Biochemical evaluation of wound healing efficacy of cold plasma-conditioned media under different operational conditions

Wound healing is a dynamic and intricate biological process crucial for tissue repair and regeneration. This study explores the potential therapeutic impact of non-thermal plasma generated by a hand-held cold atmospheric pressure plasma jet (C-APPJ) source on fibroblast cells (NIH/3T3) in vitro. The sequential phases of wound healing—inflammation, cell proliferation, and tissue remodelling, were assessed in the context of cell migration and oxidative stress dynamics. Typically, plasma generates a mixture of several reactive oxygen/nitrogen (ROS/RNS) species. The present study investigates the safety and efficacy of C-APPJ under distinct operating conditions (argon (GI) and argon + nitrogen (GII)) and exposure times (1 min and 3 min). Cell viability assays confirmed the non-cytotoxic nature of the cold plasma conditioned medium. The levels of ROS/RNS and malondialdehyde (biomarker of oxidative stress) in the plasma-treated samples remained comparable with the control fibroblast cells grown in normal media, suggesting the favourable modulation of ROS by the cellular antioxidant mechanisms. Accelerated wound-closure rates from 6th hour to 24th hour in all the treated groups ranged from 38.76% to 45.66%, when compared to 34.25% in the control cells. Substantial cell migration leading to 51.59% of wound closure was recorded in the argon + nitrogen (GII) group exposed for 3 min. Taken together, the potential of cold plasma to effectively heal wounds without causing prolonged oxidative stress and chronic inflammation is implicated. These outcomes suggest scope for clinical application of C-APPJ as safe and cost-effective treatment of wounds (ulcers, burns, diabetic foot) and wound disinfection.