Wound healing is a complex, multi-phase process involving numerous cellular and molecular mechanisms. Chronic wounds, exacerbated by conditions such as diabetes or bacterial biofilms, pose significant clinical challenges, increasing healthcare costs. This study aimed to develop biogenic nano-hydrogels (BgNH) incorporating silver nanoparticles synthesized from medicinal plant extracts. The efficacy of these hydrogels was tested in wound healing, antibiofilm, and antivirulence assays, particularly targeting Pseudomonas aeruginosa. Results revealed that BgNH exhibited significant antibiofilm activity, with 96% inhibition and 89.93% biofilm dispersal, while demonstrating minimal cytotoxicity at lower concentrations. Structural analysis confirmed an amorphous, porous matrix facilitating controlled nanoparticle release. In vivo studies demonstrated a 25.23% improvement in wound healing compared to untreated controls, highlighting BgNH’s potential as a therapeutic wound dressing with both antimicrobial and healing-promoting properties. This study concludes that BgNH offers a promising alternative to conventional wound care treatments, with further research needed to optimize formulations for clinical use.