Profiling inflammation and tissue injury markers in perfusate and bronchoalveolar lavage fluid during human ex vivo lung perfusion.
Andreasson Anders S I,Karamanou Danai M,Gillespie Colin S,Özalp Faruk,Butt Tanveer,Hill Paul,Jiwa Kasim,Walden Hannah R,Green Nicola J,Borthwick Lee A,Clark Stephen C,Pauli Henning,Gould Kate F,Corris Paul A,Ali Simi,Dark John H,Fisher Andrew J
European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery
Objectives:Availability of donor lungs suitable for transplant falls short of current demand and contributes to waiting list mortality. Ex vivo lung perfusion (EVLP) offers the opportunity to objectively assess and recondition organs unsuitable for immediate transplant. Identifying robust biomarkers that can stratify donor lungs during EVLP to use or non-use or for specific interventions could further improve its clinical impact. Methods:In this pilot study, 16 consecutive donor lungs unsuitable for immediate transplant were assessed by EVLP. Key inflammatory mediators and tissue injury markers were measured in serial perfusate samples collected hourly and in bronchoalveolar lavage fluid (BALF) collected before and after EVLP. Levels were compared between donor lungs that met criteria for transplant and those that did not. Results:Seven of the 16 donor lungs (44%) improved during EVLP and were transplanted with uniformly good outcomes. Tissue and vascular injury markers lactate dehydrogenase, HMGB-1 and Syndecan-1 were significantly lower in perfusate from transplanted lungs. A model combining IL-1β and IL-8 concentrations in perfusate could predict final EVLP outcome after 2 h assessment. In addition, perfusate IL-1β concentrations showed an inverse correlation to recipient oxygenation 24 h post-transplant. Conclusions:This study confirms the feasibility of using inflammation and tissue injury markers in perfusate and BALF to identify donor lungs most likely to improve for successful transplant during clinical EVLP. These results support examining this issue in a larger study.
Amelioration of lung ischemia‑reperfusion injury by JNK and p38 small interfering RNAs in rat pulmonary microvascular endothelial cells in an ischemia‑reperfusion injury lung transplantation model.
Wang Juan,Tan Jing,Liu Yanhong,Song Linlin,Li Di,Cui Xiaoguang
Molecular medicine reports
The inhibition of mitogen‑activated protein kinases (MAPKs), including c‑Jun NH2‑terminal protein kinase (JNK), p38 MAPK (p38) and extracellular signal‑regulated protein kinase 1/2 (ERK1/2), have an important effect on lung ischemia‑reperfusion injury (IRI) during lung transplantation (LT). However, the way in which combined MAPK inhibition exerts optimal protective effects on lung IRI remains to be elucidated. Therefore, the present study evaluated the therapeutic efficacy of the inhibition of MAPKs in rat pulmonary microvascular endothelial cells (PMVECs) in an IRI model of LT. The rat PMVECs were transfected with small interfering RNAs (siRNAs) against JNK, p38 or ERK1/2. Cotransfection was performed with siRNAs against JNK and p38 in the J+p group, JNK and ERK1/2 in the J+E group, p38 and ERK1/2 in the p+E group, or all three in the J+p+E group. Non‑targeting (NT) siRNA was used as a control. The PMVECs were then treated to induce IRI, and the levels of inflammation, apoptosis and oxidative stress were detected. Differences between compared groups were determined using Tukey's honest significant difference test. In all groups, silencing of the MAPKs was shown to attenuate inflammation, apoptosis and oxidative stress to differing extents, compared with the NT group. The J+p and J+p+E groups showed lower levels of interleukin (IL)‑1β, IL‑6 and malondialdehyde, a lower percentage of early‑apoptotic cells, and higher superoxide dismutase (SOD) activity, compared with the other groups. No significant differences were observed in the inflammatory response, SOD activity or early apoptosis between the J+p and J+p+E groups. These findings suggested that the dual inhibition of JNK and p38 led to maximal amelioration of lung IRI in the PMVECs of the IRI model of LT, which occurred through anti‑inflammatory, anti‑oxidative and anti‑apoptotic mechanisms.
Technetium-99m-labeled macroaggregated albumin lung perfusion scan for diagnosis of hepatopulmonary syndrome: A prospective study comparing brain uptake and whole-body uptake.
Zhao He,Tsauo Jiaywei,Zhang Xiao-Wu,Ma Huai-Yuan,Weng Ning-Na,Tang Gong-Shun,Li Xiao
World journal of gastroenterology
BACKGROUND:Hepatopulmonary syndrome (HPS) is an arterial oxygenation defect induced by intrapulmonary vascular dilatation (IPVD) in the setting of liver disease and/or portal hypertension. This syndrome occurs most often in cirrhotic patients (4%-32%) and has been shown to be detrimental to functional status, quality of life, and survival. The diagnosis of HPS in the setting of liver disease and/or portal hypertension requires the demonstration of IPVD (., diffuse or localized abnormally dilated pulmonary capillaries and pulmonary and pleural arteriovenous communications) and arterial oxygenation defects, preferably by contrast-enhanced echocardiography and measurement of the alveolar-arterial oxygen gradient, respectively. AIM:To compare brain and whole-body uptake of technetium for diagnosing HPS. METHODS:Sixty-nine patients with chronic liver disease and/or portal hypertension were prospectively included. Brain uptake and whole-body uptake were calculated using the geometric mean of technetium counts in the brain and lungs and in the entire body and lungs, respectively. RESULTS:Thirty-two (46%) patients had IPVD as detected by contrast-enhanced echocardiography. The demographics and clinical characteristics of the patients with and without IPVD were not significantly different with the exception of the creatinine level (0.71 ± 0.18 mg/dL 0.83 ± 0.23 mg/dL; = 0.041), alveolar-arterial oxygen gradient (23.2 ± 13.3 mmHg 16.4 ± 14.1 mmHg; = 0.043), and arterial partial pressure of oxygen (81.0 ± 12.1 mmHg 90.1 ± 12.8 mmHg; = 0.004). Whole-body uptake was significantly higher in patients with IPVD than in patients without IPVD (48.0% ± 6.1% 40.1% ± 8.1%; = 0.001). The area under the curve of whole-body uptake for detecting IPVD was significantly higher than that of brain uptake (0.75 0.54; = 0.025). The optimal cut-off values of brain uptake and whole-body uptake for detecting IPVD were 5.7% and 42.5%, respectively, based on Youden's index. The sensitivity, specificity, and accuracy of brain uptake > 5.7% and whole-body uptake > 42.5% for detecting IPVD were 23%, 89%, and 59% and 100%, 52%, and 74%, respectively. CONCLUSION:Whole-body uptake is superior to brain uptake for diagnosing HPS.
Fibrosing Mediastiniti: a Rare Cause of Unilateral Absent Lung Perfusion on a V/Q Scan.
