PMID- 30858536
OWN - NLM
STAT- MEDLINE
DCOM- 20200924
LR  - 20231006
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Mar 11
TI  - Label-Free Multi Parameter Optical Interrogation of Endothelial Activation in 
      Single Cells using a Lab on a Disc Platform.
PG  - 4157
LID - 10.1038/s41598-019-40612-8 [doi]
LID - 4157
AB  - Cellular activation and inflammation leading to endothelial dysfunction is 
      associated with cardiovascular disease (CVD). We investigated whether a single 
      cell label-free multi parameter optical interrogation system can detect 
      endothelial cell and endothelial progenitor cell (EPC) activation in vitro and ex 
      vivo, respectively. Cultured human endothelial cells were exposed to increasing 
      concentrations of tumour necrosis factor alpha (TNF-alpha) or lipopolysaccharide 
      (LPS) before endothelial activation was validated using fluorescence-activated 
      cell sorting (FACS) analysis of inflammatory marker expression (PECAM-1, 
      E-selectin and ICAM-1). A centrifugal microfluidic system and V-cup array was 
      used to capture individual cells before optical measurement of light scattering, 
      immunocytofluorescence, auto-fluorescence (AF) and cell morphology was 
      determined. In vitro, TNF-alpha promoted specific changes to the refractive index and 
      cell morphology of individual cells concomitant with enhanced photon activity of 
      fluorescently labelled inflammatory markers and increased auto-fluorescence (AF) 
      intensity at three different wavelengths, an effect blocked by inhibition of 
      downstream signalling with Ikappabeta. Ex vivo, there was a significant increase in EPC 
      number and AF intensity of individual EPCs from CVD patients concomitant with 
      enhanced PECAM-1 expression when compared to normal controls. This novel 
      label-free 'lab on a disc' (LoaD) platform can successfully detect endothelial 
      activation in response to inflammatory stimuli in vitro and ex vivo.
FAU - King, Damien
AU  - King D
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland.
FAU - Glynn, MacDara
AU  - Glynn M
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland.
FAU - Cindric, Sandra
AU  - Cindric S
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland.
FAU - Kernan, David
AU  - Kernan D
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland.
FAU - O'Connell, Triona
AU  - O'Connell T
AD  - Dublin City University, School of Biotechnology, Irish Science Separation 
      Cluster, Dublin, Ireland.
FAU - Hakimjavadi, Roya
AU  - Hakimjavadi R
AD  - Dublin City University, School of Biotechnology, Vascular Biology & Therapeutics, 
      Dublin, Ireland.
FAU - Kearney, Sinead
AU  - Kearney S
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland.
FAU - Ackermann, Tobias
AU  - Ackermann T
AD  - Dublin City University, School of Biotechnology, Vascular Biology & Therapeutics, 
      Dublin, Ireland.
FAU - Berbel, Xavier Munoz
AU  - Berbel XM
AUID- ORCID: 0000-0002-6447-5756
AD  - Centre Nacional de Microelectronica, Campus UAB, Barcelona, Spain.
FAU - Llobera, Andreu
AU  - Llobera A
AD  - Centre Nacional de Microelectronica, Campus UAB, Barcelona, Spain.
FAU - Simonsen, Ulf
AU  - Simonsen U
AUID- ORCID: 0000-0002-2169-7666
AD  - Aarhus University, Department of Biomedicine, Aarhus, Denmark.
FAU - Laursen, Britt E
AU  - Laursen BE
AD  - Aarhus University, Department of Biomedicine, Aarhus, Denmark.
FAU - Redmond, Eileen M
AU  - Redmond EM
AD  - University of Rochester, Dept Surgery Rochester, New York, United States.
FAU - Cahill, Paul A
AU  - Cahill PA
AUID- ORCID: 0000-0002-5385-6502
AD  - Dublin City University, School of Biotechnology, Vascular Biology & Therapeutics, 
      Dublin, Ireland.
FAU - Ducree, Jens
AU  - Ducree J
AUID- ORCID: 0000-0002-0366-1897
AD  - Dublin City University, School of Physical Sciences, National Centre for Sensor 
      Research, Dublin, Ireland. jens.ducree@dcu.ie.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190311
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (E-Selectin)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Platelet Endothelial Cell Adhesion Molecule-1)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 126547-89-5 (Intercellular Adhesion Molecule-1)
SB  - IM
MH  - Cell Shape
MH  - E-Selectin/genetics/metabolism
MH  - Flow Cytometry/instrumentation/*methods
MH  - Human Umbilical Vein Endothelial Cells/*cytology/drug effects/metabolism
MH  - Humans
MH  - Intercellular Adhesion Molecule-1/genetics/metabolism
MH  - Lipopolysaccharides/pharmacology
MH  - Platelet Endothelial Cell Adhesion Molecule-1/genetics/metabolism
MH  - Tumor Necrosis Factor-alpha/pharmacology
PMC - PMC6411894
COIS- The authors declare no competing interests.
EDAT- 2019/03/13 06:00
MHDA- 2020/09/25 06:00
PMCR- 2019/03/11
CRDT- 2019/03/13 06:00
PHST- 2018/03/07 00:00 [received]
PHST- 2019/01/31 00:00 [accepted]
PHST- 2019/03/13 06:00 [entrez]
PHST- 2019/03/13 06:00 [pubmed]
PHST- 2020/09/25 06:00 [medline]
PHST- 2019/03/11 00:00 [pmc-release]
AID - 10.1038/s41598-019-40612-8 [pii]
AID - 40612 [pii]
AID - 10.1038/s41598-019-40612-8 [doi]
PST - epublish
SO  - Sci Rep. 2019 Mar 11;9(1):4157. doi: 10.1038/s41598-019-40612-8.