Poly(ɛ-caprolactone) modified with fusion protein containing self-assembled hydrophobin and functional peptide for selective capture of human blood outgrowth endothelial cells
| Autor: | Baolong Niu, Yujian Huang, Mingqiang Qiao, Shuren Feng, Deling Kong, Dandan Wang, Haijin Xu, Zefang Wang, Suai Zhang |
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| Rok vydání: | 2013 |
| Předmět: |
Hydrophobin
Polyesters Recombinant Fusion Proteins Genetic Vectors Cell Separation Pichia Pichia pastoris Colloid and Surface Chemistry Tissue engineering Amphiphile Physical and Theoretical Chemistry Progenitor cell Chromatography High Pressure Liquid Grifola frondosa Tissue Scaffolds biology Chemistry Photoelectron Spectroscopy Endothelial Cells Surfaces and Interfaces General Medicine biology.organism_classification Fusion protein Biochemistry Biophysics Indicators and Reagents Peptides Oligopeptides Fetal bovine serum Grifola Biotechnology |
| Zdroj: | Colloids and Surfaces B: Biointerfaces. 101:361-369 |
| ISSN: | 0927-7765 |
| Popis: | Human blood outgrowth endothelial cells (HBOECs)-specific binding peptide, TPSLEQRTVYAK (TPS), was proposed to be applied on autologous cell therapy for treating cardiovascular diseases. Hydrophobins, as a family of self-assembly proteins originated from fungi, have demonstrated unique characteristics to modulate surface properties of other materials coated with these amphiphilic proteins in previous studies. In this report, a fusion protein which was composed of class I hydrophobin HGFI originated from Grifola frondosa and functional peptide TPS was expressed by Pichia pastoris expression system. Then, we purified this fusion protein by ultrafiltration and reverse-phase high performance liquid chromatography. Water contact angle, X-ray photoelectron spectroscopy measurements indicated that the surface properties of hydrophobin were greatly preserved by this fusion protein while comparing with wild HGFI. Cell binding assay showed that this fusion protein demonstrated specific binding property to HBOECs while coating on biodegradable poly(ε-caprolactone) (PCL) grafts in the presence of fetal bovine serum, whereas HGFI-coated PCL non-selectively enhanced all types of cells attachments. Methylthiazol tetrazolium assay was employed to verify the cytocompatibility of this fusion protein-based material. This work presented a new perspective to apply hydrophobin in tissue engineering and regenerative medicine and provided an alternative approach to study endothelial progenitor cells. |
| Databáze: | OpenAIRE |
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