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ORIGINAL ARTICLE
Year : 2014  |  Volume : 62  |  Issue : 4  |  Page : 429-436

Effects of triamcinolone acetonide on human trabecular meshwork cells in vitro


1 Department of Ophthalmology, Gavin Herbert Eye Institute, University of California, Irvine, CA, USA; Lotus Eye Care Hospital, Coimbatore, TN, India
2 Department of Ophthalmology, Gavin Herbert Eye Institute, University of California, Irvine, CA, USA; Department of Ophthalmology, Clinical and Academic, Great Ormond Street Hospital for Children and King's College Hospital, London
3 Department of Ophthalmology, Gavin Herbert Eye Institute, University of California, Irvine, CA, USA
4 Ocular Genetics and Genomics Lab, CHUL Research Center, Quebec City, QC, Canada

Correspondence Address:
Baruch D Kuppermann
Department of Ophthalmology, University of California Irvine, 118, MedSurge I, Irvine, CA 92697, USA

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Source of Support: Supported by the Discovery Eye Foundation, Henry L. Guenther Foundation, The Iris and B. Gerald Cantor Foundation, The Skirball Molecular Ophthalmology Program, Poly and Michael Smith Foundation, Research to Prevent Blindness Foundation., Conflict of Interest: None


DOI: 10.4103/0301-4738.121143

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Aim: To study the effects of triamcinolone acetonide (TA) on cultured human trabecular meshwork (HTM) cells. Materials and 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.


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