Goldbach Alyssa R,Pascarella Suzanne,Dadpravarar Simin
Nuclear medicine and molecular imaging
We report a case of a 29-year-old female with a history of asthma, post-partum ARDS, and pulmonary hypertension who presents with severe shortness of breath. The patient describes her shortness of breath as progressive over the past 10 years. Chest radiography and CT angiography of the thorax showed findings consistent with fibrosing mediastinitis with severe stenosis of the left main pulmonary artery. This resulted in appearance of unilateral absent left lung perfusion on quantitative Tc-99-MAA perfusion and Xe-133 ventilation (V/Q) scan.
Oxymatrine on Hsp90a expression and apoptosis in a model of lung ischemia-reperfusion injury.
Zhu Bing,Yang Jianru,Chen Sifeng,Zhang Pei,Shen Lin,Li Xiaolong,Li Jing
Experimental and therapeutic medicine
The protective effects of oxymatrine (OMT) on apoptosis and heat shock protein 90a (Hsp90a) expression in a rabbit model of lung ischemia-reperfusion injury (LIRI) were investigated. The model of LIRI was established in rabbits and they were randomly divided into two groups: The control group (group C, n=10), and experimental group (further divided into groups E1, n=10; and group E2, n=10), to measure the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) activity in lung tissue homogenates at several time points (T, 0 min; T, 60 min; T, 120 min; T, 180 min; and T, 240 min), and to measures changes in lung tissue wet/dry weight ratio (W/D), apoptosis index (AI), and Hsp90a expression and organization at T, T and T. Comparing group C with groups E1 and E2, the levels of SOD activity and MDA were not significantly different at T and T (P>0.05); W/D ratio and AI were significantly higher than in groups E1 and E2 (P<0.05, P<0.01); 120 min after LIR, MDA, W/D ratio, and AI were lower than in groups E1 and E2 (P<0.05, P<0.01). MDA, W/D ratio and AI were lower in E2 than in E1 (P<0.05), and SOD and Hsp90a expression increased (P<0.05). The ultrastructure in group E showed less injury compared with group C. In conclusion, by scavenging oxygen free radicals, OMT can inhibit apoptosis, increase Hsp90a expression, and reduce the injury caused by lung ischemia reperfusion.
Beware Cold Agglutinins in Organ Donors! Ex Vivo Lung Perfusion From an Uncontrolled Donation After Circulatory-Determination-of-Death Donor With a Cold Agglutinin: A Case Report.
Venkataraman A,Blackwell J W,Funkhouser W K,Birchard K R,Beamer S E,Simmons W T,Randell S H,Egan T M
BACKGROUND:We began to recover lungs from uncontrolled donation after circulatory determination of death to assess for transplant suitability by means of ex vivo lung perfusion (EVLP) and computerized tomographic (CT) scan. Our first case had a cold agglutinin with an interesting outcome. CASE REPORT:A 60-year-old man collapsed at home and was pronounced dead by Emergency Medical Services personnel. Next-of-kin consented to lung retrieval, and the decedent was ventilated and transported. Lungs were flushed with cold Perfadex, removed, and stored cold. The lungs did not flush well. Medical history revealed a recent hemolytic anemia and a known cold agglutinin. Warm nonventilated ischemia time was 51 minutes. O-ventilated ischemia time was 141 minutes. Total cold ischemia time was 6.5 hours. At cannulation for EVLP, established clots were retrieved from both pulmonary arteries. At initiation of EVLP with Steen solution, tiny red aggregates were observed initially. With warming, the aggregates disappeared and the perfusate became red. After 1 hour, EVLP was stopped because of florid pulmonary edema. The lungs were cooled to 20°C; tiny red aggregates formed again in the perfusate. Ex vivo CT scan showed areas of pulmonary edema and a pyramidal right middle lobe opacity. Dissection showed multiple pulmonary emboli-the likely cause of death. However, histology showed agglutinated red blood cells in the microvasculature in pre- and post-EVLP biopsies, which may have contributed to inadequate parenchymal preservation. CONCLUSIONS:Organ donors with cold agglutinins may not be suitable owing to the impact of hypothermic preservation.
Evaluation of differential pulmonary perfusion using 99mTc macroaggregated albumin after the Fontan procedure.
Talwar Sachin,Sankhyan Lakshmi,Patel Chetan,Sreenivas Vishnubhatla,Choudhary Shiv Kumar,Airan Balram
Interactive cardiovascular and thoracic surgery
OBJECTIVES:The Fontan procedure [total cavopulmonary connection (TCPC)] is the final palliation for patients with univentricular physiology. We studied differential perfusion ratio and percentage uptake of a radiotracer in different zones of each lung following TCPC. METHODS:Between July 2015 and June 2017, 45 patients underwent 99mTc macroaggregated albumin lung perfusion scan at a mean follow-up period of 49.3 ± SD 26.1 days following TCPC. Differential perfusion ratio and percentage uptake of the radiotracer in the upper, middle and lower zones of each lung were calculated. RESULTS:Post-foot injection [inferior vena cava (IVC) injection], preferential flow to the lungs was as follows: left lung (n = 13, 30.2%), right lung (n = 13, 30.2%) and uniformly to both lungs (n= 17, 39.6%). Post-arm injection [superior vena cava (SVC) injection], preferential flow to the lungs was as follows: left lung (n = 13, 30.2%), right lung (n = 22, 51.2%) and uniformly to both lungs (n= 8, 18.6%). The middle zone was perfused the most in both lungs. Total lower zone mean perfusion was higher than the upper zone following both SVC injection and IVC injection (34.1 ± SD 5.3% vs 17. ± SD 4.1% and 33 ± SD 5.0% vs 17.5 ± SD 4.1%, respectively). In patients with bilateral SVC, post-IVC injection, 6 (75%) patients had preferential flow to the right lung, whereas post-SVC injection, preferential flow to the left lung was visualized in 7 (87.5%) patients. CONCLUSIONS:Following TCPC, IVC blood was distributed uniformly in both lungs. SVC blood preferentially perfused the right lung. The middle zone was perfused the most in both lungs.
Cobra Venom Factor-induced complement depletion protects against lung ischemia reperfusion injury through alleviating blood-air barrier damage.
Haihua Chang,Wei Wang,Kun Huang,Yuanli Liao,Fei Lin
The purpose of this study was to study whether complement depletion induced by pretreatment with Cobra Venom Factor (CVF) could protect against lung ischemia reperfusion injury (LIRI) in a rat model and explore its molecular mechanisms. Adult Sprague-Dawley rats were randomly assigned to five groups (n = 6): Control group, Sham-operated group, I/R group, CVF group, I/R + CVF group. CVF (50 μg/kg) was injected through the tail vein 24 h before anesthesia. Lung ischemia reperfusion (I/R) was induced by clamping the left hilus pulmonis for 60 minutes followed by 4 hours of reperfusion. Measurement of complement activity, pathohistological lung injury score, inflammatory mediators, pulmonary permeability, pulmonary edema, integrity of tight junction and blood-air barrier were performed. The results showed that pretreatment with CVF significantly reduced complement activity in plasma and BALF. Evaluation in histomorphology showed that complement depletion induced by CVF significantly alleviated the damage of lung tissues and inhibited inflammatory response in lung tissues and BALF. Furthermore, CVF pretreatment had the function of ameliorating pulmonary permeability and preserving integrity of tight junctions in IR condition. In conclusion, our results indicated that complement depletion induced by CVF could inhibit I/R-induced inflammatory response and alleviate lung I/R injury. The mechanisms of its protective effects might be ameliorated blood-air barrier damage.
