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Effect of the electric field on the organization of the cytoskeleton of endothelial cells

STSM by Veselina Uzunova, Researcher

Period of mission: from 01/03/2013 to 31/05/2013

Home institution: Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, Bulgaria

Host institution: University of Primorska, Faculty of Health Sciences, Isola, Slovenia

The aim of the experimental work was to study the effect of the electric field, both independently and in combination with bleomycin on the organization of the cytoskeleton of endothelial cells. During this STSM, we studied the degree of cell membrane permeability as a function of the applied high intensity electrical pulses, and how electroporation (EP) affects endothelial cytoskeletal protein reorganisation and cell survival as part of monolayer integrity. Two endothelial cell lines were used (HMEC-1 and HUVEC), that were seeded in 8-chambers slides. Electroporation was performed with plate electrodes perpendicular to the monolayer, without bleomycin or in the presence of 300 nM bleomycin or 100 µM propidium iodide (PI) to determine the degree of cell membrane permeability, after which the treated cells were incubated for 2 or 24 hrs. Next, we performed immunofluorescent staining for cytoskeleton proteins (F-actin, β-tubulin and vimentin) as well as MTS assays for cell viability and survival. We concluded that the EP of an adherent endothelial monolayer, induced changes in cytoskeleton structures. Higher electric fields caused even more rapid disruption of the cytoskeleton and the process was voltage-dependent. At EP ≥ 200V/cm the number of affected cells increased and the monolayers were clearly affected. The cytostatic bleomycin enhanced these effects, most likely because (1) acting on its intracellular target (reached due to electropores produced in the membrane) it induces apoptosis, inhibits cell proliferation and has direct cytotoxic effect, and (2) has specific influence on cell adhesion. Both situations EP/ECT by affecting cell viability and leading to low cell survival may produce in vivo the destruction of the tumor blood vessels.

Oral presentation(s) originating from this mission

Event Scientific session of PhD students and young scientists „Biomedicine and quality of life“, The 145-anniversary of Bulgarian Academy of Sciences
Location Sofia/Bulgaria
Period

02/10/2014

Authors Veselina Uzunova, Cecil Meulenberg, Maja Cemazar, Rumiana Tzoneva
Title Effect of monophasic electric pulses both independently and in combination with bleomycin on the cytoskeleton organization in endothelial cells

 


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