Karen Harrington

Use of a human corneal epithelial cell line for screening the safety of contact lens care solutions in vitro.

Purpose. Sodium fluorescein permeability assay, alamarBlue assay, and scanning electron microscopy were performed to study the effects of various contact lens disinfecting multipurpose solutions (MPS) on the integrity of the ocular surface epithelium by using corneal epithelial cells. Methods. The sodium fluorescein permeability and alamarBlue activity of monolayer cultures of human corneal epithelial cells were compared after exposure to ReNu MultiPlus, OPTI-FREE Express, AQuify 5 Minute, SOLO-care Plus With Aqualube, and Complete Moisture Plus contact lens care solutions for 15 minutes. Additional cell monolayers were prepared for each treatment and were analyzed with a scanning electron microscope after a 10-minute exposure. Results. The sodium fluorescein permeability assay, alamarBlue assay, and scanning electron microscopy showed that OPTI-FREE Express was significantly more damaging to the human corneal epithelial cell monolayer than ReNu MultiPlus, SOLO-care Plus With Aqualube, Complete Moisture Plus, and AQuify 5 Minute contact lens solutions. Cell monolayers treated with OPTI-FREE Express were more permeable to sodium fluorescein and showed lower metabolic activity than cell monolayers treated with the other multipurpose solutions. Conclusions. This experiment shows that ReNu MultiPlus, SOLO-care Plus With Aqualube, Complete Moisture Plus, and AQuify 5 Minute contact lens solutions have a minimal effect on human corneal epithelial cells in culture, whereas OPTI-FREE Express has a higher negative effect on tight junctions, cell membranes, and overall metabolism of these human corneal epithelial cells.

Comparison of the effect of multipurpose contact lens solutions on the viability of cultured corneal epithelial cells

PURPOSE To determine the effect of four marketed multipurpose contact lens solutions (MPSs) on corneal epithelial cell viability. METHODS Comparison of the effect of MPS A (Renu MultiPlus, Bausch & Lomb), MPS B (OPTI-FREE Express, Alcon), MPS C (AQuify, CibaVision), and MPS D (OPTI-FREE RepleniSH, Alcon) on cell viability was performed by quantifying cellular ATP content, resazurin reduction, and lactate dehydrogenase (LDH) release in transformed human corneal epithelial cells (HCEpiC) and primary bovine corneal epithelial cells (BCEpiC). RESULTS Significant reductions in cellular ATP content were observed at 40% solution and above with both MPS B and MPS D, compared to at 100% only for MPS A and MPS C, and similar results were obtained in BCEpiC. Effects on resazurin reduction were also less in HCEpiC exposed to increasing doses of MPS A and MPS C than in cells exposed to MPS B and MPS D. After 15 min, HCEpiC viability measured by both resazurin reduction and cellular ATP levels was significantly lower for cells exposed to MPS B, MPS D, and MPS C, while HCEpiC exposed to MPS A were not affected. MPS B and MPS D reduced cell viability more than MPS A and MPS C over a 2-h time course in both HCEpiC and BCEpiC. CONCLUSIONS Both MPS B and MPS D can cause large decreases in the viability of cultured corneal epithelial cells even with just a 2h exposure at multiple doses. Significant reduction in cell viability is evident at brief 15-30 min exposures. In contrast, MPS A and MPS C have significantly less effect on the cell viability of corneal epithelial cells at multiple doses, after these short exposure times.

Effect of a novel multipurpose contact lens solution on human corneal epithelial barrier function

PURPOSE To explore the effect of a novel multipurpose contact lens solution (MPS) on the junction protein distribution and barrier function of cultured human corneal epithelial cell monolayers. METHODS Cultured human corneal epithelial cells (HCEpiC) were exposed to a novel MPS (MPS A; Biotrue™ multi-purpose solution, Bausch & Lomb Incorporated) at 50%, 75% and 100% for 10 or 30 min. Four commercially available MPS products, MPS B (AQuify, Ciba Vision), MPS C (COMPLETE MPS Easy Rub, AMO), MPS D (OPTI-FREE Express, Alcon) and MPS E (OPTI-FREE RepleniSH, Alcon) were tested in parallel. Tight junction structure and integrity were evaluated by confocal microscopy using ZO-1 antibody and scanning microscopy (SEM). Quantitative evaluation of MPSs on epithelial barrier function was determined by measuring transepithelial electrical resistance (TEER) across HCEpiC grown on Transwell Clear permeable supports and on electric cell-substrate impedance sensing (ECIS) electrode arrays. RESULTS Overall after exposure to the three concentrations (50%, 75%, and 100%) of MPS A, ZO-1 distribution and fluorescent intensity on the cell surface appeared similar to the media control with continuous tight junctions and clear intercellular junctions. At all measured time points after exposure to MPS A (50% or 75%) there was also no effect on the TEER using both resistance methodologies, and SEM showed that MPS A appeared similar to the Hanks balanced salt solution (HBSS) control. In cells exposed to MPS D there was a dose-dependent change in the distribution of ZO-1, some cell detachment, and a decrease in monolayer resistance at all time points measured. Ultrastructurally, MPS D caused gross changes, including damage to cell junctions and plasma membranes. To a lesser extent, the remaining three commercial MPS products demonstrated some effects on tight junction ZO-1 distribution and/or TEER. CONCLUSIONS Based on the in vitro measurements of tight junction protein expression, monolayer integrity, and transepithelial electrical resistance, the novel multipurpose contact lens solution (MPS A) did not alter corneal barrier function as compared to media, PBS or HBSS control. Clinical significance of the observed differences in epithelial barrier function among the MPSs tested needs further investigation.