Impact of two different prostaglandin preparations on human corneal epithelial cells (HCE-2) in in vitro conditions

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Abstract

Purpose

To investigate impact of two antiglaucoma prostaglandin preparations on human corneal epithelial cells in in vitro conditions.

Methods

Human corneal epithelial cells (HCE-2), cultured in standard conditions, were treated with two commercially available preparations of prostaglandins: latanoprost and tafluprost. Treatment was applied in two time points – parallel with cells plating (day 0) or after 48 hours from plating (day 2). Medium containing drugs with % concentration of 0.1, 0.3, 1, 3, 10 was changed every second day. As negative control standard medium was used, as positive – medium containing 3% methanol. After 48 hours of culture, living culture was stained with erythrosine to visualize dead cells. After fixation, immunostaining for Ki67 antigen and ubiquitin (proteasomal marker) was performed.

Results

In latanoprost group “day 0”, concentrations <1% affected cells attaching and proliferation rate, >1% made cells totally unable to attach. In tafluprost group “day 0”, concentrations <3% have shown no visible effect on cells, for concentrations >3%, fluctuations in cell shape appeared. In group “day 2” latanoprost treatment caused massive cells detachment for concentrations >1%. For lower concentration induced decrease of cells proliferation. Tafluprost treatment in this group showed no visible changes comparing with negative control, except 10% concentration, where induced mild cells detachment. In both groups treatment decreased expression of Ki67 and increased cellular ubiquitin (much more expressed in latanoprost group).

Conclusion

Latanoprost preparation revealed to be more harmful considering proliferation, proteolysis and toxicity for HCE-2 cells in comparison to tafluprost.

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