Microfluidic assay of circulating endothelial cells in coronary artery disease patients with angina pectoris

• Published in: PloS one Vol. 12; no. 7; p. e0181249
• Main Authors: Chen, Shuiyu, Sun, Yukun, Neoh, Kuang Hong, Chen, Anqi, Li, Weiju, Yang, Xiaorui, Han, Ray P S
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 13.07.2017; Public Library of Science (PLoS)
• Subjects: Aged; Angina; Angina pectoris; Angina Pectoris - blood; Angina Pectoris - complications; Angina Pectoris - diagnosis; Assaying; Atherosclerosis; Beads; Biology and Life Sciences; Biomarkers; Biomarkers - blood; Blood Cell Count; Blood Cells - cytology; Cardiology; Cardiovascular disease; Case-Control Studies; CD45 antigen; Cell growth; Cell Separation - methods; Cells, Cultured; Coronary artery; Coronary artery disease; Coronary Artery Disease - blood; Coronary Artery Disease - complications; Coronary Artery Disease - diagnosis; Coronary heart disease; Coronary vessels; Cytometry; Damage assessment; Diagnostic systems; Emergency medical care; Endothelial cells; Endothelial Cells - cytology; Endothelium, Vascular - cytology; Engineering and Technology; Engineering schools; Enumeration; Female; Flow cytometry; Glass substrates; Heart attacks; Heart diseases; Human Umbilical Vein Endothelial Cells - cytology; Humans; Identification; Male; Medical imaging; Medicine and Health Sciences; Microfluidics; Microfluidics - methods; Middle Aged; Patients; Peripheral blood; Physical Sciences; Prognosis; Research and Analysis Methods; Risk; Silicon wafers; Subgroups; Task forces; Vascular endothelial growth factor; Beijing China; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0181249

Circulating endothelial cells (CECs) are widely reported as a promising biomarker of endothelial damage/dysfunction in coronary artery disease (CAD). The two popular methods of CEC quantification include the use of immunomagnetic beads separation (IB) and flow cytometry analysis (FC); however, they suffer from two main shortcomings that affect their diagnostic and prognostic responses: non-specific bindings of magnetic beads to non-target cells and a high degree of variability in rare cell identification, respectively. We designed a microfluidic chip with spatially staggered micropillars for the efficient harvesting of CECs with intact cellular morphology in an attempt to revisit the diagnostic goal of CEC counts in CAD patients with angina pectoris. A label-free microfluidic assay that involved an in-situ enumeration and immunofluorescent identification (DAPI+/CD146+/VEGFR1+/CD45-) of CECs was carried out to assess the CEC count in human peripheral blood samples. A total of 55 CAD patients with angina pectoris [16 with chronic stable angina (CSA) and 39 with unstable angina (UA)], together with 15 heathy controls (HCs) were enrolled in the study. CEC counts are significantly higher in both CSA and UA groups compared to the HC group [respective medians of 6.9, 10.0 and 1.5 cells/ml (p < 0.01)]. Further, a significant elevation of CEC count was observed in the three UA subgroups [low risk (5.3) vs. intermediate risk (10.8) vs. high risk (18.0) cells/ml, p < 0.001) classified in accordance to the TIMI NSTEMI/UA risk score system. From the receiver-operating characteristic curve analysis, the AUCs for distinguishing CSA and UA from HC were 0.867 and 0.938, respectively. The corresponding sensitivities were 87.5% and 84.6% and the specificities were 66.7% and 86.7%, respectively. Our microfluidic assay system is efficient and stable for CEC capture and enumeration. The results showed that the CEC count has the potential to be a promising clinical biomarker for the assessment of endothelial damage/dysfunction in CAD patients with angina pectoris.