June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Efficacy of Amphotericin B in Preventing Fungal Growth in Corneal Preservation Media After Frozen Storage
Author Affiliations & Notes
  • Doowon Huh
    Weill Cornell Medical College, New York, New York, United States
  • Khoa D. Tran
    Lions VisionGift, Oregon, United States
  • Megan Straiko
    Lions VisionGift, Oregon, United States
  • Christopher Sales
    Weill Cornell Medical College, New York, New York, United States
  • Footnotes
    Commercial Relationships   Doowon Huh, None; Khoa Tran, None; Megan Straiko, None; Christopher Sales, None
  • Footnotes
    Support  EBAA Richard Lindstrom Research Grant
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 418. doi:
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      Doowon Huh, Khoa D. Tran, Megan Straiko, Christopher Sales; Efficacy of Amphotericin B in Preventing Fungal Growth in Corneal Preservation Media After Frozen Storage. Invest. Ophthalmol. Vis. Sci. 2020;61(7):418.

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

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Abstract

Purpose : Antifungals are not used in the preservation of donor corneas despite the serious complications of a postkeratoplasty fungal infection. This study investigates the efficacy of amphotericin B supplementation following long term frozen storage in reducing growth of Candida albicans for future incorporation into corneal tissue preservation by eye banks.

Methods : Triplicate Optisol-GS samples were inoculated with 106 colony-forming units (CFUs) of C. albicans. Each set of triplicate cultures were supplemented with 2.5 µg/ml of amphotericin B that was either freshly resuspended and never frozen, frozen overnight at -20 °C and thawed, or frozen at -20 °C for four weeks and thawed. These cultures were stored at 4 °C with aliquots taken at 0, 6, 24, and 72 hours for quantification of CFUs. The efficacy of each preparation of amphotericin B in reducing C. albicans growth was assessed at different time points for potential future use at eye banks.

Results : A 1.33 log10 CFU reduction resulted after 6 hours of supplementation with 2.5 µg/ml of amphotericin B that was freshly resuspended. Comparatively, a 1.31 and 1.18 log10 CFU reduction resulted with amphotericin B frozen overnight and thawed or frozen for four weeks and thawed, respectively. At 72 hours, fresh amphotericin B and overnight freeze-thawed amphotericin B showed a 2.72 and 2.64 log10 CFU reduction, respectively. Amphotericin B that had been frozen for four weeks demonstrated a 2.18 log10 CFU reduction at this same time point.

Conclusions : Our results show that a 2.5 µg/ml supplement of amphotericin B frozen for four weeks at -20 °C storage leads to > 90% reduction in growth of C. albicans by 6 hours and > 99% reduction in growth by 72 hours. Supplementation of amphotericin B frozen for long periods of time maintain high efficacy in reducing fungal growth in Optisol-GS media.

This is a 2020 ARVO Annual Meeting abstract.

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