Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation.

Abstract

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

• Authors: Caitlin O'Rourke, Georgia Shelton, Joshua D Hutcheson, Megan F Burke, Trejeeve Martyn, Timothy E Thayer, Hannah R Shakartzi, Mary D Buswell, Robert E Tainsh, Binglan Yu, Aranya Bagchi, David K Rhee, Connie Wu, Matthias Derwall, Emmanuel S Buys, Paul B Yu, Kenneth D Bloch, Elena Aikawa, Donald B Bloch, Rajeev Malhotra
• Journal: Journal of visualized experiments : JoVE (J Vis Exp) Issue 111 (05 31 2016) ISSN: 1940-087X [Electronic] United States
• PMID: 27284788 (Publication Type: Journal Article, Video-Audio Media)

Topics

• Animals
• Aortic Diseases (diagnostic imaging, etiology, metabolism)
• Atherosclerosis (diagnostic imaging, etiology, metabolism)
• Calcinosis (diagnostic imaging)
• Humans
• Image Interpretation, Computer-Assisted
• Inflammation
• Mice
• Muscle, Smooth, Vascular (diagnostic imaging, metabolism, pathology)