September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Reducing the concentration and irradiation time of rose bengal-mediated photodynamic antimicrobial therapy for inhibition of fungal keratitis isolates
Author Affiliations & Notes
  • Alejandro Arboleda
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Nidhi Relhan
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Heather Ann Durkee
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Mariela C Aguilar
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Karam AlRahman Alawa
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Francisco Halili
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Cornelis Rowaan
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Guillermo Amescua
    Anne Bates Leach Eye Hospital, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Harry W Flynn
    Anne Bates Leach Eye Hospital, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Darlene Miller
    Ocular Microbiology Laboratory, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
  • Jean-Marie A Parel
    Ophthalmic Biophysics Center, Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, United States
    CHU Sart-Tillman, Department of Ophthalmology, University of Liege, Liege, Belgium
  • Footnotes
    Commercial Relationships   Alejandro Arboleda, None; Nidhi Relhan, None; Heather Durkee, None; Mariela Aguilar, None; Karam Alawa, None; Francisco Halili, None; Cornelis Rowaan, None; Guillermo Amescua, None; Harry Flynn, None; Darlene Miller, None; Jean-Marie Parel, None
  • Footnotes
    Support  This work was supported in part by: Florida Lions Eye Bank, Edward D. and Janet K. Robson Foundation. Drs. KR Olsen and ME Hildebrandt, NIH center grant P30EY14801, Drs. Raksha Urs and Aaron Furtado, Research to Prevent Blindness, Brien Holden Vision Institute, Henri and Flore Lesieur Foundation (JMP).Technical support provided by: Alex Gonzalez, Juan Silgado, Victor Hernandez, Carolina de Freitas, and Adriana Henao-Pink.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 2342. doi:
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    • Get Citation

      Alejandro Arboleda, Nidhi Relhan, Heather Ann Durkee, Mariela C Aguilar, Karam AlRahman Alawa, Francisco Halili, Cornelis Rowaan, Guillermo Amescua, Harry W Flynn, Darlene Miller, Jean-Marie A Parel; Reducing the concentration and irradiation time of rose bengal-mediated photodynamic antimicrobial therapy for inhibition of fungal keratitis isolates. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2342.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Fungal keratitis is a potentially blinding condition that is difficult to manage clinically with the currently available antimycotics. One emerging alternate treatment is photodynamic antimicrobial therapy (PDAT). In this study we explore the efficacy of different parameters for rose bengal-mediated PDAT to inhibit growth of four fungal isolates from patients with infectious keratitis.

Methods : Four fungi (Fusarium solani, Candida albicans, Purpureocillium lilacinum (Paecilomyces lilacinus), and Pseudoallescheria boydii) were isolated from patients with confirmed fungal keratitis and grown on Sabouraud-Dextrose agar plates. A suspension of each isolate was made using sterile, deionized water and was subsequently mixed with appropriate photosensitizer solution according to experimental group and plated in triplicate onto nutrient agar. Water was used as a control to compare to experimental groups treated with rose bengal at varying concentrations (0.1%, 0.05%, & 0.01%) and irradiated for 5.4 J/cm2 total exposure. An additional test was performed with 0.1% rose bengal irradiated for a half the time resulting in a reduced exposure of 2.7 J/cm2. Green light irradiation was performed using a circular array (47 mm diameter) of 518 nm light emitting diodes. Plates were placed in a 30°C non-CO2 incubator and photographed at 72 hours. Fungi growth was evaluated using Labview-based software.

Results : At 72 hours, Rose bengal-mediated PDAT with 0.05% concentration completely inhibited the growth of the four fungal isolates within central 47mm area. 0.01% rose bengal PDAT inhibited for Pseudoallescheria boydii and incompletely for Candida albicans and Purpureocillium lilacinum. All four fungal isolates showed central inhibition zone corresponding to the diameter of the irradiation source head in 0.1% rose bengal reduced time PDAT (2.7 J/cm2).

Conclusions : Rose bengal-mediated PDAT inhibited Fusarium solani, Candida albicans, Purpureocillium lilacinum, and Pseudoallescheria boydii keratitis isolates in the irradiated area. These in vitro results demonstrate the potential strength of rose bengal-mediated PDAT as an alternative treatment modality of fungal keratitis.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

Rose bengal-mediated PDAT using reduced concentrations of photosensitizing agent.

Rose bengal-mediated PDAT using reduced concentrations of photosensitizing agent.

 

Rose bengal-mediated PDAT using 0.1% rose bengal and reduced energy

Rose bengal-mediated PDAT using 0.1% rose bengal and reduced energy

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