The development of heparan sulfate within the glycocalyx
AdvisorLong, David S.
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A key to protecting vessel health occurs at the interface between circulating blood and endothelial cells. Strategically located at this interface is the endothelial glycocalyx, the first “line of protection” for blood vessels. The glycocalyx is a thin carbohydrate-rich layer of macromolecules that contain a variety of proteoglycans and glycosaminoglycans (GAGs). The glycocalyx is not only protective but is involved in a range of biological processes from nitric oxide production to cell-cell communication. While modulating numerous biological processes, the spatial and temporal distribution and composition of the glycocalyx can vary between a healthy and diseased state. Since cultured endothelial cells may not display the same glycocalyx as the in vivo, the aim of this work was to confirm the presence of the most prevalent GAG, heparan sulfate, and image the development of heparan sulfate expressed on human microvascular cells type 1. This cell type was selected specifically for this project in that it can give a better understanding as to how this structure reacts to cardiovascular disease conditions. Heparan sulfate was successfully confirmed and proven to encompass the human microvascular cell type 1 over the course of time from three days post seeding to seven and a half days post seeding. Heparan sulfate was shown to be present by enzymatic degradation. Microscopy results show that the glycocalyx has the potential of being a surrogate and therapeutic marker for CVD as it does degrade when exposed to fluid shear stress which exhibits similar conditions to that of cardiovascular disease.. Future work can involve exposing this cell line to a fluid shear stress and imaging the degradation pattern of the glycocalyx. Successful completion of these experiments could encourage efforts to a possible therapeutic solution.
Thesis (M.S.)-- Wichita State University, College of Engineering, Dept. of Biomedical Engineering