Modulating the toxic effects of benzalkonium chloride on human corneal epithelial cells in vitro

Abstract

Purpose : Many topical ophthalmic medications have documented ocular surface toxicity attributed to preservatives, most commonly benzalkonium chloride (BAK). The main purpose of this study is to determine the critical cytotoxic BAK concentration for telomerized- immortalized Human Corneal Epithelial Cells (HCECs) using a series of concentrations and different exposure time, and find a way to prevent or treat this damage.

Methods : HCECs was plated in 96-well plates and incubated for 1 day. Culture medium was then replaced by basal media with different BAK concentrations (0.0005% to 0.005% in 0.0005 increment steps), incubated for 5, 10, or 15 minutes. In another set of experiments, the cells were incubated in a specific conditioned media 30 minutes before BAK application. Cell proliferation/viability was assessed by MTT assay.

Results : BAK treatment time of 10 minutes was selected. BAK toxic concentration was 0.0015. Our results suggest that conditioned media has some degrees of protective effects against BAK ocular toxicity. HCECs pre-treated with the specific conditioned media demonstrated 71% cell viability in comparison to 56% for un-conditioned media.

Conclusions : These results indicate that BAK induced cell damage might be reduced using a specified conditioned media. This may be used clinically to decrease the adverse effect of topical eye drops.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.