Improvement of lung ischemia-reperfusion injury by inhibition of microRNA-155 via reductions in neuroinflammation and oxidative stress of vagal afferent nerve.
Zhou Yan,Zhang Lianjie,Guan Jingjing,Yin Xin
Lung ischemia-reperfusion injury (LIRI) is a common clinical concern. As the injury occurs, the pulmonary afferent nerves play a key role in regulating respiratory functions under pathophysiological conditions. The present study was to examine the effects of inhibiting microRNA-155 on the levels of proinflammatory cytokines and products of oxidative stress in the pulmonary vagal afferent nerves and the commissural nucleus of the solitary tract (cNTS) after LIRI. A rat model of LIRI was used. ELISA method was employed to examine proinflammatory cytokines, namely, IL-1β, IL-6 and TNF-α; and key biomarkers of oxidative stress, 8-isoprostaglandin F2α (8-iso PGF2α) and 8-hydroxy-2'-deoxyguanosine (8-OHdG). In results, in the process of LIRI, the levels of microRNA-155 were amplified in the vagal afferent nerves and cNTS, and this was accompanied with increases of IL-1β, IL-6 and TNF-α; and 8-iso PGF2α and 8-OHdG. Application of microRNA-155 inhibitor, but not its scramble, attenuated the elevation of proinflammatory cytokines and amplification of 8-iso PGF2α and 8-OHdG in those nerve tissues. In conclusion, we observed the abnormalities in the pulmonary afferent pathways at the levels of the peripheral nerves and brainstem, which is likely to affect respiratory functions as LIRI occurs. Our data suggest that blocking microRNA-155 signal pathways plays a beneficial role in regulating LIRI via inhibiting responses of neuroinflammation and oxidative stress signal pathways to LIRI.
A Porcine Lung Perfusion Model To Investigate Bacterial Pathogenesis.
Dumigan Amy,Fitzgerald Marianne,Santos Joana Sá-Pessoa Graca,Hamid Umar,O'Kane Cecilia M,McAuley Danny F,Bengoechea Jose A
The use of animal infection models is essential to understand microbial pathogenesis and to develop and test treatments. Insects and two-dimensional (2D) and 3D tissue models are increasingly being used as surrogates for mammalian models. However, there are concerns about whether these models recapitulate the complexity of host-pathogen interactions. In this study, we developed the lung perfusion (EVLP) model of infection using porcine lungs to investigate -triggered pneumonia as a model of respiratory infections. The porcine EVLP model recapitulates features of -induced pneumonia lung injury. This model is also useful to assess the pathogenic potential of , as we observed that the attenuated capsule mutant strain caused less pathological tissue damage with a concomitant decrease in the bacterial burden compared to that in lungs infected with the wild type. The porcine EVLP model allows assessment of inflammatory responses following infection; similar to the case with the mouse pneumonia model, we observed an increase of in the lungs infected with the wild type and an increase of in lungs infected with the capsule mutant. This model also allows monitoring of phenotypes at the single-cell level. Wild-type skews macrophages toward an M2-like state. experiments probing pig bone marrow-derived macrophages uncovered the role for the M2 transcriptional factor STAT6 and that -induced expression is controlled by p38 and extracellular signal-regulated kinase (ERK). -induced macrophage polarization is dependent on the capsule. Together, the findings of this study support the utility of the EVLP model using pig lungs as a platform to investigate the infection biology of respiratory pathogens. The implementation of infection models that approximate human disease is essential to understand infections and for testing new therapies before they enter into clinical stages. Rodents are used in most preclinical studies, although the differences between mice and humans have fueled the conclusion that murine studies are unreliable predictors of human outcomes. In this study, we have developed a whole-lung porcine model of infection using the lung perfusion (EVLP) system established to recondition human lungs for transplant. As a proof of principle, we provide evidence demonstrating that infection of the porcine EVLP with the human pathogen recapitulates the known features of -triggered pneumonia. Moreover, our data revealed that the porcine EVLP model is useful to reveal features of the virulence of , including the manipulation of immune cells. Together, the findings of this study support the utility of the EVLP model using pig lungs as a surrogate host for assessing respiratory infections.
Pyrrolidine dithiocarbamate administered during ex-vivo lung perfusion promotes rehabilitation of injured donor rat lungs obtained after prolonged warm ischemia.
Francioli Cyril,Wang Xingyu,Parapanov Roumen,Abdelnour Etienne,Lugrin Jérôme,Gronchi Fabrizio,Perentes Jean,Eckert Philippe,Ris Hans-Beat,Piquilloud Lise,Krueger Thorsten,Liaudet Lucas
Damaged lung grafts obtained after circulatory death (DCD lungs) and warm ischemia may be at high risk of reperfusion injury after transplantation. Such lungs could be pharmacologically reconditioned using ex-vivo lung perfusion (EVLP). Since acute inflammation related to the activation of nuclear factor kappaB (NF-κB) is instrumental in lung reperfusion injury, we hypothesized that DCD lungs might be treated during EVLP by pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB. Rat lungs exposed to 1h warm ischemia and 2 h cold ischemia were subjected to EVLP during 4h, in absence (CTRL group, N = 6) or in presence of PDTC (2.5g/L, PDTC group, N = 6). Static pulmonary compliance (SPC), peak airway pressure (PAWP), pulmonary vascular resistance (PVR), and oxygenation capacity were determined during EVLP. After EVLP, we measured the weight gain of the heart-lung block (edema), and the concentration of LDH (cell damage), proteins (permeability edema) and of the cytokines IL-6, TNF-α and CINC-1 in bronchoalveolar lavage (BAL), and we evaluated NF-κB activation by the degree of phosphorylation and degradation of its inhibitor IκBα in lung tissue. In CTRL, we found significant NF-κB activation, lung edema, and a massive release of LDH, proteins and cytokines. SPC significantly decreased, PAWP and PVR increased, while oxygenation tended to decrease. Treatment with PDTC during EVLP inhibited NF-κB activation, did not influence LDH release, but markedly reduced lung edema and protein concentration in BAL, suppressed TNFα and IL-6 release, and abrogated the changes in SPC, PAWP and PVR, with unchanged oxygenation. In conclusion, suppression of innate immune activation during EVLP using the NF-κB inhibitor PDTC promotes significant improvement of damaged rat DCD lungs. Future studies will determine if such rehabilitated lungs are suitable for in vivo transplantation.
Prognostic biological factors of radiation pneumonitis after stereotactic body radiation therapy combined with pulmonary perfusion imaging.
