Evaluation of caries-associated virulence of biofilms from Candida albicans isolated from saliva of pediatric patients with sickle-cell anemia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito do óleo essencial de Melaleuca alternifolia e de seu principal componente Terpinen-4-ol sobre isolados clínicos de Candida albicans resistentes

Pós-graduação em Ciências Odontológicas - FOAR

Biofilms of Candida albicans serotypes A and B differ in their sensitivity to photodynamic therapy

Candida albicans is classified into different serotypes according to cell wall mannan composition and cell surface hydrophobicity. Since the effectiveness of photodynamic therapy (PDT) depends on the cell wall structure of microorganisms, the objective of this study was to compare the sensitivity of in vitro biofilms of C. albicans serotypes A and B to antimicrobial PDT. Reference strains of C. albicans serotype A (ATCC 36801) and serotype B (ATCC 36802) were used for the assays. A gallium-aluminum-arsenide laser (660 nm) was used as the light source and methylene blue (300 mu M) as the photosensitizer. After biofilm formation on the bottom of a 96-well microplate for 48 h, each Candida strain was submitted to assays: PDT consisting of laser and photosensitizer application (L + P+), laser application alone (L + P-), photosensitizer application alone (L-P+), and application of saline as control (L-P-). After treatment, biofilm cells were scraped off and transferred to tubes containing PBS. The content of the tubes was homogenized, diluted, and seeded onto Sabouraud agar plates to determine the number of colony-forming units (CFU/mL). The results were compared by analysis of variance and Tukey test (p < 0.05). The two strains studied were sensitive to PDT (L + P+), with a log reduction of 0.49 for serotype A and of 2.34 for serotype B. Laser application alone only reduced serotype B cells (0.53 log), and the use of the photosensitizer alone had no effect on the strains tested. It can be concluded that in vitro biofilms of C. albicans serotype B were more sensitive to PDT.

SURFACE ROUGHNESS AND CANDIDA ALBICANS BIOFILM FORMATION ON A RELINE RESIN AFTER LONG-TERM CHEMICAL DISINFECTION AND TOOTHBRUSHING

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antimicrobial photodynamic therapy in rat experimental candidiasis: evaluation of pathogenicity factors of Candida albicans

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise do tempo de ação de medicações intracanais sobre candida albicans, enterococcus faecalis, escherichia cole em canais radiculares

The purpose of this study was to evaluate in vitro the antimicrobial activity of intracanal medications against Candida albicans, Enterococcus faecalis, Escherichia coli present in root canals. It was used 24 single root human teeths, contaminated for 28 days and prepared with physiological saline solution as irrigation solution. The teeth were divided into 2 groups according to the intracanal medication used: 1) 2% gel chlorhexidine, 2) sterile and pyrogen free physiological saline solution. Samples were taken of the root canals immediately after instrumentation, 7 days after intracanal medication and 7 days after removal of intracanal medication. For all samples the antimicrobial activity was performed by plating method. All results were submitted to Mann-Whitney and Dunn's test with significance of 5%. There was significant reduction of microorganisms after instrumentation and the intracanal medication of 2% gel chlorhexidine completely eliminated C. albicans and E. coli, and significantly reduced E. faecalis. It was concluded that 2% gel chlorhexidine as intracanal medication for 7 days was effective on microorganisms

Candida albicans: genotyping methods and clade related phenotypic characteristics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Composition and antifungal activity against Candida albicans, C. parapsilopisis, Candida krusei and Cryptococcus neoformans of essential oils from leaves of Piper and Peperomia species

This study was addressed to investigate the composition and antifungal activity of essential oils from leaves of Piperaceae species (Piper aduncum, Piper amalago, Piper cernuum, Piper diospyrifolium, Piper crassinervium, Piper gaudichaudianum, Piper solmsianum, Piper regnellii, Piper tuberculatum, Piper umbelata and Peperomia obtusifolia) against Candida albicans, C. parapsilosis, C. krusei and Cryptococcus neoformans. The essential oils from P. aduncum, P. gaudichaudianum and P. solmsianum showed the highest antifungal activity against Cryptococcus neoformans with the MIC of 62.5, 62.5 and 3.9 mg.mL-1, respectively. The oil from P. gaudichaudianum showed activity against C. krusei with MIC of 31.25 mg.mL-1.

Antifungal activity of extracts and isolated compounds from Buchenavia tomentosa on Candida albicans and non-albicans

Aim: This study aimed to evaluate the antifungal activity of Buchenavia tomentosa extract and bioactive compounds on six Candida species. Materials & methods: The antimicrobial activity of extract was evaluated using standard strains and clinical isolates. Cytotoxicity was tested in order to evaluate cell damage caused by the extract. Extract was chemically characterized and the antifungal activity of its compounds was evaluated. Results: Extract showed antifungal activity on Candida species. Candida non-albicans were more susceptible than Candida albicans. Low cytotoxicity for extract was observed. The isolated compounds presented antifungal activity at least against one Candida spp. and all compounds presented antifungal effect on Candida glabrata. Conclusion: Extracts from Buchenavia tomentosa showed promising antifungal activity on Candida species with low cytotoxicity. Gallic acid, corilagin and ellagic acid showed promising inhibitory activity on Candida glabrata.

