Dual-enzyme cascade protein hydrogel membrane orchestrates metabolism-immunity coupling for diabetic wound repair.

Abstract

Diabetic wounds present a formidable clinical challenge due to a self-amplifying network of hyperglycemia-driven microenvironmental dysregulation, including hypoxia, oxidative stress, chronic inflammation, and persistent infection. Existing therapies fail to integrate mechanical support with dynamic, holistic regulation of this complex milieu, leaving intertwined biological and material challenges unresolved. Here, we report a multifunctional protein hydrogel membrane, GOX@MnO₂/CLP-EGF@BSA (GM/CEB), designed to actively remodel the diabetic wound niche. The hydrogel is constructed via topological chain entanglement of bovine serum albumin (BSA), incorporating a genetically encoded collagen-like protein fused with epidermal growth factor (CLP-EGF) for sustained regenerative signaling, and embedding a GOX@MnO₂ dual-enzyme cascade that converts glucose into oxygen while scavenging reactive oxygen species. This hierarchical, protein-based hydrogel membrane is mechanically robust and biodegradable, enabling continuous, integrated modulation of the hyperglycemia-driven pathological microenvironment. In a S. aureus-infected diabetic wound model, GM/CEB significantly accelerated healing by alleviating hypoxia and promoting macrophage polarization. Concurrently, GM/CEB activated EGFR-associated signaling pathways (PI3K/AKT/mTOR), thereby enhancing tissue regeneration and restoring dermal architecture. Taken together, this hybrid multifunctional protein-based hydrogel membrane, characterized by high toughness and a fully proteinaceous matrix, represents a new generation of protein-based biomaterials, holding strong potential as a skin substitute to provide an integrated platform for infection control, immune modulation, and functional tissue regeneration in complex wound repair. STATEMENT OF SIGNIFICANCE: Chronic wounds represent a paradigmatic failure of biomaterials design, where static scaffolds cannot adapt to dynamically evolving pathological microenvironments. Here, we introduce a protein-based hydrogel membrane that departs from conventional crosslinked systems by leveraging topological chain entanglement as the primary structural principle, coupled with an embedded enzyme-like cascade to actively reprogram the wound milieu. This integration enables simultaneous mechanical resilience and spatiotemporally adaptive regulation of oxidative stress, immunity, and regeneration. By shifting from passive support to active microenvironmental control, this work establishes a new design paradigm for protein biomaterials and advances the conceptual foundation for treating complex tissue pathologies.