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Published Online
on October 25, 2007

Arteriosclerosis, Thrombosis, and Vascular Biology. 2007
Published online before print October 25, 2007, doi: 10.1161/ATVBAHA.107.145466
A more recent version of this article appeared on January 1, 2008
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Submitted on July 24, 2007
Accepted on October 17, 2007

Magnetic Resonance Imaging of Endothelial Adhesion Molecules in Mouse Atherosclerosis Using Dual-Targeted Microparticles of Iron Oxide

Martina A. McAteer ; Jurgen E. Schneider ; Ziad A. Ali ; Nicholas Warrick ; Christina A. Bursill ; Constantin von zur Muhlen ; David R. Greaves ; Stefan Neubauer ; Keith M. Channon ; and Robin P. Choudhury *

From the Department of Cardiovascular Medicine (M.A.M., J.E.S., Z.A.A., N.W., C.A.B., D.R.G., S.N., K.M.C., R.P.C.) and Sir William Dunn School of Pathology (C.v.z.M.), Oxford, University of Oxford, United Kingdom.

* To whom correspondence should be addressed. E-mail: robin.choudhury{at}cardiov.ox.ac.uk.

Objective—Microparticles of iron oxide (MPIO) distort magnetic field creating marked contrast effects far exceeding their physical size. We hypothesized that antibody-conjugated MPIO would enable MRI (MRI) of endothelial cell adhesion molecules in mouse atherosclerosis.

Methods and Results—MPIO (4.5 µm) were conjugated to monoclonal antibodies against vascular cell adhesion molecule-1 (VCAM–MPIO) or P-selectin (P-selectin–MPIO). In vitro, VCAM–MPIO bound, in dose-dependent manner, to tumor necrosis factor (TNF)-{alpha} stimulated sEND-1 endothelial cells, as quantified by light microscopy (R2=0.94, P=0.03) and by MRI (R2=0.98, P=0.01). VCAM–MPIO binding was blocked by preincubation with soluble VCAM-1. To mimic leukocyte binding, MPIO targeting both VCAM-1 and P-selectin were administered in apolipoprotein E-/- mice. By light microscopy, dual-targeted MPIO binding to endothelium overlying aortic root atherosclerosis was 5- to 7-fold more than P-selectin–MPIO (P<0.05) or VCAM–MPIO (P<0.01) alone. Dual-targeted MPIO, injected intravenously in vivo bound aortic root endothelium and were quantifiable by MRI ex vivo (3.5-fold increase versus control; P<0.01). MPIO were well-tolerated in vivo, with sequestration in the spleen after 24 hours.

Conclusions—Dual-ligand MPIO bound to endothelium over atherosclerosis in vivo, under flow conditions. MPIO may provide a functional MRI probe for detecting endothelial-specific markers in a range of vascular pathologies.
Key words: microparticles of iron oxide • atherosclerosis • magnetic resonance imaging • molecular imaging




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