| Summary: | Introduction: The purpose of this study was to assess the antifungal activity of silver nanoparticles (AgNPs) in combination with calcium hydroxide (Ca(OH)<sub>2</sub>) against <i>Candida albicans</i> (<i>C. albicans</i>). Methods: AgNPs was mixed with pure Ca(OH)<sub>2</sub> powder in an aqueous base. A standard suspension (1 × 10<sup>8</sup> bacterial cells/mL) of <i>C. albicans</i> was prepared in a 96-well plate and incubated on shaker at 37 °C in 100% humidity to allow fungal biofilm formation in infected dentin slices (<i>n</i> = 98). The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of AgNPs alone or with Ca(OH)<sub>2</sub> were determined. The samples were separately placed in 24-well tissue culture plates and divided into three experimental groups (0.03, 0.04, and 0.06) and three control groups; negative (saline) and positive chlorhexidine gel and Ca(OH)<sub>2</sub>. Quantitative measurements of fungal activity by XTT colorimetric assay and qualitative measurements using confocal laser microscopy and scanning electron microscopy were performed. Results: The cell viability of <i>C. albicans</i> in the experimental groups was significantly reduced compared to the negative control group. The combination of (AgNPs (0.04%) and Ca(OH)<sub>2</sub>) was the most potent against <i>C. albicans</i>. Conclusions: The findings demonstrated that combining silver nanoparticles with Ca(OH)<sub>2</sub> was more effective against <i>C. albicans</i> biofilm compared to Ca(OH)<sub>2</sub> alone, suggesting a combing effect.
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