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rhAPC reduces the endothelial cell permeability via a decrease of contractile tensions induced by endothelial cells

  • All cells generate contractile tension. This strain is crucial for mechanically controlling the cell shape, function and survival. In this study, the CellDrum technology quantifying cell's (the cellular) mechanical tension on a pico-scale was used to investigate the effect of lipopolysaccharide (LPS) on human aortic endothelial cell (HAoEC) tension. The LPS effect during gram-negative sepsis on endothelial cells is cell contraction causing endothelium permeability increase. The aim was to finding out whether recombinant activated protein C (rhAPC) would reverse the endothelial cell response in an in-vitro sepsis model. In this study, the established in-vitro sepsis model was confirmed by interleukin 6 (IL-6) levels at the proteomic and genomic levels by ELISA, real time-PCR and reactive oxygen species (ROS) activation by florescence staining. The thrombin cellular contraction effect on endothelial cells was used as a positive control when the CellDrum technology was applied. Additionally, the Ras homolog gene family, member A (RhoA) mRNA expression level was checked by real time-PCR to support contractile tension results. According to contractile tension results, the mechanical predominance of actin stress fibers was a reason of the increased endothelial contractile tension leading to enhanced endothelium contractility and thus permeability enhancement. The originality of this data supports firstly the basic measurement principles of the CellDrum technology and secondly that rhAPC has a beneficial effect on sepsis influenced cellular tension. The technology presented here is promising for future high-throughput cellular tension analysis that will help identify pathological contractile tension responses of cells and prove further cell in-vitro models.

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Metadaten
Author:Eylem Kurulgan Demirci, Taylan Demirci, Peter Linder, Jürgen Trzewik, Jessica Ricarda Gierkowski, Matthias Gossmann, Peter Kayser, Dariusz Porst, Ilya Digel, Gerhard Artmann, Aysegül Temiz Artmann
ISBN:1347-4421
Parent Title (English):Journal of Bioscience and Bioengineering. 114 (2012), H. 2
Publisher:-
Document Type:Article
Language:English
Year of Completion:2012
Date of the Publication (Server):2012/12/18
First Page:212
Last Page:219
Link:http://dx.doi.org/10.1016/j.jbiosc.2012.03.019
Zugriffsart:campus
Institutes:FH Aachen / Fachbereich Medizintechnik und Technomathematik
FH Aachen / IfB - Institut für Bioengineering