Endothelial cells anchoring by functionalized yeast polypeptide.

Abstract

The immobilization and proliferation of endothelial cells on the surface of engineered tissues requires the development of new biomaterials that can mimic the anchoring and signaling functions of basement membrane. Here, we report a modified polypeptide from yeast translation termination factor protein that can self-assembly into nanofibers and improve endothelial cell adhesion by its functional motif. The polypeptide (YNNNLQGYQAGFQ) is a beta sheet forming sequence, but it is noninfectious in mammalian tissue because of the absence of substrate protein for propagation. The prion-derived polypeptide was extended at the amino terminal with a short sequence motif from laminin I (YIGSR), and the resultant polypeptide retained self-assembly propensity. Both circular dichroism (CD) measurement and molecular dynamics simulation suggest the assembled nanofibers consists mainly beta sheet structure. The 3D porous hydrogel formed by the modified polypeptide was evaluated as a coating material for vascular tissue engineering. In static culture system, the polypeptide scaffold improved the morphology of endothelial cells and confluency of cell monolayer. In the dynamic bioreactor (pulsatile vascular deformation at 5%), the polypeptide scaffold anchored 3-fold higher number of endothelial cells, which exhibited normal nitric oxide release function. These results suggest that prion-derived polypeptides have high self-assembling and motif integrating capacities. These unique properties can be utilized to build up biomaterials with robust porous structure as well as functionalized motifs for cell enrichment.