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Plasma-activated hydrogels have promising application prospects in the field of transdermal penetration therapy, such as subcutaneous tumours, due to their long-lasting, slow-release properties and ability to be easily applied to skin. In this study, surface air plasma was used to prepare plasma-activated Aristoflex AVC hydrogel (PAH) and plasma-activated water (PAW) by irradiation. The characteristics of PAH and PAW were further studied in terms of reactive oxygen species (ROS) storage, transdermal permeation, and in vitro anticancer effects in an artificial skin model. The experimental results showed that PAH had a prolonged storage time of ROS and more transdermal permeation of ROS through artificial skin in Franz diffusion cell after 30 h compared to PAW, which means PAH could be superior to PAW for long-term applications on the skin surface. The in vitro anticancer effects of PAH in an artificial skin model were also better than those of PAW, reducing the viability of A375 cells without causing damage to HaCaT cells. Our study shows that PAH is superior to PAW in body-surface coating applications and can be used for subcutaneous anticancer applications. [ABSTRACT FROM AUTHOR]