title: Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells.
abstract: The purpose of the study was to devise a fast, reliable and sensitive cell viability assay for assessment of acute cytotoxicity on human corneal epithelial cells by using a clinically relevant exposure time. Acute cytotoxic effects of the pharmaceutical excipients benzalkonium chloride (BAC), macrogolglycerol hydroxystearate (MGHS40), polysorbate 80 (PS80) and marketed ophthalmic formulations (Lumigan(®), Monoprost(®), Taflotan(®), Travatan(®), Xalatan(®)) containing these excipients were tested. Human corneal epithelial cell (HCE-T) viability was assessed by measuring the reduction of resazurin to highly fluorescent resorufin. Expression of the tight junction proteins in HCE-T cells were characterized by immunofluorescence staining. Presence of tight junction proteins in HCE-T cells was demonstrated. BAC preserved ophthalmic formulations showed concentration-dependent and time-dependent cytotoxicity to human corneal epithelium. In contrast, no acute cytotoxicity of non-ionic stabilizing/solubilizing excipients (MGSH40 and PS80) or ophthalmic formulation containing these excipients was observed. Marketed ophthalmic formulations used for glaucoma medication show differential toxicity on human corneal epithelial cells. The present study revealed that BAC-preserved ophthalmic formulations were able to induce acute cytotoxic effects even during a clinically relevant exposure time, which was not observed with MGSH40 and PS80 excipients or ophthalmic formulations containing these excipients.
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland. Electronic address: jenni@experimentica.com.
authors.forename: Jenni J
authors.initials: JJ
authors.lastname: Hakkarainen
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland; School of Pharmacy, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland.
authors.forename: Mika
authors.initials: M
authors.lastname: Reinisalo
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland; State Research Institute for Innovative Medicine, Vilnius, Lithuania.
authors.forename: Symantas
authors.initials: S
authors.lastname: Ragauskas
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland.
authors.forename: Aila
authors.initials: A
authors.lastname: Seppänen
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland; Department of Ophthalmology, Stritch School of Medicine, Loyola University Chicago, 2160 S. First Ave, Maywood, IL 60153, USA; K&P Scientific LLC, P.O. Box 1432, Hines, IL 60141, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Kaja
authors.affiliation: R&D department, Experimentica Ltd., P.O. Box 1199, FI-70211 Kuopio, Finland; Department of Ophthalmology, SILK, School of Medicine, University of Tampere, Biokatu 14, FI-33520 Tampere, Finland.
authors.forename: Giedrius
authors.initials: G
authors.lastname: Kalesnykas
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D003315:Cornea / Q000187:drug effects / Q000378:metabolism; D003603:Cytotoxins / Q000008:administration & dosage / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004339:Drug Compounding / Q000379:methods; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D005079:Excipients / Q000633:toxicity; D006801:Humans; D008297:Male; D009883:Ophthalmic Solutions / Q000008:administration & dosage / Q000633:toxicity*; D011310:Preservatives, Pharmaceutical / Q000008:administration & dosage / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar
chemical_list: D003603:Cytotoxins; D005079:Excipients; D009883:Ophthalmic Solutions; D011310:Preservatives, Pharmaceutical
keywords: Cornea; Cytotoxicity; Ophthalmic formulation; Pharmaceutical excipient