Developing antibacterial HB43 peptide-loaded chitosan nanoparticles for biofilm treatment

Biofilm-associated infections on medical devices are challenging to treat. Therefore, innovative treatment approaches are needed to penetrate biofilms and eliminate bacteria. With this study, we developed chitosan nanoparticles (CNPs) encapsulating the antibacterial peptide HB43 at increasing CNP/peptide ratios (from 1 to 4 % for P1-CNP, P2-CNP, and P4-CNP, respectively) using the ion gelation method. Our goal was to enhance antibacterial drug delivery inside a methicillin-resistant Staphylococcus aureus (MRSA) biofilm. Our analysis showed a direct correlation between the encapsulation efficacy of HB43 and the physical properties of the CNPs, such as size and zeta potential. P1-CNP was identified as the optimal formulation, characterized by its small size, high encapsulation efficiency, and cationic surface charge. Release studies indicated that HB43 was released in a sustained manner particularly under acidic conditions, which enhanced therapeutic efficacy. We tested the P1-CNP in culture media with pH levels of 7.4 and 5.5 to assess the pH responsiveness of the CNPs and mimic the infection environment. Both conditions showed that the HB43 loaded-CNPs effectively reduced bacterial populations in a dose-dependent manner, with up to a 99.99 % reduction in bacterial load. This study offers a promising new strategy for managing biofilm-associated infections and addressing antibiotic resistance by using CNPs loaded with HB43.