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1 Medical Physics, University of Amsterdam, Amsterdam, The Netherlands
* To whom correspondence should be addressed. E-mail: h.vink{at}amc.uva.nl.
The endothelial glycocalyx shields the vascular wall from direct exposure to flowing blood, contributes to the vascular permeability barrier and its anti-adhesive properties, and stimulates endothelial release of nitric oxide (NO) by mechanotransducing fluid shear stresses. It was hypothesized that perturbation of the endothelial glycocalyx contributes to the increased vulnerability of the arterial wall exposed to atherogenic risk factors. The endothelial glycocalyx and intima media ratios (IMR) were studied at two distinct sites of the murine carotid artery, i.e., at a low risk region proximal to the common carotid arterial bifurcation and at the high risk sinus region of the internal carotid branch. Glycocalyx and IMR were studied in control C57Bl/6J mice (n = 6) and in age matched C57Bl/6J/apoE*3-Leiden mice (n = 6) exposed to an atherogenic diet. Electron micrographs of the endothelial glycocalyx stained with Alcian blue and acridine orange revealed significantly thinner glycocalyces and greater IMR at the carotid sinus region of C57Bl/6J mice than in the common carotid artery (glycocalyx: 73 ± 36 nm vs. 399 ± 174 nm (P < 0.05); IMR: 0.096 ± 0.045 vs. 0.044 ± 0.023 (P < 0.05); sinus vs. common region). Exposure of age matched C57Bl/6J/apoE*3-Leiden mice to an atherogenic diet for 6 weeks decreased common region glycocalyx dimension (399 ± 174 nm vs. 100 ± 27 nm, P < 0.05; normal- vs. high fat diet) and increased common region IMR (0.044 ± 0.023 vs. 0.071 ± 0.024, P < 0.05, normal- vs. high fat diet). The greater IMR at the sinus area and diet induced increases in common area IMR were due to greater and increased dimension of the intima layer in the absence of significant changes in the dimension of the media layer. Furthermore, atherogenic diet increased the thickness of the endothelial cells at the sinus region (0.53 ± 0.28 µm vs. 0.85 ± 0.49 µm; normal vs. atherogenic, P < 0.05), but not at the common region (0.62 ± 0.32 µm vs. 0.66 ± 0.37 µm; normal vs. atherogenic). It is concluded that both regional and diet induced increases in atherogenic risk are associated by smaller glycocalyx dimensions and greater IMR. Exposure of the high risk sinus area to an additional atherogenic challenge results in endothelial thickening and excessive swelling of the subendothelial space, in line with the proposed hypothesis that vascular sites with diminished glycocalyx are more vulnerable to pro-inflammatory and atherosclerotic sequelae.
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