TY - JOUR
T1 - Dual nanocarrier of chlorhexidine and fluconazole: physicochemical characterization and effects on microcosm biofilms and oral keratinocytes
AU - Araujo, Heitor Ceolin
AU - Pessan, Juliano Pelim
AU - Caldeirão, Anne Caroline Morais
AU - Sampaio, Caio
AU - Oliveira, Sandra Helena Penha
AU - Sales, Douglas Henrique
AU - Teixeira, Silvio Rainho
AU - Constantino, Carlos José Leopoldo
AU - Delbem, Alberto Carlos Botazzo
AU - Oliveira, Sandra Helena Penha
AU - Ramage, Gordon
AU - Monteiro, Douglas Roberto
N1 - Funding Information:
The authors express their gratitude to nChemi Engenharia de Materiais (São Carlos, São Paulo, Brazil) for providing the colloidal suspension of iron oxide nanoparticles used in this study. The authors also thank LabMicro – FCT/Unesp ( School of Technology and Applied Sciences (FCT), São Paulo State University (Unesp), Presidente Prudente/São Paulo, Brazil; and FAPESP , grant number 2014/11408–3 ) for technical assistance with the confocal images.
Funding Information:
This study was supported by São Paulo Research Foundation (FAPESP , Brazil; grant number 2017/24416–2 ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq , Brazil; grant number 404721/2016–8 ) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES , Brazil; scholarship to the first author and Finance Code 001 ).
PY - 2023/11
Y1 - 2023/11
N2 - Objectives: This study assembled and characterized a dual nanocarrier of chlorhexidine (CHX) and fluconazole (FLZ), and evaluated its antibiofilm and cytotoxic effects. Methods: CHX and FLZ were added to iron oxide nanoparticles (IONPs) previously coated by chitosan (CS) and characterized by physical-chemical analyses. Biofilms from human saliva supplemented with Candida species were grown (72 h) on glass discs and treated (24 h) with IONPs-CS carrying CHX (at 39, 78, or 156 µg/mL) and FLZ (at 156, 312, or 624 µg/mL) in three growing associations. IONPs and CS alone, and 156 µg/mL CHX + 624 µg/mL FLZ (CHX156-FLZ624) were tested as controls. Next, microbiological analyses were performed. The viability of human oral keratinocytes (NOKsi lineage) was also determined (MTT reduction assay). Data were submitted to ANOVA or Kruskal-Wallis, followed by Fisher's LSD or Tukey's tests (α=0.05). Results: Nanocarriers with spherical-like shape and diameter around 6 nm were assembled, without compromising the crystalline property and stability of IONPs. Nanocarrier at the highest concentrations was the most effective in reducing colony-forming units of Streptococcus mutans, Lactobacillus spp., Candida albicans, and Candida glabrata. The other carriers and CHX156-FLZ624 showed similar antibiofilm effects, and significantly reduced lactic acid production (p<0.001). Also, a dose-dependent cytotoxic effect against oral keratinocytes was observed for the dual nanocarrier. IONPs-CS-CHX-FLZ and CHX-FLZ significantly reduced keratinocyte viability at CHX and FLZ concentrations ≥7.8 and 31.25 µg/mL, respectively (p<0.05). Conclusion: The nanotherapy developed outperformed the effect of the combination CHX-FLZ on microcosm biofilms, without increasing the cytotoxic effect of the antimicrobials administered. Clinical Significance: The dual nanocarrier is a promising topically-applied therapy for the management of oral candidiasis considering that its higher antibiofilm effects allow the use of lower concentrations of antimicrobials than those found in commercial products.
AB - Objectives: This study assembled and characterized a dual nanocarrier of chlorhexidine (CHX) and fluconazole (FLZ), and evaluated its antibiofilm and cytotoxic effects. Methods: CHX and FLZ were added to iron oxide nanoparticles (IONPs) previously coated by chitosan (CS) and characterized by physical-chemical analyses. Biofilms from human saliva supplemented with Candida species were grown (72 h) on glass discs and treated (24 h) with IONPs-CS carrying CHX (at 39, 78, or 156 µg/mL) and FLZ (at 156, 312, or 624 µg/mL) in three growing associations. IONPs and CS alone, and 156 µg/mL CHX + 624 µg/mL FLZ (CHX156-FLZ624) were tested as controls. Next, microbiological analyses were performed. The viability of human oral keratinocytes (NOKsi lineage) was also determined (MTT reduction assay). Data were submitted to ANOVA or Kruskal-Wallis, followed by Fisher's LSD or Tukey's tests (α=0.05). Results: Nanocarriers with spherical-like shape and diameter around 6 nm were assembled, without compromising the crystalline property and stability of IONPs. Nanocarrier at the highest concentrations was the most effective in reducing colony-forming units of Streptococcus mutans, Lactobacillus spp., Candida albicans, and Candida glabrata. The other carriers and CHX156-FLZ624 showed similar antibiofilm effects, and significantly reduced lactic acid production (p<0.001). Also, a dose-dependent cytotoxic effect against oral keratinocytes was observed for the dual nanocarrier. IONPs-CS-CHX-FLZ and CHX-FLZ significantly reduced keratinocyte viability at CHX and FLZ concentrations ≥7.8 and 31.25 µg/mL, respectively (p<0.05). Conclusion: The nanotherapy developed outperformed the effect of the combination CHX-FLZ on microcosm biofilms, without increasing the cytotoxic effect of the antimicrobials administered. Clinical Significance: The dual nanocarrier is a promising topically-applied therapy for the management of oral candidiasis considering that its higher antibiofilm effects allow the use of lower concentrations of antimicrobials than those found in commercial products.
KW - Biofilms
KW - Candida
KW - Chlorhexidine
KW - Cytotoxicity
KW - Fluconazole
KW - Iron oxide nanoparticles
U2 - 10.1016/j.jdent.2023.104699
DO - 10.1016/j.jdent.2023.104699
M3 - Article
AN - SCOPUS:85171268448
SN - 0300-5712
VL - 138
JO - Journal of Dentistry
JF - Journal of Dentistry
M1 - 104699
ER -