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. 2012 Jan 16:12:6.
doi: 10.1186/1472-6882-12-6.

Cranberry proanthocyanidins inhibit the adherence properties of Candida albicans and cytokine secretion by oral epithelial cells

Mark Feldman  1 , Shinichi Tanabe, Amy Howell, Daniel Grenier
Affiliations

Cranberry proanthocyanidins inhibit the adherence properties of Candida albicans and cytokine secretion by oral epithelial cells

Mark Feldman et al. BMC Complement Altern Med. .

Abstract

Background: Oral candidiasis is a common fungal disease mainly caused by Candida albicans. The aim of this study was to investigate the effects of A-type cranberry proanthocyanidins (AC-PACs) on pathogenic properties of C. albicans as well as on the inflammatory response of oral epithelial cells induced by this oral pathogen.

Methods: Microplate dilution assays were performed to determine the effect of AC-PACs on C. albicans growth as well as biofilm formation stained with crystal violet. Adhesion of FITC-labeled C. albicans to oral epithelial cells and to acrylic resin disks was monitored by fluorometry. The effects of AC-PACs on C. albicans-induced cytokine secretion, nuclear factor-kappa B (NF-κB) p65 activation and kinase phosphorylation in oral epithelial cells were determined by immunological assays.

Results: Although AC-PACs did not affect growth of C. albicans, it prevented biofilm formation and reduced adherence of C. albicans to oral epithelial cells and saliva-coated acrylic resin discs. In addition, AC-PACs significantly decreased the secretion of IL-8 and IL-6 by oral epithelial cells stimulated with C. albicans. This anti-inflammatory effect was associated with reduced activation of NF-κB p65 and phosphorylation of specific signal intracellular kinases.

Conclusion: AC-PACs by affecting the adherence properties of C. albicans and attenuating the inflammatory response induced by this pathogen represent potential novel therapeutic agents for the prevention/treatment of oral candidiasis.

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Figures

Figure 1
Figure 1
Cranberry proanthocyanidins showing the presence of A-type linkages.
Figure 2
Figure 2
Effect of AC-PACs on C. albicans biofilm formation. Panel A: C. albicans biofilms were quantified by staining with crystal violet. Assays were done in triplicate and the means ± SD from three independent experiments were calculated. A value of 100% was assigned to the biofilm formed in the absence of AC-PACs. *, significantly lower than the value for the untreated control (P < 0.05). Panels B and C: Phase contrast microscopy of biofilms formed in the presence (B) or absence (C) of AC-PACs (25 μg/ml).
Figure 3
Figure 3
Effect of AC-PACs on C. albicans biofilm desorption. Newly formed biofilms (48 h) of C. albicans biofilms were treated (30 and 120 min) with AC-PACs prior to determine biofilm biomass by staining with crystal violet. Assays were done in triplicate and the means ± SD from three independent experiments were calculated. A value of 100% was assigned to the preformed biofilm unexposed to AC-PACs. *, significantly lower than the value for the unexposed control (P < 0.05).
Figure 4
Figure 4
Effect of AC-PACs on adherence of C. albicans to oral epithelial cells. Panel A: FITC-labeled C. albicans adhered to epithelial cells were quantified by measuring fluorescence in a microplate reader. Assays were done in triplicate and the means ± SD from three independent experiments were calculated. A value of 100% was assigned to C. albicans adhered to epithelial cells not treated with AC-PACs. *, significantly lower than the value for the untreated control (P < 0.05). Panels B to E: Image processing was performed using fluorescence microscope. Images of C. albicans adhered to untreated or AC-PACs-treated epithelial cells in fluorescence mode (B and C, respectively) and in phase-contrast mode (D and E, respectively).
Figure 5
Figure 5
Effect of AC-PACs on adherence of C. albicans to acrylic resin disks. FITC-labeled C. albicans adhered to saliva-coated acrylic resin disks were quantified by measuring fluorescence in a microplate reader. Assays were done in triplicate and the means ± SD from three independent experiments were calculated. Data were performed as relative fluorescence units (RFU) obtained from the disks incubated with C. albicans and AC-PACs, and compared to control disks incubated with C. albicans alone. *, significantly lower than the value for the control (P < 0.05).
Figure 6
Figure 6
Effect of AC-PACs on IL-6 and IL-8 secretion by C. albicans (MOI of 3 or 15)-stimulated oral epithelial cells. IL-6 (Panel A) and IL-8 (Panel B) concentrations in the cell-free supernatants were determined by ELISA. Assays were run in triplicate, and the means ± SD from three independent assays were calculated. *, significantly higher than the value for the unstimulated (C. albicans) control (P < 0.05); †, significantly lower than the value for the untreated (AC-PACs) control (P < 0.05).
Figure 7
Figure 7
Effect of AC-PACs on NF-κB p65 activation by C. albicans (MOI of 15)-stimulated oral epithelial cells. The activation of NF-κB p65 in the cellular extract was determined by an ELISA-based assay. Assays were run in triplicate, and the means ± SD from three independent assays were calculated. *, significantly higher than the value for the unstimulated (C. albicans) control (P < 0.05); †, significantly lower than the value for the untreated (AC-PACs) control (P < 0.05).
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