Abstract

Chronic wounds in diabetic patients remain a significant clinical challenge due to neuropathy, vascular dysfunction, and persistent inflammation. In this study, we engineered a multifunctional wound dressing by incorporating conditioned media (CM) including secretome derived from three cell sources, natural killer (NK) cells, adipose-derived mesenchymal stem cells (ADMSCs), and umbilical cord mesenchymal stem cells (UCMSCs) into a silk fibroin-glycidyl methacrylate (SF-GMA) hydrogel scaffold. The formed SF-GMA hydrogel exhibited a porous, hydrated architecture with an optimal curing time of 15 s, a compressive modulus of 101 ± 2 KPa and favorable biocompatibility. Cytokine profiling revealed that all CM types were rich in pro-regenerative factors. In vitro assays showed that CM promoted endothelial tube formation and enhanced fibroblast and keratinocyte migration under hyperglycemic conditions. In a streptozotocin-induced diabetic mouse model, CM-loaded SF-GMA hydrogels significantly accelerated wound closure (90 ± 8.21%) compared to controls (30 ± 5.20%) by day 14. Histological analysis revealed improved re-epithelialization, granulation tissue formation, and collagen deposition, while immunofluorescence indicated enhanced neovascularization and reduced inflammation. Notably, ADMSC-CM most strongly promoted angiogenesis, whereas NK-CM enhanced epithelial migration. Overall, CM@SF-GMA hydrogel promoted diabetic wound healing by combining structural support with sustained delivery of regenerative factors, and immunomodulation. This integrated strategy offers a promising platform for treating chronic wounds in diabetic patients.