BME Masters Project: Cecilia Berniac, “Effect of Pro-Inflammatory Cytokines, TNF-α and Interleukin-6, on Endothelial Glycocalyx”

MS Project Defense   

“Effect of Pro-Inflammatory Cytokines, TNF-α and Interleukin-6, on Endothelial Glycocalyx” 

Cecilia Berniac 

Friday, March 29, 2024 

 Via Zoom 

2pm – 2:30pm 

Abstract: This study investigated the pivotal role of inflammation in endothelial cell (EC) responses, specifically focusing on Heparan Sulfate (HS) within the glycocalyx (GCX). The GCX, a network of membrane-bound proteoglycans and glycoproteins, serves as a crucial mechanosensor, maintaining vascular homeostasis. Although Von Willebrand Factor (VWF) and P-Selectin aren't primary GCX components, they play essential roles in hemostasis. VWF, a glycoprotein, facilitates platelet adhesion and promotes aggregation, while P-Selectin, an adhesion molecule promotes platelet recruitment to vascular injury sites through surface expression on activated endothelial cells and platelets.  

In pathological conditions induced by pro-inflammatory cytokines like TNF-α and IL-6, ECs undergo changes, including increased adhesion molecule expression, disturbing physiological balance and potentially elevating the risk of abnormal blood clotting. This research explores the impact of these cytokines on VWF, P-Selectin, and HS expression in ECs.  

The experimental approach involved culturing human lung microvascular endothelial cells (HLMVECs) on glass slides and treating them with TNF-α and IL-6 to simulate an inflammatory microenvironment. Staining for HS allowed visualization and assessment of the glycocalyx, a key GCX component. Additionally, VWF and P-Selectin staining aided in quantifying their expression and distribution on endothelial cells. Confocal microscopy quantified the Corrected Total Cell Fluorescence (CTCF) of VWF and P-Selectin receptors, ensuring an accurate quantification of specific signals and providing insights into GCX changes.   

Findings highlighted the impact of inflammation on glycocalyx integrity, with HS degradation significantly affecting GCX integrity. P-Selectin expression remained unaffected by the pro-inflammatory environment, emphasizing the complexity of EC responses to inflammation. Increased VWF expression suggested a potential shift towards a pro-thrombotic state, indicating VWF as a valuable vascular health indicator.  

Future investigations may use Atomic Force Microscopy (AFM) to explore P-Selectin and VWF expression and binding forces on endothelial cell surfaces, enhancing the precision of findings and contributing to a comprehensive understanding of the intricate interplay between inflammation, glycocalyx components, and coagulation dynamics. 

For Zoom Link, please email June Norton jnorton@wpi.edu