Enhancement of endothelial cell adhesion through treatment with CEACAM6 or TNF-α [Medical College of Wisconsin]
610,000 Americans die of heart disease every year and over 1 million Americans undergo coronary interventions each year. Risks of these coronary interventions include thrombosis, embolization, infection, and re-narrowing of the artery. However, it is currently believed that improving the adhesion capabilities of endothelial cells may significantly reduce the associated risks of coronary interventions, especially with regards to cardiovascular stent and graft placement procedures. In the present study, we investigated a method to biochemically facilitate endothelial cell adhesion. Using the Reactome database, we identified biochemical methods/targets which could be modified to promote endothelial cell adhesion. Here we describe a potential method of enhancing endothelial cell adhesion through controlled application of two treatment conditions: Carcinoembryonic Antigen-related Cell Adhesion Molecule 6 (CEACAM6) with Fibronectin-1 (CEACAM6+, FN1+) and Tumor Necrosis Factor-α (TNF-α) with Fibronectin-1 (TNF-α+, FN1+). DAPI stained porcine blood outgrowth endothelial cells (BOECs) were exposed to these treatments or a control treatment (FN1+) for one hour. Each treatment/control condition was then placed into a validated parallel plate flow chamber (PPFC) apparatus (flow rate accuracy and laminar flow were both validated) and exposed to a shear stress of 15 dyn/cm2 (to replicate in vivo shear stress conditions) for 30 minutes using saline solution. Fluorescent images were taken of all replicates before and after shear stress testing in order to determine the effects of treatments on cell adhesion. Statistical analysis of preliminary trials indicated that the treatment groups demonstrated a trend towards higher percentage cell retention (or lower percentage cell loss) with 82.3%, 70.2%, and 50.2% average cell retention for TNF-α, CEACAM6, and the control, respectively; however, statistical significance could not be reached using n=2 and p<0.05. Preliminary results of the TNF-α treatment are promising. In the future, an experiment with a larger sample size and a longer shear stress exposure time (e.g. 1 hour) is needed.
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