Enhancement of Endothelial Cell Retention on ePTFE Vascular Constructs by siRNA-Mediated SHP-1 or SHP-2 Gene Silencing

Brandon J. Tefft*, Adrian M. Kopacz, Wing K Liu, Shu Qian Liu

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Insufficient retention of endothelial cells to the luminal surface of small-diameter tissue engineered vascular grafts has precluded clinical translation. This study utilized a gene silencing strategy to enhance the adhesion strength of vascular endothelial cells to fibronectin-coated expanded polytetrafluoroethylene, a common vascular graft material. SHP-1 and SHP-2 are both phosphatases known to inhibit the formation of focal contacts. By employing siRNA to knockdown the expression of SHP-1 or SHP-2, we observed a significant improvement in cell retention following 6 h of pulsatile fluid shear stress. At an average fluid shear stress of 15 dyn/cm2, cell retention was improved from approximately 30% for control groups to approximately 70 and 85% for cells treated with SHP-1 and SHP-2 specific siRNA, respectively (n = 8 for all groups). We also observed that treatment with SHP-1 or SHP-2 specific siRNA caused a modest increase in focal contact density and did not cause a significant effect on the expression of the endothelial cell markers VEGFR-2, VE-cadherin, and PECAM-1. These findings indicate that molecular modulation may be an effective strategy for improving the retention of endothelial cells within tissue engineered vascular grafts, which may in turn improve their clinical performance.

Original languageEnglish (US)
Pages (from-to)507-516
Number of pages10
JournalCellular and Molecular Bioengineering
Volume8
Issue number3
DOIs
StatePublished - Sep 21 2015

Fingerprint

Endothelial Cells
Endothelial cells
Gene Silencing
Small Interfering RNA
Blood Vessels
Enhancement
Genes
Grafts
Blood Vessel Prosthesis
Gene
Focal Adhesions
Shear Stress
Shear stress
Cell
Fibronectin
Contact
Tissue
CD31 Antigens
Fluid
Vascular Endothelial Growth Factor Receptor-2

Keywords

  • Endothelialization
  • Gene knockdown
  • RNAi
  • Shear stress
  • Vascular graft

ASJC Scopus subject areas

  • Modeling and Simulation
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

@article{f0ba399aef30454eaa507e09ed4a65c6,
title = "Enhancement of Endothelial Cell Retention on ePTFE Vascular Constructs by siRNA-Mediated SHP-1 or SHP-2 Gene Silencing",
abstract = "Insufficient retention of endothelial cells to the luminal surface of small-diameter tissue engineered vascular grafts has precluded clinical translation. This study utilized a gene silencing strategy to enhance the adhesion strength of vascular endothelial cells to fibronectin-coated expanded polytetrafluoroethylene, a common vascular graft material. SHP-1 and SHP-2 are both phosphatases known to inhibit the formation of focal contacts. By employing siRNA to knockdown the expression of SHP-1 or SHP-2, we observed a significant improvement in cell retention following 6 h of pulsatile fluid shear stress. At an average fluid shear stress of 15 dyn/cm2, cell retention was improved from approximately 30{\%} for control groups to approximately 70 and 85{\%} for cells treated with SHP-1 and SHP-2 specific siRNA, respectively (n = 8 for all groups). We also observed that treatment with SHP-1 or SHP-2 specific siRNA caused a modest increase in focal contact density and did not cause a significant effect on the expression of the endothelial cell markers VEGFR-2, VE-cadherin, and PECAM-1. These findings indicate that molecular modulation may be an effective strategy for improving the retention of endothelial cells within tissue engineered vascular grafts, which may in turn improve their clinical performance.",
keywords = "Endothelialization, Gene knockdown, RNAi, Shear stress, Vascular graft",
author = "Tefft, {Brandon J.} and Kopacz, {Adrian M.} and Liu, {Wing K} and Liu, {Shu Qian}",
year = "2015",
month = "9",
day = "21",
doi = "10.1007/s12195-015-0392-9",
language = "English (US)",
volume = "8",
pages = "507--516",
journal = "Cellular and Molecular Bioengineering",
issn = "1865-5025",
publisher = "Springer New York",
number = "3",

}

Enhancement of Endothelial Cell Retention on ePTFE Vascular Constructs by siRNA-Mediated SHP-1 or SHP-2 Gene Silencing. / Tefft, Brandon J.; Kopacz, Adrian M.; Liu, Wing K; Liu, Shu Qian.

