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Inducing intracellular ros and cellular redox without cell death in mesenchymal cells using microsecondpulsed DBD plasma
Conference proceeding

Inducing intracellular ros and cellular redox without cell death in mesenchymal cells using microsecondpulsed DBD plasma

N Shainsky, G Friedman, G Fridman, A Fridman, M. J Steinbeck and T. A Freema
2012 Abstracts IEEE International Conference on Plasma Science, 7E-7
Jul 2012

Abstract

Biomedical engineering Biomembranes Educational institutions Electrical engineering Fluorescence Plasmas USA Councils
Intracellular reactive oxygen species (ROS) is a known activator of cell signaling, however too much ROS causes cell death. Our goal was to determine the optimal microsecond-pulsed Dielectric Barrier Discharge plasma (DBD-plasma) treatment dose (J/cm 2 ) in which ROS/redox responses would potentially activate cell function but not cell death. In this study, we applied DBD-plasma to mesenchymal cells and evaluated intracellular ROS generation, redox, mitochondrial membrane potential and the Live/Dead response to increasing levels of DBD-plasma. Using fluorescent indicators to measure superoxide anion (O 2 - .) (MitoSOX™ Red), reduced glutathione (GSH) (ThiolTracker™ Violet) and mitochondrial membrane potential (ΔΨm) (MitoTracker® Red), we show that DBD-plasma doses of 1-5 J/cm 2 stimulated an immediate (minutes) increase in O 2 - levels with a concomitant decrease in reduced thiol. The mitosox fluorescence intensity increased with DBD-plasma treatments until 1 J/cm 2 , after which levels plateaued regardless of increased dose. Reduced thiol fluorescent intensity showed an immediate 50% decrease regardless of DBD-plasma treatment dose. A 20% decrease in mitochondrial membrane potential occurred at 2 hrs post-DBD-plasma treatments 1 - 5 J/cm 2 , with significant cell death observed (fluorescent Live/Dead assay; syto- 13/propidium iodide) at levels of 2.5 J/cm 2 and greater. Taken together, these results provide a range in which DBD (0-2.0 J/cm 2 ) intracellular ROS generation and cellular redox change with little effect on cell viability. Understanding the optimal treatment levels DBD-plasma interactions is essential for developing DBD-plasma treatments to induce directed cell function such as; proliferation, differentiation and targeted gene expression.

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