A Stage-Adaptive Injectable Thermogelling Hydrogel Platform for Local Neutralization of Clostridium α-Toxin Using MgO Nanoparticles and mRNA-Encoded α-Toxin Inhibitors

Clostridial wound infections represent a severe and rapidly progressive clinical threat, driven primarily by secreted enzymatic virulence factors rather than bacterial burden alone. Among these, Clostridium α-toxin (phospholipase C) plays a central role in membrane disruption, vascular compromise, and accelerated tissue necrosis, particularly in high-risk wound populations such as diabetic foot ulcers, pressure injuries, and contaminated surgical wounds. This preprint presents a conceptual translational framework for stage-adaptive, localized neutralization of α-toxin activity using an injectable thermogelling hydrogel platform. The proposed system is designed to intervene during early, pre-necrotic stages of wound risk, complementing—rather than replacing—standard surgical debridement and systemic antimicrobial therapy. The platform integrates three coordinated components: Injectable thermogelling hydrogel that transitions from liquid to semi-solid at physiological temperature, enabling conformal filling and sustained retention within irregular wound geometries; Embedded magnesium oxide (MgO) nanoparticles providing non-antibiotic antimicrobial activity and biofilm disruption; Stage-adaptive mRNA-encoded enzyme inhibitors designed for localized, transient expression to functionally neutralize α-toxin without systemic exposure or immune activation. Design and optimization are guided by the Rami Five-Checkpoint Framework, adapted for enzyme neutralization applications: target selection and validation, sequence optimization, RNA structural control, immunogenicity minimization, and cost/manufacturability considerations. The framework emphasizes prevention of enzyme-mediated tissue destruction rather than treatment of established gas gangrene. This manuscript does not report experimental or clinical results. Instead, it serves as a mechanism-driven design blueprint outlining biological rationale, delivery architecture, safety considerations, regulatory pathways, and clinical integration strategies for future preclinical development. The work is intended for researchers and clinicians in wound care, biomaterials, infectious disease, and translational mRNA therapeutics, and is provided as an early-stage research framework rather than a validated medical intervention.