Ex Vivo Lung Perfusion in the Rat: Detailed Procedure and Videos

Bassani, Giulia Alessandra; Lonati, Caterina; Brambilla, Daniela; Rapido, Francesca; Valenza, Franco; Gatti, Stefano

doi:10.1371/journal.pone.0167898

Cited by 24 publications

(30 citation statements)

References 31 publications

(38 reference statements)

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“…On this basis, we opted to broadly explore the biologic effects of EVLP in a preclinical setting using our reliable EVLP rat model which could enable ex vivo perfusion without the use of anti-inflammatory treatments (17).…”

Section: Discussionmentioning

confidence: 99%

“…Ex vivo perfusion was performed using the isolated lung perfusion system (Hugo Sachs Elektronik, Harvard Apparatus, March-Hugstetten, Germany; Fig. 2) according to the procedure implemented in our laboratory and as previously described (17).…”

Section: Evlp Protocolmentioning

confidence: 99%

“…Research was based on a rat model that was implemented in our laboratory that allows prolonged ex vivo perfusion without the use of anti-inflammatory drugs (17). The main focus was the identification of significant EVLP-induced changes that occur in the lung transcriptional profile and signaling pathways.…”

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confidence: 99%

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Influence of ex vivo perfusion on the biomolecular profile of rat lungs

et al. 2018

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Despite increasing clinical adoption, biologic influences of ex vivo lung perfusion (EVLP) remain insufficiently elucidated. The aim of the current study was to investigate biomolecular changes induced by EVLP in rat lungs. EVLP was maintained for 180 min. Hyaluronan, mediators, and cells were assessed in the perfusate. Gene expression, signaling pathways, and ATP content were investigated in lung tissue. EVLP induced the release of medium-high molecular weight hyaluronan and transcription of hyaluronan synthases ( P< 0.001). Increasing concentrations of inflammatory mediators were detected in the perfusate ( P < 0.001). Perfused lungs exhibited a distinctive transcriptional signature compared with organs examined before or after surgery/procurement ( P = 0.003). Up-regulated genes were involved in inflammation and its regulation, apoptosis/survival, heat shock, and oxidative stress response ( q = 0). Down-regulated genes were related to lymphocyte function ( q = 0). The NF-κB, signal transducer and activator of transcription 3, ERK1/2, p38, Akt, and stress-activated protein kinase/JNK signaling pathways were modulated by EVLP ( P < 0.05). Most of these biomolecular changes were examined and confirmed in additional experiments that were performed in lungs procured from donation after cardiocirculatory death after 180 min of warm ischemia. The current study demonstrates that EVLP broadly affects the lung biomolecular phenotype. These findings improve our comprehension of the effects exerted by the procedure and encourage additional research in preclinical models to implement therapeutic interventions.-Lonati, C., Bassani, G. A., Brambilla, D., Leonardi, P., Carlin, A., Faversani, A., Gatti, S., Valenza, F. Influence of ex vivo perfusion on the biomolecular profile of rat lungs.

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Section: Discussionmentioning

confidence: 99%

Section: Evlp Protocolmentioning

confidence: 99%

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Influence of ex vivo perfusion on the biomolecular profile of rat lungs

et al. 2018

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“…EVLP began to be applied in combination with regenerative medicine advancement, such as stem cell biology in these 2-3 years [30]. Animal models of EVLP have been developed and helped to explore more novel experimental platforms that can be used to do translational research on pulmonary physiology and transplant medicine [31].…”

Section: Experimental Intervention On Donor Lungs Based On the Evlp Pmentioning

confidence: 99%

Evolving Trend of EVLP: Advancements and Emerging Pathways

Jiao¹

2019

SN Compr. Clin. Med.

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Lung transplant augmentation in the waiting list is impeded by donor lung shortage with a low percentage of procurement. Ex vivo lung perfusion (EVLP) was recognized to have the capacity of providing a platform to preserve, assess, recondition, and even mange existing diseases in rejected donor lungs to expand the donor pool. This review aims to provide the most advanced experimental evidence of EVLP application and perspective as a platform providing qualified donor lungs. Literature review using PubMed, Medscape, Clinical Trials, and Cochrane databases was performed. Data was collected and analyzed. From a view point of publisher, current research centers with their paper contribution volume are also presented to illustrate the future trend of EVLP academically. EVLP incubated donor lungs could have a comparable outcome with conventional transplants. Further interventional studies have shown growing interest in and expectations on EVLP platform as pathways to determine graft quality, to bridge a critical phase of the initial rejected organs and to predict prognosis or complications in an early stage. EVLP is also considered as a tissue regeneration translational research environment with multidisciplinary potential to enhance the recovery and rehabilitation after surgery and advance pulmonary medicine. Ongoing clinical trials and accelerated procedure modulation with commercialization of EVLP are leading to Bcentralization^organ preparation model, opening a new era of lifechange health innovation.

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“…Fluid exchange depends on the balance of hydrostatic and oncotic pressure differences between the lumen of the microvasculature and the interstitial space, and the selective permeability of the endothelium to molecules of different sizes, as described in the classic Starling hypothesis (27). The essential roles of the normal electrolyte composition, pH, nutrient, hormonal and colloid osmotic pressure of the perfusion solution in maintaining vascular function have been well characterized in the classic isolated organ preparations such as the isolated heart, lung, and kidney, and more recently in ex vivo perfusion protocols used to recondition donor lungs before transplantation (6,16,22,25,27,35). However, these isolated organ preparations develop edema after relatively brief periods (hours).…”

Section: Introductionmentioning

confidence: 99%

Pulmonary endothelial permeability and tissue fluid balance depend on the viscosity of the perfusion solution

Rowan

Rochfort

Piouceau

et al. 2018

American Journal of Physiology-Lung Cellular and Molecular Phys

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Fluid filtration in the pulmonary microcirculation depends on the hydrostatic and oncotic pressure gradients across the endothelium and the selective permeability of the endothelial barrier. Maintaining normal fluid balance depends both on specific properties of the endothelium and of the perfusing blood. Although some of the essential properties of blood needed to prevent excessive fluid leak have been identified and characterized, our understanding of these remains incomplete. The role of perfusate viscosity in maintaining normal fluid exchange has not previously been examined. We prepared a high-viscosity perfusion solution (HVS) with a relative viscosity of 2.5, i.e., within the range displayed by blood flowing in vessels of different diameters in vivo (1.5-4.0). Perfusion of isolated murine lungs with HVS significantly reduced the rate of edema formation compared with perfusion with a standard solution (SS), which had a lower viscosity similar to plasma (relative viscosity 1.5). HVS did not alter capillary filtration pressure. Increased endothelial shear stress produced by increasing flow rates of SS, to mimic the increased shear stress produced by HVS, did not reduce edema formation. HVS significantly reduced extravasation of Evans blue-labeled albumin compared with SS, indicating that it attenuated endothelial leak. These findings demonstrate for the first time that the viscosity of the solution perfusing the pulmonary microcirculation is an important physiological property contributing to the maintenance of normal fluid exchange. This has significant implications for our understanding of fluid homeostasis in the healthy lung, edema formation in disease, and reconditioning of donor organs for transplantation.

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Ex Vivo Lung Perfusion in the Rat: Detailed Procedure and Videos

Cited by 24 publications

References 31 publications

Influence of ex vivo perfusion on the biomolecular profile of rat lungs

Influence of ex vivo perfusion on the biomolecular profile of rat lungs

Evolving Trend of EVLP: Advancements and Emerging Pathways

Pulmonary endothelial permeability and tissue fluid balance depend on the viscosity of the perfusion solution

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