In: Cellular and Molecular Bioengineering, Vol. 8, No. 3, 21.09.2015, p. 507-516.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Enhancement of Endothelial Cell Retention on ePTFE Vascular Constructs by siRNA-Mediated SHP-1 or SHP-2 Gene Silencing

AU - Tefft, Brandon J.

AU - Kopacz, Adrian M.

AU - Liu, Wing K

AU - Liu, Shu Qian

PY - 2015/9/21

Y1 - 2015/9/21

N2 - Insufficient retention of endothelial cells to the luminal surface of small-diameter tissue engineered vascular grafts has precluded clinical translation. This study utilized a gene silencing strategy to enhance the adhesion strength of vascular endothelial cells to fibronectin-coated expanded polytetrafluoroethylene, a common vascular graft material. SHP-1 and SHP-2 are both phosphatases known to inhibit the formation of focal contacts. By employing siRNA to knockdown the expression of SHP-1 or SHP-2, we observed a significant improvement in cell retention following 6 h of pulsatile fluid shear stress. At an average fluid shear stress of 15 dyn/cm2, cell retention was improved from approximately 30% for control groups to approximately 70 and 85% for cells treated with SHP-1 and SHP-2 specific siRNA, respectively (n = 8 for all groups). We also observed that treatment with SHP-1 or SHP-2 specific siRNA caused a modest increase in focal contact density and did not cause a significant effect on the expression of the endothelial cell markers VEGFR-2, VE-cadherin, and PECAM-1. These findings indicate that molecular modulation may be an effective strategy for improving the retention of endothelial cells within tissue engineered vascular grafts, which may in turn improve their clinical performance.

AB - Insufficient retention of endothelial cells to the luminal surface of small-diameter tissue engineered vascular grafts has precluded clinical translation. This study utilized a gene silencing strategy to enhance the adhesion strength of vascular endothelial cells to fibronectin-coated expanded polytetrafluoroethylene, a common vascular graft material. SHP-1 and SHP-2 are both phosphatases known to inhibit the formation of focal contacts. By employing siRNA to knockdown the expression of SHP-1 or SHP-2, we observed a significant improvement in cell retention following 6 h of pulsatile fluid shear stress. At an average fluid shear stress of 15 dyn/cm2, cell retention was improved from approximately 30% for control groups to approximately 70 and 85% for cells treated with SHP-1 and SHP-2 specific siRNA, respectively (n = 8 for all groups). We also observed that treatment with SHP-1 or SHP-2 specific siRNA caused a modest increase in focal contact density and did not cause a significant effect on the expression of the endothelial cell markers VEGFR-2, VE-cadherin, and PECAM-1. These findings indicate that molecular modulation may be an effective strategy for improving the retention of endothelial cells within tissue engineered vascular grafts, which may in turn improve their clinical performance.

KW - Endothelialization

KW - Gene knockdown

KW - RNAi

KW - Shear stress

KW - Vascular graft

UR - http://www.scopus.com/inward/record.url?scp=84939568497&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84939568497&partnerID=8YFLogxK

U2 - 10.1007/s12195-015-0392-9

DO - 10.1007/s12195-015-0392-9

M3 - Article

AN - SCOPUS:84939568497

VL - 8

SP - 507

EP - 516

JO - Cellular and Molecular Bioengineering

JF - Cellular and Molecular Bioengineering

SN - 1865-5025

IS - 3

ER -