Endothelial Glycocalyx Structure in the Intact Carotid Artery: A Two-Photon Laser Scanning Microscopy Study

Sietze Reitsma*, Mirjam G. A. Oude Egbrink, Hans Vink, Bernard M. van den Berg, Valeria Lima Passos, Wim Engels, Dick W. Slaaf, Marc A. M. J. van Zandvoort

*Corresponding author for this work

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Abstract

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.
Original languageEnglish
Pages (from-to)297-306
JournalJournal of Vascular Research
Volume48
Issue number4
DOIs
Publication statusPublished - 2011

Keywords

  • Endothelial glycocalyx
  • Endothelial surface layer
  • Multiphoton microscopy
  • Lectin
  • Image analysis

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