March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
A Quantitative In Vivo Assay For Measuring Tnf-α Induced Retinal Vascular Permeability
Author Affiliations & Notes
  • Mike Lin
    Retina Research, Alcon Research Ltd, Fort Worth, Texas
  • Footnotes
    Commercial Relationships  Mike Lin, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 2963. doi:
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      Mike Lin; A Quantitative In Vivo Assay For Measuring Tnf-α Induced Retinal Vascular Permeability. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2963.

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Abstract

Purpose: : Purpose: Blood-retinal barrier breakdown occurs in many ocular diseases associated with inflammation. The effect of inflammatory cytokine Tumor necrosis factor (TNF)-α on retinal vascular permeability was investigated in adult rats, and a quantitative in vivo assay was established.

Methods: : Methods: Adult Sprague Dawley rats were intravitreally challenged with a range of doses (10-1000 ng) of recombinant human (rh) TNF-α. At several time points post TNF-α stimulation, retinal vascular permeability was evaluated by measuring the extravasated vascular tracer Evan’s Blue dye in retinal tissue normalized with plasma Evan’s Blue concentration and retina weight. The optimum amount and duration of rhTNF-α stimulation were used in later inhibitor study, in which two TNF-α specific inhibitors, AL-50738 and AL-88974, were given intravitreally 24 hrs prior to TNF-α injection. In addition, the retinal protein level of Vascular Endothelial Growth Factor (VEGF) after TNF-α treatment was measured by Elisa.

Results: : Results: rh TNF-α dose-dependently induced retinal vascular leakage in rats, with robust leakage observed at 48-72 hours after 300-1000 ng TNF-α intravitreal injection. Two specific TNF-α inhibitors, ESBA105 and ESBA1622 efficaciously prevented TNF-α induced retinal vascular leakage. No significant change in retinal VEGF protein level was observed after recombinant human or rat TNF-α treatment.

Conclusions: : Conclusions: The rat retinal vasculature reproducibly responded to human TNF-α stimulation in a VEGF-independent manner. Evan’s Blue measurement can quantitatively measure TNF-α induced retinal vascular leakage. This in vivo model can be further used for evaluating anti-TNF-α agents.

Keywords: growth factors/growth factor receptors • retina • drug toxicity/drug effects 
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