Mechanisms underlying Benzyl alcohol cytotoxicity (triamcinolone acetonide preservative) in human retinal pigment epithelial cells

Yi Sheng Chang, Chiou Feng Lin, Chao Liang Wu, Pao Ying Kuo, Fong Sen Wu, Chi Chang Shieh, Po Wu Gean, Shur Tzu Chen, Muh Shy Chen, Wen Chuan Wu, Ming Hong Tai, Sung Huei Tseng

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13 Citations (Scopus)

Abstract

Purpose. Benzyl alcohol (BA) is the preservative in triamcinolone acetonide (TA) suspensions, which are used in treating vitreoretinal diseases and during surgery. This paper investigates the molecular mechanisms and signaling pathways underlying BA toxicity in human retinal pigment epithelial (RPE) cells. Methods. Cultured human RPE cells from the ARPE-19 cell line were exposed to culture medium alone (control) or with BA (0.0225, 0.225, 0.9, 3, or 9 mg/mL) for up to 6 hours. BA toxicity was assessed by TUNEL assay, propidium iodide/annexin V-FITC staining and flow cytometry, caspase activation assay, caspase and apoptosis inhibition assays, mitochondrial transmembrane potential by rhodamine staining and flow cytometry, reactive oxygen species by chemiluminescence, and apoptosis-inducing factor staining. Results. BA caused RPE cell death not only by necrosis but also by apoptosis, evidenced by exposure to 9 mg/mL BA for 6 hours leading to 19.0% early apoptotic cells and 64.2% apoptotic necrotic cells. Apoptotic signaling involved the immediate production of reactive oxygen species, activation of caspase-8, impairment of the mitochondrial transmembrane potential, and further activation of caspase-9 and -3. In addition, BA induced translocation of apoptosis-inducing factor into the nucleus, indicating caspase-independent apoptosis. Conclusions. BA leads to necrosis of RPE cells and triggers mitochondrial apoptosis through both caspase-dependent and - independent pathways. Extreme caution is suggested in the intraocular use of TA suspensions and meticulous evaluation before adoption of BA as a preservative in the future development of ophthalmic formulations.

Original languageEnglish
Pages (from-to)4214-4222
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number7
DOIs
Publication statusPublished - Jun 2011
Externally publishedYes

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

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  • Cite this

    Chang, Y. S., Lin, C. F., Wu, C. L., Kuo, P. Y., Wu, F. S., Shieh, C. C., Gean, P. W., Chen, S. T., Chen, M. S., Wu, W. C., Tai, M. H., & Tseng, S. H. (2011). Mechanisms underlying Benzyl alcohol cytotoxicity (triamcinolone acetonide preservative) in human retinal pigment epithelial cells. Investigative Ophthalmology and Visual Science, 52(7), 4214-4222. https://doi.org/10.1167/iovs.10-6058