Document Detail

Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media.
MedLine Citation:
PMID:  22910297     Owner:  NLM     Status:  MEDLINE    
Nanosecond pulsed electric field (nsPEF) is a novel modality for permeabilization of membranous structures and intracellular delivery of xenobiotics. We hypothesized that oxidative effects of nsPEF could be a separate primary mechanism responsible for bioeffects. ROS production in cultured cells and media exposed to 300-ns PEF (1-13 kV/cm) was assessed by oxidation of 2',7'-dichlorodihydrofluoresein (H(2)DCF), dihidroethidium (DHE), or Amplex Red. When a suspension of H(2)DCF-loaded cells was subjected to nsPEF, the yield of fluorescent 2',7'-dichlorofluorescein (DCF) increased proportionally to the pulse number and cell density. DCF emission increased with time after exposure in nsPEF-sensitive Jurkat cells, but remained stable in nsPEF-resistant U937 cells. In cell-free media, nsPEF facilitated the conversion of H(2)DCF into DCF. This effect was not related to heating and was reduced by catalase, but not by mannitol or superoxide dismutase. Formation of H(2)O(2) in nsPEF-treated media was confirmed by increased oxidation of Amplex Red. ROS increase within individual cells exposed to nsPEF was visualized by oxidation of DHE. We conclude that nsPEF can generate both extracellular (electrochemical) and intracellular ROS, including H(2)O(2) and possibly other species. Therefore, bioeffects of nsPEF are not limited to electropermeabilization; concurrent ROS formation may lead to cell stimulation and/or oxidative cell damage.
Olga N Pakhomova; Vera A Khorokhorina; Angela M Bowman; Raminta Rodaitė-Riševičienė; Gintautas Saulis; Shu Xiao; Andrei G Pakhomov
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-08-15
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  527     ISSN:  1096-0384     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-09-26     Completed Date:  2012-12-03     Revised Date:  2013-11-06    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  United States    
Other Details:
Languages:  eng     Pagination:  55-64     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA 23508, USA.
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MeSH Terms
CHO Cells
Cell Line
Cell Membrane / metabolism
Cell Membrane Permeability*
Cell Survival
Cell-Free System / metabolism
Electroporation* / methods
Fluoresceins / analysis,  metabolism
Fluorescent Dyes / analysis,  metabolism
Hydrogen Peroxide / metabolism
Jurkat Cells
Oxidative Stress*
Reactive Oxygen Species / metabolism*
Grant Support
Reg. No./Substance:
0/Fluoresceins; 0/Fluorescent Dyes; 0/Reactive Oxygen Species; 76-54-0/2',7'-dichlorofluorescein; 7722-84-1/Hydrogen Peroxide

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