title: Glucose modulates antimicrobial photodynamic inactivation of Candida albicans in biofilms.
abstract: Candida albicans biofilm is a main cause of infections associated with medical devices such as catheters, contact lens and artificial joint prosthesis. The current treatment comprises antifungal chemotherapy that presents low success rates. Photodynamic inactivation (PDI) involves the combination of a photosensitizing compound (PS) and light to generate oxidative stress that has demonstrated effective antimicrobial activity against a broad-spectrum of pathogens, including C. albicans. This fungus senses glucose inducing an upregulation of membrane transporters that can facilitate PS uptake into the cell. The aim of this study was to evaluate the effects of glucose on methylene blue (MB) uptake and its influence on PDI efficiency when combined to a red LED with central wavelength at λ=660nm. C. albicans biofilms were grown on hydrogel disks. Prior to PDI assays, MB uptake tests were performed with and without glucose-sensitization. In this system, the optimum PS administration was determined as 500μM of MB in contact with the biofilm during 30min before irradiation. Irradiation was performed during 3, 6, 9, 12, 15 and 18min with irradiance of 127.3mW/cm2. Our results showed that glucose was able to increase MB uptake in C. albicans cells. In addition, PDI without glucose showed a higher viability reduction until 6min; after 9min, glucose group demonstrated a significant decrease in cell viability when compared to glucose-free group. Taken together, our data suggest that glucose is capable to enhance MB uptake and modulate photodynamic inactivation of C. albicans biofilm.
authors.affiliation: Center for Lasers and Applications, IPEN-CNEN/SP, 05508-000, São Paulo, SP, Brazil.
authors.forename: Luis Cláudio
authors.initials: LC
authors.lastname: Suzuki
authors.affiliation: The Engineering, Modelling and Applied Social Science Department, UFABC, 09210-580, Santo André, SP, Brazil.
authors.forename: Ilka Tiemy
authors.initials: IT
authors.lastname: Kato
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, UNINOVE, 01504-001, São Paulo, SP, Brazil; Dentistry School, UNINOVE, 01504-001, São Paulo, SP, Brazil.
authors.forename: Renato Araujo
authors.initials: RA
authors.lastname: Prates
authors.affiliation: Center for Lasers and Applications, IPEN-CNEN/SP, 05508-000, São Paulo, SP, Brazil; Department of Microbiology, ICB/USP, 05508-000, São Paulo, SP, Brazil; Department of Clinical Analysis, FCF/USP, 05508-000, São Paulo, SP, Brazil.
authors.forename: Caetano Padial
authors.initials: CP
authors.lastname: Sabino
authors.affiliation: Center for Lasers and Applications, IPEN-CNEN/SP, 05508-000, São Paulo, SP, Brazil.
authors.forename: Tania Mateus
authors.initials: TM
authors.lastname: Yoshimura
authors.affiliation: Postgraduate Program in Biophotonics Applied to Health Sciences, UNINOVE, 01504-001, São Paulo, SP, Brazil; Dentistry School, UNINOVE, 01504-001, São Paulo, SP, Brazil.
authors.forename: Tamires Oliveira
authors.initials: TO
authors.lastname: Silva
authors.affiliation: Center for Lasers and Applications, IPEN-CNEN/SP, 05508-000, São Paulo, SP, Brazil. Electronic address: marthasr@usp.br.
authors.forename: Martha Simões
authors.initials: MS
authors.lastname: Ribeiro
mesh_terms: D001431:Bacteriological Techniques; D018441:Biofilms / Q000187:drug effects*; D002176:Candida albicans / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D008751:Methylene Blue / Q000493:pharmacokinetics* / Q000494:pharmacology; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000493:pharmacokinetics* / Q000494:pharmacology
chemical_list: D017319:Photosensitizing Agents; D005947:Glucose; D008751:Methylene Blue
keywords: Methylene blue; PDI; Photosensitizer uptake; Red LED; Yeast inactivation