title: Effects of triamcinolone acetonide on human trabecular meshwork cells in vitro.
abstract: OBJECTIVE
  To study the effects of triamcinolone acetonide (TA) on cultured human trabecular meshwork (HTM) cells.
  
  
  METHODS
  HTM cells were cultured and treated with 125, 250, 500 and 1000 μg/mL concentration of TA for 24 h. The cells were treated with both crystalline TA (TA-C) (commercial preparation) and solubilized TA (TA-S). Cell viability was measured by a trypan blue dye exclusion test. The activity of caspse-3/7 was measured by a fluorescence caspase kit and DNA laddering was evaluated by electrophoresis on 3% agarose gel. Levels of lactate dehydrogenase (LDH) were assessed with LDH cytotoxicity assay kit-II.
  
  
  RESULTS
  Mean cell viabilities of HTM cells after 24 h exposure to TA-C 125, 250, 500, and 1000 μg/mL were 75.4 ±2.45% (P < 0.0001), 49.43 ± 1.85% (P < 0.0001), 17.07 ± 2.39% (P < 0.0001), and 3.7 ± 0.9% (P < 0.0001), respectively, compared with the untreated HTM cells 92.49 ± 1.21%. The mean cell viabilities with 125, 250, 500, and 1000 μg/mL of TA-S were 94.47 ± 1.60% (P > 0.05), 90.13 ± 0.40% (P < 0.01), 85.57 ± 0.47% (P < 0.001), and 71.67 ± 3.30% (P < 0.0001), respectively, compared to DMSO-equivalent cultures. Untreated HTM control had a cell viability of 96.57 ± 1.98%. DMSO-treated controls of 125, 250, 500, and 1000 μg/mL had a cell viability of 94.73 ± 0.57%, 96.97 ± 1.08%, 93.97 ± 1.85%, and 97.27 ± 1.15%, respectively. There was no increase of caspase-3/7 activity in cultures treated with either TA-C or TA-S. DNA laddering showed no bands in the TA-C or TA-S treated cultures. There were significantly higher LDH release rates at all concentrations of TA-C compared to TA-S.
  
  
  CONCLUSIONS
  Results show that the effect of TA-C and TA-S on HTM cells is due to cell death by necrosis at all concentrations except 125 μg/mL of TA-S. Elevated levels of LDH confirmed necrotic cell death. Our study also infers the relative safety of TA-S over TA-C.
authors.forename: Ashish
authors.initials: A
authors.lastname: Sharma
authors.forename: A Jayaprakash
authors.initials: AJ
authors.lastname: Patil
authors.forename: Navin
authors.initials: N
authors.lastname: Gupta
authors.forename: M F
authors.initials: MF
authors.lastname: Estrago-Franco
authors.forename: Saffar
authors.initials: S
authors.lastname: Mansoor
authors.forename: Vincent
authors.initials: V
authors.lastname: Raymond
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Kenney
authors.affiliation: Department of Ophthalmology, Gavin Herbert Eye Institute, University of California, Irvine, CA, USA, .
authors.forename: Baruch D
authors.initials: BD
authors.lastname: Kuppermann
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D014129:Trabecular Meshwork / Q000166:cytology* / Q000187:drug effects; D014222:Triamcinolone Acetonide / Q000494:pharmacology*
chemical_list: D005938:Glucocorticoids; D014222:Triamcinolone Acetonide