The structural stability of the endothelial glycocalyx after enzymatic removal of glycosaminoglycans

• Published in: PloS one 2012-08, Vol.7 (8), p.e43168-e43168
• Main Authors: Zeng, Ye, Ebong, Eno E, Fu, Bingmei M, Tarbell, John M
• Format: Article
• Language: English
• Subjects: Adsorption; Albumin; Albumins - chemistry; Animals; Atherosclerosis; Binding Sites; Biology; Biomedical engineering; Carotid arteries; Chondroitin sulfate; Chondroitin Sulfates - chemistry; Chondroitinases and Chondroitin Lyases - metabolism; Collapse; Confocal microscopy; Endothelial cells; Endothelium; Endothelium - metabolism; Enzymes; Glucose; Glycocalyx - physiology; Glycosaminoglycans; Glycosaminoglycans - chemistry; Heparan sulfate; Heparitin Sulfate - chemistry; Hyaluronic acid; Hyaluronic Acid - chemistry; Hyaluronoglucosaminidase - metabolism; Medicine; Microscopy; Microscopy, Fluorescence - methods; Plasma proteins; Polysaccharide-Lyases - metabolism; Protein Binding; Proteins; Rats; Shear stress; Stability analysis; Stem cells; Structural stability; Sulfates
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0043168

It is widely believed that glycosaminoglycans (GAGs) and bound plasma proteins form an interconnected gel-like structure on the surface of endothelial cells (the endothelial glycocalyx layer-EGL) that is stabilized by the interaction of its components. However, the structural organization of GAGs and proteins and the contribution of individual components to the stability of the EGL are largely unknown. To evaluate the hypothesis that the interconnected gel-like glycocalyx would collapse when individual GAG components were almost completely removed by a specific enzyme. Using confocal microscopy, we observed that the coverage and thickness of heparan sulfate (HS), chondroitin sulfate (CS), hyaluronic acid (HA), and adsorbed albumin were similar, and that the thicknesses of individual GAGs were spatially nonuniform. The individual GAGs were degraded by specific enzymes in a dose-dependent manner, and decreased much more in coverage than in thickness. Removal of HS or HA did not result in cleavage or collapse of any of the remaining components. Simultaneous removal of CS and HA by chondroitinase did not affect HS, but did reduce adsorbed albumin, although the effect was not large. All GAGs and adsorbed proteins are well inter-mixed within the structure of the EGL, but the GAG components do not interact with one another. The GAG components do provide binding sites for albumin. Our results provide a new view of the organization of the endothelial glycocalyx layer and provide the first demonstration of the interaction between individual GAG components.