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Analysis of in situ and ex vivo αVß3 integrin expression during experimental carotid atherogenesis

Authors Yao Y, Jiang Y, Sheng Z, Zhang Y, An Y, Yan F, Ma G, Liu N, Teng G, Cheng Z

Received 9 November 2011

Accepted for publication 15 December 2011

Published 8 February 2012 Volume 2012:7 Pages 641—649

DOI https://doi.org/10.2147/IJN.S28065

Review by Single-blind

Peer reviewer comments 4

Yuyu Yao1, Yibo Jiang1,3, Zulong Sheng1, Yi Zhang2, Yanli An2, Fengdi Yan1, Genshan Ma1, Naifeng Liu1, Gaojun Teng2, Zhen Cheng4
1Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu, People’s Republic of China; 2Jiangsu Key Lab of Molecular and Function Imaging, Department of Radiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, People’s Republic of China; 3Department of Cardiology, Taixing Hospital, Yangzhou University, Taixing, People's Republic of China; 4Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Bio-X Program, Stanford University School of Medicine, Stanford, CA, USA

Objective: Mural inflammation has been shown to contribute to the development of plaque, with the αVß3 integrin highly expressed in atherosclerotic plaques. We herein examined αVß3 integrin expression as a function of carotid atherosclerosis formation in the apolipoprotein E-deficient (apoE–/–) mouse.
Methods and results: Constrictive collars were placed around the left common carotid arteries of apo E–/– mice maintained on a high-fat diet (n = 14). Before and 21 days following collar placement, in vivo serial magnetic resonance imaging (MRI) measurements of the carotid aortic diameter were performed using a 7T magnetic resonance (MR) scanner. Near-infrared fluorescence (NIRF) imaging was performed (n = 6) using an in vivo imaging system 0–24 hours following administration of 1.0 nmol c(RGDyK)-Cy5.5 via the tail vein. A competition experiment was performed by the co-injection of a saturating dose of bicyclic RGD peptide H-Glu[cyclo(Arg-Gly-Asp-D-Tyr-Lys)]2 (n = 3). Following image acquisition and sacrifice at 24 hours after injection, carotid arteries were harvested for histological analyses. Neointima formation and arterial remodeling in the carotid arteries of apoE–/– mice were induced by the placement of a constrictive collar. Significantly greater fluorescent signals were obtained from constrictive collar left common carotid arteries as compared to uninvolved aortic segments in constrictive collar mice. Binding to stenotic lesions was efficiently blocked in competition experiments. Immunostaining confirmed the presence of mural αVß3 integrin expression in macrophages in the neointima. Signal intensity increased in a macrophage density-dependent fashion in the stenotic segments.
Conclusion: Mural αVß3 integrin expression, as determined using RGD-Cy5.5 near-infrared optical imaging, was increased in carotid arteries with constrictive collars in experimental mice. This expression can estimate the macrophage-bound inflammatory activity of atherosclerotic lesions.

Keywords: near-infrared fluorescence (NIRF), macrophage, αVß3 integrin, carotid atherogenesis

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