Abstract
Purpose :
Chronic use of preserved topical ophthalmic medications is a known cause for ocular surface disease and associated inflammation. The purpose of this study is to assess the cytotoxicity profile, ocular surface images and expression of inflammatory mediators in human conjunctival cell lines after Latanoprost+Benzalkonium Chloride (BAK) treatment administered by MAP technology as a microdose vs. a standard drop.
Methods :
Human conjunctival epithelial cell line (HCjE) were exposed to four drug delivery modalities: Preserved Latanoprost Drops (BAK+ drops), Non-preserved Latanoprost Drops (BAK- drops), Microdosed Preserved Latanoprost (Microdosed BAK+), and Control. Cell-based assays were conducted to assess cell viability, cytotoxicity, apoptosis, and ATP/ROS generation. Visual cells analysis was carried out with confocal microscope under brightfield conditions. RT-qPCR and ELISA were conducted to assess gene and protein expression of common cytokine and chemokine inflammatory mediators.
Results :
HCjE cells treated with Microdosed BAK+ showed no significant difference (p>0.05) in cell viability, cytotoxicity, apoptosis, and metabolic activity compared to those treated with BAK- drops and control. On Brightfield microscopy Microdosed BAK+ treated cells showed a normal cellular mosaic pattern and higher survivability, whereas those treated with BAK+ drops showed Microblebs, Microcysts, Apoptic bodies and cell loss. HCjE cells treated with Microdosed BAK+ showed significantly lower gene expression for IL6 (p<0.05) and CCL 2 (p<0.01) and lower protein expression of IL-6 (p<0.01) compared to cells treated with BAK+ drops.
Conclusions :
This study suggests that microdosed delivery of preserved Latanoprost has a cellular toxicity profile on par with a non-preserved drop and Control. Cells treated with the microdosed form of preserved Latanoprost exhibited a significantly better toxicity profile compared to cells treated with the drop form of the same drug. Drug delivery by MAP technology performed better than drops to significantly lessen upregulation of typical proinflammatory mediators seen in response to the use of BAK in preserved medication for glaucoma.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.