pH‐Responsive Protein‐Polycation Nanocarriers for Efficient Eradication of Bacterial Biofilms and Intracellular Bacteria

Bacterial biofilms and intracellular pathogens pose significant challenges in eradication, often leading to persistent infections that are difficult to treat. To address this issue, the hydrophobic biofilm dispersant D-tyrosine is encapsulated within protein-polycation nanoparticles, designed using a mannose-terminated cationic polymer and concanavalin through electrostatic interactions. Thermodynamic studies reveal that free mannosyl groups on the nanoparticle surface promote spontaneous binding to receptor molecules mimicking those on bacterial biofilms and host cells. Under mildly acidic conditions, the nanoparticles reduce in size from 550 to ≈48 nm within 2 h, releasing 76% of encapsulated D-tyrosine. The combination of mannose targeting, particle size reduction, and controlled D-tyrosine release enable the nanoparticles to eliminate 70%-80% of the Pseudomonas aeruginosa and Staphylococcus aureus biofilm biomass at minimum bactericidal concentration (MBC) and 2MBC while eradicating 8 log of bacteria embedded within the biofilm. In an intracellular Pseudomonas aeruginosa infection model using RAW 264.7 macrophages, the nanoparticles at 2MBC eliminate over 95% of the intracellular bacteria without inducing an increase in the inflammatory cytokine interleukin-6. These protein-polycation nanoparticles, which activate their antimicrobial properties under acidic conditions, efficiently penetrate bacterial biofilms and host cell barriers via their mannose-rich surface, offering a promising strategy for the treatment of persistent infections.