Biofilm formation on textiles presents significant challenges in healthcare, industry, and daily use, contributing to microbial contamination, infections, and material degradation. Among biofilm-forming pathogens, Pseudomonas aeruginosa is particularly concerning due to its multidrug resistance and ability to evade conventional antibiotics, significantly increasing healthcare burden. To address this issue, we developed medical textiles digitally printed with multimodal silver–chitosan–acylase nanoparticles (AgCS@AC NPs). Fabrics endowed with antimicrobial and quorum-quenching properties inhibited bacterial growth and biofilm formation against P. aeruginosa. The enzyme acylase disrupted the quorum-sensing process of bacterial communication and prevented biofilm development, while the AgCS component of the NPs provided antimicrobial efficacy. Cytotoxicity assays confirmed that printed AgCS@AC NPs did not compromise human fibroblast or keratinocyte viability, ensuring biocompatibility. The eco-friendly, scalable, and versatile digital printing technology was innovatively validated for producing enzyme-enabled nanocomposite antimicrobial textiles for medical applications.