Toxicological evaluation of preservative-containing and preservative-free topical prostaglandin analogs on a 3D-reconstituted corneal epithelium system
Résumé
Purpose: Using an established three-dimensional (3D) toxicological model based on reconstituted human corneal epithelium (HCE), this study investigated the tolerability of four topical intraocular pressure (IOP)-lowering agents: the commercial solutions of benzalkonium chloride (BAC)-containing 0.005% latanoprost, 0.004% travoprost, 0.03% bimatoprost containing 0.02%, 0.015%, and 0.005% BAC, respectively and the preservative-free (PF) tafluprost. Solutions of 0.01% and 0.02% BAC alone were also evaluated for comparison. Methods: The 3D-HCEs were treated with either solutions for 24 hours followed or not by a 24-hour recovery period. We used a modified MTT procedure to assess cell viability in the HCE. Frozen sections of HCE were analyzed using fluorescence microscopy for the evaluation of apoptosis (TUNEL), inflammation (ICAM-1), and proliferation (Ki67). Corneal epithelial tight junctions (occludin and ZO-1) were also assessed by en-face confocal microscopy in response to the different eye drops. Results: The MTT test revealed that the cytotoxicity of antiglaucoma eye drops was primarily related to the concentration of their common BAC preservative (0.02%BAC-latanoprost > 0.015%BAC-travoprost > 0.005%BAC-bimatoprost). PF-tafluprost did not induce obvious cytotoxicity and showed the least expression of inflammatory or apoptotic markers and revealed preservation of membrane immunostaining of tight junction proteins in comparison with BAC-containing solutions. Conclusion: The toxicological model of 3D reconstructed corneal epithelia model confirmed the ocular surface cytotoxicity of BAC-containing antiglaucomatous solutions. Compared to the formulations containing the toxic preservative BAC, PF-tafluprost was well tolerated without inducing significant corneal epithelium deterioration.
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