Antibody-Functionalized Polymer Nanoparticles for Targeted Antibiotic Delivery in Models of Pathogenic Bacteria Infecting Human Macrophages.

Abstract

The efficacy of antibody-functionalized poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles (NPs), prepared by nanoprecipitation, carrying rifampicin (RIF) against planktonic, sessile, and intracellular Staphylococcus aureus and Escherichia coli is reported here. A biotinylated anti-S. aureus polyclonal antibody, which binds to structural antigens of the whole bacterium, was functionalized on the surface of RIF-loaded PLGA-based NPs by using the high-affinity avidin-biotin complex. This general strategy allows the binding of commercially available biotinylated antibodies. Coculture models of S. aureus ATCC 25923 and Escherichia coli S17 were used to demonstrate the preferential selectivity of the antibody-functionalized NPs against the Gram-positive bacterium only. At 0.2 μg/mL, complete S. aureus eradication was observed for the antibody-functionalized RIF-loaded NPs, whereas only a 5-log reduction was observed for the nontargeted RIF-loaded NPs. S. aureus is a commensal facultative pathogen having part of its live cycle intracellularly in both phagocytic and nonphagocytic cells. Those intracellular bacterial persisters, named small colony variants, have been postulated as reservoirs of relapsed episodes of infection and consequent treatment failure. At 0.5 μg/mL, the RIF-loaded NPs reduced in 2-log intracellular S. aureus-infecting human macrophages. The ability of those antibody-functionalized nanoparticles to prevent biofilm formation or to reduce the bacterial burden in already-formed mature biofilms is also reported here using S. aureus and E. coli single and cocultured biofilms. In the prevention of S. aureus biofilm formation, the antibody-functionalized NPs exerted a superior inhibition of bacterial growth (up to 2 logs) compared to the nonfunctionalized ones. This study demonstrates the selectivity of the synthesized immunonanoparticles and their antimicrobial efficacy in different scenarios, including planktonic cultures, sessile conditions, and even against intracellular infective pathogens.