Capture of endothelial cells under flow using immobilized vascular endothelial growth factor

Randall J Smith Jr; Maxwell T Koobatian; Aref Shahini; Daniel D Swartz; Stelios T Andreadis

doi:10.1016/j.biomaterials.2015.02.025

Capture of endothelial cells under flow using immobilized vascular endothelial growth factor

Biomaterials. 2015 May:51:303-312. doi: 10.1016/j.biomaterials.2015.02.025. Epub 2015 Feb 21.

Authors

Randall J Smith Jr¹, Maxwell T Koobatian², Aref Shahini³, Daniel D Swartz⁴, Stelios T Andreadis⁵

Affiliations

¹ Department of Biomedical Engineering, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA.
² Department of Physiology and Biophysics, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA.
³ Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA.
⁴ Department of Physiology and Biophysics, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA; Department of Pediatrics, Women and Children's Hospital of Buffalo, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA; Center of Excellence in Bioinformatics and Life Sciences, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA.
⁵ Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA; Department of Biomedical Engineering, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA; Center of Excellence in Bioinformatics and Life Sciences, University at Buffalo, State University of New York, Amherst, NY 14260-4200, USA. Electronic address: sandread@buffalo.edu.

Abstract

We demonstrate the ability of immobilized vascular endothelial growth factor (VEGF) to capture endothelial cells (EC) with high specificity under fluid flow. To this end, we engineered a surface consisting of heparin bound to poly-l-lysine to permit immobilization of VEGF through the C-terminal heparin-binding domain. The immobilized growth factor retained its biological activity as shown by proliferation of EC and prolonged activation of KDR signaling. Using a microfluidic device we assessed the ability to capture EC under a range of shear stresses from low (0.5 dyne/cm(2)) to physiological (15 dyne/cm(2)). Capture was significant for all shear stresses tested. Immobilized VEGF was highly selective for EC as evidenced by significant capture of human umbilical vein and ovine pulmonary artery EC but no capture of human dermal fibroblasts, human hair follicle derived mesenchymal stem cells, or mouse fibroblasts. Further, VEGF could capture EC from mixtures with non-EC under low and high shear conditions as well as from complex fluids like whole human blood under high shear. Our findings may have far reaching implications, as they suggest that VEGF could be used to promote endothelialization of vascular grafts or neovascularization of implanted tissues by rare but continuously circulating EC.

Keywords: Immobilized growth factor; Shear stress; VEGF; Vascular grafts.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Cell Proliferation / drug effects
Human Umbilical Vein Endothelial Cells / cytology*
Human Umbilical Vein Endothelial Cells / drug effects
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Immobilized Proteins / pharmacology*
Lab-On-A-Chip Devices
Mice
NIH 3T3 Cells
Receptors, Vascular Endothelial Growth Factor / metabolism
Rheology*
Sheep
Stress, Mechanical
Vascular Endothelial Growth Factor A / pharmacology*

Substances

Immobilized Proteins
Vascular Endothelial Growth Factor A
Receptors, Vascular Endothelial Growth Factor

Grants and funding

R01 HL086582/HL/NHLBI NIH HHS/United States