title: A collaborative evaluation of the cytotoxicity of two surfactants by using the human corneal epithelial cell line and the WST-1 test.
abstract: This study was undertaken to investigate the use of the in vitro test WST-1, an assay of cell proliferation and viability, for a preliminary safety evaluation of topical ophthalmic preparations. The cytotoxicity of two surfactants, benzalkonium chloride (BAC) and polyoxyethylene-20-stearyl ether (Brij78, PSE) was independently investigated in four laboratories in the EU by using an immortalized human corneal epithelial (HCE) cell line. The HCE cells were exposed to BAC and PSE for 5 min, 15 min, and 1 hour, and the results of the HCE-WST-1 tests were collected and compared. After one-hour exposure, the EC(50) values in BAC-treated cells in the presence of serum ranged between 0.0650 +/- 0.0284 (mean +/- SD) mM, and those in the absence of serum 0.0296 +/- 0.0081 mM. The corresponding values for PSE were 0.0581 +/-.0300 mM and 0.0228 +/-.0063 mM. There were variations in the results between different laboratories, with coefficients of variation ranging from 31 to 121%, mean 58%. The use of one-hour exposure time is to be preferred, and the elimination of serum in the culture medium is recommended to avoid both underestimation of toxic effects and variability of the test results.
authors.affiliation: Medical School, University of Tampere, Finland. anne.huhtala@uta.fi
authors.forename: Anne
authors.initials: A
authors.lastname: Huhtala
authors.forename: Päivi
authors.initials: P
authors.lastname: Alajuuma
authors.forename: Susi
authors.initials: S
authors.lastname: Burgalassi
authors.forename: Patrizia
authors.initials: P
authors.lastname: Chetoni
authors.forename: Horst
authors.initials: H
authors.lastname: Diehl
authors.forename: Maria
authors.initials: M
authors.lastname: Engelke
authors.forename: Marios
authors.initials: M
authors.lastname: Marselos
authors.forename: Daniela
authors.initials: D
authors.lastname: Monti
authors.forename: Periklis
authors.initials: P
authors.lastname: Pappas
authors.forename: M Fabrizio
authors.initials: MF
authors.lastname: Saettone
authors.forename: Lotta
authors.initials: L
authors.lastname: Salminen
authors.forename: Marianthi
authors.initials: M
authors.lastname: Sotiropoulou
authors.forename: Hanna
authors.initials: H
authors.lastname: Tähti
authors.forename: Hannu
authors.initials: H
authors.lastname: Uusitalo
authors.forename: Michaela
authors.initials: M
authors.lastname: Zorn-Kruppa
mesh_terms: D001548:Benzalkonium Compounds / Q000506:poisoning*; D001769:Blood; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004728:Endothelium, Corneal / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D011092:Polyethylene Glycols / Q000506:poisoning*; D013501:Surface-Active Agents / Q000506:poisoning*; D013997:Time Factors
chemical_list: D001548:Benzalkonium Compounds; D016895:Culture Media, Serum-Free; D013501:Surface-Active Agents; D011092:Polyethylene Glycols; C043444:octadecyl polyoxyethylene ether