Abstract
Purpose :
Drug accumulation of systemic medications has the potential to induce unwanted ocular effects, including light sensitivity, pain, or corneal edema that are due to inflammation and/or cytotoxicity. Current studies utilize animal models which are not suitable for rapid drug screening, have poor species extrapolation and standardization. There is a need for physiologically relevant, human primary corneal epithelial tissue models to address ocular safety for the evaluation of new drug formulations.
Methods :
We have utilized an in vitro 3D human corneal epithelial tissue model, EpiCorneal, to analyze the effects of common drugs with known adverse ocular side effects. EpiCorneal tissues are comprised of normal human corneal epithelial cells, express site-specific mucins and tight junctions and attain morphology, barrier properties (TEER>900±200Ω*cm2), and gene expression similar to the in vivo human cornea. The effect of physiologically relevant concentrations of Chlorpromazine hydrochloride (CPZ), a common psychotropic agent; Hydroxychloroquine sulfate (HCQ), an anti-inflammatory and anti-malaria drug (HCQ); Fosamax (FOS), a common anti-osteoporosis agent; and Alfuzosin hydrochloride (ALF), an alpha-adrenergic blocker, were investigated. Effects on tissue viability (MTT assay), barrier function (Transepithelial electrical resistance, TEER), histology, LDH and cytokine release were studied.
Results :
EpiCorneal tissues, incubated at the air-liquid-interface, were exposed to various doses of the drugs from the basolateral side (through the medium) for up to 72h. Barrier function analysis was the most rapid and sensitive test. For CPZ-treated tissues the lowest dose to cause a significant decline in barrier function was 12.5 µM at 24h (67.4%), for HCQ -18.52 µg/ml at 24h (67.4%), for FOS - 0.1 µg/ml at 48h (57.8%), and for ALF – 0.01 µg/ml at 72h (73.3%).Treatment-specific changes in tissue viability, morphology, and LDH and cytokine release were also observed.
Conclusions :
EpiCorneal tissue model has been very useful for evaluating formulations with negligible irritation potential. It will model systemic drug exposure, generate rapid response, avoid species extrapolation, be more cost-effective and more reproducible than animal methods, and will facilitate drug discovery by allowing screening and optimization of pharmaceuticals prior to clinical studies.
This is a 2020 ARVO Annual Meeting abstract.