Abstract
The purpose of this study was to investigate whether dietary carbohydrates can modulate the development of Candida albicans biofilms on the denture material surface. Poly (methyl methacrylate) acrylic resin discs were fabricated and had their surface roughness measured. Biofilms of C. albicans ATCC 90028 were developed on saliva-coated specimens in culture medium without (control) or with carbohydrate supplementation by starch, starch+sucrose, glucose, or sucrose for 72 h. The cell count, metabolic activity, biovolume, average thickness, and roughness coefficient were evaluated at the adhesion phase (1.5 h) and after 24, 48, and 72 h. The secretion of proteinases and phospholipases, cell surface energy, and production of extra/intracellular polysaccharides were analyzed after 72 h of biofilm development. Data were analyzed by one- and two-way ANOVA followed by Tukey’s test at 5% significance level. In the early stages of colonization (adhesion and 24 h), the glucose group showed the highest cell counts and metabolic activity among the groups (p<0.05). After maturation (48 and 72 h), biofilms exposed to glucose, sucrose, or starch+sucrose showed higher cell counts and metabolic activity than the control and starch groups (p<0.001). Compared to the control group, biofilms developed on starch or starch+sucrose had more proteinase activity (p<0.001), whereas biofilms developed on glucose or sucrose had more phospholipase activity (p<0.05). Exposure to starch+sucrose increased the production of extracellular and intracellular polysaccharides (p<0.05). Biofilms developed on starch or without carbohydrate supplementation presented cells with more hydrophobic behavior compared to the other groups. Confocal images showed hyphae forms on biofilms exposed to starch or starch+sucrose. Within the conditions studied, it can be concluded that dietary carbohydrates can modulate biofilm development on the denture surface by affecting virulence factors and structural features.
Highlights
The availability of nutrient sources plays an important role in the pathogenesis of fungal infections [1,2]
Carbohydrates consumed in the diet are the primary and preferred nutrient sources for Candida albicans [3,4,5], the main pathogen related to Candida-associated denture stomatitis (CADS) [6]
Biofilms developed on starch secreted the highest amount of proteinase (p,0.001), followed by biofilms developed on starch+sucrose (Fig. 1C)
Summary
The availability of nutrient sources plays an important role in the pathogenesis of fungal infections [1,2]. The first step in the dynamics of CADS pathogenesis is the adhesion of C. albicans to the denture surface and epithelial cells, followed by cell multiplication, organization, and secretion of the extracellular matrix (ECM). This process results in the formation of a three-dimensional (3D) structure known as biofilm [9]. The production of organic acids during carbohydrate hydrolysis [13] could activate other virulence factors, such as the yeast-hyphae transition and the secretion of hydrolytic enzymes (e.g., phospholipases and proteases) [10,14]. These virulence factors are mainly related to the invasion of host tissues, which aggravates the inflammatory response of the oral mucosa [15,16]
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