Dynamics of biofilm formation and the interaction between candida albicans and methicillin-susceptible (MSSA) and -resistant staphylococcus aureus (MRSA)

Coordenação de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)

Genotoxic effect of photodynamic therapy mediated by curcumin on Candida albicans

Photodynamic therapy (PDT) is a promising method for localized and specific inactivation of fungi and bacteria. A nontoxic light-sensitive compound is taken up by cells, which are then exposed selectively to light, which activates toxicity of the compound. We investigated the potential of sublethal PDT using light-sensitive curcumin (CUR) in combination with blue (455 nm) light to promote reactive oxygen species (ROS) formation in the form of singlet oxygen and DNA damage of Candida albicans. Surprisingly, CUR-mediated PDT but also light alone caused significantly longer comet tails, an indication of DNA damage of C. albicans when compared with the negative control. The intracellular ROS production was also significantly higher for the group treated only with light. However, PDT compared to blue light alone significantly slowed DNA repair. Comet tails decreased during 30 min visualized as a 90% reduction in length in the absence of light for cells treated with light alone, while comet tails of cells treated with PDT only diminished in size about 45%. These results indicate that complex mechanisms may result in PDT in a way that should be considered when choosing the photosensitive compound and other aspects of the treatment design.

Effect of human whole saliva on the in vitro adhesion of Candida albicans to a denture base acrylic resin: a focus on collection and preparation of saliva samples

Objective In studies on Candida albicans adhesion to surfaces, diverse protocols have been used for collection and preparation of saliva samples. Thus, this study investigated whether variations in the centrifugation parameters and number of donors of saliva would influence the adhesion of C. albicans to a denture base resin. Methods Resin acrylic samples (n = 72) were made and then divided into four groups: (a) control – specimens were left without preconditioning in saliva; (b) three experimental groups, in which the specimens were preconditioned with saliva collected from 15 volunteers and centrifuged at 12 000 g for 5 min (G1); from 15 volunteers and centrifuged at 18 000 g for 30 min (G2); and from one volunteer and centrifuged at 12 000 g for 5 min (G3). Candida adhesion was evaluated by both the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction method and crystal violet staining. Data were analyzed by one-way analyses of variance (P = 0.05). Results For XTT reduction assay, groups G2, G3, and control were not significantly different, whereas group G1 showed significantly higher absorbance value than control. For crystal violet staining there were no significant differences among all groups. Conclusion Variations in the centrifugation parameters and number of donors of saliva may influence C. albicans adhesion to denture base resins.

Effect of surface roughness on the hydrophobicity of a denture-base acrylic resin and Candida albicans colonization

Aim The aim of the present study was to evaluate the effect of surface roughness (roughness average [Ra] μm) on the hydrophobicity of a denture-base acrylic resin and the initial adherence and biofilm formation of Candida albicans (C. albicans). Methods Disk-shaped specimens were divided into six groups: Ra 0.05, Ra 0.2, Ra 0.4, Ra 0.8, Ra 1.5, and Ra 3.0. Water contact angles (WCA) were measured, and the specimens incubated with C. albicans for 90 min (initial adherence, n = 108) or 48 h (biofilm formation, n = 108). Adhered and biofilm cells were evaluated by c.f.u./mL and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), and the correlation between the two methods was evaluated. The surface of the specimens and cells (adhered and biofilm) were also analyzed by scanning electron microscopy (SEM). Results Groups Ra 0.05 and 3.0 exhibited the lowest (~75°) and the highest (~100°) WCA mean values, respectively. For both initial adherence and biofilm formation, no statistically-significant differences were observed among all groups, as determined by c.f.u./mL and XTT. A positive correlation between these two methods was found. SEM analysis showed the presence of scratches and valleys on the acrylic specimens and densely-packed yeast cells covering the entire surface. Conclusions Roughness significantly increased hydrophobicity (WCA), but had no effect on the number and metabolic activity of adherent and biofilm cells of C. albicans.

Atividade antimicrobiana de enxaguatórios bucais sem álcool à base de clorexidina sobre candida albicans

The aim of this study was to evaluate the antimicrobial activity of alcohol-free mouthwashes on Candida albicans. Twenty clinical isolates of C. albicans and one reference strain (ATCC 18804) were evaluated after exposure to two 0.12% chlorhexidine-based and alcohol-free (“Ca” and “Or”) in comparison to gluconate chlorhexidine with ethanol (positive control). The maximum inhibitory dilution (MID) and maximum fugal dilution (MFD) were determined by the microdilution method. Twelve serial dilutions (from 50 to 0.02%) were prepared in duplicate. Then, 100 µL of C. albicans suspension (106 cells.mL–1) were added to the wells. After incubation (37 °C/24 hours), MID was determined by reading the optical density. For MFD determination, the content of the wells were plated on Saouraud agar. For MID, there were no differences between groups Or and control, but Ca group showed a MID statistically higher (Kruskal-Wallis, p = 0.0012). For MFD, there were no differences between Ca and control (Mann-Whitney test, p = 0.1631). It can be concluded that Ca group showed a fungicid activity against C. albicans similar to the control, but lower fungistatic activity when compared to the control. Group Or showed only a fungistatic action similar to control.