Guo Leiming,Ding Gaofeng,Xu Wencai,Lu Yufei,Ge Hong,Jiang Yue,Chen Xijuan,Li Yin
Experimental and therapeutic medicine
Radiation pneumonitis (RP) is one of the most common dose-limiting toxicity syndromes in patients with thoracic malignant tumors receiving radiotherapy. The present study aimed to identify biological factors for the prediction of RP. Pulmonary perfusion imaging is capable of reflecting the differential functional activity of various regions of the lung, and in the present study, radiotherapy plans that were established on the basis that pulmonary perfusion images have high biological conformality, which may identify regions vulnerable to RP to spare them from radiation. A total of 46 patients with non-small cell lung cancer (NSCLC), exhibiting high and low levels of apurinic/apyrimidinic endonuclease-1 (Ape-1), intercellular adhesion molecule (ICAM)-1 and interleukin (IL)-17A prior to treatment, with SBRT with respective cut-off values of 4.2, 3.0 and 5.1 µg/l were stratified into groups A and B. Patients received radiation doses within the margin of the planning target volume. Stereotactic body radiation therapy (SBRT) was used for the treatment of NSCLC and single-photon emission computed tomography pulmonary perfusion imaging was used to assess all patients for the presence of RP. Furthermore, the serum levels of Ape-1, ICAM-1 and IL-17A were examined by ELISA. Prior to SBRT, perfusion images indicated that no RP was present in any of the patients, and 23 patients had high levels of Ape-1, ICAM-1 and IL-17A. After SBRT, 22 out of 23 patients in group A (95.65%) presented with RP and 1 patient (4.35%) had no RP. In group B, 6 out of 23 patients (26.09%) had RP and 17 patients (73.91%) had no RP after SBRT. The difference between the two groups in the incidence of RP was significant (P=1.66×10 <0.05). In conclusion, high levels of Ape-1, ICAM-1 and IL-17A are associated with an increased risk of RP. A further analysis should be performed in the future to verify whether these factors have significant prognostic value.
Pushing the limits of reconditioning: extended normothermic lung perfusion in an extended criteria donor.
Schiavon Marco,Zampieri Davide,Marulli Giuseppe,Verderi Enrico,Rebusso Alessandro,Comacchio Giovanni Maria,Nicotra Samuele,Loy Monica,Lunardi Francesca,Feltracco Paolo,Calabrese Fiorella,Cozzi Emanuele,Rea Federico
Journal of thoracic disease
Short-interval exposure to ambient fine particulate matter (PM2.5) exacerbates the susceptibility of pulmonary damage in setting of lung ischemia-reperfusion injury in rodent: Pharmacomodulation of melatonin.
Lee Fan-Yen,Lee Mel S,Wallace Christopher Glenn,Huang Chi-Ruei,Chu Chi-Hsiang,Wen Zhi-Hong,Huang Jhih-Hong,Chen Xue-Sheng,Wang Chia C,Yip Hon-Kan
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
This study tested the hypothesis that exposure to ambient fine particulate matter (PM) pollution increased susceptibility of rat lung to damage from acute ischemia-reperfusion (IR) injury that was reversed by melatonin (Mel) treatment. Male-adult SD rats (n = 30) were categorized into group 1 (normal control), group 2 (PM only), group 3 (IR only at day 8 after PM exposure), group 4 (PM + IR) and group 5 (PM + IR + Mel), and all animals were sacrificed by day 10 after PM exposure. Oxygen saturation (%) was significantly higher in group 1 than in other groups and significantly lower in group 4 than in groups 2, 3 and 5 but it did not differ among the latter three groups (p < 0.01). Pulmonary protein expressions of inflammation (MMP-9/TNF-α/NF-kB), oxidative stress (NOX-1/NOX-2/oxidized protein), apoptosis (mitochondrial-Bax/caspase-3/PARP) and fibrosis were lowest in group 1, highest in group 4, significantly higher in group 5 than in groups 2 and 3 (all p < 0.0001), but they did not differ between groups 2 and 3. Inflammatory cell infiltration in lung parenchyma, specific inflammatory cell surface markers (CD14+, F4/88+), allergic inflammatory cells (IgE+, eosinophil+), number of goblet cells, thickness of tracheal epithelial layer and fibrotic area exhibited an identical pattern of protein expressions to inflammation among the five groups (all p < 0.0001). In conclusion, lung parenchymal damage and a rigorous inflammatory response were identified in rodent even with short-term PM exposure.
Hepatopulmonary Syndrome with Right-to-left Shunt in Cirrhotic Patients Using Macro-Aggregated Albumin Lung Perfusion Scan: Comparison with Contrast Echocardiography and Association with Clinical Data
Alipour Zeynab,Armin Abbas,Mohamadi Sudabeh,Tabib Seyed Masoud,Azizmohammadi Zahra,Gholamrezanezhad Ali,Assadi Majid
Molecular imaging and radionuclide therapy
Objectives:The diagnosis of hepatopulmonary syndrome (HPS) which is a common complication in cirrhotic patients is still subject to debate. This study investigated the association of clinical findings with HPS in cirrhotic patients using macro-aggregated albumin lung perfusion scan (Tc-MAA lung scintigraphy). In addition, comparison between Tc-MAA lung scintigraphy and contrast echocardiography (CEE) in detection of HPS was also performed. Methods:In this study, 27 patients with cirrhosis underwent Tc-MAA lung scintigraphy and contrast echocardiography comparison CEE and the frequency of HPS was assessed in them and also was compared across the other variables. Results:The Tc-MAA lung scintigraphy showed HPS in 13 patients (48.1%) while CEE demonstrated HPS in 5 patients with cirrhosis (18.51%). HPS was mild in 40.74% (11/27) of the patients, and severe in only 2 patients. There was no relationship between gender, disease duration, having diagnosis of disease previously, pulmonary symptoms and Child-Pugh score variations and HPS (p>0.05). Comparison of hemodynamic indices, arterial blood gas analysis and laboratory indices between patients with and without HPS was also non-significant (p value >0.05). Among coagulation factors assessed in cirrhotic patients, we found only significant correlation between HPS and prothrombin time (p<0.05). Conclusion:HPS, particularly its mild form, is noted in a great number of patients with cirrhosis using Tc-MAA lung scintigraphy. Because of its technical ease, and possibility to obtain objective quantitative information, Tc-MAA lung scintigraphy can be complementary to other diagnostic methods in the evaluation of HPS assessment, although additional studies are needed.
Comparison of Two Different Segmentation Methods on Planar Lung Perfusion Scan with Reference to Quantitative Value on SPECT/CT.
