Atmospheric pressure plasma jet treatment evokes transient oxidative stress in HaCaT keratinocytes and influences cell physiology

Cell Biol Int. 2014 Apr;38(4):412-25. doi: 10.1002/cbin.10200. Epub 2014 Feb 10.

Abstract

Modern non-thermal atmospheric pressure plasma sources enable controllable interaction with biological systems. Their future applications - e.g. wound management - are based on their unique mixture of reactive components sparking both stimulatory as well as inhibitory processes. To gain detailed understanding of plasma-cell interaction and with respect to risk awareness, key mechanisms need to be identified. This study focuses on the impact of an argon non-thermal atmospheric pressure plasma jet (kINPen 09) on human HaCaT keratinocytes. With increasing duration, cell viability decreased. In accordance, cells accumulated in G2/M phase within the following 24 h. DNA single-strand breaks were detected immediately after treatment and receded in the aftermath, returning to control levels after 24 h. No directly plasma-related DNA double-strand breaks were detected over the same time. Concurrently, DNA synthesis decreased. Coincident with treatment time, an increase in intracellular 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCFDA) conversion increased reactive oxygen species (ROS) levels. The radical scavenging activity of culture medium crucially influenced these effects. Thus, ROS changed DNA integrity, and the effectiveness of cellular defence mechanisms characterises the interaction of non-thermal plasma and eukaryotic cells. Effects were time-dependent, indicating an active response of the eukaryotic cells. Hence, a stimulation of eukaryotic cells using short-term non-thermal plasma treatment seems possible, eg in the context of chronic wound care. Long-term plasma treatments stopped in cell proliferation and apoptosis, which might be relevant in controlling neoplastic conditions.

Keywords: DNA damage; non-thermal atmospheric pressure plasma; plasma medicine; radical scavenging activity; single cell electrophoresis assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Caspase 3 / metabolism
  • Cell Line
  • Cell Survival / drug effects
  • DNA / metabolism
  • DNA Breaks, Single-Stranded / drug effects
  • Fluoresceins / metabolism
  • G2 Phase Cell Cycle Checkpoints / drug effects
  • Humans
  • Keratinocytes / cytology
  • Keratinocytes / metabolism*
  • M Phase Cell Cycle Checkpoints / drug effects
  • Oxidative Stress / drug effects*
  • Plasma Gases / pharmacology*
  • Reactive Oxygen Species / metabolism

Substances

  • 2',7'-dichlorodihydrofluorescein diacetate
  • Fluoresceins
  • Plasma Gases
  • Reactive Oxygen Species
  • DNA
  • Caspase 3