AIMS:The ocular surface is the very first barrier between the visual system and external environment. It protects the eye from the exposure to various light sources that significantly emit in blue spectrum. However, the impact of blue light on the ocular surface has been poorly explored so far. In this study, we investigated in vitro the phototoxicity of blue light illumination in human epithelial cells of the ocular surface. We worked either in basal conditions or under hyperosmolar stress, in order to mimic dry eye disease (DED) that is the most common disease involving the ocular surface.RESULTS:Corneal and conjunctival epithelial cells suffered the most from violet-blue light but also from longer-wave blue light. Exposure to blue wavebands significantly decreased cellular viability, impacted on cellular morphology and provoked reactive oxygen species (ROS) over-production. Conjunctival epithelial cell line had a greater photosensitivity than the corneal epithelial one. Hyperosmolar stress potentiated the blue light phototoxicity, increasing inflammation, altering mitochondrial membrane potential, and triggering the glutathione-based antioxidant system.INNOVATION:In human epithelial corneal and conjunctival cells of the ocular surface, we demonstrated the harmful impact of blue light on viability, redox state and inflammation processes, which was modified by hyperosmolarity.CONCLUSION:Blue light induced cell death and significant ROS production, and altered the expression of inflammatory genes and operation of the cellular defensive system. We established for the first time that hyperosmolar stress impacted phototoxicity, further suggesting that DED patients might be more sensitive to blue light ocular toxicity.