Abstract
This study aimed to investigate the effect of ozone ultrafine bubble water (OUFBW) on the formation and growth of Candida albicans (C. albicans) biofilms and surface properties of denture base resins. OUFBWs were prepared under concentrations of 6 (OUFBW6), 9 (OUFBW9), and 11 ppm (OUFBW11). Phosphate buffered saline and ozone-free electrolyte aqueous solutions (OFEAS) were used as controls. Acrylic resin discs were made according to manufacturer instructions, and C. albicans was initially cultured on the discs for 1.5 h. A colony forming unit (CFU) assay was performed by soaking the discs in OUFBW for 5 min after forming a 24-h C. albicans biofilm. The discs after initial attachment for 1.5 h were immersed in OUFBW and then cultured for 0, 3, and 5 h. CFUs were subsequently evaluated at each time point. Moreover, a viability assay, scanning electron microscopy (SEM), Alamar Blue assay, and quantitative real-time polymerase chain reaction (qRT-PCR) test were performed. To investigate the long-term effects of OUFBW on acrylic resin surface properties, Vickers hardness (VH) and surface roughness (Ra) were measured. We found that OUFBW9 and OUFBW11 significantly degraded the formed 24-h biofilm. The time point CFU assay showed that C. albicans biofilm formation was significantly inhibited due to OUFBW11 exposure. Interestingly, fluorescence microscopy revealed that almost living cells were observed in all groups. In SEM images, the OUFBW group had lesser number of fungi and the amount of non-three-dimensional biofilm than the control group. In the Alamar Blue assay, OUFBW11 was found to suppress Candida metabolic function. The qRT-PCR test showed that OUFBW down-regulated ALS1 and ALS3 expression regarding cell-cell, cell-material adhesion, and biofilm formation. Additionally, VH and Ra were not significantly different between the two groups. Overall, our data suggest that OUFBW suppressed C. albicans growth and biofilm formation on polymethyl methacrylate without impairing surface properties.
Highlights
Oral candidiasis is one of the most common oral infections in patients receiving immunotherapy and in older individuals with weak immunity [1]
The number of colony forming unit (CFU) was significantly lower in the OUFBW9 and OUFBW11 groups than in the control group (Fig 1)
There were no significant differences in the number of CFUs between the control and ozone-free electrolyte aqueous solutions (OFEAS) groups
Summary
Oral candidiasis is one of the most common oral infections in patients receiving immunotherapy and in older individuals with weak immunity [1]. Oral candidiasis related to wearing dentures is called denture stomatitis (DS) [2, 3]. Candida albicans (C. albicans), a pathogenic fungus abundant in denture plaque, is highly associated with DS [4, 5]. C. albicans on the denture surface adheres to other bacteria and acts as a scaffold for biofilms [6]. The adherence and subsequent increase of C. albicans on the denture surface accelerate biofilm formation [7]. Biofilms have a stronger drug resistance than free-floating microorganisms [8]. Preventing the adherence and increase of C. albicans on dentures and suppressing denture biofilm formation early is the first step in preventing DS
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