Abstract
Purpose: :
To study the in vitro effects of Trivaris (Allergan, Inc.) on human retinal pigment epithelial (ARPE-19) cells and Müller cells (MIO-M1).
Methods: :
ARPE-19 cells and MIO-M1 cells were treated for 24 hours with 1000, 500, 200 or 100 µg/mL of Trivaris or hydrogel control. Cell viability (CV) was measured using trypan blue dye exclusion assay, mitochondrial membrane potential (ΔΨm) was measured using JC-1 assay, and caspase-3/7 activity was measured to determine apoptosis.
Results: :
There was a significant dose-dependent decrease in cell viability of ARPE-19 and MIO-M1 cells when treated with 1000 µg/mL (clinical dose), 500 µg/mL or 200 µg/mL Trivaris compared to hydrogel controls (ARPE-19 cells 32.9±0.05, 39.5±1.05, and 51.3±0.6 respectively versus 87.2±0.7, 87.4±0.5, and 91.2±1.05 respectively; MIO-M1 cells 13.3±0.7, 27.9±0.7, 48.9±1.75 versus 89.3±0.4, 86.2±1.3, 84.5±0.5). ΔΨm was decreased and caspase 3/7 activity increased in ARPE-19 and MIO-M1 cells treated with Trivaris 1000 µg/mL, 500 µg/mL and 200 µg/mL compared to hydrogel controls (ΔΨm ARPE-19 cells 1.3±0.01, 2.2±0.2, 3.8± 0.3 versus 5.7±0.09, 7.8±0.42, 8±0.4; ΔΨm MIO-M1 cells 0.7±0.05, 0.6±0.1, 1.0±0.06 versus 1.8±0.03, 2.3±0.03, 3.6±0.06; caspase 3/7 activity ARPE-19 cells 18800±893, 16260±66.9, 11800±425.2 versus 10440±282.4, 6956±140.9, 4519±34.65; caspase 3/7 MIO-M1 cells 26850±4794, 18090±3348, 13530±1029 versus 9157±293, 7216±59.3, 6927±363.8). Cells treated with 100 µg/mL Trivaris were not significantly affected: CV 71.3±0.6 for ARPE-19 cells and 74.9±3.4 for MIO-M1 cells versus 81.2±1.05 and 82.5±2.5 respectively; p>0.05.
Conclusions: :
Trivaris is a preservative-free triamcinolone acetonide formulation in a hydrogel vehicle approved by FDA for intraocular use. Our in vitro study shows that Trivaris exhibits cytotoxicity at doses comparable to other triamcinolone acetonide preparations such as Kenalog, Triesence, and preservative free Triamcinolone Acetonide.
Keywords: retinal culture • retinal degenerations: cell biology • Muller cells