A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus

Endothelial dysfunction is one of the common complications in type-2 diabetes mellitus (T2DM) known to increase the risk of cardiovascular diseases[1, 2]. Quantification of circulating endothelial cells (CECs), endothelial progenitor cells (EPC) and endothelial micro-particles (EMP) in peripheral bl...

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Main Author: Chua, Yong Liang
Other Authors: School of Chemical and Biomedical Engineering
Format: Final Year Project
Language:English
Published: 2014
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Online Access:http://hdl.handle.net/10356/61578
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-615782023-03-03T15:40:52Z A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus Chua, Yong Liang School of Chemical and Biomedical Engineering Luo Qian Kathy DRNTU::Engineering::Bioengineering Endothelial dysfunction is one of the common complications in type-2 diabetes mellitus (T2DM) known to increase the risk of cardiovascular diseases[1, 2]. Quantification of circulating endothelial cells (CECs), endothelial progenitor cells (EPC) and endothelial micro-particles (EMP) in peripheral blood is an essential indicator for the assessment of endothelial dysfunction and could serve as potential biomarkers for the risk assessment of vascular related diseases[3]. To achieve this, a rigorous method in flow cytometric analysis is important to ensure consistency and reliability of data. This study focused on the development of flow cytometric protocols for the quantification of CEC, EMP and EPC in type 2 diabetes mellitus patients and to determine correlations between variables of endothelial dysfunction to HbA1c and hyperglycemia. Treatment were given to patient to see any improvement for CEC, EMP and EPC population in endothelial dysfunction. Blood samples from 17 patients were analyzed at the interval of 4 months to allow treatment to take effect. The sample were analyzed phenotypically by flow cytometry for the expression of stem cell marker (CD34) and endothelial cell antigens (CD146, KDR/VEGFR-2), and enumeration of EPC, CEC and EMP. Three-color flow cytometric protocols used for the quantification of CECs and EMPs while the quantification of EPCs were based on the ISHAGE protocol. Correlation analysis was performed to find out if there was a relationship between endothelial function and blood sugar levels. From the experiment, HbA1c correlate to endothelial function as measured by the EPCs and CECs in this study. Quantification of CEC, EPC and EMP based on a negative control gating strategy was a more reliable and reproducible approach, and can be used to establish a clear relationship between endothelial dysfunction and common indicators in diabetes. The methods established in this study provide the potential use in enumeration of endothelial cells for monitoring T2DM patients. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2014-06-16T03:21:02Z 2014-06-16T03:21:02Z 2014 2014 Final Year Project (FYP) http://hdl.handle.net/10356/61578 en Nanyang Technological University 44 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Engineering::Bioengineering
spellingShingle DRNTU::Engineering::Bioengineering
Chua, Yong Liang
A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
description Endothelial dysfunction is one of the common complications in type-2 diabetes mellitus (T2DM) known to increase the risk of cardiovascular diseases[1, 2]. Quantification of circulating endothelial cells (CECs), endothelial progenitor cells (EPC) and endothelial micro-particles (EMP) in peripheral blood is an essential indicator for the assessment of endothelial dysfunction and could serve as potential biomarkers for the risk assessment of vascular related diseases[3]. To achieve this, a rigorous method in flow cytometric analysis is important to ensure consistency and reliability of data. This study focused on the development of flow cytometric protocols for the quantification of CEC, EMP and EPC in type 2 diabetes mellitus patients and to determine correlations between variables of endothelial dysfunction to HbA1c and hyperglycemia. Treatment were given to patient to see any improvement for CEC, EMP and EPC population in endothelial dysfunction. Blood samples from 17 patients were analyzed at the interval of 4 months to allow treatment to take effect. The sample were analyzed phenotypically by flow cytometry for the expression of stem cell marker (CD34) and endothelial cell antigens (CD146, KDR/VEGFR-2), and enumeration of EPC, CEC and EMP. Three-color flow cytometric protocols used for the quantification of CECs and EMPs while the quantification of EPCs were based on the ISHAGE protocol. Correlation analysis was performed to find out if there was a relationship between endothelial function and blood sugar levels. From the experiment, HbA1c correlate to endothelial function as measured by the EPCs and CECs in this study. Quantification of CEC, EPC and EMP based on a negative control gating strategy was a more reliable and reproducible approach, and can be used to establish a clear relationship between endothelial dysfunction and common indicators in diabetes. The methods established in this study provide the potential use in enumeration of endothelial cells for monitoring T2DM patients.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Chua, Yong Liang
format Final Year Project
author Chua, Yong Liang
author_sort Chua, Yong Liang
title A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
title_short A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
title_full A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
title_fullStr A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
title_full_unstemmed A flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
title_sort flow cytometric approach to evaluate endothelial dysfunction in patients with type 2 diabetes mellitus
publishDate 2014
url http://hdl.handle.net/10356/61578
_version_ 1759858040882855936