Evaluation of the toxicity of triamcinolone acetonide and dexamethasone sodium phosphate on human lens epithelial cells (HLE B-3)

J Ocul Pharmacol Ther. 2011 Jun;27(3):265-71. doi: 10.1089/jop.2010.0120. Epub 2011 May 16.

Abstract

Purpose: The purpose of this study was to compare the in vitro effects of triamcinolone acetonide (TA) and dexamethasone sodium phosphate (DEX) on human lens epithelial cells (HLE B-3).

Methods: HLE B-3 cells were exposed for 24 h to commercially available TA (c-TA) and dimethylsulfoxide-solubilized TA (s-TA). The cells were treated with 1,000 (clinical dose), 750, 500, 200, and 100 μg/mL concentrations of c-TA, s-TA, and supernatant for 24 h. The cells were also treated with DEX at 2, 1, 0.5, 0.2, 0.1 (clinical dose), and 0.05 mg/mL. Cell viability, caspase-3/7 activity, and DNA fragmentation analyses were performed.

Results: The mean cell viabilities of HLE B-3 after exposure to c-TA at 1,000, 750, 500, 200, and 100 μg/mL were significantly reduced compared with control untreated cells. The s-TA also significantly reduced cell viability at 1,000, 750, and 500 μg/mL compared with dimethylsulfoxide control. The supernatant did not reduce cell viability. Caspase-3/7 activity significantly increased after treatment with c-TA and s-TA. DNA laddering revealed bands at 200 bp intervals with both c-TA at≥100 μg/mL and s-TA at ≥500 μg/mL. The cell viabilities of HLE B-3 after 24 h exposure to DEX were significantly reduced at 2 and 1 mg/mL but not at lower concentrations tested. Caspase-3/7 activities in HLE B-3 cells were not increased significantly after treatment with DEX at any dose tested. DNA laddering did not reveal any band at any dose tested.

Conclusion: This study showed that TA at its clinical dose (1,000 μg/mL) in both commercial preparation and solubilized forms decrease HLE B-3 cell viability through an apoptotic pathway. DEX at its clinical dose (0.1 mg/mL) does not decrease cell viability or cause any increase of caspase-3/7 activity. This study suggests that for long-term sustained-release devices, DEX may be less damaging to human lens cells than TA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents / chemistry
  • Anti-Inflammatory Agents / toxicity*
  • Apoptosis / drug effects
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Line
  • Cell Survival / drug effects
  • DNA Fragmentation / drug effects
  • Delayed-Action Preparations / chemistry
  • Delayed-Action Preparations / toxicity
  • Dexamethasone / analogs & derivatives*
  • Dexamethasone / toxicity
  • Dimethyl Sulfoxide / chemistry
  • Epithelial Cells / drug effects*
  • Glucocorticoids / chemistry
  • Glucocorticoids / toxicity*
  • Humans
  • Lens, Crystalline / drug effects*
  • Osmolar Concentration
  • Pharmaceutical Vehicles / chemistry
  • Solubility
  • Triamcinolone Acetonide / chemistry
  • Triamcinolone Acetonide / toxicity*

Substances

  • Anti-Inflammatory Agents
  • Delayed-Action Preparations
  • Glucocorticoids
  • Pharmaceutical Vehicles
  • dexamethasone 21-phosphate
  • Dexamethasone
  • CASP3 protein, human
  • CASP7 protein, human
  • Caspase 3
  • Caspase 7
  • Triamcinolone Acetonide
  • Dimethyl Sulfoxide