Abstract
Purpose: :
To develop a method to evaluate in vitro cytotoxic effects of contact lenses (CL) soaked in multi-purpose contact lens care solutions (MPS) on the morphology, viability, and barrier function of monolayer and stratified cultures of human corneal limbal epithelial (HCLE) cells.
Methods: :
Acuvue Oasys® CL soaked in one of 3 MPS (A,B,C) for 24 hr were placed on cells for 30, 60, or 120 min. Monolayer cultures: Cell morphology following treatment and after 24 hr of recovery in culture medium was observed using Hoffman modulation contrast optics. Effects of MPS on cell viability were determined using a live/dead cell assay with flow cytometry and fluorescence microscopy. Stratified cultures: Cells were grown on permeable membrane supports and exposed to CL with MPS. Morphology was observed. Viability was determined with a live/dead cell assay and fluorescence microscopy. Effects on barrier function were determined by measuring fluorescein permeability and trans-epithelial electrical resistance (TEER).
Results: :
Monolayer cultures:
Conclusions: :
A method has been described for evaluating interactions of HCLE cells with CL soaked in MPS using several cytotoxicity tests. Time and MPS-dependent adverse effects were detected using monolayer cultures; however, multilayer cultures may be a better model of the stratified corneal epithelium since CL and MPS known to be safe in clinical use had no adverse effects on multilayer cells. Further development of the model will include studies of additional lens types, lens cycling times and measurement of uptake and release of MPS components.
Keywords: contact lens • cornea: epithelium • ocular irritancy/toxicity testing