Novel anti-inflammatory liposomal formulation for the pre-ocular tear film: In?vitro and ex?vivo functionality studies in corneal epithelial cells

摘要

Abstract In ocular surface inflammatory diseases, such as dry eye disease, long-term symptom relief requires targeting the inflammation itself rather than treating only the surface-associated dryness with artificial tears. Therefore, we included an anti-inflammatory agent in an unpreserved liposome-based (LP) formulation used as artificial tears. Our aim was to characterize and study its in?vitro and ex?vivo cell uptake and functionality. Human corneal epithelial (HCE) cells were used to study MPA-LP-induced effects after 60?min of exposure, using blank LP and non-LP MPA formulations as controls. A fluorescent labeled LP formulation was used to determine uptake by HCE cells and localization in ex?vivo porcine corneas. The LP formulation complied with the required physicochemical properties and had no cytotoxicity on HCE cells after 60?min of exposure. HCE cells showed LP-associated fluorescence at 24, 48, and 72?h after 60?min of exposure, and the LP-associated fluorescence was uniformly distributed throughout the porcine corneal epithelium immediately after 5?min of exposure. MPA-LP increased protein expression and nuclear translocation of progesterone receptor in comparison with controls as determined by Western blotting and immunofluorescence. Moreover, MPA-LP significantly reduced the cell proliferation rate and IL-6 and IL-8 production 48?h after the exposure period, as determined by the alamarBlue assay and ELISA, respectively. None of these effects were evident in blank LP-exposed cells and non-LP MPA formulation reduced only IL-6 production. Our results suggest that the LP-based formulation, used to replenish the lipids of the tear film, can be loaded with anti-inflammatory agents that can be delivered into the cells and activate specific drug receptors. These agents can reduce inflammatory cytokine production and may be effective in the treatment of inflammatory processes associated with ocular surface diseases. Graphical abstract Display Omitted Highlights ? Medroxyprogesterone was incorporated into a liposomal artificial tear formulation. ? Encapsulated drug readily penetrates corneal tissue in an ex?vivo porcine model. ? Encapsulated drug exerts anti-inflammatory effects in human corneal cells in?vitro . ? The novel liposomal artificial tears enhance the effect of the drug in?vitro .