Assessment of fungal viability after long-wave ultraviolet light irradiation combined with riboflavin administration
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Assessment of fungal viability after long-wave ultraviolet light irradiation combined with riboflavin administration Renata T. Kashiwabuchi & Fabio R. S. Carvalho & Yasin A. Khan & Flavio Hirai & Mauro S. Campos & Peter J. McDonnell
Received: 2 October 2012 / Revised: 2 November 2012 / Accepted: 5 November 2012 / Published online: 25 November 2012 # Springer-Verlag Berlin Heidelberg 2012
Abstract Background Corneal collagen cross-linking (CXL), a technique that combines riboflavin administration with longwave ultraviolet light irradiation, was primarily developed to increase the biomechanical strength of collagen fibrils of the cornea to avoid the progression of keratoconus. Recently, this method has been proposed to treat selected cases of infectious keratitis. Methods To test the protocol used for progressive keratoconus in infectious keratitis, Candida albicans, and Fusarium solani, strains were exposed to irradiation using a wavelength of 365 nm at a power density of 3 mW/cm2 for 30 min in the presence of riboflavin photosensitizer. All experiments were performed in triplicate. Qualitative and quantitative measurements of fungal viability used plate cultures and an automated trypan blue dye exclusion method respectively. Fungal cell diameter was also assessed in all groups. Statistical analyses were performed using the triplicate values of each experimental condition. Results Experimental findings of photodynamic therapy applied to the cell inactivation of both yeasts and filamentous All authors have full control of all primary data, and we agree to allow Graefe’s Archive for Clinical and Experimental Ophthalmology to review our data upon request. R. T. Kashiwabuchi : F. R. S. Carvalho : F. Hirai : M. S. Campos Department of Ophthalmology, Paulista School of Medicine – Federal University of Sao Paulo – UNIFESP, Sao Paulo, Brazil Y. A. Khan : P. J. McDonnell The Wilmer Ophthalmological Institute, The Johns Hopkins University, School of Medicine, Baltimore, MD, USA R. T. Kashiwabuchi (*) Rua Botucatu, 820; Vila Clementino, Sao Paulo, SP, Brazil 04023-062 e-mail: [email protected]
fungi were compared with control groups. Qualitative results were corroborated with quantitative findings which showed no statistical significance between challenged samples (experimental groups) and the control group (p-value0 1). In comparison with a control group of live cells, statistical significance was observed when riboflavin solution alone had an effect on the morphologic size of filamentous fungi, while ultraviolet light irradiation alone showed a slight decrease in the cell structure of C. albicans. Conclusions The impact of long-wave ultraviolet combined with riboflavin photosensitizer showed no antifungal effect on C. albicans and F. solani. The significant decrease in cell morphology of both filamentous fungi and yeasts submitted to photosensitizing riboflavin and exposure to ultraviolet light, respectively, may be promising in the development and standardization of alternatives for fungal cell inact
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