Suh Minseok,Kang Yeon-Koo,Ha Seunggyun,Kim Yong-Il,Paeng Jin Chul,Cheon Gi Jeong,Park Samina,Kim Young Tae,Lee Dong Soo,Kim E Edmund,Chung June-Key
Nuclear medicine and molecular imaging
PURPOSE:Until now, there was no single standardized regional segmentation method of planar lung perfusion scan. We compared planar scan based two segmentation methods, which are frequently used in the Society of Nuclear Medicine, with reference to the lung perfusion single photon emission computed tomography (SPECT)/computed tomography (CT) derived values in lung cancer patients. METHODS:Fifty-five lung cancer patients (male:female, 37:18; age, 67.8 ± 10.7 years) were evaluated. The patients underwent planar scan and SPECT/CT after injection of technetium-99 m macroaggregated albumin (Tc-99 m-MAA). The % uptake and predicted postoperative percentage forced expiratory volume in 1 s (ppoFEV1%) derived from both posterior oblique (PO) and anterior posterior (AP) methods were compared with SPECT/CT derived parameters. Concordance analysis, paired comparison, reproducibility analysis and spearman correlation analysis were conducted. RESULTS:The % uptake derived from PO method showed higher concordance with SPECT/CT derived % uptake in every lobe compared to AP method. Both methods showed significantly different lobar distribution of % uptake compared to SPECT/CT. For the target region, ppoFEV1% measured from PO method showed higher concordance with SPECT/CT, but lower reproducibility compared to AP method. Preliminary data revealed that every method significantly correlated with actual postoperative FEV1%, with SPECT/CT showing the best correlation. CONCLUSION:The PO method derived values showed better concordance with SPECT/CT compared to the AP method. Both PO and AP methods showed significantly different lobar distribution compared to SPECT/CT. In clinical practice such difference according to different methods and lobes should be considered for more accurate postoperative lung function prediction.
Cellular and acellular ex vivo lung perfusion preserve functional lung ultrastructure in a large animal model: a stereological study.
Steinmeyer Jasmin,Becker Simon,Avsar Murat,Salman Jawad,Höffler Klaus,Haverich Axel,Warnecke Gregor,Mühlfeld Christian,Ochs Matthias,Schnapper-Isl Anke
BACKGROUND:Ex vivo lung perfusion (EVLP) is used by an increasing number of transplant centres. It is still controversial whether an acellular or cellular (erythrocyte enriched) perfusate is preferable. The aim of this paper was to evaluate whether acellular (aEVLP) or cellular EVLP (cEVLP) preserves functional lung ultrastructure better and to generate a hypothesis regarding possible underlying mechanisms. METHODS:Lungs of 20 pigs were assigned to 4 groups: control, ischaemia (24 h), aEVLP and cEVLP (both EVLP groups: 24 h ischaemia + 12 h EVLP). After experimental procedures, whole lungs were perfusion fixed, samples for light and electron microscopic stereology were taken, and ventilation, diffusion and perfusion related parameters were estimated. RESULTS:Lung structure was well preserved in all groups. Lungs had less atelectasis and higher air content after EVLP. No significant group differences were found in alveolar septum composition or blood-air barrier thickness. Small amounts of intraalveolar oedema were detected in both EVLP groups but significantly more in aEVLP than in cEVLP. CONCLUSIONS:Both EVLP protocols supported lungs well for up to 12 h and could largely prevent ischaemia ex vivo reperfusion associated lung injury. In both EVLP groups, oedema volume remained below the level of functional relevance. The group difference in oedema formation was possibly due to inferior septal perfusion in aEVLP.
Lung ischemia reperfusion injury: the therapeutic role of dipeptidyl peptidase 4 inhibition.
Beckers Paul A J,Gielis Jan F,Van Schil Paul E,Adriaensen Dirk
Annals of translational medicine
Dipeptidyl peptidase 4 (DPP4) is a cell surface protease that has been reported to play a role in glucose homeostasis, cancer, HIV, autoimmunity, immunology and inflammation. A role for DPP4 in ischemia-reperfusion injury (IRI) in the heart has been established. Dipeptidyl peptidase 4 inhibition (DPP4i) appeared to decrease infarct size, improves cardiac function and promotes myocardial regeneration. Lung ischemia reperfusion injury is caused by a complex mechanism in which macrophages and neutrophils play an important role. Generation of reactive oxygen species (ROS), uncoupling of nitric oxide synthase (NOS), activation of nuclear factor-κB (NF-κB), activation of nicotinamide adenine dinucleotide phosphate metabolism, and generation of pro-inflammatory cytokines lead to acute lung injury (ALI). In this review we present the current knowledge on DPP4 as a target to treat IRI in the lung. We also provide evidence of the roles of the DPP4 substrates glucagon-like peptide 1 (GLP-1), vasoactive intestinal peptide (VIP) and stromal cell-derived factor-1α (SDF-1α) in protection against oxidative stress through activation of the mitogen-activated protein kinase (MAPK) 1/2 and phosphatidylinositol 3'-kinase (PI3K)/Akt signal transduction pathways.
Pulmonary Perfusion Using Intrabronchial Capnography in Pulmonary Artery Stenosis.
Nishine Hiroki,Muraoka Hiromi,Inoue Takeo,Miyazawa Teruomi,Mineshita Masamichi
Respiration; international review of thoracic diseases
Stenting at the flow-limiting segment can improve the ventilation-perfusion ratio in patients with central airway stenosis. However, there is no quantitative examination for assessing the perfusion status during interventional bronchoscopy. Intrabronchial capnography can estimate regional gas exchange by measuring carbon dioxide concentration. We herein report a case of bilateral bronchial stenosis where stenting was able to improve ventilation-perfusion ratio using intrabronchial capnography. A 44-year-old man was admitted to our institution with orthopnea. Chest computed tomography showed an extrinsic compression at the bilateral main bronchus and right pulmonary artery due to a mediastinal mass. After introduction of general anesthesia, arterial oxygen tension suddenly decreased in the supine position. After initial stenting, an increase was seen in ventilation at the right lung; however, a ventilation-perfusion mismatch occurred due to an increase in dead-space ventilation at the right pulmonary artery stenosis. Intrabronchial capnography was an effective modality to confirm the regional perfusion status during interventional bronchoscopy in real time.
Netrin-1 reduces lung ischemia-reperfusion injury by increasing the proportion of regulatory T cells.
Chen Zhili,Chen Yuxi,Zhou Jue,Li Yong,Gong Changyao,Wang Xiaobo
The Journal of international medical research
OBJECTIVE:Inflammation is the primary mechanism of lung ischemia-reperfusion injury (LIRI) and neurologic factors can regulate inflammatory immune responses. Netrin-1 is an axonal guidance molecule, but whether Netrin-1 plays a role in LIRI remains unclear. METHODS:A mouse model of LIRI was established. Immunohistochemistry was used to detect expression of Netrin-1 and to enumerate macrophages and T cells in lung tissue. The proportion of regulatory T cells (Tregs) was assessed by flow cytometry. Levels of apoptosis were assessed by terminal deoxynucleotidyl transferase dUTP nick end staining. RESULTS:Numbers of macrophages and T cells in the lung tissues of mice with LIRI were elevated, while expression of netrin-1 was significantly decreased. Flow cytometry showed that the proportion of Tregs in mice with LIRI was significantly decreased. The proportion of Tregs among lymphocytes was positively correlated with netrin-1 expression. experiments showed that netrin-1 promoted an increase in Treg proportion through the A2b receptor. Animal experiments showed that netrin-1 could inhibit apoptosis and reduce T cell and macrophage infiltration by increasing the proportion of Tregs, ultimately reducing LIRI. Treg depletion using an anti-CD25 monoclonal antibody blocked the effects of netrin-1. CONCLUSION:Netrin-1 reduced LIRI by increasing the proportion of Tregs.
