Intracellular bacteria-driven macrophage glycolysis in diabetic infected wounds: A hydrogel-based nanotherapy for targeting intracellular pathogens and modulating immune dysregulation

Diabetic infected wounds (DIWs) are a significant clinical challenge, characterized by suboptimal treatment outcomes due to prolonged healing times, frequent recurrences, and high amputation rates. Recent studies have highlighted the role of intracellular bacteria, particularly Staphylococcus aureus , in driving macrophage dysfunction and sustaining chronic inflammation through glycolytic activation. To address this metabolic reprogramming, we utilized single-cell RNA sequencing to analyze the cellular composition of diabetic infected wounds and identified a distinct M1 macrophage subpopulation with upregulated glycolytic function. We developed a hydrogel formulation encapsulating cefalosporin-modified Atractylodes macrocephala extracellular vesicles to target intracellular pathogens and modulate macrophage polarization. The hydrogel demonstrated excellent biocompatibility, efficacy in eradicating intracellular bacteria, capacity to modulate macrophage polarization and metabolic reprogramming, and ability to promote angiogenesis and skin healing. In vitro and in vivo experiments confirmed its dual therapeutic effect of symptom relief and root cause resolution. This study provides insights into the crosstalk between intracellular bacteria and macrophages in DIWs and offers a promising therapeutic strategy for targeting intracellular pathogens and modulating immune dysregulation. • Single-cell RNA sequencing reveals intracellular bacteria driving M1 macrophage glycolysis in diabetic wounds. • A hydrogel encapsulating antibiotic-modified plant EVs is developed for enhanced intracellular delivery and macrophage reprogramming. • In vitro and in vivo studies demonstrate the hydrogel's efficacy in eradicating bacteria, promoting healing, and achieving dual therapeutic effects.