title: rhAPC reduces the endothelial cell permeability via a decrease of contractile tensions induced by endothelial cells.
abstract: 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.
authors.affiliation: Aachen University of Applied Sciences, Institute of Bioengineering, Laboratory of Cell Biophysics, Juelich, Germany. eylemkurulgan@gmail.com
authors.forename: Eylem Kurulgan
authors.initials: EK
authors.lastname: Demirci
authors.forename: Taylan
authors.initials: T
authors.lastname: Demirci
authors.forename: Peter
authors.initials: P
authors.lastname: Linder
authors.forename: Juergen
authors.initials: J
authors.lastname: Trzewik
authors.forename: Jessica Ricarda
authors.initials: JR
authors.lastname: Gierkowski
authors.forename: Matthias
authors.initials: M
authors.lastname: Gossmann
authors.forename: Peter
authors.initials: P
authors.lastname: Kayser
authors.forename: Dariusz
authors.initials: D
authors.lastname: Porst
authors.forename: Ilya
authors.initials: I
authors.lastname: Digel
authors.forename: Gerhard Michael
authors.initials: GM
authors.lastname: Artmann
authors.forename: Aysegul Temiz
authors.initials: AT
authors.lastname: Artmann
mesh_terms: D000199:Actins / Q000378:metabolism; D001011:Aorta / Q000166:cytology; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology; D010539:Permeability / Q000187:drug effects*; D011486:Protein C / Q000494:pharmacology* / Q000627:therapeutic use; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology / Q000627:therapeutic use; D018805:Sepsis / Q000188:drug therapy / Q000378:metabolism; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology; D020742:rhoA GTP-Binding Protein / Q000235:genetics
chemical_list: D000199:Actins; D015850:Interleukin-6; D008070:Lipopolysaccharides; D011486:Protein C; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; C066737:RHOA protein, human; D013917:Thrombin; D020742:rhoA GTP-Binding Protein; C421124:drotrecogin alfa activated