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A more recent version of this article appeared on August 1, 2008
 (Investigative Ophthalmology and Visual Science. )
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-1592
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Article

Antimicrobial Efficacy of Riboflavin/UVA Combination (365 nm) in Vitro for Bacterial and Fungal Isolates: a Potential New Treatment for Infectious Keratitis

Suy Anne Reboucas Martins 1*, Juan Castro-Combs 1, Guillermo Noguera 1, Walter Camacho 1, Priscila Wittmann 1, Rhonda Walther 1, Marisol del Valle Cano 1, James Dick 1, and Ashley Behrens 1

1 The Wilmer Ophthalmological Institute, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States

* To whom correspondence should be addressed. E-mail: suyreboucas{at}gmail.com.


  Abstract

Purpose: To demonstrate antimicrobial properties of Riboflavin/UVA (365 nm) against common pathogens. Methods: One group of bacteria [P. aeruginosa (PA), S. aureus (SA), S. epidermidis (SE)] was tested using Kirby-Bauer disks with: a) empty disc (Control - C); b) Riboflavin 0.1% (B2); c) riboflavin 0.1% previously activated by UVA (B2’); d) UVA alone (UVA); e) group b + additional UVA exposure (UVA+B2), and f) group c + additional UVA exposure (UVA+B2’). Additionally, another group of microbes was tested using the same approach: methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Pseudomonas aeruginosa (MDRPA), drug resistant Streptococcus pneumoniae (DRSP), and Candida albicans (CA). Mean growth inhibition zone (GIZ) in mm2 was measured around disks. The Mean Standard Deviation (MSD) was calculated to be 3.65 when {alpha}=0.01. If Mean Deviation (MD) > MSD, this indicates a significant difference. Results: In the first group, the GIZ was significantly greater after UVA (MD=14.30), UVA+B2 (MD=39.61), and UVA+B2’ (MD=40.45) when compared to C, B2 and B2’. UVA alone was less effective than UVA+B2 (MD=25.31) and UVA+B2’ (26.15). The second group demonstrated increased GIZ in UVA (MD=6.98), UVA+B2 (MD=17.80), and UVA+B2’ (MD=21.15) when compared to C, B2 and B2’. UVA alone was less effective against the second group of bacteria than UVA+B2 (MD=10.82) and UVA+B2’ (MD=14.17). CA did not show any GIZ after treatment. Conclusion: Riboflavin/UVA treatment was effective against SA, SE, PA, MRSA, MDRPA, and DRSP, but ineffective on CA. This opens a potential new treatment for microbial keratitis in the future.

Key Words: cornea, infection, ultraviolet rays, riboflavin, disk diffusion antimicrobial test






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