May 2006
Volume 47, Issue 13
ARVO Annual Meeting Abstract  |   May 2006
NSAID–Induced Swelling of Peroxide–Stressed Mitochondria in vitro
Author Affiliations & Notes
  • G. Graff
    Ophthalmic Product Research, Alcon, Fort Worth, TX
  • J.–Y. Wei
    Ophthalmic Product Research, Alcon, Fort Worth, TX
  • Footnotes
    Commercial Relationships  G. Graff, Alcon Research, Ltd., E; J. Wei, Alcon Research, LTD., E.
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 2760. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      G. Graff, J.–Y. Wei; NSAID–Induced Swelling of Peroxide–Stressed Mitochondria in vitro . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2760.

      Download citation file:

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

  • Supplements

Purpose: : Assess the in vitro effect of nepafenac and selected NSAIDs on mitochondria under normal and peroxide–stress conditions.

Methods: : Mitochondria were prepared from livers of male Sprague Dawley rats according to Broekemeier at al. and were resuspended in ice–cold, iso–osmotic HEPES buffer (3.0 mM), supplemented with mannitol (207 mM), and sucrose (63mM) (pH 7.4). An appropriate aliquot of the mitochondrial preparation was added to 2.95 mL of iso–osmotic, HEPES (3.0 mM) buffer containing sodium succinate (10 mM), and rotenone (1 nmol/mg protein) (pH 7.4). Mitochondrial swelling was monitored at 540 nm with an OLIS/Aminco DW2c dual–beam spectrophotometer. Swelling (positive control) was induced by addition of CaCl2 (60 nmol/mg mitochondrial protein) and tertiary butylhydroperoxide (t–BHP;150 µM).

Results: : Exposure of unstressed mitochondria to concentrations of 100 µM of nepafenac, amfenac, bromfenac, ketorolac, ibuprofen and diclofenac in iso–osmotic buffer did not promote swelling as assessed by light scattering. However, when stressed with t–BHP (which by itself does not promote swelling) mitochondria underwent rapid swelling upon addition of i.e., diclofenac. The rate and extent of organelle swelling increased with increasing concentrations of NSAID added. This behavior was qualitatively similar for every NSAID tested, including ibuprofen, amfenac, bromfenac, dicofenac, and ketorolac. The order of potency for induction of half–maximal swelling was bromfenac (EC50 = 9 µM) = amfenac (EC50 = 10 µM) > diclofenac (EC50 = 19 µM) = ibuprofen (EC50 = 22 µM) > ketorolac (EC50 = 529 µM). Notably, nepafenac, the carboxamide derivative and prodrug form of amfenac, did not promote mitochondrial swelling even at the highest concentration tested (i.e., 600 µM).

Conclusions: : Classical NSAIDs with free carboxylic acid function were innocuous to unstressed, isolated liver mitochondria, but caused concentration–dependent swelling in the presence of low concentrations of peroxide. Nepafenac, the carboxamide derivative and prodrug form of amfenac, is unique among clinically used ocular NSAIDs in that it had no detectable toxic effects on mitochondria, even when stressed with peroxide.

Keywords: mitochondria • wound healing 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.