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Shear stress-conditioned, endothelial cell-seeded vascular grafts: improved cell adherence in response to in vitro
1Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Background:
Prosthetic vascular grafts with adherent endothelial cell monolayers may prove useful for small-caliber vessel bypass. However, endothelial cells adhere poorly to prosthetic graft material, and they are stripped when exposed to in vivo shear stress. This study sought to determine whether in vitro shear stress conditioning improves endothelial cell adhesion and decreases thrombogenicity of endothelial cell-seeded grafts.
Methods:
The lumens of 1.5 mm (inside diameter) spun polyurethane polymer vascular grafts were seeded with bovine aortic endothelial cells and cultured in vitro for 6 days with or without continuous laminar shear stress, first at 1 to 2 dynes/cm2 for 3 days, then at approximately 25 dynes/cm2 for 3 days. Grafts preconditioned by shear stress and the static control grafts were then exposed to arterial shear stress at 25 dynes/cm2 for 25 seconds. The number of dislodged cells was counted, and the grafts were examined by light and scanning electron microscopy. Whole blood clotting time in the grafts was also determined.
Results:
Exposure of grafts to acute shear stress dislodged 1.35 x 10(6) +/- 0.44 x 10(6) cells from static grafts compared with 1.05 x 10(4) +/- 0.16 x 10(4) cells from grafts preconditioned by shear stress. By light and electron microscopy an intact endothelial monolayer was observed to cover the lumen of shear stress-conditioned grafts, whereas few cells remained on the luminal surface of grafts not previously exposed to shear stress. The clotting time in shear stress-conditioned grafts was significantly prolonged in relation to grafts not exposed to shear stress.
Conclusions:
These findings show that endothelial cell adhesion and retention on vascular grafts in vitro is markedly enhanced by preconditioning the seeded endothelial cell monolayer with long-term shear stress. Consequently, vascular grafts containing shear stress-conditioned endothelial monolayers are less thrombogenic in vitro than small-caliber vascular grafts without intact endothelial cell monolayers.