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Journal of Tissue Science and Engineering

ISSN: 2157-7552

Open Access

Surface-Bound Vascular Endothelial Growth Factor Promotes Prolonged Activation of Endothelial Cells: A New Technology for Capturing Endothelial Progenitor Cells by Intravascular Stents

Abstract

Shotoku Tagawa, Takehisa Matsuda, Tatsuki Aomizu, Makoto Kuwana, Hiroshi Ohtake, Go Watanabe and Masakazu Yamagishi

In -situ selective capture of the Endothelial Progenitor Cell (EPC) in the arterial blood stream can provide antithrombogenic and -cell proliferation potential to implanted intravascular stents. Therefore, we defined molecular mechanisms of EPC activation associated with stent surface-bound proteins. We sought suitable bound protein to capture and proliferate EPCs. Then, to determine whether and how long the surface-bound protein activates intracellular signal-transduction pathways of endothelial cells through its receptor, we studied the phosphorylation of key intracellular macromolecules including Vascular Endothelial Growth Factor (VEGF) Receptor (R)-2 (VEGFR-2), focal adhesion protein-tyrosine kinase, Akt, and extracellular signal-regulated kinase in human umbilical vein endothelial cells. We found the most suitable surface-bound protein was VEGF. Phosphorylation of these macromolecules continued for a long time up to 72 hours. Under these conditions, quantitative RT-PCR revealed time-dependent up-regulation of the mRNAs encoding three major extracellular matrix macromolecules, collagen IV, laminin-5, and fibronectin. Immuno histo- chemical analysis revealed that these macromolecules were secreted on the basal sides of adherent cells over time and that within a few days after initial adhesion occurred, deposition of these macromolecules shut down the EC adhesion. These results demonstrate that activation of the VEGF-VEGFR intracellular signaling pathway is significant for in situ EPC capture technology for intravascular stents, although further in vivo studies should be done to confirm these processes.

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Citations: 807

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