Chitosan/cyclodextrin surface-adsorbed naringenin-loaded nanocapsules enhance bacterial quorum quenching and anti-biofilm activities.

Pathogenic bacteria use quorum sensing (QS), a cell-to-cell communication process that relies on small signaling molecules, to regulate the genetic expression that leads to several essential virulence factors such as bioluminescence, biofilm formation, bacterial motility, among other. Naringenin (NAR), a bitter and colorless flavanone ubiquitous in herbs and fruits, has been shown to inhibit QS activity in P. aeruginosa by decreasing the production of pyocyanin and elastase. In this study, to evaluate the anti-QS activity of naringenin against an E. coli Top 10 biosensor, we developed a novel core-corona nanocapsule formulation comprising surface co-adsorbed β-cyclodextrin (Captisol®) and chitosan loaded with NAR. The results showed that both the nanocapsule (NC) and nanoemulsion (NE) formulations, NAR payload associated with high efficiency , namely ~ 92.88 and ~ 67.98%, respectively. These formulations remained stable for 24 h and showed a biphasic controlled release profile in bacterial M9 medium. Captisol® was effectively immobilized on the NC's surface, resulting in a surface charge inversion from positive (~ + 42 mV) to negative (~ -32 mV) ζ-potential. The biosensor assay revealed that NC outperformed NE in quenching the QS response and the incorporation of naringenin at the NC's multifunctional surface enhanced this bioactivity. Cytotoxicity assays showed that when NAR was associated in NC (188 µM) it was not cytotoxic to Caco2 cells, by contrast with its free form, thus highlighting the cytoprotective effect of the developed formulation. Biofilm formation was inhibited up to ~ 60% in NAR-loaded NC (188 μM), indicating the synergistic effect of positively charged chitosan with the bioactivity of NAR while harnessing the NC's high surface area-to-volume ratio.