Abstract:The escalating threat of healthcare-associated infections highlights the urgent need for biocompatible antibacterial materials that effectively combat drug-resistant pathogens. In this study, we present a novel fabrication method for triple-helical recombinant collagen (THRC)-silver hybrid nanofibers, specifically designed for anti-methicillin-resistant staphylococcus aureus (MRSA) applications. Utilizing a silver-mediated crosslinking strategy, we harness a low-power 38 W lamp to enable silver ions (Ag+) to mediate crosslinking across various proteins. Mechanistic insights reveal the pivotal role of nine amino acids in facilitating this reaction. The THRC maintains its native structure, forming well-ordered nanofibers, while other globular proteins form a distinctive network-like structure. THRC also serves as a reducing and dispersing agent, facilitating the in situ synthesis of highly dispersed silver nanoparticles (AgNPs) (∼7 nm in diameter) within the nanofibers. Systematic investigation of the reaction conditions between THRC and Ag+ demonstrates the versatility of this novel approach for nanofiber fabrication. The incorporation of AgNPs imparts exceptional antibacterial activity to the THRC/AgNPs nanofibers, exhibiting a minimum inhibitory concentration of 19.2 mg l−1 and a minimum bactericidal concentration of 153.6 mg l−1 against MRSA. This innovative approach holds significant potential for developing antibacterial protein-based biomaterials for infection management in wound healing and other biomedical applications.