Abstract
Purpose: :
to evaluate the influence of hyaluronic acid (HA) -based preparations on cultured human corneal epithelial cells morphology, ultrastructure and behavior in model of protection of cultured human epithelial cells against desiccation
Methods: :
Desiccation protection of 5 commercially available hyaluronan containing artificial tear drops was tested using cultured human corneal cells in confluent monolayer. On confluent cell growth the cultures were wetted for 5 min with 5 various hyaluronan-based preparations of artificial tears The cell cultures were exposed to a constant air flow for 0, 5, 15, 30 and 45 minutes. Cells were observed by means of light and scanning electron microscope. Also, p63 and caspase 3 presence was measured by fluorescence immuno-stainings. Finally, the viability assay was performed to evaluate protection capability of applied substances.
Results: :
Cell survival rates after 0, 5, 15, 30, 45 min were (0.96, 0.94, 0.34, 0.10, 0.01) for Hialeye 0,2%, (0.95, 0.88, 0.09, 0.0, 0.0) for Hialeye 0,4%, (0.95, 0.92, 0.43, 0.07, 0.0) for Starazolin Hydrobalance, (0.99, 0.96, 0.49, 0.31, 0.11) for Hyabak, (0.98, 0.94, 0.44, 0.26, 0.04) for Oxyal and (0.99, 0.80, 0.01, 0.0, 0.0) for PBS as a contol. Hyabak and Oxyal showed a significantly better protective effect after a drying period of 30 and 45 min. Significant toxic effect resulted in membrane damage, shape changes, protein expression disorders and high apoptosis rate was induced by BAK preserved product with 0,4% HA.
Conclusions: :
Preservatives-free HA based product assures in this model a significant better protection for cultured human epithelial cells against desiccation in comparison to preserved preparations.Preparations with lower HA concentration (below 0,2%) ensured significantly better protection on cultured human corneal cells against desiccation.Whether these in vitro results are conferrable to the efficacy of artificial tear drops in vivo has to be evaluated in clinical trials.