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Trehalose-Based Eye Drops Preserve Viability and Functionality of Cultured Human Corneal Epithelial Cells during Desiccation

机译:基于海藻糖的眼药水在干燥过程中保留了培养的人角膜上皮细胞的活力和功能

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摘要

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.
机译:本文比较了在培养的人角膜上皮细胞实验干燥过程中,与市售制剂相比,基于海藻糖的滴眼液对细胞的保护作用。将培养的人角膜上皮细胞(hCEC)与干眼症治疗中常用的7种不同的市售药物一起孵育,然后在通风橱中于空气中进行干燥试验5、15、30和45分钟。细胞活力通过活/死荧光测定法定量,而凋亡细胞的存在通过对活性胱天蛋白酶3蛋白进行免疫荧光染色来估计。使用中性红染色评估膜功能的保留,而通过肌动蛋白丝,细胞核和p63蛋白的荧光染色确定适当形态和表型的保留。基于海藻糖的滴眼液在防止细胞因干燥而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。

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