Leishmanicidal efficacy of cold atmospheric multiple plasma jet against Leishmania major in a murine model: effects on parasite burden, cytokine profiles, and wound healing

Cutaneous leishmaniasis (CL) is a neglected tropical disease with limited effective treatments options due to drug toxicity, emerging resistance, and complex therapeutic regimens. The Cold Atmospheric Multiple Plasma Jet (CAMPJ) has recently gained attention as a novel therapeutic modality because it generates reactive oxygen and nitrogen species, with antimicrobial and immunomodulatory activity. This study evaluates the therapeutic efficacy of CAMPJ in a BALB/c mouse model of CL, focusing on its leishmanicidal, wound-healing, and immunomodulatory effects. Female BALB/c mice were inoculated with Leishmania major promastigotes and, after lesion development, assigned to six groups: topical amphotericin B (positive control), CAMPJ (treatment for 5, 10, or 15) minutes, untreated control, and air-treated control. CAMPJ and control treatments were administered twice a week for 21 days, while amphotericin B was applied daily. Lesion progression was monitored weekly using Digitizer software, parasite burden was quantified via Leishman-Donovan Units (LDU), and cytokine levels (IFN-γ and IL-4) were measured by ELISA. CAMPJ treatment produced a significant reduction in lesion size compared with negative controls ( p < 0.05), with the 15-minute exposure yielding the greatest improvement. Splenic parasite burden decreased significantly across all CAMPJ-treated groups, again with the 15 min group showing the lowest LDU values. Notably, 15 min CAMPJ exposure promoted a Th1-biased immune response, characterized by elevated IFN-γ and reduced IL-4 contrasting with the Th2 dominant profile observed in untreated controls. These findings indicate that CAMPJ exhibits strong leishmanicidal activity and beneficial immunomodulatory effects, supporting its potential as an alternative or adjunct therapy for CL.