Endothelial glycocalyx thickness and platelet-vessel wall interactions during atherogenesis

Sietze Reitsma; Mirjam G. A. oude Egbrink; Viviane V. Th. Heijnen; Remco T. A. Megens; Wim Engels; Hans Vink; Dick W. Slaaf; Marc A. M. J. van Zandvoort

doi:10.1160/TH11-02-0133

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 106(05): 939-946
DOI: 10.1160/TH11-02-0133

Cardiovascular Biology and Cell Signalling

Schattauer GmbH

Endothelial glycocalyx thickness and platelet-vessel wall interactions during atherogenesis

Sietze Reitsma

¹Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

,

Mirjam G. A. oude Egbrink

²Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

,

Viviane V. Th. Heijnen

²Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

,

Remco T. A. Megens

¹Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

³Institute for Cardiovascular Prevention, Ludwig-Maximilians University Munich, Munich, Germany

,

Wim Engels

¹Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

,

Hans Vink

²Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

,

Dick W. Slaaf

¹Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

⁴Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, the Netherlands

,

Marc A. M. J. van Zandvoort

¹Department of Biomedical Engineering, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, the Netherlands

⁵Institute for Molecular Cardiovascular Research (IMCAR), Universitätsklinikum Aachen, Aachen, Germany

› Author Affiliations

Abstract

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Summary

The endothelial glycocalyx (EG), the luminal cover of endothelial cells, is considered to be atheroprotective. During atherogenesis, platelets adhere to the vessel wall, possibly triggered by simultaneous EG modulation. It was the objective of this study to investigate both EG thickness and platelet-vessel wall interactions during atherogenesis in the same experimental model. Intravital fluorescence microscopy was used to study platelet-vessel wall interactions in vivo in common carotid arteries and bifurcations of C57bl6/J (B6) and apolipoprotein E knock-out (ApoE-/-) mice (age 7 – 31 weeks). At the same locations, EG thickness was determined ex vivo using two-photon laser scanning microscopy. In ApoE-/- bifurcations the overall median level of adhesion was 48 platelets/mm2 (interquartile range: 16 – 80), which was significantly higher than in B6 bifurcations (0 (0 – 16), p = 0.001). This difference appeared to result from a significant age-dependent increase in ApoE-/- mice, while no such change was observed in B6 mice. At the same time, the EG in ApoE-/- bifurcations was significantly thinner than in B6 bifurcations (2.2 vs. 2.5 μm, respectively; p < 0.05). This resulted from the fact that in B6 bifurcations EG thickness increased with age (from 2.4 μm in young mice to 3.0 μm in aged ones), while in bifurcations of ApoE-/- mice this growth appeared to be absent (2.2 μm at all ages). During atherogenesis, platelet adhesion to the wall of the carotid artery bifurcation increases significantly. At the same location, EG growth with age is hampered. Therefore, glycocalyx-reinforcing strategies could possibly ameliorate atherosclerosis.

