Hill-Bator Aneta, Misiuk-Hojło Marta, Marycz Krzysztof, Grzesiak Jakub
Department of Ophthalmology, Medical University Wroclaw, Borowska 213, 50-556 Wroclaw, Poland.
Electron Microscopy Laboratory, University of Environmental and Life Sciences in Wroclaw, Kozuchowska 5b, 51-631 Wroclaw, Poland.
Biomed Res Int. 2014;2014:292139. doi: 10.1155/2014/292139. Epub 2014 Jun 8.
This paper presents the evaluation of cytoprotective ability of trehalose-based eye drops in comparison with commercially available preparations during the experimental desiccation of cultured human corneal epithelial cells. Cultured human corneal epithelial cells (hCEC) underwent incubation with 7 different, commercially available medicaments used commonly in dry eye syndrome treatment, followed by desiccation trial performed on air under the flow hood for 5, 15, 30, and 45 minutes. Cell viability was quantified by live/dead fluorescent assay, while the presence of apoptotic cells was estimated by immunofluorescent staining for active caspase 3 protein. The preservation of membrane functions was evaluated using neutral red staining, while the preservation of proper morphology and phenotype was determined by fluorescent staining for actin filaments, nuclei, and p63 protein. The trehalose-based eye drops showed the highest efficiency in prevention of cell death from desiccation; moreover, this preparation preserved the normal cellular morphology, functions of cell membrane, and proliferative activity more effectively than other tested medicaments.
本文介绍了在培养的人角膜上皮细胞实验性干燥过程中，与市售制剂相比，海藻糖滴眼液的细胞保护能力评估。将培养的人角膜上皮细胞（hCEC）与7种常用于治疗干眼综合征的市售药物一起孵育，然后在通风橱的空气中进行5、15、30和45分钟的干燥试验。通过活/死荧光测定法定量细胞活力，同时通过对活性半胱天冬酶3蛋白进行免疫荧光染色来估计凋亡细胞的存在。使用中性红染色评估膜功能的保存情况，而通过对肌动蛋白丝、细胞核和p63蛋白进行荧光染色来确定正常形态和表型的保存情况。海藻糖滴眼液在预防干燥引起的细胞死亡方面显示出最高的效率；此外，该制剂比其他测试药物更有效地保留了正常的细胞形态、细胞膜功能和增殖活性。
