分层对角膜上皮细胞表面特性的影响。
Effect of Stratification on Surface Properties of Corneal Epithelial Cells.
作者信息
Yáñez-Soto Bernardo, Leonard Brian C, Raghunathan Vijay Krishna, Abbott Nicholas L, Murphy Christopher J
机构信息
Instituto de Física Manuel Sandoval Vallarta, Universidad Autónoma de San Luis Potosí, San Luis Potosí, San Luis Potosí, México 2Department of Veterinary Surgical and Radiological Sciences, School of Veterinary Medicine, University of California Davis, Da.
William R. Pritchard Veterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California Davis, Davis, California, United States.
出版信息
Invest Ophthalmol Vis Sci. 2015 Dec;56(13):8340-8. doi: 10.1167/iovs.15-17468.
PURPOSE
The purpose of this study was to determine the influence of mucin expression in an immortalized human corneal epithelial cell line (hTCEpi) on the surface properties of cells, such as wettability, contact angle, and surface heterogeneity.
METHODS
hTCEpi cells were cultured to confluence in serum-free medium. The medium was then replaced by stratification medium to induce mucin biosynthesis. The mucin expression profile was analyzed using quantitative PCR and Western blotting. Contact angles were measured using a two-immiscible liquid method, and contact angle hysteresis was evaluated by tilting the apparatus and recording advancing and receding contact angles. The spatial distribution of mucins was evaluated with fluorescently labeled lectin.
RESULTS
hTCEpi cells expressed the three main ocular mucins (MUC1, MUC4, and MUC16) with a maximum between days 1 and 3 of the stratification process. Upon stratification, cells caused a very significant increase in contact angle hysteresis, suggesting the development of spatially discrete and heterogeneously distributed surface features, defined by topography and/or chemical functionality. Although atomic force microscopy measurements showed no formation of appreciable topographic features on the surface of the cells, we observed a significant increase in surface chemical heterogeneity.
CONCLUSIONS
The surface chemical heterogeneity of the corneal epithelium may influence the dynamic behavior of tear film by "pinning" the contact line between the cellular surface and aqueous tear film. Engineering the surface properties of corneal epithelium could potentially lead to novel treatments in dry eye disease.
目的
本研究旨在确定永生化人角膜上皮细胞系（hTCEpi）中粘蛋白表达对细胞表面特性（如润湿性、接触角和表面异质性）的影响。
方法
将hTCEpi细胞在无血清培养基中培养至汇合。然后用分层培养基替换培养基以诱导粘蛋白生物合成。使用定量PCR和蛋白质印迹分析粘蛋白表达谱。使用两种不混溶液体法测量接触角，并通过倾斜仪器并记录前进和后退接触角来评估接触角滞后。用荧光标记的凝集素评估粘蛋白的空间分布。
结果
hTCEpi细胞表达三种主要的眼部粘蛋白（MUC1、MUC4和MUC16），在分层过程的第1天至第3天达到最大值。分层后，细胞导致接触角滞后非常显著增加，表明由地形和/或化学功能定义的空间离散和异质分布的表面特征的发展。尽管原子力显微镜测量显示细胞表面没有形成明显的地形特征，但我们观察到表面化学异质性显著增加。
结论
角膜上皮的表面化学异质性可能通过“固定”细胞表面与水性泪膜之间的接触线来影响泪膜的动态行为。设计角膜上皮的表面特性可能会为干眼症带来新的治疗方法。
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