Hconepics

Two different human cell lines were employed to assess the in vitro tolerance of the developed preparations. Immortalized human corneal epithelial cells (HCECs; Evercyte GmbH, Vienna, Austria) were maintained at 37 °C under 5% CO₂ in a humid atmosphere (95%). The medium was changed every 48–72 h. HCECs were kept in an EpiLife^® cell-culture medium (Life Technologies, Madrid, Spain) with EDGS^® 1X (Life Technologies, Madrid, Spain) and penicillin–streptomycin 1% (Life Technologies, Madrid, Spain) as supplementation. Likewise, immortalized human conjunctival epithelial cells (HConEpiCs; Innoprot, Bizkaia, Spain) were maintained under the same temperature, humidity, and CO₂ conditions, while the medium was changed every 48 h. HConEpiCs were cultured using the IM-Ocular Epithelial Cell Medium Kit (Innoprot, Bizkaia, Spain). To ensure correct cell attachment and maintenance, the flasks were coated with collagen I (1 mg/mL) (Innoprot, Bizkaia, Spain).