Abstract 13124: Arterial Sampling in Patients Undergoing Coronary Angiography Using a Percutaneous Radial Artery Approach: a New Method for Isolation and Molecular Characterization of Human Arterial Endothelial Cells
Background: Limited availability of endothelial cells (ECs) is a major constraint when investigating the cellular mechanisms of arterial endothelial dysfunction in patients with cardiovascular disease. This study aimed to describe a novel approach that allows collection and molecular characterization of arterial ECs in patients undergoing coronary angiography using a percutaneous radial artery approach.
Methods: Eight consecutive patients (all males, age 64±14 years) undergoing elective coronary angiography using a radial artery approach were studied. Arterial ECs were collected using an endovascular wire which was inserted through a 5 or 6 French sheath, while a tourniquet cuff was inflated around the arm (above systolic pressure) to prevent detachment of the ECs from the wire by the pulsatile arterial flow. Two thirds of the sample underwent EC separation (using magnetic beads coated with EC specific antibodies), mRNA amplification, and Affymetrix HG-U133 Plus 2.0 Microarray analysis. One third of the sample underwent fixation, and quantitative immunofluorescence analysis using Image J. Levels of vonWillebrand factor (vWF), a marker of hypercoagulability, and endothelial nitric oxide synthase (eNOS) protein expression were measured and compared in 6 patients with angiographic evidence of coronary artery disease (CAD) and in 2 patients without angiographic evidence of CAD.
Results: The amplified mRNA product ranged from 33 to 117μg and was adequate for microarray analysis in all cases. Arterial ECs ranged from 34 to 625 cells per slide (total of four slides) and was adequate for quantitative immunofluorescence analysis in all cases. Endothelial vWF expression was 2.4 fold higher in patients with CAD when compared with those without CAD; eNOS expression was similar in both groups.
Conclusion: Reliable sampling of radial artery ECs coupled with gene and protein expression profiling is feasible in patients undergoing percutaneous coronary angiography. This novel method provides a new and more effective approach for the investigation of the cellular mechanisms of arterial endothelial dysfunction in patients with cardiovascular disease.
- © 2013 by American Heart Association, Inc.