Fucoidan coated ciprofloxacin loaded chitosan nanoparticles for the treatment of intracellular and biofilm infections of Salmonella.

Salmonella infections and their gallstone associated biofilm infections are difficult to treat due to poor penetration of antibiotics into the intracellular compartments of macrophages and within biofilms. Here we developed ciprofloxacin loaded chitosan nanoparticles (cCNPs) and fucoidan (Fu) coated cCNPs (Fu-cCNPs). Characterizations of these nanoparticles were carried out using Dynamic Light Scattering‎, Transmission electron microscopy and Fourier transform infrared spectroscopy. The prepared cCNPs and Fu-cCNPs have the size range of 124±7nm and 320±18nm, respectively. Both nanoparticles were found to be non-hemolytic and cytocompatible. In vitro sustained release of ciprofloxacin was observed from both cCNPs and Fu-cCNPs over a period of 2 weeks. The antimicrobial activity of cCNPs and Fu-cCNPs was tested under in vitro and in vivo conditions. The intracellular anti-Salmonella activity of Fu-cCNPs was 2 fold higher than cCNPs and 6 fold higher than ciprofloxacin alone. Fluorescence microscopic images confirmed enhanced delivery of Fu-cCNPs than the cCNPs within the intracellular compartment of macrophages. Both cCNPs and Fu-cCNPs are found to be equally effective in dispersing Salmonella Paratyphi A gallstone biofilms. The in vivo antibacterial activities of Fu-cCNPs were superior to cCNPs which we have validated using Salmonella Paratyphi A infected Drosophila melanogaster fly model. Our overall results showed that (1) Fu-cCNPs are more effective in eradicating Salmonella infections than cCNPs; (2) both cCNPs and Fu-cCNPs were equally effective in dispersing Salmonella gallstone biofilms.