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A novel nonradioactive method to evaluate vascular barrier breakdown and leakage
George Trichonas1, Akrivi Manola, Yuki Morizane
1Retina Service, Angiogenesis Laboratory Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Investigative Ophthalmology & Visual Science
|October 31, 2009
Summary
This study introduces a new, nonradioactive assay using biotinylated bovine serum albumin (bBSA) to measure vascular leakage, offering a sensitive method for assessing blood-retinal barrier breakdown in rodents.
Area of Science:
- Biomedical Science
- Ophthalmology
- Physiology
Background:
- Vascular permeability is crucial in ocular diseases.
- Current methods for assessing vascular leakage can be limited.
- A sensitive, nonradioactive assay is needed for retinal vascular permeability studies.
Purpose of the Study:
- To develop and validate a novel, sensitive, nonradioactive assay for quantifying vascular leakage.
- To assess the assay's utility in measuring blood-retinal barrier (BRB) breakdown in rats and mice.
Main Methods:
- Induction of vascular barrier breakdown using vascular endothelial growth factor (VEGF), lipopolysaccharide (LPS), or diabetes.
- Administration of biotinylated bovine serum albumin (bBSA) as a tracer.
- Detection of extravasated bBSA via immunoprecipitation, Western blot, or ELISA, with normalization to plasma concentration and circulation time.
Main Results:
- VEGF injection increased BRB breakdown 2.5-fold in rats.
- LPS induced significant vascular leakage in both injected and contralateral eyes.
- Diabetic rats showed a threefold increase in vascular leakage.
- LPS in mice increased leakage in retina (1.7-fold), brain (1.5-fold), and kidney (1.3-fold).
Conclusions:
- Biotinylated bovine serum albumin (bBSA) is an effective, nonradioactive alternative for quantifying vascular leakage.
- The bBSA assay is suitable for assessing blood-retinal barrier breakdown.
- This method is cost-effective, easy to perform, and adaptable for mouse models.
