Purchase this article with an account.
A. Hafezi-Moghadam, S. Miyahara, L. Almulki, K. Noda, T. Nakazawa, T. Hisatomi, K. L. Thomas, J. W. Miller, S. Masli, E. S. Gragoudas; In vivo Imaging of Endothelial Injury in the Choriocapillaris During Endotoxin-Induced Uveitis. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3450.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Early diagnosis of ocular inflammation may prevent vision loss. Therefore, clinical outcomes could be improved, if inflammation could be detected prior to structural damage. We aim to develop a non-invasive technique for molecular imaging of endothelial injury in the choroidal vessels of live animals.
Rats were injected with 100µg of LPS to cause endotoxin-induced uveitis (EIU). Recombinant P-selectin glycoprotein ligand-1 immunoglobulin (YSPSL, Y's Therapeutics, Inc.) was conjugated to fluorescent microspheres (2µm diam.), and PSGL-1 site densities were determined by flow cytometry. Microspheres were injected systemically into anesthetized rats, and their rolling and adhesion in the choroidal vessels were investigated under physiologic flow by scanning laser ophthalmoscopy (SLO). Furthermore, the expression of P-selectin in the choroidal vessels was measured by reverse transcription-PCR at different time points after LPS.
YSPSL conjugated microspheres rolled (4±1) and firmly adhered (14±1) to the choroidal endothelium in normal animals (n=6) at baseline levels, resembling constitutive leukocyte endothelial interaction. However, in EIU animals the rolling numbers significantly increased, with a peak 4-10h after LPS (13±2, n=6, P<0.01). Similarly, the adhesion numbers peaked at 4h after LPS in the temporal areas of the choroidal microcirculation (9.5-fold, 133±10, n=6, P<0.01), while there was a biphasic peak at 4h (6.9-fold, P<0.01) and 36h (5.5-fold, P<0.01) after LPS around the optic disk. Consistent with our functional data, choroidal P-selectin mRNA expression in these animals peaked at 4 and 36h after LPS.
We introduce a novel non-invasive method for detection of choroidal endothelial surface antigens in vivo, allowing quantitative assessment of endothelial injury. Though the bio-safety of these microspheres in humans is currently unknown, this technique could be further developed to diagnose subclinical signs of ocular inflammatory diseases such as diabetic retinopathy, uveitis, or AMD.
This PDF is available to Subscribers Only