Cytostatic and cytotoxic effects of 5‐fluorouracil on human corneal epithelial cells and keratocytes

• Published in: Acta ophthalmologica (Oxford, England) Vol. 88; no. s246
• Main Authors: MIDENA, E, LAZZARINI, D, PARROZZANI, R, VIOLATO, D, FREGONA, I
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.09.2010
• ISSN: 1755-375X; 1755-3768
• DOI: 10.1111/j.1755-3768.2010.4363.x

Purpose To investigate the effects of different 5‐Fluorouracil (5‐FU) concentrations, exposure times and application techniques on in vitro cultured human corneal cells. Methods Human corneal epithelial cells (HCEC) and human corneal keratocytes (HCK) cultures were exposed to different 5‐FU concentration (0.025% to 1%) and incubation times (single 5’ to 2 hrs). The cytostatic effect was evaluated as ratio of inhibition of migration towards controls. The cytotoxic effect evaluation included both contrast phase microscopic observations, and viability measures performed using a MTT[3‐(4,5‐dimethylthiazo‐2‐yl)‐2,5‐diphenyltetrazolium bromide; thiazolyl blue] colorimetric assay. The results were expressed as ratio of optical density (OD) reduction 24 hrs after exposures. Results Cytostatic effect is time and dose dependent. Minimal inhibiting dose (ID50) was 0,55% after 1 hour incubation for HCEC; ID50 was 0,5% after 2 hrs incubation for HCK . A 100% inhibitory effect was never observed at any concentration or incubation interval. No cytotoxic changes were observed at 5‐FU < 1%; 5‐FU 1% showed time‐dependent cytotoxic changes just in HCEC cultures. MTT analysis showed no OD reduction at 5‐FU concentration < 1%, whereas 1% 5‐FU showed OD reduction < 50% at any tested exposure time. HCEC showed higher reduction in OD than HCK. Conclusion 5‐FU shows no definite signs of irreversible toxicity to normal cultured corneal epithelial and keratocyte cells at examined concentrations and exposure time.