Abstract
The coating of surfaces with bio-functional proteins is a promising strategy for the creation of highly biocompatible medical implants. Bio-functional proteins from the extracellular matrix (ECM) provide effective surface functions for controlling cellular behavior. We have previously screened bio-functional tripeptides for feasibility of mass production with the aim of identifying those that are medically useful, such as cell-selective peptides. In this work, we focused on the screening of tripeptides that selectively accumulate collagen type IV (Col IV), an ECM protein that accelerates the re-endothelialization of medical implants. A SPOT peptide microarray was selected for screening owing to its unique cellulose membrane platform, which can mimic fibrous scaffolds used in regenerative medicine. However, since the library size on the SPOT microarray was limited, physicochemical clustering was used to provide broader variation than that of random peptide selection. Using the custom focused microarray of 500 selected peptides, we assayed the relative binding rates of tripeptides to Col IV, collagen type I (Col I), and albumin. We discovered a cluster of Col IV-selective adhesion peptides that exhibit bio-safety with endothelial cells. The results from this study can be used to improve the screening of regeneration-enhancing peptides.
Highlights
In cases of long-term implantation of medical devices used to treat cardiovascular diseases, there is a critical risk of restenosis caused by thrombosis and neointimal hyperplasia [1]
We compared the binding rates and relative selectivities of the representative peptides to three extracellular matrix (ECM) proteins: collagen type IV (Col IV), collagen type I (Col I), and Alb
Since cluster 21 is not a large cluster (2% of 8000 peptides), the fact that we identified Col IV-selective adhesion peptides in the first screen indicates that our clustering-assisted library design did enhance screening efficiency
Summary
In cases of long-term implantation of medical devices used to treat cardiovascular diseases, there is a critical risk of restenosis caused by thrombosis and neointimal hyperplasia [1]. Accelerated endothelialization is among the most effective means of preventing restenosis, thereby reducing the risks associated with vascular implants. Endothelialization involves reorganizing the damaged tissue around the implant area. The rapid and precise re-organization of vascular smooth muscle cells and fibroblasts is important for reducing the risk of neointimal hyperplasia. Accelerated re-endothelialization can be achieved on the surface of an implant using two different tissue-engineering approaches. The first involves seeding and culturing of endothelial cells (ECs) directly on implant surfaces, while the second involves enhancing the adhesion and growth of ECs through material surface design, frequently by using a bimolecular (protein or peptide) coating
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