The atmospheric pressure helium plasma jet driven by pulsed dc voltage was utilized to treat human lung cancer cells in vitro. The properties of plasma plume were adjusted by the injection type and flow rate of additive oxygen gas in atmospheric pressure helium plasma jet. The plasma characteristics such as plume length, electric current and optical emission spectra (OES) were measured at different flow rates of additive oxygen to helium. The plasma plume length and total current decreased with an increase in the additive oxygen flow rate. The electron excitation temperature estimated by the Boltzmann plot from several excited helium emission lines increased slightly with the additive oxygen flow. The oxygen atom density in the gas phase estimated by actinometry utilizing argon was observed to increase with the additive oxygen flow. The concentration of intracellular reactive oxygen species (ROS) measured by fluorescence assay was found to be not exactly proportional to that of extracellular ROS (measured by OES), but both correlated considerably. It was also observed that the expression levels of p53 and the phospho-p53 were enhanced in the presence of additive oxygen flow compared with those from the pure helium plasma treatment.

The effects of atmospheric pressure plasma jet on cancer cells (human lung carcinoma cells) and normal cells (embryonic kidney cells and bronchial epithelial cells) were investigated. Using a detection dye, the production of intracellular reactive oxygen species (ROS) was found to be increased in plasma-treated cells compared to non-treated and gas flow-treated cells. A significant overproduction of ROS and a reduction in cell viability were induced by plasma exposure on cancer cells. Normal cells were observed to be less affected by the plasma-mediated ROS, and cell viability was less changed. The selective effect on cancer and normal cells provides a promising prospect of cold plasma as a cancer therapy.

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Atmospheric pressure plasma jets employing nitrogen, helium, or argon gases driven by low-frequency (several tens of kilohertz) ac voltage and pulsed dc voltage were fabricated and characterized. The changes in discharge current, optical emission intensities from reactive radicals, gas temperature, and plume length of plasma jets with the control parameters were measured and compared. The control parameters include applied voltage, working gas, and gas flow rate. As an application to plasma-cancer cell interactions, the effects of atmospheric pressure plasma jet on the morphology and intracellular reactive oxygen species (ROS) level of human lung adenocarcinoma cell (A549) and human bladder cancer cell (EJ) were explored. The experimental results show that the plasma can effectively control the intracellular concentrations of ROS. Although there exist slight differences in the production of ROS, helium, argon, or nitrogen plasma jets are found to be useful in enhancing the intracellular ROS concentrations in cancer cells

An atmospheric-pressure plasma jet array source driven by a pulsed bipolar wave of several tens of kilohertz was designed and characterized. This source was assembled with each individual module. Thus, it is possible to adjust the area of treatment by arranging the number of jets. It has the potential to greatly enhance the scale of surface treatment over that of a single plasma jet. Multiple plasma plumes were generated and propagated to grounded-surface relatively uniformly without an auxiliary circuit. Due to the large separation between each jet, plasma bullets propagate through their individual gas channels. The transition from the bullet to the continuous modes occurred at an applied voltage of 5.5 kV pp . The interaction between adjacent plasma plumes and behaviors of primary bullet and subbullet are investigated with intensified charge coupled device camera images.Index Terms-Atmospheric-pressure plasma jet, nonthermal plasma, plasma bullets, plasma jet array.

An atmospheric pressure plasma jet array source driven by a pulsed bipolar voltage of several tens of kilohertz was characterized and utilized for cancer cell treatment. Electrical and optical emission characteristics of plasma jet array were obtained as functions of the applied voltage, gas flow rate, and pulse frequency, and the optimal operating parameters were obtained. The emission intensities from reactive oxygen and nitrogen species (RONS) in a gas phase, such as OH, NO, H, and O, were measured by optical emission spectroscopy. In the plasma-liquid interaction experiment, the OH concentration was measured using the TA solution. The nitrite concentration in the plasma treated media (DW, HBSS, and DMEM) was measured using Griess reaction assay. Human skin cancer cells injected with plasma treated liquid were observed for changes in cell viability using the MTS assay. The results demonstrate that the plasma jet array source can be a good candidate for delivering RONS to liquid for plasma activated medium (PAM).

Cold atmospheric plasma (CAP) that generates reactive oxygen species (ROS) has received considerable scientific attentions as a new type of anticancer. In particular, an indirect treatment method of inducing cancer cell death through plasma-activated medium (PAM), rather than direct plasma treatment has been well established. Although various cell death pathways such as apoptosis, necroptosis, and autophagy have been suggested to be involved in PAM-induced cell death, the involvement of ferroptosis, another type of cell death regulated by lipid ROS is largely unknown. This study reports, that PAM promotes cell death via ferroptosis in human lung cancer cells, and PAM increases intracellular and lipid ROS, thereby resulting in mitochondrial dysfunction. The treatment of cells with N-acetylcysteine, an ROS scavenging agent, or ferrostatin-1, a ferroptosis inhibitor, protects cells against PAM-induced cell death. Interestingly, ferroptosis suppressor protein 1 (FSP1) is downregulated upon PAM treatment. Furthermore, the treatment of cells with iFSP1, an inhibitor of FSP1, further enhances PAM-induced ferroptosis. Finally, this study demonstrates that PAM inhibits tumor growth in a xenograft model with an increase in 4-hydroxynoneal and PTGS2, a byproduct of lipid peroxidation, and a decrease in FSP1 expression. This study will provide new insights into the underlying mechanism and therapeutic strategies of PAM-mediated cancer treatment.

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