Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Department of Ophthalmology and Visual Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Curr Eye Res. 2022 Sep;47(9):1259-1265. doi: 10.1080/02713683.2022.2091147. Epub 2022 Jun 27.
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.
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.
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.
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.
评估两性霉素 B(AmB)在 Optisol™-GS 角膜保存液中的毒性对角膜上皮细胞(CEC)形态和迁移能力的影响。
从雄性日本白兔中取出硬性角膜缘,然后在 4°C 下保存在含有 0µg/ml AmB(对照组)和 2.5、5、25 和 50µg/ml AmB(AmB 组;每组四只眼睛)的 Optisol™-GS 中。保存 7 天后,通过苏木精-伊红染色、免疫组织化学染色(ZO-1)和末端脱氧核苷酸转移酶 dUTP 缺口末端标记(TUNEL)检测评估 CEC 形态。此外,为了评估 CEC 迁移能力,从一个保存的角膜中取出三个角膜块(每个 6-8×3mm)进行 24 小时培养,然后测量 CEC 迁移到基质表面的区域(中央区域的 2mm)。
在 5、25 和 50µg/ml 的 AmB 作用下,所有保存的角膜中均观察到 CEC 的变形和空泡化。在 AmB 浓度为 25 和 50µg/ml 时,ZO-1 表达明显减少。TUNEL 标记指数在 AmB 浓度≥5µg/ml 时明显增加。与对照组相比,AmB 浓度为 25 和 50µg/ml 时,CEC 迁移呈剂量依赖性抑制。
在 Optisol™-GS 中添加 AmB 可能对 CEC 有毒性,并抑制其在浓度≥5µg/ml 时的迁移。AmB 浓度为 2.5µg/ml 时可被认为对用于角膜上皮移植手术的供体角膜组织的保存是安全的。