Enhancing endothelial cell retention on ePTFE constructs by siRNA mediated SHP-1 gene silencing

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

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Polymeric vascular grafts hold great promise for vascular reconstruction, but the lack of endothelial cells renders these grafts susceptible to intimal hyperplasia and restenosis, precluding widespread clinical applications. The purpose of this study is to establish a stable endothelium on expanded polytetrafluoroethylene (ePTFE) membrane by small interfering RNA (siRNA)-induced suppression of the cell adhesion inhibitor SH2 domaincontaining protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were treated with scrambled siRNA as a control or SHP-1 specific siRNA. Treated cells were seeded onto fibronectin-coated ePTFE scaffolds and exposed to a physiological range of pulsatile fluid shear stresses for 1 h in a variable-width parallel plate flow chamber. Retention of cells was measured and compared between various shear stress levels and between groups treated with scrambled siRNA and SHP-1 specific siRNA. HUVECs seeded on ePTFE membrane exhibited shear stress-dependent retention. Exposure to physiological shear stress (10 dyn/cm2) induced a reduction in the retention of scrambled siRNA treated cells from 100% to 85% at 1 h. Increased shear stress (20 dyn/cm2) further reduced retention of scrambled siRNA treated cells to 55% at 1 h. SHP-1 knockdown mediated by siRNA enhanced endothelial cell retention from approximately 60% to 85% after 1 h of exposure to average shear stresses in the range of 15-30 dyn/cm2. This study demonstrates that siRNA-mediated gene silencing may be an effective strategy for improving the retention of endothelial cells within vascular grafts.

Original languageEnglish (US)
JournalJournal of Nanotechnology in Engineering and Medicine
Volume2
Issue number1
DOIs
StatePublished - Feb 1 2011

Fingerprint

Protein Phosphatase 1
Protein Tyrosine Phosphatases
Phosphatases
Endothelial cells
Polytetrafluoroethylene
Gene Silencing
RNA
Polytetrafluoroethylenes
Small Interfering RNA
Endothelial Cells
Genes
Proteins
Shear stress
Grafts
Human Umbilical Vein Endothelial Cells
Cells
Transplants
Blood Vessels
Tunica Intima
Membranes

Keywords

  • Cell adhesion
  • SHP-1
  • SiRNA
  • Tissue engineering
  • Vascular regeneration

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Materials Science(all)
  • Medicine(all)

Cite this

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abstract = "Polymeric vascular grafts hold great promise for vascular reconstruction, but the lack of endothelial cells renders these grafts susceptible to intimal hyperplasia and restenosis, precluding widespread clinical applications. The purpose of this study is to establish a stable endothelium on expanded polytetrafluoroethylene (ePTFE) membrane by small interfering RNA (siRNA)-induced suppression of the cell adhesion inhibitor SH2 domaincontaining protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were treated with scrambled siRNA as a control or SHP-1 specific siRNA. Treated cells were seeded onto fibronectin-coated ePTFE scaffolds and exposed to a physiological range of pulsatile fluid shear stresses for 1 h in a variable-width parallel plate flow chamber. Retention of cells was measured and compared between various shear stress levels and between groups treated with scrambled siRNA and SHP-1 specific siRNA. HUVECs seeded on ePTFE membrane exhibited shear stress-dependent retention. Exposure to physiological shear stress (10 dyn/cm2) induced a reduction in the retention of scrambled siRNA treated cells from 100{\%} to 85{\%} at 1 h. Increased shear stress (20 dyn/cm2) further reduced retention of scrambled siRNA treated cells to 55{\%} at 1 h. SHP-1 knockdown mediated by siRNA enhanced endothelial cell retention from approximately 60{\%} to 85{\%} after 1 h of exposure to average shear stresses in the range of 15-30 dyn/cm2. This study demonstrates that siRNA-mediated gene silencing may be an effective strategy for improving the retention of endothelial cells within vascular grafts.",
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Enhancing endothelial cell retention on ePTFE constructs by siRNA mediated SHP-1 gene silencing. / Tefft, Brandon J.; Kopacz, Adrian M.; Liu, Wing K; Liu, Shu Qian.

In: Journal of Nanotechnology in Engineering and Medicine, Vol. 2, No. 1, 01.02.2011.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Enhancing endothelial cell retention on ePTFE constructs by siRNA mediated SHP-1 gene silencing

AU - Tefft, Brandon J.

AU - Kopacz, Adrian M.

AU - Liu, Wing K

AU - Liu, Shu Qian

PY - 2011/2/1

Y1 - 2011/2/1

N2 - Polymeric vascular grafts hold great promise for vascular reconstruction, but the lack of endothelial cells renders these grafts susceptible to intimal hyperplasia and restenosis, precluding widespread clinical applications. The purpose of this study is to establish a stable endothelium on expanded polytetrafluoroethylene (ePTFE) membrane by small interfering RNA (siRNA)-induced suppression of the cell adhesion inhibitor SH2 domaincontaining protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were treated with scrambled siRNA as a control or SHP-1 specific siRNA. Treated cells were seeded onto fibronectin-coated ePTFE scaffolds and exposed to a physiological range of pulsatile fluid shear stresses for 1 h in a variable-width parallel plate flow chamber. Retention of cells was measured and compared between various shear stress levels and between groups treated with scrambled siRNA and SHP-1 specific siRNA. HUVECs seeded on ePTFE membrane exhibited shear stress-dependent retention. Exposure to physiological shear stress (10 dyn/cm2) induced a reduction in the retention of scrambled siRNA treated cells from 100% to 85% at 1 h. Increased shear stress (20 dyn/cm2) further reduced retention of scrambled siRNA treated cells to 55% at 1 h. SHP-1 knockdown mediated by siRNA enhanced endothelial cell retention from approximately 60% to 85% after 1 h of exposure to average shear stresses in the range of 15-30 dyn/cm2. This study demonstrates that siRNA-mediated gene silencing may be an effective strategy for improving the retention of endothelial cells within vascular grafts.

AB - Polymeric vascular grafts hold great promise for vascular reconstruction, but the lack of endothelial cells renders these grafts susceptible to intimal hyperplasia and restenosis, precluding widespread clinical applications. The purpose of this study is to establish a stable endothelium on expanded polytetrafluoroethylene (ePTFE) membrane by small interfering RNA (siRNA)-induced suppression of the cell adhesion inhibitor SH2 domaincontaining protein tyrosine phosphatase-1 (SHP-1). Human umbilical vein endothelial cells (HUVECs) were treated with scrambled siRNA as a control or SHP-1 specific siRNA. Treated cells were seeded onto fibronectin-coated ePTFE scaffolds and exposed to a physiological range of pulsatile fluid shear stresses for 1 h in a variable-width parallel plate flow chamber. Retention of cells was measured and compared between various shear stress levels and between groups treated with scrambled siRNA and SHP-1 specific siRNA. HUVECs seeded on ePTFE membrane exhibited shear stress-dependent retention. Exposure to physiological shear stress (10 dyn/cm2) induced a reduction in the retention of scrambled siRNA treated cells from 100% to 85% at 1 h. Increased shear stress (20 dyn/cm2) further reduced retention of scrambled siRNA treated cells to 55% at 1 h. SHP-1 knockdown mediated by siRNA enhanced endothelial cell retention from approximately 60% to 85% after 1 h of exposure to average shear stresses in the range of 15-30 dyn/cm2. This study demonstrates that siRNA-mediated gene silencing may be an effective strategy for improving the retention of endothelial cells within vascular grafts.

KW - Cell adhesion

KW - SHP-1

KW - SiRNA

KW - Tissue engineering

KW - Vascular regeneration

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