Biofilm infection will cause chronic inflammation and hinder the normal healing process of wound. Here, based on the self-assembly of three designed amphiphilic pentapeptides named EK, GG, and DR, pH-switchable antibacterial hydrogels with amphiphilic fiber network are used for the eradication of biofilms and the rescue of delayed healing in infected wounds. These pentapeptides-based hydrogels exhibit an acidic pHswitchable antimicrobial effect and are biocompatible at neutral pH. Additionally, supramolecular nanofiber networks with physical cross-linking with thermosensitive polymers (PNIPAm) and loaded antibacterial oregano oil are further developed. In vitro experiments indicate that the antimicrobial activity of hydrogels comes from the disassembly of acidic pH-dependent nanofiber network and activated release of pentapeptides and oregano oil, which achieves synergistic biofilm eradication. Remarkably, DRbased supramolecular hydrogel improves the healing efficiency of the full-thickness wound of skin in vivo, which is manifested by increased wound closure rate, reduced inflammatory response, faster angiogenesis, and collagen deposition in the wound, exhibiting great potential as wound dressing. The proposed synergistic strategy of inhibiting biofilm formation and activating healing may provide an efficient method for the treatment of clinically infected wounds.