Contribution of electric fields and active species in nanosecond pulsed DBD plasma treatment for stimulation of murine mesenchymal C3H10T1/2 cells

Summary form only given. High dose non-thermal nanosecond pulsed dielectric barrier discharge (nspDBD) plasma treatment of mammalian cells can result in cell death. Alternatively, low dose DBD can stimulate cellular proliferation and differentiation [1, 2]. While these results highlight the great potential of this technology, the ability to tailor nspDBD treatments to achieve an effective and predictable cellular response would be a great advancement for the field of Plasma Medicine. With this goal in mind, we have attempted to separate the individual components of nspDBD plasma (electric field, radiation, charged and neutral species) and measure cellular response to each. First, to determine the correct treatment range, we investigated the effect of nspDBD plasma on cell death using the murine mesenchymal cell line C3H10T1/2. nspDBD plasma was applied to the cells and viability was evaluated one hour after treatment using the fluorescent indicator, propidium iodide (membrane permeable in dead cells). In subsequent experiments, each plasma component was selectively removed and cell viability was measured. Electric field was measured by submersing the electrode in cell media to eliminate the formation of plasma. To evaluate the effects of charged and neutral species, a grounded copper grid was inserted between the high voltage electrode and the cells. Results showed no statistical difference in cell viability between control (live untreated) cells and cells subjected to electric field alone. However, when the effect of charged and neutral species were evaluated, a significant increase in cell death was present at one hour post-treatment. These results suggest electric field is not the major contributor to cell death during nspDBD treatment and that charges and short-lived neutral species play a greater role. On-going experiments includes measuring intracellular reactive oxygen and nitrogen species in addition to cell death and the effects of radiation. This study wi- l advance our understanding of plasma-cell interaction and lead to the development of a tunable plasma device to direct cells toward specific, predictable processes.