Adiponectin protects against lung ischemia-reperfusion injury in rats with type 2 diabetes mellitus.
Li Di,Song Lin-Lin,Wang Juan,Meng Chao,Cui Xiao-Guang
Molecular medicine reports
Adiponectin (APN) has been associated with the pathogenesis of acute brain, liver and heart injury. However, the role of APN in lung ischemia-reperfusion injury (LIRI) in diabetes mellitus remains unclear. To investigate this, the present study evaluated the effects of APN on lung dysfunction and pathological alterations in rats with type 2 diabetes mellitus via lung ischemia/reperfusion (I/R). The lung‑protective effects of APN globular domain (gAPN) in rats with type 2 diabetes mellitus were also investigated by measuring the oxygenation index, inflammatory cytokines, lung edema, histopathology, oxidative stress, apoptosis and the protein expression levels of phosphorylated 5'adenosine monophosphate‑activated protein kinase (p‑AMPK), endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS). The results of the present study demonstrated that the diabetes mellitus rats + I/R (DIR) group exhibited greater concentrations of tumor necrosis factor‑α and interleukin‑6, and increases in the wet‑weight to dry‑weight ratio, lung injury score, oxidative stress and cellular apoptosis. These effects were accompanied by lower pulmonary oxygenation compared with the normal rat + I/R (NIR) group (P<0.05). Additionally, all of these alterations were attenuated in the NIR + gAPN and DIR + gAPN groups compared with in the NIR and DIR groups, respectively. In the DIR group, the expression levels of p‑AMPK/AMPK and eNOS were significantly downregulated, and the levels of iNOS were upregulated, compared with those of the NIR group. Treatment with APN activated AMPK, increased eNOS expression and attenuated iNOS expression. The results of the present study demonstrated that APN exerted protective effects against LIRI via its anti‑inflammatory, antioxidative stress and anti‑apoptotic activities. These protective effects of APN were eliminated in rats with type 2 diabetes mellitus, in which LIRI was exacerbated. The present study indicated that APN may be a potential therapeutic agent for LIRI in type 2 diabetes mellitus.
Imaging of pulmonary perfusion using subtraction CT angiography is feasible in clinical practice.
Grob Dagmar,Oostveen Luuk J,Prokop Mathias,Schaefer-Prokop Cornelia M,Sechopoulos Ioannis,Brink Monique
Subtraction computed tomography (SCT) is a technique that uses software-based motion correction between an unenhanced and an enhanced CT scan for obtaining the iodine distribution in the pulmonary parenchyma. This technique has been implemented in clinical practice for the evaluation of lung perfusion in CT pulmonary angiography (CTPA) in patients with suspicion of acute and chronic pulmonary embolism, with acceptable radiation dose. This paper discusses the technical principles, clinical interpretation, benefits and limitations of arterial subtraction CTPA. KEY POINTS: • SCT uses motion correction and image subtraction between an unenhanced and an enhanced CT scan to obtain iodine distribution in the pulmonary parenchyma. • SCT could have an added value in detection of pulmonary embolism. • SCT requires only software implementation, making it potentially more widely available for patient care than dual-energy CT.
Maresin 1 Ameliorates Lung Ischemia/Reperfusion Injury by Suppressing Oxidative Stress via Activation of the Nrf-2-Mediated HO-1 Signaling Pathway.
Sun Quanchao,Wu You,Zhao Feng,Wang Jianjun
Oxidative medicine and cellular longevity
Lung ischemia/reperfusion (I/R) injury occurs in various clinical conditions and heavily damaged lung function. Oxidative stress reaction and antioxidant enzymes play a pivotal role in the etiopathogenesis of lung I/R injury. In the current study, we investigated the impact of Maresin 1 on lung I/R injury and explored the possible mechanism involved in this process. MaR 1 ameliorated I/R-induced lung injury score, wet/dry weight ratio, myeloperoxidase, tumor necrosis factor, bronchoalveolar lavage fluid (BALF) leukocyte count, BALF neutrophil ratio, and pulmonary permeability index levels in lung tissue. MaR 1 significantly reduced ROS, methane dicarboxylic aldehyde, and 15-F2t-isoprostane generation and restored antioxidative enzyme (superoxide dismutase, glutathione peroxidase, and catalase) activities. Administration of MaR 1 improved the expression of nuclear Nrf-2 and cytosolic HO-1 in I/R-treated lung tissue. Furthermore, we also found that the protective effects of MaR 1 on lung tissue injury and oxidative stress were reversed by HO-1 activity inhibitor, Znpp-IX. Nrf-2 transcription factor inhibitor, brusatol, significantly decreased MaR 1-induced nuclear Nrf-2 and cytosolic HO-1 expression. In conclusion, these results indicate that MaR 1 protects against lung I/R injury through suppressing oxidative stress. The mechanism is partially explained by activation of the Nrf-2-mediated HO-1 signaling pathway.
Normothermic ex vivo lung perfusion: Does the indication impact organ utilization and patient outcomes after transplantation?
Cypel Marcelo,Yeung Jonathan C,Donahoe Laura,Chen Manyin,Zamel Ricardo,Hoetzenecker Konrad,Yasufuku Kazuhiro,de Perrot Marc,Pierre Andrew F,Waddell Thomas K,Keshavjee Shaf
The Journal of thoracic and cardiovascular surgery
BACKGROUND:Ex vivo lung perfusion (EVLP) is being increasingly applied as a method to evaluate and treat donor lungs for transplantation. However, with the previous limited worldwide experience, no studies have been able to evaluate the impact of indication for EVLP on organ utilization rates and recipient outcomes after lung transplantation (LTx). We examined these outcomes in a large-cohort, single-center series of clinical EVLP cases. METHODS:All EVLP procedures performed at our institution between October 2008 and December 2017 were examined. The EVLPs were divided into 4 groups based on the indication for the procedure: group 1, high-risk brain death donors (HR-BDD); group 2, standard-risk donation after cardiac death (S-DCD); group 3, high-risk donation after cardiac death (HR-DCD); and group 4, logistics (LOGISTICS, the need for prolongation of preservation time or organ retrieval by a different transplantation team). RESULTS:During the study period, a total of 1106 lung transplants were performed in our institution. In this period, 372 EVLPs were performed, 255 (69%) of which were accepted for transplantation, resulting in 262 transplants. Utilization rates were 70% (140 of 198) for group 1, 82% (40 of 49) for group 2, 63% (69 of 109) for group 3, and 81% (13 of 16) for group 4 (P = .42, Fisher's exact test). Recipient age (P = .27) and medical diagnosis (P = .31) were not different across the 4 groups. Kaplan-Meier survival by EVLP indication group demonstrated no differences. Thirty-day mortality was 2.1% in group 1, 5% in group 2, 2.9% in group 3, and 0% in group 4 (P = .87, Fisher's exact test). The median days of mechanical ventilation, intensive care unit stay, and hospital stay were 2, 4, and 21 in group 1; 2, 3, and 21 in group 2; 3, 5, and 28 in group 3; and 2, 4, and 17 in group 4 (P = .29, .17, and .09, respectively, Kruskal-Wallis rank-sum test). CONCLUSIONS:Clinical implementation of EVLP has allowed our program to expand the annual lung transplantation activity by 70% in this time period. It has improved confidence in the utilization of DCD lungs and BDD lungs, with an average 70% utilization of post-EVLP treated donor lungs with excellent outcomes, while addressing significant challenges in donor lung assessment and the logistics of "real-life" clinical lung transplantation.
