Abstract:

BACKGROUND: Previous studies have shown that vascular endothelial cells were seed in grafts to enhance endothelialization rapidly and early, which can improve graft fluency. But the adhesion of seeded endothelial cells is often poor in single-layer. They are easily washed off under the blood flow.
OBJECTIVE: To improve cell retention on the graft with dual cell seeding technique, adding a layer of smooth muscle cells (SMCs) between the graft and vascular endothelial cells.
METHODS: Rabbit endothelial cells were cultured by standard enzyme separation technique and cell culture technique. Rabbit smooth muscle cells were isolated and cultured using tissue explant method. The lacz and green fluorescent porotein genes were used to transfer endothelial cells and smooth muscle cells. Polytetrafluoroethylene grafts pretreated with fibronectin were seeded with smooth muscle cell followed by endothelial cells in two-layer cells group. Vascular endothelial cells implantation served as controls. Vascular prosthesis was connected to perfusion models for 2 hours. The density of endothelial cells was calculated prior to and following perfusion. In vivo experiment, cell retention rate was measured following vascular prosthesis was inserted into the rabbit for 7 days.
RESULTS AND CONCLUSION: 2 hours following perfusion, cell retention rate was (0.39±0.04) in endothelial cells group, and (0.73±0.07) in the two-lay cells group. The cell retention rate was greater in the two-layer cells group compared with endothelial cells group (P < 0.01). 7 days following vascular prosthesis implantation, the cell retention rate was (0.63±0.10) in the endothelial cells group, and (0.95±0.06) in the two-layer cells group (P < 0.05). Results indicated that using smooth muscle cell as a media layer between endothelial cells and graft surface can improve the retention of seeded endothelial cells in vitro and vivo.