Document Type

Article

Publication Date

2012

Publication Title

Archives of Biochemistry and Biophysics

Volume

527

Issue

1

Pages

55-64

DOI

10.1016/j.abb.2012.08.004

Abstract

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 (H2DCF), dihidroethidium (DHE), or Amplex Red. When a suspension of H2DCF-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 H2DCF into DCF. This effect was not related to heating and was reduced by catalase, but not by mannitol or superoxide dismutase. Formation of H2O2 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 H2O2 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.

Comments

NOTE: This is the author's manuscript version of a work that was published in Archives of Biochemistry and Biophysics. The final version was published as:

Pakhomova, O. N., Khorokhorina, V. A., Bowman, A. M., Rodaite-Riseviciene, R., Saulis, G., Xiao, S., & Pakhomov, A. G. (2012). Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media. Archives of Biochemistry and Biophysics, 527(1), 55-64. doi:10.1016/j.abb.2012.08.004

Original Publication Citation

Pakhomova, O. N., Khorokhorina, V. A., Bowman, A. M., Rodaite-Riseviciene, R., Saulis, G., Xiao, S., & Pakhomov, A. G. (2012). Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media. Archives of Biochemistry and Biophysics, 527(1), 55-64. doi:10.1016/j.abb.2012.08.004

ORCID

0000-0003-4950-4130 (Pakhomova), 0000-0002-0375-8373 (Xiao)

Download

Share

COinS