Endothelial dysfunction of bypass graft: direct comparison of in Vitro and in vivo models of ischemia-reperfusion injury

Background Although, ischemia/reperfusion induced vascular dysfunction has been widely described, no comparative study of in vivo- and in vitro-models exist. In this study, we provide a direct comparison between models (A) ischemic storage and in-vitro reoxygenation (B) ischemic storage and in vitro...

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Main Authors: Veres, Gábor (Author) , Hegedűs, Péter (Author) , Barnucz, Enikő (Author) , Zöller, Raphael (Author) , Klein, Stephanie (Author) , Schmidt, Harald (Author) , Radovits, Tamás (Author) , Korkmaz-İçöz, Sevil (Author) , Karck, Matthias (Author) , Szabó, Gábor (Author)
Format: Article (Journal)
Language:English
Published: April 15, 2015
In: PLOS ONE
Year: 2015, Volume: 10, Issue: 4
ISSN:1932-6203
DOI:10.1371/journal.pone.0124025
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1371/journal.pone.0124025
Verlag, lizenzpflichtig, Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0124025
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Author Notes:Gábor Veres, Péter Hegedűs, Enikő Barnucz, Raphael Zöller, Stephanie Klein, Harald Schmidt, Tamás Radovits, Sevil Korkmaz, Matthias Karck, Gábor Szabó
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Summary:Background Although, ischemia/reperfusion induced vascular dysfunction has been widely described, no comparative study of in vivo- and in vitro-models exist. In this study, we provide a direct comparison between models (A) ischemic storage and in-vitro reoxygenation (B) ischemic storage and in vitro reperfusion (C) ischemic storage and in-vivo reperfusion. Methods and Results Aortic arches from rats were stored for 2 hours in saline. Arches were then (A) in vitro reoxygenated (B) in vitro incubated in hypochlorite for 30 minutes (C) in vivo reperfused after heterotransplantation (2, 24 hours and 7 days reperfusion). Endothelium-dependent and independent vasorelaxations were assessed in organ bath. DNA strand breaks were assessed by TUNEL-method, mRNA expressions (caspase-3, bax, bcl-2, eNOS) by quantitative real-time PCR, proteins by Western blot analysis and the expression of CD-31 by immunochemistry. Endothelium-dependent maximal relaxation was drastically reduced in the in-vivo models compared to ischemic storage and in-vitro reperfusion group, and no difference showed between ischemic storage and control group. CD31-staining showed significantly lower endothelium surface ratio in-vivo, which correlated with TUNEL-positive ratio. Increased mRNA and protein levels of pro- and anti-apoptotic gens indicated a significantly higher damage in the in-vivo models. Conclusion Even short-period of ischemia induces severe endothelial damage (in-vivo reperfusion model). In-vitro models of ischemia-reperfusion injury can be limitedly suited for reliable investigations. Time course of endothelial stunning is also described.
Item Description:Gesehen am 09.07.2020
Physical Description:Online Resource
ISSN:1932-6203
DOI:10.1371/journal.pone.0124025