A collaborative evaluation of the cytotoxicity of two surfactants by using the human corneal epithelial cell line and the WST-1 test.
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.
Huhtala
Anne
A
Medical School, University of Tampere, Finland. anne.huhtala@uta.fi
Alajuuma
Päivi
P
Burgalassi
Susi
S
Chetoni
Patrizia
P
Diehl
Horst
H
Engelke
Maria
M
Marselos
Marios
M
Monti
Daniela
D
Pappas
Periklis
P
Saettone
M Fabrizio
MF
Salminen
Lotta
L
Sotiropoulou
Marianthi
M
Tähti
Hanna
H
Uusitalo
Hannu
H
Zorn-Kruppa
Michaela
M
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
D001548:Benzalkonium Compounds; D016895:Culture Media, Serum-Free; D013501:Surface-Active Agents; D011092:Polyethylene Glycols; C043444:octadecyl polyoxyethylene ether