PURPOSE We aimed to investigate the toxicity of 6 fixed-combination drugs for glaucoma therapy using human corneal epithelial sheets (HCES). STUDY DESIGN Experimental. MATERIALS AND METHODS We used 6 kinds of commercially available fixed-combination drugs: latanoprost/carteolol (LAT/CAR), latanoprost/timolol (LAT/TIM), tafluprost/timolol (TAF/TIM), travoprost/timolol (TRA/TIM), brinzolamide/timolol (BRZ/TIM), and dorzolamide/timolol (DRZ/TIM) including different preservatives. The cell viability and barrier function of the HCES after exposure to the eye drops for 10 or 30 minutes were assessed using the WST-1 assay and transepithelial electrical resistance (TEER) measurements, respectively. The HCES were also evaluated using hematoxylin and eosin (HE) staining and transmission electron microscopy. RESULTS The cell viability significantly decreased in the HCES treated with LAT/TIM or DRZ/TIM after 10 and 30 minutes and in those treated with BRZ/TIM after 30 minutes. The barrier function increased significantly in the HCES treated with LAT/CAR. Histologically, the HCES were damaged after treatment with LAT/TIM, BRZ/TIM, or DRZ/TIM for 30 minutes. Transmission electron microscopy indicated narrow intercellular spaces and multiple intercellular junctions in the HCES treated with LAT/CAR, TAF/TIM, or TRA/TIM. The HCES treated with DRZ/TIM, BRZ/TIM, or LAT/TIM contained cytoplasmic vacuoles and collapsed cellular structures. CONCLUSION Glaucoma fixed-combination eye drops demonstrated a different toxic effect on the cell viability, barrier function, and morphologic changes of HCES.

中文翻译：

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使用培养的人角膜上皮片的青光眼固定组合滴眼液的毒性概况。

目的我们旨在研究使用人角膜上皮片（HCES）治疗青光眼的6种固定组合药物的毒性。研究设计实验。材料与方法我们使用了6种市售固定组合药物：拉坦前列素/心得乐（LAT / CAR），拉坦前列素/替莫洛尔（LAT / TIM），他氟前列素/替莫洛尔（TAF / TIM），特拉沃前列素/替莫洛尔（TRA / TIM） ，溴苯甲酰胺/噻吗洛尔（BRZ / TIM）和多佐酰胺/噻吗洛尔（DRZ / TIM），包括不同的防腐剂。分别使用WST-1分析和经上皮电阻（TEER）测量，评估了HCES在滴眼液中暴露10或30分钟后的细胞活力和屏障功能。还使用苏木精和曙红（HE）染色和透射电子显微镜对HCES进行了评估。结果在10和30分钟后，用LAT / TIM或DRZ / TIM处理的HCES和在30分钟后用BRZ / TIM处理的HCES中，细胞活力显着降低。在用LAT / CAR治疗的HCES中，屏障功能显着增加。从组织学上讲，用LAT / TIM，BRZ / TIM或DRZ / TIM处理30分钟后，HCES受损。透射电子显微镜显示，用LAT / CAR，TAF / TIM或TRA / TIM处理的HCES中狭窄的细胞间空间和多个细胞间连接。用DRZ / TIM，BRZ / TIM或LAT / TIM处理的HCES含有细胞质液泡和塌陷的细胞结构。结论青光眼固定组合滴眼液对HCES的细胞活力，屏障功能和形态变化表现出不同的毒性作用。