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Abstract #105906 Published in IGR 23-3
Cytotoxic, Apoptotic, and Oxidative Effects of Preserved and Preservative-Free Brimonidine in a Corneal Epithelial Cell Line
Kucukoduk A; Durmus IM; Aksoy M; Karakurt SJournal of Ocular Pharmacology and Therapeutics 2022; 38: 576-583
This study aims to compare the cytotoxic, apoptotic, and oxidative effects of preserved and preservative-free forms of brimonidine 0.15% on the human corneal epithelial cell (HCEC) line. Time-dependent cytotoxicity studies were performed with the Alamar Blue method. For apoptotic studies, PE Annexin V and 7-amino-actinomycin (7-AAD) staining and flow cytometry were performed. Messenger RNA (mRNA) expressions of , and caspase-3, -9, -12, and protein expressions of and were evaluated by quantitative real-time polymerase chain reaction and Western blot method, respectively. Cell viability was 76.4% with the preserved solution and 36.05% with the preservative-free solution at the fifth minute. No significant difference was observed with either solution at the 15-min mark, whereas cell viability did not change significantly after 1 h. In the apoptosis evaluation, it was observed that the preservative-free solution increased the early apoptotic activity to a greater degree ( < 0.05). Preservative-free solution also induced gene expression of proapoptotic , caspase-9 and -12, and protein expression of Bax while reducing the protein expression of anti-apoptotic ( < 0.0001). Preserved solution induced only the gene expression of caspase-12, and reduced the protein expression of Bcl-2 ( < 0.0001). No significant difference was observed in the reactive oxygen species (ROS) levels of either solution compared with the control group ( > 0.05). It was demonstrated that the preserved solution is less cytotoxic to the HCEC line in the early period, has less early apoptotic activity, and does not significantly increase ROS levels.
Department of Ophthalmology, Faculty of Medicine, KTO Karatay University, Konya, Turkey.
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