Silver Nanoparticle-Embedded Carbon Nitride: Antifungal Activity on Candida albicans and Toxicity toward Animal Cells.

The development of engineered nanomaterials has been considered a promising strategy to control oral infections. In this study, silver-embedded carbon nitrides (Ag@g-CN) were synthesized and tested against Candida albicans, investigating their antifungal action and biocompatibility in animal cells. Ag@g-CN was synthesized by a simple one-pot thermal polymerization technique and characterized by various analytical techniques. X-ray diffraction (XRD) analysis revealed slight alterations in the crystal structure of g-CN upon the incorporation of Ag. Fourier transform infrared (FT-IR) spectroscopy confirmed the presence of Ag-N bonds, indicating successful silver incorporation and potential interactions with g-CN's amino groups. UV-vis spectroscopy demonstrated a red shift in the absorption edge of Ag@g-CN compared with g-CN, attributed to the surface plasmon resonance effect of silver nanoparticles. Field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) confirmed the 2D layered sheet like morphology of both materials. The Ag 3d peaks found in X-ray photoelectron spectroscopy (XPS) confirmed the presence of metallic Ag0 nanoparticles in Ag@g-CN. The Ag@g-CN materials exhibited high antifungal activity against reference and oral clinical strains of C. albicans, with minimal inhibitory concentration (MIC) ranges between 16-256 µg/mL. The mechanism of Ag@g-CN on C. albicans was attributed to the disruption of the membrane integrity and disturbance of the biofilm. In addition, the Ag@g-CN material showed good biocompatibility in the fibroblastic cell line and in Galleria mellonella, with no apparent cytotoxicity observed at a concentration up to 1000 µg/mL. These findings demonstrate the potential of the Ag@g-CN material as an effective and safe antifungal agent for the treatment of oral fungal infections.

Amphotericin B-loaded natural latex dressing for treating Candida albicans wound infections using Galleria mellonella model.

Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL-1) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL-1) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections.

Enhancing effect of chitosan on methylene blue-mediated photodynamic therapy against C. albicans: A study in planktonic growth, biofilms, and persister cells.

Chitosan (CS) is a natural polymer extracted from the exoskeleton of crustaceans. Due to its cationic structure, CS has been studied as a possible enhancer of antimicrobial photodynamic therapy (aPDT). The objective was to evaluate the association of CS with methylene blue (MB)-mediated aPDT on Candida albicans, investigating its effects on planktonic growth, biofilms, and cells persistent to fluconazole. The ability of CS to interfere with MB absorption by Candida cells was also evaluated. For the assays, planktonic cells of C. albicans were cultivated for 24 h, and the biofilms were formed for 48 h. For the induction of persister cells, C. albicans was cultivated with high concentration of fluconazole for 48 h. Treatments were performed with MB, CS or MB+CS, followed by irradiation with LED (660 nm ). As results, aPDT with MB (300 µm) reduced the planktonic cells by 1.6 log10 CFU, while the MB+CS association led to a reduction of 4.8 log10 CFU. For aPDT in biofilms, there was a microbial reduction of 2.9 log10 CFU for the treatment with MB (600 µm) and 5.3 log10 CFU for MB+CS. In relation to persister cells, the fungal reductions were 0.4 log10 CFU for MB and 1.5 log10 CFU for MB+CS. In the absorption assays, the penetration of MB into Candida cells was increased in the presence of CS. It was concluded that CS enhanced the antimicrobial activity of aPDT in planktonic growth, biofilms, and persister cells of C. albicans, probably by facilitating the penetration of MB into fungal cells.

Antimicrobial effects of photodynamic therapy with Fotoenticine on Streptococcus mutans isolated from dental caries.

Photodynamic therapy (PDT) is a promising strategy to control cariogenic pathogens, such as Streptococcus mutans. Seeking to reach the total bacterial elimination from dental surfaces, novel photosensitizers have been investigated, such as Fotoenticine (FTC) derived from chlorin e6. The objective of this study was to investigate the photodynamic effects of FTC against several clinical strains of S. mutans. Clinical isolates were obtained from patients with active carious lesions, identified by molecular analysis and subjected to PDT using laser irradiation (660 nm and 39.5 J/cm2) in planktonic and biofilm stages. We identified 11 S. mutans strains from cervical, occlusal and proximal caries. PDT mediated by FTC has totally eliminated the S. mutans cells in planktonic growth for all analyzed strains. In biofilms, PDT with FTC reached statistically significant reductions compared with the non-treated control group, at 5.4, 5.5 and 6.5 Log10 (CFU/mL), respectively, for the strains from proximal, occlusal and cervical caries. The scanning electron microscopy evaluations confirmed that PDT mediated by FTC was able to disaggregate and kill the S. mutans cells adhered to enamel surface, suggesting its potential to disinfect the dental tissues.

Photodynamic therapy mediated by chlorin-type photosensitizers against Streptococcus mutans biofilms.

Photodynamic therapy (PDT) can be used for the control of oral pathogens and different photosensitizers (PS) have been investigated. This study evaluated the efficacy of PDT against Streptococcus mutans biofilms using two second-generation PS derived from chlorin: Photoditazine® (PDZ) and Fotoenticine® (FTC). These PS were compared to methylene blue (MB), a dye with proven antimicrobial activity against S. mutans. Suspensions of S. mutans were cultured in contact with bovine tooth disks for biofilm formation. After 48 h, the biofilms were treated with PDZ (0.6 mg/mL), FTC (0.6 mg/mL) or MB (1 mg/mL) and submitted to laser irradiation (660 nm, 50 mW/cm2). The biofilms were quantified by the determination of CFU/mL count and analyzed by scanning electron microscopy (SEM). All PS used for PDT reduced the number of S. mutans, with a statistically significant difference compared to the untreated groups. PDT achieved microbial reductions of 4 log with MB and 6 log with PDZ, while the use of FTC resulted in the complete elimination of S. mutans biofilms. SEM analysis confirmed the CFU/mL results, showing that all PS, particularly FTC, were able to detach the biofilms and to eliminate the bacteria. In conclusion, PDT mediated by chlorin-type PS exhibited greater antimicrobial activity against S. mutans than MB-mediated PDT, indicating that these PS can be useful for the control of dental caries.