Liquid crystalline lipid nanoparticles improve the antibacterial activity of tobramycin and vancomycin against intracellular Pseudomonas aeruginosa and Staphylococcus aureus

The intracellular survival of bacteria is a significant challenge in the fight against antimicrobial resistance. Currently available antibiotics suffer from limited penetration across host cell membranes, resulting in suboptimal treatment against the internalised bacteria. Liquid crystalline nanoparticles (LCNP) are gaining significant research interest in promoting the cellular uptake of therapeutics due to their fusogenic properties; however, they have not been reported for targeting intracellular bacteria. Herein, the cellular internalisation of LCNPs in RAW 264.7 macrophages and A549 epithelial cells was investigated and optimized through the incorporation of a cationic lipid, dimethyldioctadecylammonium bromide (DDAB). LCNPs displayed a honeycomb-like structure, while the inclusion of DDAB resulted into an onion-like organisation with larger internal pores. Cationic LCNPs enhanced the cellular uptake in both cells, reaching up to similar to 90% uptake in cells. Further, LCNPs were encapsulated with tobramycin or vancomycin to improve their activity against intracellular gram-negative, Pseudomonas aeruginosa (P. aeruginosa) and gram-positive, Staphylococcus aureus (S. aureus) bacteria. The enhanced cellular uptake of cationic LCNP resulted in significant reduction of intracellular bacterial load (up to 90% reduction), compared to antibiotic dosed in its free form; with reduced performance observed for epithelial cells infected with S. aureus. Specifically engineered LCNP can re-sensitise antibiotics against both intracellular Gram positive and negative bacteria in diverse cell lines.