May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Cytotoxicity of Triamcinolone Acetonide on Human Retinal Pigment Epithelial Cells in vitro
Author Affiliations & Notes
  • J. Zhang
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • M. Zou
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • Y. Liu
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • F. Lu
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
    Yale Eye Center, Yale University School of Medicine, New Haven, CT
  • M. Zhang
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • M. Zhang
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • L. Ma
    Dept. of Ophthalmology, West China Hospital, Sichuan University, Chengdu, China
  • Footnotes
    Commercial Relationships  J. Zhang, None; M. Zou, None; Y. Liu, None; F. Lu, None; M. Zhang, None; M. Zhang, None; L. Ma, None.
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 5120. doi:
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      J. Zhang, M. Zou, Y. Liu, F. Lu, M. Zhang, M. Zhang, L. Ma; Cytotoxicity of Triamcinolone Acetonide on Human Retinal Pigment Epithelial Cells in vitro . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5120.

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

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Abstract

Purpose: : To study the influence of triamcinolone acetonide (TA) on the growth and proliferation of human retinal pigment epithelial (RPE) cells, and to observe the morphological and ultrastructural changes of RPE cells induced by TA.

Methods: : Normal human RPE cells were cultured in vitro, and identified through observing the morphologic and cytochemical staining of keratin. TA (0.4 mg/ml,0.2 mg/ml,0.1 mg/ml,0.05 mg/ml,0.025mg/ml) was added to RPE cultures on day 0 and then subsequently for 1,3,5 and 7 days. The amount of cell proliferations with or without TA treatment was determined through MTT assay. All samples were read in triplicate. Since the 50% inhibition ratio of TA on day 3 was about 0.3mg/ml, so with 0.3mg/ml TA at day 3, we observed the changes of cell cycle with Flow Cytometry (FCM) analysis, and the morphological and ultrastructural changes through inverted phase–contrast microscope and transmission electron microscope (TEM) respectively.

Results: : Normal RPE growth curve was similar to an "S" type, which contained the adaptative phase in first 2 days, then into logarithmic growth phase, and finally into platform phase at about the 7th day. TA at the dose ranging from 0.4 to 0.025mg/ml caused a significant reduction in cell proliferations in a dose–dependent manner. And the cells treated with 0.3mg/ml TA compared to the control were decreased 10.6% in S phase (P<0.05).Under light microscope, TA–treated cells were less than the control group, and their shape was irregular with many prominences and cellular vacuoles, even some debris. Under the TEM, cell chromatin was maldistributed and cytoplasm was cavitated, even some necrosis cells.

Conclusions: : TA inhibites the karyomitosis by decrease cells in S phase, and eventually inhibites the proliferation; while the cellular necrosis of induced by TA, which might indicate the cytotoxicity of TA on RPE in vitro.

Keywords: drug toxicity/drug effects • retinal pigment epithelium • apoptosis/cell death 
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