Abstract

Atmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo. These effects are based on the biochemistry of the reactive oxygen and nitrogen species generated by plasmas in physiological liquids, referred to as plasma-conditioned liquids. Plasma-conditioned media are efficient in the generation of reactive species, inducing selective cancer cell death. However, the concentration of reactive species generated by plasma in the cell culture media of different cell types can be highly variable, complicating the ability to draw precise conclusions due to the differential sensitivity of different cells to reactive species. Here, we compared the effects of direct and indirect plasma treatment on non-malignant bone cells (hOBs and hMSCs) and bone cancer cells (SaOs-2s and MG63s) by treating the cells directly or exposing them to previously treated cell culture medium. Biological effects were correlated with the concentrations of reactive species generated in the liquid. A linear increase in reactive species in the cell culture medium was observed with increased plasma treatment time independent of the volume treated. Values up to 700 µM for H2O2 and 140 µM of NO2− were attained in 2 mL after 15 min of plasma treatment in AdvDMEM cell culture media. Selectivity towards bone cancer cells was observed after both direct and indirect plasma treatments, leading to a decrease in bone cancer cell viability at 72 h to 30% for the longest plasma treatment times while maintaining the survival of non-malignant cells. Therefore, plasma-conditioned media may represent the basis for a potentially novel non-invasive technique for bone cancer therapy.

Highlights

  • Atmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo

  • Treating different cell culture media resulted in the generation of different amounts of reactive oxygen and nitrogen species (RONS) by plasma, which prevented us from drawing definitive conclusions on the sensitivity of the two cell types to the reactive species generated by plasma

  • The results clearly show that both non-malignant cell lines remained fully viable (Fig. 3a,b), as the viability of human bone marrow-derived mesenchymal stem cells (hMSCs) was greater than 80% at all treatment times human osteoblast primary cells (hOBs) exposed to direct plasma initially displayed stimulated cell proliferation with respect to control—which is referenced at 100% cell viability—(Fig. 3a) following atmospheric pressure plasma jet (APPJ) treatment after 24 h of incubation (135 ± 2% for 1.5 min) which was mitigated with incubation time to values close to the control cells (110 ± 10% for 1.5 min at 72 h). hMSCs were more sensitive to direct plasma treatment, exhibiting a small reduction in viability to values between 85 and 100% (Fig. 3b)

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Summary

Introduction

Atmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo. APPJ has the potential to affect cells through complex biochemical processes with the ability to selectively kill cancer cells without affecting non-malignant cells, i.e., the surrounding ­tissues[6,7,8] This effect is primarily attributed to the reactive oxygen and nitrogen species (RONS) generated by APPJ and is responsible for disturbing the cellular metabolic ­environment[9,10]. As cells are surrounded by biological media, i.e., blood in in vivo or cell culture media in vitro, interactions between APPJ and liquids occur during treatment, leading to biological effects Based on these findings, plasma-conditioned media were investigated in this work as a potential vehicle of RONS transport, avoiding the effects of visible UV radiation, electric fields and thermal heating from APPJs, which can disturb the cellular environment. The potential for the use of plasma-conditioned media is studied in this work as a potential tool for future non-invasive therapy for bone cancer

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