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Toxicity of Amphotericin B in Rabbit Corneal Epithelial Cells Stored in Optisol™-GS: Corneal Epithelial Cell
Kohei Harada1,2, Hideki Fukuoka1, Yuriko Ban1
1Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Purpose:
To evaluate the toxicity of Amphotericin B (AmB) in Optisol™-GS Corneal Storage Media (Bausch & Lomb) on corneal epithelial cell (CEC) morphology and migration ability.
Methods:
Sclerocorneal strips were removed from male Japanese white rabbits, and then stored at 4 °C in Optisol™-GS containing 0 µg/ml of AmB (control group) and 2.5, 5, 25, and 50 µg/ml of AmB (AmB groups; four eyes per group). After 7 days of storage, CEC morphology was evaluated by hematoxylin-eosin staining, immunohistochemical staining (ZO-1), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Moreover, to evaluate CEC migration ability, three corneal blocks (6-8 × 3 mm each) from one preserved cornea were cultured for 24 h, and the area of CEC migration (2 mm at the central region) onto the stromal surface was then measured.
Results:
At 5, 25, and 50 µg/ml of AmB, deformation and vacuolation of CECs were observed in all preserved corneas. ZO-1 expression was significantly reduced in corneas preserved at AmB concentrations of 25 and 50 µg/ml. TUNEL Labeling Index was significantly increased at AmB concentrations of ≥5 µg/ml. CEC migration was inhibited in a dose-dependent manner at AmB concentrations of 25 and 50 µg/ml compared to the control group.
Conclusions:
The addition of AmB to Optisol™-GS can be toxic to CECs and inhibit their migration at a concentration of ≥5 µg/ml. AmB at a concentration of 2.5 µg/ml can be considered safe for the preservation of donor corneal tissue used in corneal epithelial transplantation surgery.
Insights
Amphotericin B (AmB) in corneal storage media is toxic to corneal epithelial cells (CECs) at concentrations of 5 µg/ml and above. However, AmB at 2.5 µg/ml is safe for preserving donor corneal tissue for transplantation.
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Corneal transplantation is a common procedure for vision restoration.
- Preservation of donor corneal tissue is crucial for successful transplantation.
- Optisol™-GS is a widely used corneal storage medium.
Purpose of the Study:
- To assess the toxicity of Amphotericin B (AmB) in Optisol™-GS corneal storage media.
- To evaluate the impact of AmB on corneal epithelial cell (CEC) morphology and migration.
- To determine safe concentrations of AmB for corneal tissue preservation.
Main Methods:
- Rabbit sclerocorneal strips were stored in Optisol™-GS with varying AmB concentrations (0-50 µg/ml) for 7 days.
- CEC morphology was assessed using hematoxylin-eosin staining, ZO-1 immunohistochemistry, and TUNEL assay.
- CEC migration ability was evaluated by culturing corneal blocks and measuring migration area.
Main Results:
- AmB at 5, 25, and 50 µg/ml caused CEC deformation and vacuolation.
- ZO-1 expression decreased significantly at AmB concentrations of 25 and 50 µg/ml.
- CEC migration was inhibited in a dose-dependent manner at AmB concentrations ≥25 µg/ml, and apoptosis increased at ≥5 µg/ml.
Conclusions:
- Amphotericin B is toxic to corneal epithelial cells and inhibits their migration at concentrations ≥5 µg/ml.
- AmB at 2.5 µg/ml is considered safe for preserving donor corneal tissue.
- These findings guide the safe use of AmB in corneal storage media for transplantation.
