Objective
It is widely accepted that the presence of a glycosaminoglycan-rich glycocalyx is essential for endothelialized vasculature health; in fact, a damaged or impaired glycocalyx has been demonstrated in a number of vascular diseases. Currently, there are no methods that characterize glycocalyx functionality, thus limiting investigators’ ability to assess the role of the glycocalyx in vascular health.
Approach and Results
We have developed novel easy to use in vitro assays that directly quantify live endothelialized surface’s functional heparin weights and their anti-coagulant capacity to inactivate Factor Xa and thrombin. Using our assays, we characterized two commonly used vascular models: native rat aorta and cultured human umbilical vein endothelial cell (HUVEC) monolayer. We determined heparin contents to be about 10,000 ng/cm2 on the native aorta, and about ten-fold lower on cultured HUVECs. Interestingly HUVECs demonstrated a five-fold lower anti-coagulation capacity in inactivating both Factor Xa and thrombin relative to native aortas. We verified the validity and accuracy of the novel assays developed in this work using LC-MS analysis.
Conclusions
Our assays are of high relevance in the vascular community as they can be utilized to establish the anti-thrombogenic capacity of many different types of surfaces such as vascular grafts and transplants. This work will also advance the capacity for glycocalyx-targeting therapeutics development to treat damaged vasculatures.
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