin group showed better findings in pulmonary vascular resistance (PVR), lung edema, dynamic compliance, and oxygenation compared with the non-plasmin group. PVR and PaO2 at the end of EVLP were significantly correlated with those at the end of reperfusion (Figure). Among all, PVR during EVLP was the most useful predictive factor for the severity of IRI. At the end of reperfusion, the plasmin group showed fewer signs of histological injury. Caspase 3/7 activity in the plasmin group was lower than that in the non-plasmin group. Conclusion: We confirmed that plasmin administration during EVLP could protect the donor lungs after reperfusion. We also found that several physiological values in EVLP are predictive markers for the lung function after reperfusion.
6( 93) Apoptosis and Expression of Inducible Nitric Oxide Synthase in Normothermic Lung Perfused With Organ Care System (OCS) Compared To Standard Cold Storage Donor Lungs F. Calabrese , M. Schiavon, N. Nannini, F. Lunardi, G. Marulli, G. Di Gregorio, A. Rebusso, E. Balestro, M. Loy, F. Rea. University of Padova, Padova, Italy. Purpose: Ischemia-reperfusion (I/R) injury remains a major cause of graft dysfunction following lung transplantation. High level of NO produced by inducible NO synthase (iNOS) is detrimental in I/R, promoting ischemic apoptosis. Organ Care System Lung (OCS), a portable normothermic ex-vivo lung perfusion system, has been demonstrated a valid option for donor organ preservation. I/R tissue injury together with clinical data were evaluated in OCS compared to standard cold storage (SOC) preserved lungs. Methods: Lung fragments sampled during back table and 90 min after blood reperfusion of the first implanted lung were studied in 22 donor lungs randomly preserved with OCS (10 cases, 5 males/5 females, mean: 36 years old, range 13-49) or with SOC (12 cases, 8 males/4 females, mean: 43 years old, range 19-56). In all cases the following histological parameters were quantified: edema, blood extravasation and granulocyte margination (score 0-3). Apoptosis (by terminal deoxyribonucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling, using an apoptotic index-AI-) and iNOS (by immunohistochemistry distinguishing epithelium, macrophage and endothelium) expression were also investigated. Early clinical post-operative parameters were investigated, including PGD score, mechanical ventilation, ICU time and hospital mortality. Results: The two groups presented comparable donor (Eurotransplant donor score 6 vs 6.5) and recipient characteristics (matched for age, sex, underlying disease and urgency status). All recipients with OCS lungs recovered reasonably well and showed similar early clinical postoperative parameters.At pre-reperfusion time OCS lungs showed less AI and i-NOS expression than SOC group (AI: 1% vs 1.9% and iNOS: 135 vs 280). At post-reperfusion time AI and iNOS expression was lower in OCS than SOC group, particularly when considering endothelial cells
(AI: 1.6% vs 3.6%, iNOS: 9 vs 25). AI values and iNOS expression were directly related in both groups. No significant differences were observed for the other morphological data. Conclusion: Our study supports the beneficial effect of OCS device particularly against apoptosis iNOS related, a major damage in I/R injury. 6( 94) Image Guided Evaluation of Cellular Retention and Survival After Intramyocardial Transcatheter Based Mesenchymal Stem Cell Transplantation Into the Infarcted Pig Heart M.Y. Emmert ,1 P. Wolint,1 J. Pavicevic,1 V. Falk,1 M. Gyöngyösi,2 S.P. Hoerstrup.1 1Cardiac Surgery, University Hopsital Zurich, Zurich, Switzerland; 2Cardiology, University Hopsital Vienna, Vienna, Austria. Purpose: Cardiac cell therapy represents as a promising concept for the failing heart. However, while several delivery methods exist, the overall low retention and survival within the heart remains a major issue and numerous studies suggest a high washout particularly in the early phase after transplantation. We investigate the efficacy of transcatheter-guided, intramyocardial transplantation of mesenchymal stem cells (MSCs) in a translational porcine myocardial infarction model with particular regards to retention and survival. Methods: Allogenic porcine MSCs were characterized and labelled with iron-oxide micro-particles (MPIOs). Next, eight pigs (30-35kg) with chronic MI underwent 3D NOGA mapping-guided, transcatheter intramyocardial transplantation of 1.5x107 MSCs into the anterior-septal wall of the MI border-zone. Thereafter in-vivo cell-tracking was performed using MRI, before the hearts were harvested at different time-points (1, 3, 6, 12 and 24hours) post transplantation for post-mortem assessment. According to the 3D NOGA injection-map, tissue-samples were taken to undergo a combined MACS/FACS cell-separation approach and immunohistochemistry (IHC) to determine cell retention and survival. Results: Transplantation was successful in all animals and MRI displayed the transplanted MPIO-labelled MSCs in the anterior-septal wall (corresponding to the injection sites). At all time-points, viable MSCs could be isolated from the heart and combined MACS/FACS separation (purity up to 98%) revealed up to 11.4% of the injected MSCs (mean: 5.2±4.1%) in the respective samples. IHC further verified intramyocardial presence via positive staining for MSC-specific markers and anti-FITC targeting the MPIOs. The cells appeared to be integrated and could be detected in clusters or in the interstitial areas. Conclusion: This study provides important insights into the early efficacy of transcatheter guided intramyocardial MSC transplantation in a translational animal model. Although the overall retention appears to be relatively low, these data confirm cell survival in the early phase after transplantation. Further optimization of delivery strategies and cell application formats is mandatory to improve intramyocardial retention and survival in future cell therapy concepts. 6( 95) The Importance of Airway and Vascular Parameters in Cellular Ex Vivo Lung Perfusion Assessment of the Donor Lung T. Okamoto ,1 D.M. Wheeler,2 K.R. McCurry.3 1Transplant Center, Cleveland Clinic, Cleveland, OH; 2Cardiothoracic Anesthesia, Cleveland Clinic, Cleveland, OH; 3Cardiovascular Surgery, Cleveland Clinic, Cleveland, OH. Purpose: Ex vivo lung perfusion (EVLP) is an essential process for the evaluation of the donor lung. Currently the PaO2/FiO2 (PF) ratio is the standard in the assessment of lung function in cellular EVLP, while airway and vascular parameters have been partially utilized. The purpose of this study is to investigate the potential utility of airway and vascular parameters as a complementary indication of lung function in EVLP. Methods: Using rejected human lungs (n = 6) and domestic pig lungs (n = 11), cellular-based EVLP was performed for 2 hr. Each pair of lungs had a combination of warm ischemia and cold ischemia. PF ratio, airway parameters (peak airway pressure, plateau pressure, dynamic compliance, and static compliance), and vascular parameters (pulmonary vascular resistance and PA pressure) were measured and transplant suitability was evaluated. Out of
