Antimicrobial, antibiofilm and cytotoxic effects of a colloidal nanocarrier composed by chitosan-coated iron oxide nanoparticles loaded with chlorhexidine

Araujo, HC; da Silva, ACG; Paiao, LI; Magario, MKW; Frasnelli, SCT; Oliveira, SHP; Pessan, JP; Monteiro, DR

Abstract

Objectives: This study evaluated the antimicrobial and antibiofilm effects of a colloidal nanocarrier for chlorhexidine (CHX) on Candida glabrata and Enterococcus faecalis, as well as tested its cytotoxic effect on murine fibroblasts. Methods: Iron oxide nanoparticles (IONPs) were coated with chitosan (CS) and loaded with CHX at 31.2, 78 and 156 mu g/mL. Antimicrobial effects were assessed by determining the minimum inhibitory concentration (MIC), using the broth microdilution method, and fractional inhibitory concentration index (FICI). Preformed biofilms (48 h) were treated with different concentrations of the nanocarrier (24 h) and quantified by colony-forming units (CFUs), total biomass and metabolic activity. For cytotoxicity, the viability of L929 cells was evaluated by MTT assay after 24 and 48 h of exposure to the nanocarrier. Data were submitted to ANOVA and Fisher LSD or Tukey post-hoc tests (alpha = 0.05). Results: MIC and FICI results showed an indifferent interaction among the components of the nanocarrier for all strains evaluated. CHX alone and nanocarrier containing 156 mu g/mL CHX did not differ from each other in reducing the number of CFUs. However, the nanocarrier containing 156 mu g/mL CHX promoted the highest reductions in total biofilm biomass and metabolism, surpassing the effect of CHX alone. After 24 and 48 h of exposure, the nanocarrier reduced CHX toxicity to the L929 cell at low concentrations. Conclusion: These findings suggest that the CHX nanocarrier has potential to be used in the control of oral diseases associated with C. glabrata and E. faecalis.

Keywords: Biofilms; Candida; Chlorhexidine; Cytotoxicity; Enterococcus faecalis; Iron oxide nanoparticles

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