JTCS Click here to go to SJM website.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Alert me to new issues of the journal
Right arrow Add to Personal Folders
Right arrow Download to citation manager
Right arrow Author home page(s):
Monica Casiraghi
Thomas M. Egan
Right arrow Permission Requests
Google Scholar
Right arrow Articles by Casiraghi, M.
Right arrow Articles by Egan, T. M.
PubMed
Right arrow Articles by Casiraghi, M.
Right arrow Articles by Egan, T. M.
Related Collections
Right arrow Lung - transplantation
Right arrow Lung - basic science

J Thorac Cardiovasc Surg 2009;138:760-767
© 2009 The American Association for Thoracic Surgery

Cardiothoracic Transplantation

In vitro modeling of nonhypoxic cold ischemia–reperfusion simulating lung transplantation

Monica Casiraghi, MDa,b, Jason R. Tatreau, BSc, John B. Abano, MDa, John W. Blackwell, BSa, Larry Watson, MDa, Keith Burridge, PhDd, Scott H. Randell, PhDc, Thomas M. Egan, MD, MSca,*

a Division of Cardiothoracic Surgery, University of North Carolina at Chapel Hill, Chapel Hill, NC
b Department of Thoracic Surgery, Division of Thoracic Surgery, European Institute of Oncology, Milan, Italy
c Cystic Fibrosis/Pulmonary Research and Treatment Center and the Division of Pulmonary and Critical Care Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC
d Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC

Received for publication December 1, 2008; revisions received April 2, 2009; accepted for publication May 26, 2009.

* Address for reprints: Thomas M. Egan, MD, MSc, Division of Cardiothoracic Surgery, Department of Surgery, University of North Carolina at Chapel Hill, 3040 Burnett-Womack Bldg, CB #7065, Chapel Hill, NC 27599-7065. (Email: ltxtme{at}med.unc.edu).

Objective: Although anoxia/reoxygenation of cultured cells has been used to model lung ischemia–reperfusion injury, this does not accurately mimic events experienced by lung cells while a lung is retrieved from a donor, stored, and transplanted. We developed an in vitro model of nonhypoxic ischemia–reperfusion injury to simulate these events.

Methods: Human umbilical vein endothelial cells underwent simulated cold ischemia by replacing 37°C culture media with 4°C Perfadex (Vitrolife, Kungsbacka, Sweden) solution for 5 hours in 100% O2. Culture dishes were allowed to warm to room temperature for 1 hour (implantation), and then Perfadex solution was replaced with 37°C culture media (reperfusion).

Results: During cold ischemia, the human umbilical vein endothelial cell filamentous actin cytoskeleton quickly became rearranged, and gaps developed in the previously confluent monolayer occupying 20% of the surface area. Simulated reperfusion resulted in reorganization to a confluent monolayer. Development of gaps was not due to enhanced necrosis based on lactate dehydrogenase retention assay. Endothelial cytoskeletal rearrangement could account for early edema caused by ischemia–reperfusion injury with reperfusion. Mitogen-activated protein kinase and nuclear factor {kappa}B activation occurred with simulated reperfusion despite normoxia. Levels of the proinflammatory cytokines interleukin 6 and interleukin 8 were significantly increased in media at the end of reperfusion.

Conclusions: Exposing human umbilical vein endothelial cells to simulated cold ischemia without hypoxia causes reversible cytoskeletal alterations, activation of inflammatory pathways, and elaboration of cytokines. Because this model accurately depicts events occurring during lung transplantation, it will be useful to explore mechanisms regulating lung cell response to this unique form of ischemia–reperfusion injury.

Abbreviations and Acronyms AM = alveolar macrophage; BOS = bronchiolitis obliterans syndrome; ERK = extracellular signal-regulated kinase; HMVEC = human pulmonary microvascular endothelial cell; HUVEC = human umbilical vein endothelial cell; I{kappa}B{alpha} = inhibitor of nuclear factor {kappa}B,{alpha}; IL = interleukin; IRI = ischemia–reperfusion injury; JNK = Janus kinase; LDH = lactate dehydrogenase; LTX = lung transplantation; MAPK = mitogen-activated protein kinase; NF = nuclear factor; PBS = phosphate-buffered saline; TLR = Toll-like receptor






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
ANN THORAC SURG ASIAN CARDIOVASC THORAC ANN EUR J CARDIOTHORAC SURG
J THORAC CARDIOVASC SURG ICVTS ALL CTSNet JOURNALS
Copyright © 2009 by The American Association for Thoracic Surgery.