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H. T. Uusitalo, A. M. Huhtala; The Cytotoxic Effects of Commercial Preparations of Prostaglandin Analogues and Benzalkonium Chloride on Human Corneal Epithelial and Human Conjunctival Epithelial Cells in vitro. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3817.
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© ARVO (1962-2015); The Authors (2016-present)
To define the cytotoxic effects of the commercial preparations of prostaglandin analogues, the most commonly used drug group for glaucoma using cell culture methods. The possible adverse effects of benzalkonium chloride (BAC) containing Xalatan (0.02% BAC), Travatan (0.015% BAC), Lumigan (0.005% BAC), and BAC-free tafluprost, and correspondingly the effects of the appropriate concentrations of BAC were evaluated in vitro using human corneal epithelial (HCE) and human conjunctival epithelial (IOBA-NHC) cell lines.
Cells grown for 24 hours were exposed to eye drop concentrations of 0.1%-10% in culture medium without fetal bovine serum for one hour. Correspondingly, the cells were exposed to 0.00008%-0.005% BAC in serum-free medium for one hour. Cytotoxicity was assessed with the WST-1 assay as a colorimetric assay for cellular growth and viability and the lactate dehydrogenase (LDH) test as the measurement of cell membrane integrity.
The order of decreasing toxicity of the tested drugs, assessed with the WST-1 test was Xalatan ≥ Travatan > Lumigan ≥ tafluprost. There were statistically significant differences between the studied drugs either with IOBA-NHC or HCE cells. IOBA-NHC cells appeared to be more sensitive than HCE cells. The EC50 value of BAC in HCE cells was 0.0013% and 0.00047% in IOBA-NHC cells. In HCE cells, the tested commercial preparations of prostaglandin analogues had no effect on LDH leakage expect for Xalatan with the highest concentration tested (10%). Also in IOBA-NHC cells, LDH leakage was very minor and was statistically significant only in the case of 10% Xalatan and 3-10% Travatan.
The cytotoxic effect of Xalatan, Travatan, and Lumigan was dependent on the BAC concentration of the drug. Also, the in vitro toxicity of BAC itself was highly concentration dependent and appeared at the concentrations above those corresponding to 0.001% of BAC in ophthalmic medications. Preservative-free tafluprost was the least toxic of the drugs tested.
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