Cytometric assessment of cytostatic and cytotoxic effects of topical glaucoma medications on human epithelial corneal line cells.
BACKGROUND
The long-term-treatment of glaucoma with topical medications is associated with side effects involving cornea damage. We examined the effect of glaucoma topical medications (bimatoprost, travoprost, latanoprost, timolol, betaxolol, dorzolamide, brinzolamide, brimonidine) on growth of cells of three human epithelial corneal lines.


METHODS
The cells were cultured in 8-chamber slides, treated with different concentrations of the medications, and fixed at 24, 48, and 72 h. Cell number on slides to estimate viability and growth curves, frequency of apoptosis (FLICA and caspase-3 activation probes), and proliferation (BrdU incorporation assay) were measured by laser scanning cytometry (LSC).


RESULTS
Depending on concentration all examined medications induced cell necrosis or apoptosis and suppressed proliferation. Significant variability in proliferation and apoptosis was observed within the same cultures depending on local cell density, with cells in high density areas being more resistant. The data indicate that commonly used topical medications exert cytostatic and cytotoxic effects in cultures of corneal cells and suggest that caution should be exercised in their use, particularly, when the corneal diseases are accompanied by cell proliferation and regeneration, in long-term-treatment.


CONCLUSIONS
The present approach of using LSC makes it possible to assess and compare cytostatic and cytotoxic effects of different topical medications on the respective target cells.
Pozarowska
Dorota
D
Department of Ophthalmology, Medical University, Lublin, Poland. ppozarowski@yahoo.com
Pozarowski
Piotr
P
Darzynkiewicz
Zbigniew
Z
D000287:Administration, Topical; D000959:Antihypertensive Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects*; D005901:Glaucoma / Q000188:drug therapy*; D006801:Humans; D047569:Laser Scanning Cytometry / Q000379:methods*; D012680:Sensitivity and Specificity; D018675:Toxicity Tests
D000959:Antihypertensive Agents