Evaluation Of Toxicity Of Anti-glaucoma Drugs Using Stratified Human Cultivated Corneal Epithelial Sheets

Abstract

Purpose: : Because glaucoma patients received anti-glaucoma drugs for many years, the assessment of safe profiles of anti-glaucoma drug is important. We investigated the toxicity profiles of seven antiglaucoma drugs and benzalkonium chloride (BAK) using the stratified human cultivated corneal epithelial sheets in serum-free culture system.

Methods: : The stratified human cultivated corneal epithelial sheets in serum-free culture system were exposed to seven antiglaucoma drugs in their commercial presentation (Latanoprost® (Nitten), LatanoprostPF® (Nitten), TravatanZ® (Alcon), Tapros® (Santen), Xalatan® (Pfizer), DuoTrav® (Alcon), Xalacom® (Pfizer)) and two concentrations (0.002% and 0.02%) of benzalkonium chloride (BAK). Each solution was applied on the cell sheets for 10 minutes. The cell proliferative capacity and barrier function were examined by WST-1 and carboxyfluorescein permeability assays. Histologic evaluation of cell sheets were performed by hematoxylin and eosin (HE) stains after 1 hour, 6 hours, 24 hours application of each solution on the cell sheet.

Results: : Xalatan®, Xalacom® and 0.02%BAK resulted in significantly lower cell proliferation capacity and barrier function than control (P<0.05), but the others did not show significant difference. Cell sheets with 1 hour, 6 hours and 24 hours applications of Xalatan®, Xalacom® and 0.02%BAK were more damaged histologically than the others.

Conclusions: : Xalatan®, Xalacom® and 0.02%BAK have high toxicity in vitro, whereas Latanoprost®, LatanoprostPF®, TravatanZ®, Tapros®, DuoTrav® and 0.002%BAK has very low or no toxicity. This confirms that higher concentrations of BAK were responsible for toxicity. Our assays using the stratified human cultivated corneal epithelial sheets in serum-free culture system are promising methods in ophthalmic toxicology.

Keywords: cornea: epithelium • drug toxicity/drug effects • ocular irritancy/toxicity testing