Artículos de revistas
Exopolysaccharide Matrix Of Developed Candida Albicans Biofilms After Exposure To Antifungal Agents
Registro en:
Brazilian Dental Journal. , v. 23, n. 6, p. 716 - 722, 2012.
1036440
10.1590/S0103-64402012000600016
2-s2.0-84872978033
Autor
Da Silva W.J.
Goncalves L.M.
Seneviratne J.
Parahitiyawa N.
Samaranayake L.P.
Del Bel Cury A.A.
Institución
Resumen
This study aimed to evaluate the effects of fluconazole or nystatin exposure on developed Candida albicans biofilms regarding their exopolysaccharide matrix. The minimal inhibitory concentration (MIC) against fluconazole or nystatin was determined for C. albicans reference strain (ATCC 90028). Poly(methlymethacrylate) resin (PMMA) specimens were fabricated according to the manufacturer's instructions and had their surface roughness measured. Biofilms were developed on specimens surfaces for 48 h and after that were exposed during 24 h to fluconazole or nystatin prepared in a medium at MIC, 10 x MIC or 100 x MIC. Metabolic activity was evaluated using an XTT assay. Production of soluble and insoluble exopolysaccharide and intracellular polysaccharides was evaluated by the phenol-sulfuric method. Confocal laser scanning microscope was used to evaluate biofilm architecture and percentage of dead/live cells. Data were analyzed statistically by ANOVA and Tukey's test at 5% significance level. The presence of fluconazole or nystatin at concentrations higher than MIC results in a great reduction of metabolic activity (p<0.001). At MIC or 10 x MIC, fluconazole showed high amounts of intracellular polysaccharides (p<0.05), but did not affect the exopolysaccharide matrix (p>0.05). The exposure to nystatin also did not alter the exopolysaccharide matrix at all the tested concentrations (p>0.05). Biofilm architecture was not affected by either of the antifungal agents (p>0.05). Nystatin promoted higher proportion of dead cells (p<0.05). It may be concluded that fluconazole and nystatin above the MIC concentration reduced the metabolic activity of C. albicans biofilms; however, they were not able to alter the exopolysaccharide matrix and biofilm architecture. 23 6 716 722 Gendreau, L., Loewy, Z.G., Epidemiology and etiology of denture stomatitis (2011) J Prosthodont, 20, pp. 251-260 Tobudic, S., Kratzer, C., Lassnigg, A., Presterl, E., Antifungal susceptibility of Candida albicans in biofilms (2012) Mycoses, 55, pp. 199-204 Chaffin, W.L., Candida albicans cell wall proteins (2008) Microbiol Mol Biol Rev, 72, pp. 495-544 Seneviratne, C.J., Jin, L., Samaranayake, L.P., Biofilm lifestyle of Candida: A mini review (2008) Oral Dis, 14, pp. 582-590 Ramage, G., Rajendran, R., Sherry, L., Williams, C., Fungal biofilm resistance (2012) Int J Microbiol, , 528-521 Chandra, J., Kuhn, D.M., Mukherjee, P.K., Hoyer, L.L., McCormick, T., Ghannoum, M.A., Biofilm formation by the fungal pathogen Candida albicans: Development, architecture, and drug resistance (2001) J Bacteriol, 183, pp. 5385-5394 Konopka, K., Dorocka-Bobkowska, B., Gebremedhin, S., Duzgunes, N., Susceptibility of Candida biofilms to histatin 5 and fluconazole (2010) Antonie Van Leeuwenhoek, 97, pp. 413-417 Baillie, G.S., Douglas, L.J., Effect of growth rate on resistance of Candida albicans biofilms to antifungal agents (1998) Antimicrob Agents Chemother, 42, pp. 1900-1905 Niimi, M., Firth, N.A., Cannon, R.D., Antifungal drug resistance of oral fungi (2010) Odontology, 98, pp. 15-25 Ramage, G., Mowat, E., Jones, B., Williams, C., Lopez-Ribot, J., Our current understanding of fungal biofilms (2009) Crit Rev Microbiol, 35, pp. 340-355 Baillie, G.S., Douglas, L.J., Matrix polymers of Candida biofilms and their possible role in biofilm resistance to antifungal agents (2000) J Antimicrob Chemother, 46, pp. 397-403 Seneviratne, C.J., Jin, L.J., Samaranayake, Y.H., Samaranayake, L.P., Cell density and cell aging as factors modulating antifungal resistance of Candida albicans biofilms (2008) Antimicrob Agents Chemother, 52, pp. 3259-3266 da Silva, W.J., Seneviratne, J., Samaranayake, L.P., Del Bel Cury, A.A., Bioactivity and architecture of Candida albicans biofilms developed on poly (methyl methacrylate) resin surface (2010) J Biomed Mater Res B Appl Biomater, 94, pp. 149-156 (2008) Reference Method For Broth Dilution Antifungal Susceptibility Testing of Yeasts, Approved Standard, , CLSI, CLSI Document M27-A3, Wayne, PA:CLSI da Silva, W.J., Seneviratne, J., Parahitiyawa, N., Rosa, E.A., Samaranayake, L.P., Del Bel Cury, A.A., Improvement of XTT assay performance for studies involving Candida albicans biofilms (2008) Braz Dent J, 19, pp. 364-369 Tenuta, L.M., Ricomini, F.A.P., Del Bel Cury, A.A., Cury, J.A., Effect of sucrose on the selection of mutans streptococci and lactobacilli in dental biofilm formed in situ (2006) Caries Res, 40, pp. 546-549 Dubois, M., Gilles, K., Hamilton, J.K., Rebers, P.A., Smith, F., A colorimetric method for the determination of sugars (1951) Nature, 28, p. 167. , 168 Heydorn, A., Nielsen, A.T., Hentzer, M., Sternberg, C., Givskov, M., Ersboll, B.K., Quantification of biofilm structures by the novel computer program COMSTAT (2000) Microbiology, 146, pp. 2395-2407 Force, R.W., Nahata, M.C., Salivary concentrations of ketoconazole and fluconazole: Implications for drug efficacy in oropharyngeal and esophageal candidiasis (1995) Ann Pharmacother, 29, pp. 10-15 Gomes, P.N., da Silva, W.J., Pousa, C.C., Narvaes, E.A., Del Bel Cury, A.A., Bioactivity and cellular structure of Candida albicans and Candida glabrata biofilms grown in the presence of fluconazole (2011) Arch Oral Biol, 56, pp. 1274-1281 Ellepola, A.N., Samaranayake, L.P., Oral candidal infections and antimycotics (2000) Crit Rev Oral Biol Med, 11, pp. 172-198 Ellepola, A.N., Samaranayake, L.P., The postantifungal effect (PAFE) of antimycotics on oral C. albicans isolates and its impact on candidal adhesion (1998) Oral Dis, 4, pp. 260-267 Nett, J.E., Sanchez, H., Cain, M.T., Andes, D.R., Genetic basis of Candida biofilm resistance due to drug-sequestering matrix glucan (2010) J Infect Dis, 202, pp. 171-175 Mores, A.U., Souza, R.D., Cavalca, L., de Paula e Carvalho, A., Gursky, L.C., Rosa, R.T., Enhancement of secretory aspartyl protease production in biofilms of Candida albicans exposed to subinhibitory concentrations of fluconazole (2011) Mycoses, 54, pp. 195-201 Goncalves, L.M., Del Bel Cury, A.A., Sartoratto, A., Garcia, R.V.L., Silva, W.J., Effects of undecylenic acid released from denture liner on Candida biofilms (2012) J Dent Res, 91, pp. 985-989