Inactivation of Vaccinia Virus via Nitric Oxide-Plasma Activated Water: A Potential Way to Inactivate Mpox Virus

Mpox virus (Poxviridae family) an emerging environmental bio-contaminant caused a non-zoonotic human infection in 2022. The recent surge mirrors early COVID-19 trends, highlighting the need for effective viral inactivation to prevent outbreaks and reduce environmental risks. Our study explores an eco-friendly and non-toxic antiviral approach using nitric oxide (NO_x)-plasma activated water (PAW) for environmental decontamination. Vaccinia virus (VACV) chosen as a surrogate model due to their genetic similarities with Mpox virus (MPXV). Results demonstrated that NO_x-PAW was non-toxic to host cells and significantly reduced VACV infection in lung cell cultures. Moreover, it induced structural alterations in viral attachment proteins A27 and H3, compromising their functionality resulting in reduced binding affinity towards heparan sulfate and lowering internalization via macropinocytosis. Sequence analysis between VACV and MPXV, including receptor-binding domains, confirmed high similarity, supporting VACV's utility as a model for MPXV inactivation studies. Furthermore, in-silico analysis revealed NO_x species (NO, NO₂, NO₃ and N₂O) played crucial role in modification of surface protein by interaction with the amino acids. Overall, the study demonstrated successful VACV inactivation highlighting NO_x-PAW as a promising environmentally safe antiviral strategy for mitigating the spread of DNA viruses like MPXV in contaminated settings, contributing to proactive outbreak prevention and environmental biosafety.