Molecular mechanism of ocular surface damage: Application to an in vitro dry eye model on human corneal epithelium

[摘要] Purpose: The present study was concernedwith the development of a new experimental model of dry eye using humanreconstructed in vitro corneal epithelium (HCE). The model is based onthe use of adapted culture conditions that induce relevantmodifications at the cellular and molecular level thus mimicking dryeye. Methods: The HCE model was maintained ina controlled environmental setting (relative humidity <40% and40 °C temperature) for 24 h and up to 72 h to induce dryeye. The evolution of the dry eye condition was assessed by histology,immunohistochemistry staining, scanning electron microscopy, and geneexpression by using TaqMan gene assay technology (mucin-4 [MUC4],matrixmetallopeptidase-9[MMP9], tumor necrosis factor-α [TNF-α],anddefensin β-2 [DEFB2). The effects of different commerciallyavailable tear substitutes on the induced dry eye condition weretested. Results: This in vitro dry eye HCEmodel, that was well established within 24 h, has the characteristicfeatures of a dry eye epithelium and could be satisfactorily used forpreliminary assessment of the protective activity of some artificialtears. The transcriptional study of selected biomarkers showed anincrease in MUC4, MMP9, TNF-α, and hBD-2 (DEFB2)geneexpression.Conclusions: By using a dynamicapproach, we were able to define a biomarker gene signature of dryeye-induced effects that could be predictive of corneal damage in vivoand to discriminate the efficacy among different commercial artificialtears.

[学科分类] 生物化学/生物物理