Donor Leukocyte Trafficking and Damage-associated Molecular Pattern Expression During Ex Vivo Lung Perfusion.
Davis Robert P,Yerxa John,Gao Qimeng,Gloria Jared,Scheuermann Uwe,Song Mingqing,Zhang Min,Parker William,Lee Jaewoo,Hartwig Matthew G,Barbas Andrew S
Background:While ex vivo lung perfusion (EVLP) has become established in lung transplantation, the cellular processes occurring during this period are not yet fully understood. Prior studies demonstrated that donor leukocytes (DLs) migrate from the graft into the perfusate during EVLP, but the distribution of DLs in graft and perfusate compartments has not been characterized. Moreover, cell death of DLs has been implicated in mediating graft injury during EVLP, but the underlying mechanisms have not been elucidated. We hypothesized the following: (1) there is a nonspecific migration of DLs from the graft into perfusate and (2) cell death of DLs releases damage-associated molecular patterns (DAMPs) that contribute to the inflammatory milieu during EVLP. Methods:EVLP was performed on rat lungs for 3 hours (N = 6). At the end of EVLP, flow cytometry was used to quantify the distribution of different DL cell types in both the graft and perfusate compartments. During EVLP, the perfusate was also sampled hourly to measure levels of DAMPs and downstream inflammatory cytokines generated during EVLP. Results:At the conclusion of EVLP, there was a significantly higher proportion of T and B cells present in the perfusate compartment compared with the graft compartment. There was a time-dependent increase in extracellular DNA and tumor necrosis factor α in the perfusate during EVLP. Conclusions:T cells and B cells are enriched in the perfusate compartment during EVLP. Cell death of DLs contributes to an accumulation of DAMPs during EVLP.
Value of lung perfusion scintigraphy in patients with idiopathic pulmonary arterial hypertension: a patchy pattern to consider.
Wang Meng,Ma Rongzheng,Wu Dayong,Xiong Changming,He Jianguo,Wang Lei,Sun Xiaoxin,Fang Wei
The ventilation/perfusion lung scan is recommended to exclude chronic thromboembolic pulmonary hypertension in the diagnostic algorithm of pulmonary hypertension, but its role in pulmonary arterial hypertension (PAH) has not been well explored. We characterized the lung perfusion pattern assessed by lung perfusion scintigraphy in idiopathic PAH (IPAH) patients and evaluate the potential prognostic significance of the patchy pattern perfusion defect. A total of 318 patients with IPAH confirmed by right heart catheterization who performed lung perfusion scintigraphy were included. On lung perfusion scintigraphy, 134 patients had normal lung perfusion and 184 patients showed patchy perfusion defects. In comparison to patients with normal lung perfusion, patients with patchy perfusion defects experienced significantly higher mean pulmonary arterial pressure (58.0 ± 15.4 mmHg vs. 54.1 ± 16.2 mmHg, P = 0.027) and total pulmonary resistance (1192.6 ± 533.7 dyn·s·cm vs. 1067.2 ± 549.3 dyn·s·cm, P = 0.042). During a median follow-up period of 884.0 days, 53 patients reached the primary endpoint of all-cause mortality. On univariate Cox analysis, the patchy pattern of perfusion defect was significantly associated with the all-cause mortality (hazard ratio [HR] = 2.47, 95% confidence interval [CI] = 1.32-4.63, P = 0.005). Patients with patchy perfusion defects had a worse outcome (log-rank = 8.605, P = 0.003). On multivariate analysis, the patchy pattern remained as a significant independent predictor of the endpoint (HR = 2.30, 95% CI = 1.22-4.31, P = 0.010). IPAH patients presented with heterogeneity in lung perfusion and the patchy pattern of lung perfusion defect commonly existed. Patients with patchy pattern identified by lung perfusion scintigraphy were associated with more severe disease and worse outcome.
Ac2-26 ameliorates lung ischemia-reperfusion injury via the eNOS pathway.
Gong Jing,Ju Ying-Nan,Wang Xue-Ting,Zhu Jing-Li,Jin Zhe-Hao,Gao Wei
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie
BACKGROUND:Lung ischemia-reperfusion injury (LIRI) is a major complication after lung transplantation. Annexin A1 (AnxA1) ameliorates inflammation in various injured organs. This study aimed to determine the effects and mechanism of AnxA1 on LIRI after lung transplantation. METHODS:Thirty-two rats were randomized into sham, saline, Ac2-26 and Ac2-26/L groups. Rats in the saline, Ac2-26 and Ac2-26/L groups underwent left lung transplantation and received saline, Ac2-26, and Ac2-26/L-NIO, respectively. After 24 h of reperfusion, serum and transplanted lung tissues were examined. RESULTS:The partial pressure of oxygen (PaO) was increased in the Ac2-26 group compared to that in the saline group but was decreased by L-NIO treatment. In the Ac2-26 group, the wet-to-dry (W/D) weight ratios, total protein concentrations, proinflammatory factors and inducible nitric oxide synthase levels were notably decreased, but the concentrations of anti-inflammatory factors and endothelial nitric oxide synthase levels were significantly increased. Ac2-26 attenuated histological injury and cell apoptosis, and this improvement was reversed by L-NIO. CONCLUSIONS:Ac2-26 reduced LIRI and improved alveoli-capillary permeability by inhibiting oxygen stress, inflammation and apoptosis. The protective effect of Ac2-26 on LIRI largely depended on the endothelial nitric oxide synthase pathway.
Lung protection by inhalation of exogenous solubilized extracellular matrix.
