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KMID : 0213520130270060446
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Korean Journal of Ophthalmology
2013 Volume.27 No. 6 p.446 ~ p.453
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In Vitro Effects of Preservative-free and Preserved Prostaglandin Analogs on Primary Cultured Human Conjunctival Fibroblast Cells
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Kim Eun-Joo
Kim Yeoun-Hee
Kang Sun-Hee
Lee Kyoo-Won
Park Young-Jeung
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Abstract
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Purpose: Long-term use of topical medication is needed for glaucoma treatment. One of the most commonly prescribed
classes of hypotensive agents are prostaglandin analogs (PGs) used as both first-line monotherapy; as well as in combination therapy with other hypotensive agents. Several side effects of eye drops can be caused by preservatives. The purpose of this study was to evaluate the effects of PGs with varying concentrations of benzalkonium chloride (BAC), alternative preservatives, or no preservatives on human conjunctival fibroblast cells.
Methods: Primary human conjunctival fibroblast cells were used in these experiments. Cells were exposed to the following drugs: BAC at different concentrations, bimatoprost 0.01% (with BAC 0.02%), latanoprost 0.005% (with BAC
0.02%), tafluprost 0.0015% with/without 0.001% BAC and travoprost 0.004% (with 0.001% Polyquad) for 15 and 30
minutes. Cell cytotoxicity was evaluated by phase-contrast microscopy to monitor morphological changes of cells,
Counting Kit-8 (CCK-8) assay to cell viability, and fluorescent activated cell sorting (FACS) analysis to measure
apoptosis.
Results: BAC caused cell shrinkage and detachment from the plate in a dose-dependent manner. Morphological
changes were observed in cells treated with bimatoprost 0.01% and latanoprost 0.005%. However, mild cell shrinkage
was noted in cells treated with tafluprost 0.0015%, while a non-toxic effect was noted with travoprost 0.004% and preservative-free tafluprost 0.0015%. CCK-8 assay and FACS analysis showed all groups had a significantly decreased cell viability and higher apoptosis rate compared with the control group. However, travoprost 0.004% and preservative-free tafluprost 0.0015% showed lower cytotoxicity and apoptosis rate than other drugs.
Conclusions: This in vitro study revealed that BAC-induced cytotoxicity is dose-dependent, although it is important
to emphasize that the clinical significance of toxicity differences observed among the different PGs formulations has not yet been firmly established. Alternatively preserved or preservative-free glaucoma medications seem to be a reasonable and viable alternative to those preserved with BAC.
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KEYWORD
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Benzalkonium compounds, Conjunctiva, Pharmaceutical preservatives, Synthetic prostaglandins
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