IOVS AJP: Regulatory, Integrative and Comparative Physiology
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Originally published In Press as doi:10.1167/iovs.07-1592 on April 11, 2008
 (Investigative Ophthalmology and Visual Science. 2008;49:3402-3408.)
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.07-1592
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Antimicrobial Efficacy of Riboflavin/UVA Combination (365 nm) In Vitro for Bacterial and Fungal Isolates: A Potential New Treatment for Infectious Keratitis

Suy Anne R. Martins,1 Juan Castro Combs,1 Guillermo Noguera,1 Walter Camacho,1 Priscila Wittmann,1 Rhonda Walther,2 Marisol Cano,1 James Dick,2 and Ashley Behrens1

1From The Wilmer Ophthalmological Institute and the 2Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland.

PURPOSE. To demonstrate the antimicrobial properties of riboflavin/UVA (365 nm) against common pathogens.

METHODS. One group of bacteria (Pseudomonas aeruginosa [PA], Staphylococcus aureus [SA], and Staphylococcus epidermidis [SE]) was tested by using Kirby-Bauer discs with (1) empty disc (Control – C); (2) riboflavin 0.1% (B2); (3) riboflavin 0.1% previously activated by UVA (B2'); (4) UVA alone (UVA); (5) group 2+additional UVA exposure (UVA+B2); and (6) group 3+additional UVA exposure (UVA+B2'). In addition, another group of microbes was tested with the same approach: methicillin-resistant S. aureus (MRSA), multidrug-resistant P. aeruginosa (MDRPA), drug-resistant Streptococcus pneumoniae (DRSP), and Candida albicans (CA). The mean growth inhibition zone (GIZ) in square millimeters was measured around the discs. The mean standard deviation (MSD) was calculated to be 3.65 when {alpha} = 0.01. A mean deviation (MD) > MSD 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 with C, B2, and B2'. UVA alone was less effective than UVA+B2 (MD = 25.31) and UVA+B2' (MD = 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 with C, B2, and B2'. UVA alone was less effective against the second group of bacteria than was UVA+B2 (MD = 10.82) and UVA+B2' (MD = 14.17). CA did not show any GIZ after treatment.

CONCLUSIONS. Riboflavin/UVA was effective against SA, SE, PA, MRSA, MDRPA, and DRSP, but was ineffective on CA and has the potential for use in treatment of microbial keratitis in the future.






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