May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Effect of Hydroquinone, a Toxicant in Cigarette Smoke, on Human Retinal Pigment Epithelial Cells, Rat Retinal Neurosensory Cells and Human Microvascular Endothelial Cells in vitro
Author Affiliations & Notes
  • T. H. Huynh
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • A. Sharma
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • A. L. Gramajo
    Department of Ophthalmology, University of California, Irvine, Irvine, California
    Departamento de Oftalmologia, Fundacion VER, Cordoba, Argentina
  • A. Jayaprakash Patil
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • M. F. Estrago Franco
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • M. Chwa
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • G. M. Seigel
    Department of Ophthalmology, University at Buffalo, The State University of New York, Buffalo, New York
  • B. D. Kuppermann
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • M. C. Kenney
    Department of Ophthalmology, University of California, Irvine, Irvine, California
  • Footnotes
    Commercial Relationships  T.H. Huynh, None; A. Sharma, None; A.L. Gramajo, None; A. Jayaprakash Patil, None; M.F. Estrago Franco, None; M. Chwa, None; G.M. Seigel, None; B.D. Kuppermann, None; M.C. Kenney, None.
  • Footnotes
    Support  Discovery Eye Foundation, Henry L. Guenther Foundation, Iris and B. Gerald Cantor Foundation, Gilbert Foundation, Skirball Molecular Ophthalmology Program, Research to Prevent Blindness Foundation
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5936. doi:https://doi.org/
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      T. H. Huynh, A. Sharma, A. L. Gramajo, A. Jayaprakash Patil, M. F. Estrago Franco, M. Chwa, G. M. Seigel, B. D. Kuppermann, M. C. Kenney; Effect of Hydroquinone, a Toxicant in Cigarette Smoke, on Human Retinal Pigment Epithelial Cells, Rat Retinal Neurosensory Cells and Human Microvascular Endothelial Cells in vitro. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5936. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To determine the epidemiologic association between cigarette smoke and age-related macular degeneration, we examined the effects of Hydroquinone (HQ), a toxic element present in high concentration in cigarette-smoke related tar, on human retinal pigment epithelial cells (ARPE-19), rat neurosensory cells (R28) and human microvascular endothelial (HMVEC) cells.

Methods: : ARPE-19, R28, and HMVEC cells were grown in tissue culture and treated with four different concentrations of HQ (50µM, 100µM, 200µM, 500µM). Trypan blue cell viability assay, caspase-3/7 assay, and DNA laddering were performed to analyze the molecular effects of HQ on these cells.

Results: : The mean cell viability percentages of ARPE-19 cells after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 95.70±0.42 (P>0.05), 52.95±2.051 (P<0.001), 33.50± 3.53 (P<0.001) and 26.60±1.56 (P<0.001) respectively compared to 500µM equivalent DMSO controls 96.250±0.78.The mean cell viability percentages of R28 cells after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 67.3± 2.9 (P<0.01), 35.70±3.252 (P<0.001), 26.20± 1.13(P <0.001) and 14.15±1.20 (P<0.001) respectively compared to 500µM equivalent DMSO controls 78.56 ±2.90.The mean cell viability percentages of HMVEC after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 93.80±0.92 (P>0.05), 37.52±1.93 (P<0.001), 25.93± 2.17(P <0.001) and 11.73±2.27 (P<0.001) respectively compared to 500µM equivalent DMSO controls 91.50± 2.19.At 50µM HQ, the R28 cells showed a significant decrease of cell viability 67.3± 2.9 (P<0.01), while ARPE-19 and HMVEC cultures were similar to the DMSO treated controls. All cells showed loss of cell viability at 100µM, 200µM and 500µM HQ. There was no significant increase in caspase-3/7 activities in all three cell lines at all concentrations tested. DNA laddering did not show any banding pattern.

Conclusions: : This study suggests that R28 cells are more sensitive to HQ than human ARPE-19 and HMVEC cells. The mechanism of cell death was necrosis in all three cells lines.

Keywords: ocular irritancy/toxicity testing • photoreceptors • retinal pigment epithelium 
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