Keywords

Atherogenesis - carotid artery - endothelial glycocalyx - platelets - mice

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References

References
1 Reitsma S, Slaaf DW, Vink H. et al. The endothelial glycocalyx: composition, functions, and visualization. Pflugers Arch 2007; 454: 345-359.
2 Gouverneur M, Berg B, Nieuwdorp M. et al. Vasculoprotective properties of the endothelial glycocalyx: effects of fluid shear stress. J Intern Med 2006; 259: 393-400.
3 Nieuwdorp M, Meuwese MC, Vink H. et al. The endothelial glycocalyx: a potential barrier between health and vascular disease. Curr Opin Lipidol 2005; 16: 507-511.
4 Constantinescu A, Spaan JA, Arkenbout EK. et al. Degradation of the endothelial glycocalyx is associated with chylomicron leakage in mouse cremaster muscle microcirculation. Thromb Haemost 2011; 105: 790-801.
5 Nieuwdorp M, Mooij HL, Kroon J. et al. Endothelial glycocalyx damage coincides with microalbuminuria in type 1 diabetes. Diabetes 2006; 55: 1127-1132.
6 Mulivor AW and Lipowsky HH. Inflammation- and ischemia-induced shedding of venular glycocalyx. Am J Physiol Heart Circ Physiol 2004; 286: H1672-680.
7 van den Berg BM, Spaan JA, Rolf TM. et al. Atherogenic region and diet diminish glycocalyx dimension and increase intima-to-media ratios at murine carotid artery bifurcation. Am J Physiol Heart Circ Physiol 2006; 290: H915-920.
8 van den Berg BM, Spaan JA and Vink H. Impaired glycocalyx barrier properties contribute to enhanced intimal low-density lipoprotein accumulation at the carotid artery bifurcation in mice. Pflugers Arch 2009; 457: 1199-1206.
9 Hansson GK. Inflammation, atherosclerosis, and coronary artery disease. N Engl J Med 2005; 352: 1685-1695.
10 Libby P. Inflammation in atherosclerosis. Nature 2002; 420: 868-874.
11 Massberg S, Brand K, Gruner S. et al. A critical role of platelet adhesion in the initiation of atherosclerotic lesion formation. J Exp Med 2002; 196: 887-896.
12 Sachais BS, Turrentine T, Dawicki McKenna JM. et al. Elimination of platelet factor 4 (PF4) from platelets reduces atherosclerosis in C57Bl/6 and apoE-/- mice. Thromb Haemost 2007; 98: 1108-1113.
13 Huo Y, Schober A, Forlow SB. et al. Circulating activated platelets exacerbate atherosclerosis in mice deficient in apolipoprotein E. Nat Med 2003; 9: 61-67.
14 Koenen RR, von Hundelshausen P, Nesmelova IV. et al. Disrupting functional interactions between platelet chemokines inhibits atherosclerosis in hyperlipidemic mice. Nat Med 2009; 15: 97-103.
15 Weber C. Platelets and chemokines in atherosclerosis: partners in crime. Circ Res 2005; 96: 612-616.
16 Sandberg WJ, Otterdal K, Gullestad L. et al. The tumour necrosis factor superfamily ligand APRIL (TNFSF13) is released upon platelet activation and expressed in atherosclerosis. Thromb Haemost 2009; 102: 704-710.
17 Aidoudi S and Bikfalvi A. Interaction of PF4 (CXCL4) with the vasculature: a role in atherosclerosis and angiogenesis. Thromb Haemost 2010; 104: 941-948.
18 Nagy N, Freudenberger T, Melchior-Becker A. et al. Inhibition of hyaluronan synthesis accelerates murine atherosclerosis: novel insights into the role of hyaluronan synthesis. Circulation 2011; 122: 2313-2322.
19 Vink H, Constantinescu AA and Spaan JA. Oxidized lipoproteins degrade the endothelial surface layer: implications for platelet-endothelial cell adhesion. Circulation 2000; 101: 1500-1502.
20 Kuijpers MJ, Gilio K, Reitsma S. et al. Complementary roles of platelets and coagulation in thrombus formation on plaques acutely ruptured by targeted ultrasound treatment: a novel intravital model. J Thromb Haemost 2009; 7: 152-161.
21 van Gestel MA, Heemskerk JW, Slaaf DW. et al. Real-time detection of activation patterns in individual platelets during thromboembolism in vivo: differences between thrombus growth and embolus formation. J Vasc Res 2002; 39: 534-543.
22 Reitsma S, oude Egbrink MGA, Vink H. et al. Endothelial glycocalyx structure in the intact carotid artery: a two-photon laser scanning microscopy study. J Vasc Res 2011; 48: 297-306.
23 Slaaf DW, Tangelder GJ, Reneman RS. et al. A versatile incident illuminator for intravital microscopy. Int J Microcirc Clin Exp 1987; 6: 391-397.
24 Megens RT, Reitsma S, Schiffers PH. et al. Two-photon microscopy of vital murine elastic and muscular arteries. Combined structural and functional imaging with subcellular resolution. J Vasc Res 2007; 44: 87-98.
25 Megens RT, Oude Egbrink MG, Cleutjens JP. et al. Imaging collagen in intact viable healthy and atherosclerotic arteries using fluorescently labeled CNA35 and two-photon laser scanning microscopy. Mol Imaging 2007; 6: 247-260.
26 Becker BF, Chappell D, Bruegger D. et al. Therapeutic strategies targeting the endothelial glycocalyx: acute deficits, but great potential. Cardiovasc Res 2010; 87: 300-310.
27 Springer TA. Adhesion receptors of the immune system. Nature 1990; 346: 425-434.
28 Becker BF, Chappell D, Jacob M. Endothelial glycocalyx and coronary vascular permeability: the fringe benefit. Basic Res Cardiol 2010; 105: 687-701.
29 Maxwell MJ, Westein E, Nesbitt WS. et al. Identification of a 2-stage platelet aggregation process mediating shear-dependent thrombus formation. Blood 2007; 109: 566-576.
30 Han YF, Weinbaum S, Spaan JAE. et al. Large-deformation analysis of the elastic recoil of fibre layers in a Brinkman medium with application to the endothelial glycocalyx. Journal of Fluid Mechanics 2006; 554: 217-235.
31 Meir KS, Leitersdorf E. Atherosclerosis in the apolipoprotein-E-deficient mouse: a decade of progress. Arterioscler Thromb Vasc Biol 2004; 24: 1006-1014.
32 Ramos CL, Huo Y, Jung U. et al. Direct demonstration of P-selectin- and VCAM-1-dependent mononuclear cell rolling in early atherosclerotic lesions of apolipoprotein E-deficient mice. Circ Res 1999; 84: 1237-1244.