title: Poly(ε-caprolactone) modified with fusion protein containing self-assembled hydrophobin and functional peptide for selective capture of human blood outgrowth endothelial cells.
abstract: 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.
authors.affiliation: The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin 300071, China.
authors.forename: Yujian
authors.initials: Y
authors.lastname: Huang
authors.forename: Suai
authors.initials: S
authors.lastname: Zhang
authors.forename: Baolong
authors.initials: B
authors.lastname: Niu
authors.forename: Dandan
authors.initials: D
authors.lastname: Wang
authors.forename: Zefang
authors.initials: Z
authors.lastname: Wang
authors.forename: Shuren
authors.initials: S
authors.lastname: Feng
authors.forename: Haijin
authors.initials: H
authors.lastname: Xu
authors.forename: Deling
authors.initials: D
authors.lastname: Kong
authors.forename: Mingqiang
authors.initials: M
authors.lastname: Qiao
mesh_terms: D002469:Cell Separation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D042783:Endothelial Cells / Q000737:chemistry*; D005822:Genetic Vectors; D048691:Grifola / Q000737:chemistry; D007202:Indicators and Reagents; D009842:Oligopeptides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D056951:Photoelectron Spectroscopy; D010843:Pichia / Q000737:chemistry / Q000235:genetics; D011091:Polyesters / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D054457:Tissue Scaffolds
chemical_list: C580939:HGFI-TPS fusion protein; D007202:Indicators and Reagents; D009842:Oligopeptides; D010455:Peptides; D011091:Polyesters; D011993:Recombinant Fusion Proteins; C580940:TPS-peptide; C016240:polycaprolactone