Exploring the antibiofilm potential of chitosan nanoparticles by functional modification with chloroquine and deoxyribonuclease.

Planktonic bacteria tend to form sessile community architectures to shield resident bacteria from various environmental stresses. The formed biofilm leads to the failure of conventional antimicrobial therapy. Extracellular macromolecules, including extracellular DNA (eDNA), proteins, lipids, and polysaccharides, crosslink into gel-like structures through electrostatic forces in the mature biofilm matrix. The stereo-structural integrity and chemical inertia of the extracellular polymeric matrix result in comprehensive antimicrobial resistance to antibacterial polysaccharides. Herein, an ionic gelation method was employed to functionalize cationic chitosan nanoparticles (CSNPs) with chloroquine and deoxyribonuclease. The modification involved shifting eDNA chirality through a DNA-intercalating agent, chloroquine, and hydrolyzing an eDNA scaffold with deoxyribonuclease. The antibiofilm activity was assessed against a standard Staphylococcus aureus strain and clinical subtype isolates. Functional modifications targeting eDNA improved the chitosan anti-biofilm efficiency (residual biomass decreased from 74.2 to 90.3 % to 16.7-24.6 %) by disrupting the biofilm matrix. The functional CSNPs worked as a sensitizer prodrug, contributing to a bactericidal process of chitosan itself (cell wall damage increased from 11.38-18.16 % to 55.2-61.4 %) by dispersing the biofilm-enclosed bacteria. In vivo, the bacterial burden of infected mouse joints was reduced by 4.1 lg CFU/mL. Our results indicate the potential of this chitosan-based anti-infection strategy in biofilm-related infections.