Abstract
Purpose: :
To study the toxicity of Nepafenac ophthalmic suspension 0.1% (NEVANAC Alcon, Fort Worth, Texas) on human retinal pigment epithelial (ARPE-19) and rat neurosensory (R28) cells in vitro.
Methods: :
ARPE-19 and R28 cells were grown in tissue culture and were treated with five different concentrations of Nepafenac (100, 50, 25, 12,5 and 6,25 µg/ml) for 24 hours. The doses were chosen to bracket the intravitreal clinical dose determined by assuming 0.1 cc of the commercial preparation distributes equally through the 4 ml human vitreous volume. The calculated clinical dose is 25 µg/ml. Toxicity was determined by a trypan blue dye-exclusion assay. Detection of membrane potentials was performed using the JC-1 mitochondrial membrane potential (Δψm) detection kit.
Results: :
The mean cell viability of ARPE-19 cells after 24 hours exposed to 100, 50, 25, 12,5 and 6,25 µg/ml of Nepafenac was: 37.3±16.8(P<0.001), 89.1±5.2(P>0.05), 92.8±1.0(P>0.05), 91.9±0.9(P>0.05) and 92.1±1.3(P>0.05), respectively, compared to untreated cells that had a viability of 92.3±0.9. The ratio of red/green fluorescence (JC-1 potential) in Nepafenac exposed wells for the 100, 50, 25, 12.5 and 6.25 µg/ml was 0.68±0.03(P<0.01), 0.76±0.018 (P<0.05), 0.8±0.10(P<0.05), 0.84±0.13(P>0.05), and 0.94±0.06(P>0.05) respectively, compared to 1±0.0 in untreated ARPE-19 cells.The mean cell viability of R28 cells after 24 hours exposed to 100, 50, 25, 12.5 and 6.25 µg/ml of Nepafenac was: 54.2±7.2(P<0.001), 82.3±12.6(P>0.05), 89.9±5.0(P>0.05), 88.4±3.3(P>0.05) and 86.4±3.1(P>0.05), respectively, compared to untreated cells that had a viability of 89,7±2.5. The ratio of red/green fluorescence in Nepafenac exposed wells for the 100, 50, 25, 12.5 and 6.25 µg/ml was 1.1±0.1(P<0.001), 1.06±00.5(P<0.001), 1.16±0.05(P<0.001), 1.16±0.15(P<0.001), and 1.96±0.3(P>0.05) respectively, compared to 1.9±0.1 in untreated R28 cells.
Conclusions: :
This study suggests that Nepafenac shows significant retinal cytotoxicity finally standard clinical doses in our in vitro ARPE-19 and R28 cell culture system.
Keywords: retinal pigment epithelium • drug toxicity/drug effects • retinal culture