TY - JOUR
T1 - Endothelial Glycocalyx Structure in the Intact Carotid Artery: A Two-Photon Laser Scanning Microscopy Study
AU - Reitsma, Sietze
AU - Egbrink, Mirjam G. A. Oude
AU - Vink, Hans
AU - van den Berg, Bernard M.
AU - Passos, Valeria Lima
AU - Engels, Wim
AU - Slaaf, Dick W.
AU - van Zandvoort, Marc A. M. J.
PY - 2011
Y1 - 2011
N2 - Background: The endothelial glycocalyx (EG) is the carbohydrate-rich luminal lining of endothelial cells that mediates permeability and blood cell-vessel wall interactions. To establish an atheroprotective role of the EG, adequate imaging and quantification of its properties in intact, viable, atherogenesis-prone arteries is needed. Methods: Carotid arteries of C57Bl6/J mice (n = 22) were isolated including the bifurcation, mounted in a perfusion chamber, and perfused with fluorescent lectin wheat germ agglutinin-fluorescein isothiocyanate. The EG was visualized through the vessel wall using two-photon laser scanning microscopy. An image quantification protocol was developed to assess EG thickness, which was sensitive to hyaluronidase-induced changes. Results: In the lesion-protected common carotid artery, EG thickness was found to be 2.3 +/- 0.1 mu m (mean +/- SEM), while the surface area devoid of (wheat germ agglutinin-sensitive) EG was 8.9 +/- 4.2%. Data from the external carotid artery were similar (2.5 +/- 0.1 mu m; 9.1 +/- 5.0%). In the atherogenesis-prone internal carotid artery the EG-devoid surface area was significantly higher (27.4 +/- 5.5%, p <0.05); thickness at the remaining areas was 2.5 +/- 0.1 mu m. Conclusion: The EG can be adequately imaged and quantified using two-photon laser scanning microscopy in intact, viable mounted carotid arteries. Spatial EG differences could underlie atherogenesis.
AB - Background: The endothelial glycocalyx (EG) is the carbohydrate-rich luminal lining of endothelial cells that mediates permeability and blood cell-vessel wall interactions. To establish an atheroprotective role of the EG, adequate imaging and quantification of its properties in intact, viable, atherogenesis-prone arteries is needed. Methods: Carotid arteries of C57Bl6/J mice (n = 22) were isolated including the bifurcation, mounted in a perfusion chamber, and perfused with fluorescent lectin wheat germ agglutinin-fluorescein isothiocyanate. The EG was visualized through the vessel wall using two-photon laser scanning microscopy. An image quantification protocol was developed to assess EG thickness, which was sensitive to hyaluronidase-induced changes. Results: In the lesion-protected common carotid artery, EG thickness was found to be 2.3 +/- 0.1 mu m (mean +/- SEM), while the surface area devoid of (wheat germ agglutinin-sensitive) EG was 8.9 +/- 4.2%. Data from the external carotid artery were similar (2.5 +/- 0.1 mu m; 9.1 +/- 5.0%). In the atherogenesis-prone internal carotid artery the EG-devoid surface area was significantly higher (27.4 +/- 5.5%, p <0.05); thickness at the remaining areas was 2.5 +/- 0.1 mu m. Conclusion: The EG can be adequately imaged and quantified using two-photon laser scanning microscopy in intact, viable mounted carotid arteries. Spatial EG differences could underlie atherogenesis.
KW - Endothelial glycocalyx
KW - Endothelial surface layer
KW - Multiphoton microscopy
KW - Lectin
KW - Image analysis
U2 - 10.1159/000322176
DO - 10.1159/000322176
M3 - Article
C2 - 21273784
SN - 1018-1172
VL - 48
SP - 297
EP - 306
JO - Journal of Vascular Research
JF - Journal of Vascular Research
IS - 4
ER -