Session FT1: Plasma Medicine

Cold Atmospheric Plasma for Medicine: State of Research and Clinical Application

Basic research in plasma medicine has made excellent progress and resulted in the fundamental insights that biological effects of cold atmospheric plasmas (CAP) are significantly caused by changes of the liquid environment of cells, and are dominated by redox-active species. First CAP sources are CE-certified as medical devices. Main focus of plasma application is on wound healing and treatment of infective skin diseases. Clinical applications in this field confirm the supportive effect of cold plasma treatment in acceleration of healing of chronic wounds above all in cases where conventional treatment fails. Cancer treatment is another actual and emerging field of CAP application. The ability of CAP to kill cancer cells by induction of apoptosis has been proved in vitro. First clinical applications of CAP in palliative care of cancer are realized.\\[4pt] In collaboration with Hans-Robert Metelmann, University Medicine Greifswald; Helmut Uhlemann, Klinikum Altenburger Land GmbH Altenburg; Anke Schmidt and Kai Masur, Leibniz Institute for Plasma Science and Technology (INP Greifswald); Renate Sch\"onebeck, Neoplas Tools GmbH Greifswald; and Klaus-Dieter Weltmann, Leibniz Institute for Plasma Science and Technology (INP Greifswald).   

DNA damage in oral cancer and normal cells induced by nitrogen atmospheric pressure plasma jets

Nitrogen atmospheric pressure plasma jets (APPJs) have been shown to effectively induce DNA double strand breaks in SCC25 oral cancer cells. The APPJ source constructed in our laboratory operates based on dielectric barrier discharge. It consists of two copper electrodes alternatively wrapping around a fused silica tube with nitrogen as a feed gas. It is generally more challenging to ignite plasma in N2 atmosphere than in noble gases. However, N2 provides additional advantages such as lower costs compared to noble gases, thus this design can be beneficial for the future long-term clinical use. To compare the effects of plasma on cancer cells (SCC25) and normal cells (OKF), the cells from both types were treated at the same experimental condition for various treatment times. The effective area with different damage levels after the treatment was visualized as 3D maps. The delayed damage effects were also explored by varying the incubation times after the treatment. All of these studies are critical for a better understanding of the damage responses of cellular systems exposed to the plasma radiation, thus are useful for the development of the advanced plasma cancer therapy.   

Plasma treatment of onychomycosis

Onychomycosis or fungal infection of the toenail or fingernail is a common affliction. Approximately 10{\%} of the world's adult population is estimated to suffer from onychomycosis. Current treatment options such as topical creams, oral drugs, or laser treatments are generally limited by a variety of problems. We present results for an alternative onychomycosis treatment scheme using atmospheric pressure cold air plasmas. Using thinned cow hoof as a model nail material, we tested the ability of various plasma sources to act through the model nail to eradicate either bacteria or fungus deposited on the opposite side. Following 20 minute exposure to a surface microdischarge (SMD) device operating in room air, we observed a $\sim$ 2 log reduction of E. coli. A similar result was obtained against T. rubrum after 45 min plasma treatment. NOx species concentration penetrating through the model nail as well as uptake into the nail were measured as a function of nail thickness. We propose that these plasma-generated species, or perhaps their reaction products, are responsible for at least part of the observed anti-microbial effect. We also explore the use of ultraviolet light acting in synergy with plasma-generated chemical species.   

Responses of cells in plasma-activated medium

Plasma consists of electrons, ions, radicals, and lights, and produces various reactive species in gas and liquid phase. Cells receive various inputs from their circumstances, and induce several physiological outputs. Our goal is to clarify the relationships between plasma inputs and physiological outputs. Plasma-activated medium (PAM) is a circumstance that plasma provides cells and our previous studies suggest that PAM is a promising tool for cancer therapy. However, the mode of actions remains to be elucidated. We propose survival and proliferation signaling networks as well as redox signaling networks are key factors to understand cellular responses of PAM-treated glioblastoma cells.   

Effect of microplasma irradiation on skin barrier function

This study investigates the feasibility of atmospheric-pressure argon microplasma irradiation (AAMI) to promote drug delivery through skin. Yucatan micropig skin was used as a biological object for evaluation of in vitro percutaneous absorption. The changes in lipids, proteins and water content of the pig stratum corneum (SC) after AAMI were compared to those of a tape stripping test (TST) and plasma jet irradiation (PJI) using attenuated total reflection-Fourier transform infrared spectroscopy analysis. The significant reduction in the methylene stretching modes absorbance resulted in the disturbance in the SC lipids caused by AAMI was observed at 2850 and 2920 cm$^{-1}$. Moreover, as the result of TST, trans-epidermal water loss (TEWL) after both AAMI and PJI were also increased, that could lead to a decrease of barrier function of SC, and could enhance the transdermal absorption of drugs. Under the conditions of this study, TEWL value of 5 minutes AAMI (35.92 $\pm$ 3.48 g/m$^{2}$h) was approximately the same as that value of 10 times TST (34.30 $\pm$ 3.54 g/m$^{2}$h), that makes the effect of these manipulations on the surfaces is considered to be at the same levels. Furthermore, unlike the obtained microscopic observation from PJI, there was no thermal damage observed on the skins after AAMI.   

Investigation of selective induction of breast cancer cells to death with treatment of plasma-activated medium

The applications of plasma in medicine have much attention. We previously showed that plasma-activated medium (PAM) induced glioblastoma cells to apoptosis. However, it has not been elucidated the selectivity of PAM in detail. In this study, we investigated the selective effect of PAM on the death of human breast normal and cancer cells, MCF10A and MCF7, respectively, and observed the selective death with fluorescent microscopy. For the investigation of cell viability with PAM treatment, we prepared various PAMs according to the strengths, and treated each of cells with PAMs. Week PAM treatment only decreased the viability of MCF7 cells, while strong PAM treatment significantly affected both viabilities of MCF7 and MCF10A cells. For the fluorescent observation, we prepared the mixture of MCF7 and fluorescent-probed MCF10A cells, and seeded them. After the treatment of PAMs, the images showed that only MCF7 cells damaged in the mixture with week PAM treatment. These results suggested that a specific range existed with the selective effect in the strength of PAM.   

Antitumor effect of synergistic contribution of nitrite and hydrogen peroxide in the plasma activated medium

Non-equilibrium atmospheric pressure plasmas (NEAPP) have been attracted attention in the noble application of cancer therapy. Although good effects of the Plasma-Activated-Medium (PAM) such as the selective antitumor effect and killing effect for the anticancer agent resistant cells were reported, a mechanism of this effect has not been still clarified yet. In this study, we have investigated a contribution of the reactive nitrogen and oxygen species (RNOS) generated in PAM such as hydrogen peroxide and nitrite. Those species generated in the PAM quantitatively measured by light absorbance of commercial regent. Moreover, viable cell count after cell culture with those RNOS intentionally added medium or PAM were also measured by MTS assay. Our NEAPP source generated hydrogen peroxide and nitrite with the generation ratio of 0.35 $\mu$M/s and 9.8 $\mu$M/s. In those RNOS, hydrogen peroxide has respective antitumor effect. On the other hands, nitrite has no antitumor effect singly. But, synergistically enhance the antitumor effect of hydrogen peroxide. Moreover, this effect of those RNOS also contribute for the selectively cancer killing effect of PAM.