Surface-Bound Vascular Endothelial Growth Factor Promotes Prolonged Activation of Endothelial Cells: A New Technology for Capturing Endothelial Progenitor Cells by Intravascular Stents
- Corresponding Author:
- Masakazu Yamagishi, MD, PhD
Division of Cardiovascular Medicine
Kanazawa University Graduate School of Medicine
13-1 Takara-machi, Kanazawa, 920-8641 Ishikawa, Japan
E-mail: [email protected]
Received date: February 25, 2014; Accepted date: April 5 2014; Published date: April 7, 2014
Citation: Tagawa S, Matsuda T, Aomizu T, Kuwana M, Ohtake H, et al. (2014) Surface-Bound Vascular Endothelial Growth Factor Promotes Prolonged Activation of Endothelial Cells: A New Technology for Capturing Endothelial Progenitor Cells by Intravascular Stents. J Tissue Sci Eng 5:1000140. doi:10.4172/2157-7552.1000140
Copyright: © 2014 Tagawa S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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.