Evaluation of in vitro Effects of Trivaris (Triamcinolone Acetonide) on Human Retinal Pigment Epithelial Cells and M??ller Cells

摘要

Purpose: : To study the in vitro effects of Trivaris (Allergan, Inc.) on human retinal pigment epithelial (ARPE-19) cells and M??ller 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 (???¨m) 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). ???¨m 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 (???¨m 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; ???¨m 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; p0.05. 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.