Antifungal activity of polymethyl methacrylate/Si₃N₄ composites against Candida albicans.

Abstract

Previous studies using gram-positive and -negative bacteria demonstrated that hydrolysis of silicon nitride (Si₃N₄) in aqueous suspensions elutes nitrogen and produces gaseous ammonia while buffering pH. According to immunochemistry assays, fluorescence imaging, and in situ Raman spectroscopy, we demonstrate here that the antipathogenic surface chemistry of Si₃N₄ can be extended to polymethylmethacrylate (PMMA) by compounding it with a minor fraction (~8 vol.%) of Si₃N₄ particles without any tangible loss in bulk properties. The hydrolytic products, which were eluted from partly exposed Si₃N₄ particles at the composite surface, exhibited fungicidal action against Candida albicans. Using a specific nitrative stress sensing dye and highly resolved fluorescence micrographs, we observed in situ congestion of peroxynitrite (ONOO^-) radicals in the mitochondria of the Candida cells exposed to the PMMA/Si₃N₄ composite, while these radicals were absent in the mitochondria of identical cells exposed to monolithic PMMA. These in situ observations suggest that the surface chemistry of Si₃N₄ mimics the antifungal activity of macrophages, which concurrently produce NO radicals and superoxide anions (O₂•^-) resulting in the formation of candidacidal ONOO^-. The fungicidal properties of PMMA/Si₃N₄ composites could be used in dental appliances to inhibit the uncontrolled growth of Candida albicans and ensuing candidiasis while being synergic with chemoprophylaxis. STATEMENT OF SIGNIFICANCE: In a follow-up of previous studies of gram-positive and gram-negative bacteria, we demonstrate here that the antipathogenic surface chemistry of Si₃N₄ could be extended to polymethylmethacrylate (PMMA) containing a minor fraction (~8 vol.%) of Si₃N₄ particles without tangible loss in bulk properties. Hydrolytic products eluted from Si₃N₄ particles at the composite surface exhibited fungicidal action against Candida albicans. Highly resolved fluorescence microscopy revealed congestion of peroxynitrite (ONOO^-) radicals in the mitochondria of the Candida cells exposed to the PMMA/Si₃N₄ composite, while radicals were absent in the mitochondria of identical cells exposed to monolithic PMMA. The fungicidal properties of PMMA/Si₃N₄ composites could be used in dental appliances to inhibit uncontrolled growth of Candida albicans and ensuing candidiasis in synergy with chemoprophylaxis.