title: Addition of DNase improves the in vitro activity of antifungal drugs against Candida albicans biofilms.
abstract: Cells within Candida albicans biofilms display decreased susceptibility to most clinically used antifungal agents. We recently demonstrated that extracellular DNA (eDNA) plays an important role in biofilm integrity, as a component of the biofilm matrix. This study aimed at gaining insights into the contributions of eDNA to C. albicans biofilms antifungal susceptibility by the investigation of the impact of the combined use of deoxyribonuclease I (DNase) and antifungals to treat biofilms. Candida albicans biofilms were formed using a simple and reproducible 96-well plate-based method. The activity of the combined use of 0.13 mg l(-1) DNase and antifungals was estimated using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay and total viable counts. Herein, we report the improved efficacy of amphotericin B when in combination with DNase against C. albicans biofilms, as assessed using XTT readings and viable counts. Furthermore, although DNase increased the efficacy of caspofungin in the reduction of mitochondrial activity, no changes were observed in terms of culturable cells. Deoxyribonuclease I did not affect biofilm cells susceptibility to fluconazole. This work suggests that agents that target processes affecting the biofilm structural integrity may have potential use as adjuvants of a catheter-lock therapy.
authors.affiliation: IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Braga, Portugal.
authors.forename: Margarida
authors.initials: M
authors.lastname: Martins
authors.forename: Mariana
authors.initials: M
authors.lastname: Henriques
authors.forename: José L
authors.initials: JL
authors.lastname: Lopez-Ribot
authors.forename: Rosário
authors.initials: R
authors.lastname: Oliveira
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects*; D002176:Candida albicans / Q000187:drug effects* / Q000502:physiology; D000077336:Caspofungin; D003851:Deoxyribonucleases / Q000494:pharmacology*; D004338:Drug Combinations; D054714:Echinocandins / Q000494:pharmacology; D015725:Fluconazole / Q000494:pharmacology; D055666:Lipopeptides; D008826:Microbial Sensitivity Tests / Q000379:methods*; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D015203:Reproducibility of Results; D013778:Tetrazolium Salts / Q000737:chemistry
chemical_list: D000935:Antifungal Agents; D004338:Drug Combinations; D054714:Echinocandins; D055666:Lipopeptides; D013778:Tetrazolium Salts; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; D000666:Amphotericin B; D015725:Fluconazole; D003851:Deoxyribonucleases; D000077336:Caspofungin