Reactive oxygen species-functionalized hydrogels loaded with ginger-derived nanoparticles promote diabetic wound healing by modulating macrophage polarization

Purpose: Diabetic cutaneous injuries frequently exhibit prolonged inflammatory responses and excessive accumulation of reactive oxygen species (ROS), which collectively impede normal healing cascades. Standard therapeutic approaches often yield suboptimal healing outcomes in this patient population, highlighting the urgent need for innovative and effective interventions. In this study, we examined whether macrophage polarization induced by a ROS-responsive hydrogel loaded with ginger-derived nanoparticles enhanced diabetic wound healing and identified the mo-lecular pathways involved. Methods: In this study, a ROS-responsive hydrogel loaded with ginger-derived nanoparticles (Gdn@PT) was developed, and its morphology, ROS-triggered degradation, and drug release be-havior were thoroughly characterized. The db/db spontaneous diabetic mouse model was employed, with animals randomly assigned to three groups (n = 10 per group): control, hydrogel alone (PT), and nanoparticle-loaded hydrogel (Gdn@PT). Full-thickness skin wounds were induced on day 0, fol-lowed by the respective treatments, and wound healing was monitored for 13 days. Histological analysis, immunofluorescence, and Western blotting were conducted to assess wound closure, collagen deposition, inflammatory cytokine expression (TNF-α), and macrophage polarization markers (CD86 and CD206). In vitro, the effects of Gdn@PT on macrophage polarization were investigated in LPS-induced RAW264.7 cells. Results: Ginger-derived spherical nanoparticles were successfully isolated via tangential flow fil-tration and incorporated into ROS-responsive hydrogel matrices, yielding a biocompatible nano-composite. The therapeutic mechanism relied on the hydrogel scaffold's ROS-dependent structural degradation, which facilitated controlled release of the nanoparticulate payload within the wound microenvironment. In db/db diabetic wound models, this hydrogel significantly promoted wound healing and collagen deposition, which was accompanied by PI3K-AKT pathway activation, a prominent shift in macrophage polarization toward the M2 phenotype, decreased TNF-α expression, and upregulated Mcl-1 pro-survival mediator levels. These immunomodulatory effects were cor-roborated in vitro. Conclusion: This ROS-responsive hydrogel system, delivering ginger-derived nanoparticles, con-stitutes a promising therapeutic strategy for diabetic wound management by leveraging oxidative stress to trigger on-demand bioactive release. Notably, treatment with this system was associated with PI3K-AKT pathway activation, M2 macrophage polarization, and enhanced tissue repair, sug-gesting its clinical potential. Nevertheless, the profound efficacy observed mandates further com-prehensive investigations, including pathway-specific inhibition or genetic manipulation studies, to fully delineate the molecular mechanisms underlying these associations and optimize translational viability. Keywords: Zingiber officinale, reactive oxygen species-responsive, ginger nanoparticle bodies, diabetic care .