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Osteosarcoma (OS), a malignancy primarily affecting children and adolescents, is the most frequently encountered malignant, non-hematologic, bone tumor. Despite the gradual improvement of survival rates, the management of this disease remains problematic due to challenges such metastasis development, its heterogeneous characteristics, and resistance to cytostatic drugs. Cold atmospheric plasma (CAP), a partially ionized gas operating at near room temperature, which is comprised of free carriers, excited or neutral molecules, and active radicals capable of initiating diverse physical phenomena and chemical reactions represent a new and innovative potential solution in cancer therapy. The aim of this study was to evaluate the capacity of CAP produced by our custom-build plasma source to induce cytotoxic effects to HOS cells and to analyze post-treatment modulations of cdk1, tnf α and tp53 genes expression. Direct and indirect CAP treatments effectiveness on HOS cells were evaluated by MTT assay and the regulation of interest genes expression were carried out by RT-qPCR analysis. Cell viability analysis revealed a strong cytotoxic effect of direct CAP treatment on HOS cells, while the indirect treatment resulted in a slight decrease of cells viability. Direct CAP treatment modulates the expression of all analyzed genes, both at 2- and 24-hours posttreatment. In conclusion, direct CAP treatment produced by our custom-build plasma source have cytotoxic effects on HOS cells up to 24 hours post-treatment. Furthermore, direct CAP treatment induces cell cycle arrest of HOS cells, and the CAP-induced cell death is independent of tp53 gene. [ABSTRACT FROM AUTHOR]

Over the past decade, cold atmospheric plasma (CAP) has undergone extensive research as a promising therapeutic approach in oncology, with different treatment methods and exposure configurations being investigated and resulting in various biological effects, most of them after long exposure or treatment durations. This study aimed to evaluate the potential of a custom-made CAP generation source to produce plasma-activated medium (PAM) with cytotoxic effects and subsequently to establish the optimal exposure and treatment parameters. The exposure's electrical parameters, as well as pH and NO 2 − content of PAM were analyzed. The cytotoxic potential and optimal parameters of the treatment were established by evaluating the viability of human osteosarcoma cells (HOS cell line) and human osteoblasts (HOB cell line) treated with PAM under different conditions. Our results showed that indirect treatment with CAP presents selective dose-dependent cytotoxic effects, while the cell viability decrease was not found to be correlated with the PAM acidification due to CAP exposure. The Griess assay revealed very high and long-term stable concentrations of NO 2 − in PAM. Overall, this study presents a simple and faster method of producing PAM treatment with cytotoxic effects on HOS cells, by using a custom-built CAP source. [ABSTRACT FROM AUTHOR]