Effect of Stratification on Surface Properties of Corneal Epithelial Cells

Bernardo Yanez-Soto,Brian C. Leonard,Vijay Krishna Raghunathan,Nicholas L. Abbott,Christopher J. Murphy

Investigative Ophthalmology & Visual Science（2015）SCI 2区

Univ Autonoma San Luis Potosi | Univ Calif Davis | 3016 Engn Hall

Cited 33|Views9

Abstract

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.

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Key words

dry eye,mucins,surface heterogeneity,surface phenomena

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