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Recent advances in molecular biology and biomedical research have led to the development and utilization of new techniques and agents for treating diseases. One such technique is Cold Atmospheric Plasma (CAP), which is increasingly adopted in healthcare. Over the last two decades, CAP has attracted the attention of many biomedical scientists due to its ability to generate reactive species [1]. CAP is a non-thermal plasma that operates near room temperature and has the capacity to produce reactive oxygen and nitrogen species, enabling CAP to benefit experimental applications in wound healing, microbial inactivation, cancer treatment, and many more [2,3]. Apart from the reactive species, CAP also generates UV radiation, an electric field, charged species, and heat during its production. The plasma parameters and concentration of reactive species are device-dependent. Therefore, the design of a CAP device, its optimization, and validation are of utmost importance before biomedical applications [4].
In this study, we evaluated the pre-clinical safety and efficacy of CAP generated by an indigenously developed dielectric barrier discharge (DBD) setup. Electrical and optical characterization of the DBD-CAP device confirmed stable plasma discharge and consistent production of reactive oxygen and nitrogen species. Safety assessments were performed on mouse peritoneal cells (ex vivo study) and the skin of Wistar rats (in vivo study). Mice peritoneal cells are the primary cells used to assess safety in ex vivo studies. A safety study on mouse peritoneal cells was conducted at 24 kV with treatment times ranging from 0 to 120 seconds, and cell viability was assessed following a 24-hour incubation. A safety study was conducted on Wistar rats at 18 kV, with a treatment time of 15-45 seconds. Various blood parameters and histology of the treated skin tissue were analyzed. In contrast, treatment of human melanoma skin cancer cells (G-361) was performed as part of an anticancer study. The CAP treatment of human melanoma skin cancer cells at 24 kV with a treatment time of 0-45 s. Cell viability, intracellular ROS, and apoptosis were analyzed following 24-hour incubation. The CAP treatment on G-361 cells resulted in a substantial reduction in cell viability, an increase in apoptosis, and an increase in intracellular ROS, with increasing treatment time. Cell mortality after treatment is driven by ROS-mediated apoptosis. Together, these findings demonstrate the selective profile of the DBD-CAP device, safety in normal skin cells, and strong anti-cancer efficacy in melanoma, supporting its translational potential as a minimally invasive therapeutic option for skin malignancies.