Wu Jinglei,Ravikumar Priya,Nguyen Kytai T,Hsia Connie C W,Hong Yi
Decellularized extracellular matrix (ECM) contains complex tissue-specific components that work in concert to promote tissue repair and constructive remodeling and has been used experimentally and clinically to accelerate epithelial wound repair, leading us to hypothesize that lung-derived ECM could mitigate acute lung injury. To explore the therapeutic potential of ECM for noninvasive delivery to the lung, we decellularized and solubilized porcine lung ECM, then characterized the composition, concentration, particle size and stability of the preparation. The ECM preparation at 3.2 mg/mL with average particle size <3 μm was tested in vitro on human A549 lung epithelial cells exposed to 95% O2 for 24 hours, and in vivo by tracheal instillation or nebulization into the lungs of rats exposed intermittently or continuously to 90% O2 for a cumulative 72 hours. Our results showed that the preparation was enriched in collagen, reduced in glycosaminoglycans, and contained various bioactive molecules. Particle size was concentration-dependent. Compared to the respective controls treated with cell culture medium in vitro or saline in vivo, ECM inhalation normalized cell survival and alveolar morphology, and reduced hyperoxia-induced apoptosis and oxidative damage. This proof-of-concept study established the methodology, feasibility and therapeutic potential of exogenous solubilized ECM for pulmonary cytoprotection, possibly as an adjunct or potentiator of conventional therapy.
Immunoregulatory effects of multipotent adult progenitor cells in a porcine ex vivo lung perfusion model.
Martens An,Ordies Sofie,Vanaudenaerde Bart M,Verleden Stijn E,Vos Robin,Van Raemdonck Dirk E,Verleden Geert M,Roobrouck Valerie D,Claes Sandra,Schols Dominique,Verbeken Eric,Verfaillie Catherine M,Neyrinck Arne P
Stem cell research & therapy
BACKGROUND:Primary graft dysfunction (PGD) is considered to be the end result of an inflammatory response targeting the new lung allograft after transplant. Previous research has indicated that MAPC cell therapy might attenuate this injury by its paracrine effects on the pro-/anti-inflammatory balance. This study aims to investigate the immunoregulatory capacities of MAPC cells in PGD when administered in the airways. METHODS:Lungs of domestic pigs (n = 6/group) were subjected to 90 minutes of warm ischemia. Lungs were cold flushed, cannulated on ice and placed on EVLP for 6 hours. At the start of EVLP, 40 ml of an albumin-plasmalyte mixture was distributed in the airways (CONTR group). In the MAPC cell group, 150 million MAPC cells (ReGenesys/Athersys, Cleveland, OH, USA) were added to this mixture. At the end of EVLP, a physiological evaluation (pulmonary vascular resistance, lung compliance, PaO/FiO), wet-to-dry weight ratio (W/D) sampling and a multiplex analysis of bronchoalveolar lavage (BAL) (2 × 30 ml) was performed. RESULTS:Pulmonary vascular resistance, lung compliance, PaO/FiO and W/D were not statistically different at the end of EVLP between both groups. BAL neutrophilia was significantly reduced in the MAPC cell group. Moreover, there was a significant decrease in TNF-α, IL-1β and IFN-γ in the BAL, but not in IFN-α; whereas IL-4, IL-10 and IL-8 were below the detection limit. CONCLUSIONS:Although no physiologic effect of MAPC cell distribution in the airways was detected during EVLP, we observed a reduction in pro-inflammatory cytokines and neutrophils in BAL in the MAPC cell group. This effect on the innate immune system might play an important role in critically modifying the process of PGD after transplantation. Further experiments will have to elucidate the immunoregulatory effect of MAPC cell administration on graft function after transplantation.
Is "lung repair centre" a possible answer to organ shortage?-transplantation of left and right lung at two different centres after ex vivo lung perfusion evaluation and repair: case report.
Palleschi Alessandro,Rosso Lorenzo,Schiavon Marco,Rebusso Alessandro,Mendogni Paolo,Rea Federico,Santambrogio Luigi,Nosotti Mario,Valenza Franco
Journal of thoracic disease
Ex vivo lung perfusion (EVLP) has become a reality as a technique to evaluate and recondition lungs from marginal donors. We report the first case on the use of EVLP followed by separate transplantation in two different centres. The local organ procurement organization proposed the lungs of a 53-year-old non-smoker donor who died for cerebral haemorrhage. P/F ratio was 294 after lung recruitment manoeuvres. Oto score was 10. Two centres accepted the grafts for two single transplantations under the condition of EVLP evaluation. After usual retrieval, the bi-pulmonary block was transferred to Centre 1 and EVLP was run as previously described. At the end of the procedure the two lungs were evaluated separately and both judged suitable for transplantation. After cooling and storage on ice, the block was separated on the back table. The left lung was transplanted in a patient with pulmonary fibrosis at Centre 1; surgery was complicated by cardiac arrhythmias that required several defibrillations. The right lung was transferred on ice to Centre 2, 250 km away from Centre 1, and transplanted in a patient with idiopathic pulmonary fibrosis. Thirty months after transplantations Patient 1 and Patient 2 are both alive, in good clinical conditions. This is the first report of the separate use of lungs after EVLP for non-urgent recipients in two different centres. This experience opens the door to a new allocation model with great potentials on organ shortage. Actually, we demonstrated that the perspective of a 'lung repair centre' is feasible and effective.
Endothelial Glycocalyx Shedding Occurs during Ex Vivo Lung Perfusion: A Pilot Study.
Sladden Timothy M,Yerkovich Stephanie,Wall Douglas,Tan Maxine,Hunt William,Hill Jonathan,Smith Ian,Hopkins Peter,Chambers Daniel C
Journal of transplantation
Background:Damage to the endothelium has been established as a key pathological process in lung transplantation and ex vivo lung perfusion (EVLP), a new technology that provides a platform for the assessment of injured donor lungs. Damage to the lung endothelial glycocalyx, a structure that lines the endothelium and is integral to vascular barrier function, has been associated with lung dysfunction. We hypothesised that endothelial glycocalyx shedding occurs during EVLP and aimed to establish a porcine model to investigate the mechanism underlying glycocalyx breakdown during EVLP. Methods:Concentrations of endothelial glycocalyx breakdown products, syndecan-1, hyaluronan, heparan sulphate, and CD44, were measured using the ELISA and matrix metalloproteinase (MMP) activity by zymography in the perfusate of both human ( = 9) and porcine ( = 4) lungs undergoing EVLP. Porcine lungs underwent prolonged EVLP (up to 12 hours) with perfusion and ventilation parameters recorded hourly. Results:During human EVLP, endothelial glycocalyx breakdown products in the perfusate increased over time. Increasing MMP-2 activity over time was positively correlated with levels of syndecan-1 ( = 0.886; =0.03) and hyaluronan ( = 0.943; =0.02). In the porcine EVLP model, hyaluronan was the only glycocalyx product detectable during EVLP (1 hr: 19 (13-84) vs 12 hr: 143 (109-264) ng/ml; =0.13). Porcine hyaluronan was associated with MMP-9 activity ( = 0.83; =0.02) and also with dynamic compliance ( = 0.57; =0.03). Conclusion:Endothelial glycocalyx products accumulate during both porcine and human EVLP, and this accumulation parallels an accumulation of matrix-degrading enzyme activity. Preliminary evidence in our porcine EVLP model suggests that shedding may be related to organ function, thus warranting additional study.