Non-steroidal anti-inflammatory drugs may modulate the protease activity of Candida albicans Alessandra P. Carvalho a , Lauren C. Gursky a , Rosimeire T. Rosa a , Alinne U.M. Rymovicz a , Patrícia M.S. Campelo a , Ana Maria T. Grégio a , Cristiane Y. Koga-Ito b , Lakshman P. Samaranayake c , Edvaldo A.R. Rosa a, * a Xenobiotics Research Unity, Biological and Health Sciences Centre, The Pontifical Catholic University of Paraná, Rua Imaculada Conceição 1155, Curitiba, Brazil b Laboratory of Microbiology, Faculty of Dentistry, The State University of São Paulo, Avenida Eng Francisco José Longo 777, São José dos Campos, Brazil c Oral Biosciences Unit, Faculty of Dentistry, The University of Hong Kong, 5F/34 Hospital Road, Hong Kong, HKSAR, China article info Article history: Received 21 May 2010 Received in revised form 22 July 2010 Accepted 26 July 2010 Available online 12 August 2010 Keywords: Candida albicans NSAID Protease Biofilm Normoxia Anoxia abstract The phenotypic pressure exerted by non-steroidal anti-inflammatory drugs (NSAIDs) on autochthonous and pathogenic microbiota remains sparsely known. In this study, we investigated if some NSAIDs increment or diminish the secretion of aspartyl-proteases (Sap) by Candida albicans grown under different phenotypes and oxygen availability using a set of SAP knock-out mutants and other set for genes (EFG1 and CPH1) that codify transcription factors involved in filamentation and protease secretion. Pre- conditioned cells were grown under planktonic and biofilm phenotypes, in normoxia and anoxia, in the presence of plasma concentrations of acetylsalicylic acid, diclofenac, indomethacin, nimesulide, pirox- icam, ibuprofen, and acetaminophen. For diclofenac, indomethacin, nimesulide, and piroxicam the secretion rates of Sap by SAP1-6, EFG1 , and CPH1 mutants were similar or, even, inferior to parental wild- type strain. This suggests that neither Sap 1e6 isoenzymes nor Efg1/Cph1 pathways may be entirely responsible for protease release when exposed to these NSAIDs. Ibuprofen and acetaminophen enhanced Sap secretion rates in three environmental conditions (normoxic biofilm, normoxic planktonic and anoxic planktonic). In other hand, aspirin seems to reduce the Sap-related pathogenic behavior of can- didal biofilms. Modulation of Sap activity may occur according to candidal phenotypic state, oxygen availability, and type of NSAID to which the cells are exposed. Ó 2010 Elsevier Ltd. All rights reserved. 1. Introduction The contribution of non-steroidal anti-inflammatory drugs (NSAIDs) for the control of pain and other inflammation-related upsets is unquestionable. However, whereas some side effects are well documented, others are completely unknown. For instance, there is a large gap on the knowledge regarding to the effects of such drugs on autochthonous and pathogenic microbiota. Among the few studies that investigated such feature, Alem and Douglas [1] showed that some cyclooxygenase inhibitors, in special, aspirin (acetylsalicylic acid) may exert some desirable inhibitory effects on Candida albicans biofilms. Also, it was demonstrated that indomethacin and piroxicam reduce the production of prosta- glandin E2 (PGE2) by Paracoccidioides brasiliensis, leading to a diminished fungal growth [2]. The studies above have only focused the role of NSAIDs on fungal growth inhibitory effect. Our and few others have found that different classes of xenobiotics may modulate candidal virulence [3e6]; however, to our knowledge no other previous study had prospected the NSAIDs effects on the constitutive virulence attri- butes of C. albicans, as exodepolymerases and hemolysins. As such class of therapeutic drugs is wide-world consumed it is important to state if some side effects inherent to microbial features are prompt to occur when they are in use. The aim of this study was to investigate if some NSAIDs incre- ment or diminish the secretion of aspartyl-proteases (Sap) by C. albicans grown in planktonic and biofilm phenotypes under nor- moxia and anoxia. Also we employed one set of SAP knock-out mutants and other set for genes that codify transcription factors involved in filamentation and protease secretion processes. * Corresponding author. The Pontifical Catholic University of Paraná, Biological and Health Sciences Centre, Faculty of Dentistry, Rua Imaculada Conceição 1155, 80215-901 Curitiba, PR, Brazil. Tel.: þ55 41 3271 1497; fax: þ55 41 3271 1405. E-mail address: edvaldo.rosa@pucpr.br (E.A.R. Rosa). Contents lists available at ScienceDirect Microbial Pathogenesis journal homepage: www.elsevier.com/locate/micpath 0882-4010/$ e see front matter Ó 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.micpath.2010.07.007 Microbial Pathogenesis 49 (2010) 315e322