Osteoinduction of stem cells by collagen peptide-immobilized hydrolyzed poly(butylene succinate)/β-tricalcium phosphate scaffold for bone tissue engineering

Abstract

Bone substitute is a therapeutic approach to treat bone abnormalities. A scaffold serves mainly as osteoconductive elements. To facilitate a better biological performance, short collagen peptide was immobilized onto hydrolyzed poly(butylene succinate)/β-tricalcium phosphate (HPBSu/TCP) scaffolds. PBSu/TCP (80:20) scaffolds were fabricated by a supercritical CO technique, hydrolyzed with 0.6 M NaOH and conjugated with short collagen peptide tagged with or without red fluorescence. The surface morphology and porous structure of scaffolds were characterized by scanning electron microscopy and micro-computed tomography. Human mesenchymal stem cells were cultured onto the scaffolds and examined for osteogenic differentiation and biomineralization in vitro by means of alkaline phosphatase activity, alizarin red staining, and reverse transcription-polymerase chain reaction. The PBSu/TCP and HPBSu/TCP scaffolds were successfully prepared. Scanning electron microscopy and micro-computed tomography results showed that the porosity was distributed throughout the scaffolds with the pore sizes in the range of 250-900 µm. Fluorescence microscopy demonstrated retention of tagged short collagen peptide on the scaffold. Mesenchymal stem cells adhered and grew well on the material. Under osteogenic induction, cells cultured on the short collagen peptide -immobilized scaffold significantly produced a greater amount of alkaline phosphatase activity and positive mineralization than those cultured on the control scaffold. The present results have shown that the short collagen peptide-immobilized HPBSu/TCP scaffold enhanced osteoinduction and biomineralization of stem cell-derived osteoblasts, possibly via stimulation of alkaline phosphatase activity. This suggests the potential use of osteogenic peptide-immobilized material in bone tissue engineering for correcting bone defects.

Preparation and Characterization of Nanocomposite Scaffolds (Collagen/β-TCP/SrO) for Bone Tissue Engineering

“…The scaffolds comprising synthetic or natural biopolymers and bioceramics, such as b-tri-calcium phosphate (b-TCP) and hydroxyapatite (HA) or their composites are used as biomedical materials for bone regeneration [1,6,7]. Collagen and b-TCP are the main organic and inorganic components of natural bone and are investigated as scaffolds for bone tissue engineering [8][9][10][11]. Collagen has unique features including appropriate osteoconductivity, bioactivity, biodegradability and excellent biocompatibility [7,12].…”

Analysis of sequential dual immobilization of type I collagen and BMP-2 short peptides on hydrolyzed poly(buthylene succinate)/β-tricalcium phosphate composites for bone tissue engineering

“…The present study has shown, for the first time, that sequential immobilization of COLsp and BMP-2sp was successfully achieved, with each of the peptides being chemically bound to the HPBSu/TCP surface and the resulting surface remaining bioactive. In addition, this method helps improve our previously reported osteogenic efficacy of the HPBSu/TCP composites 5,25,26 as the currently modified surface induced in vitro mineralization by hMSCs without exogenously added osteogenic supplements.…”

“…We have previously shown that modified PBSu/TCP composites have good biocompatibility and osteoconductivity and stimulate osteogenic differentiation and mineralization, 4,5 and single COLsp immobilization onto HPBSu/TCP composites stimulated matrix mineralization. 25 It is, therefore, undoubtedly beneficial for investigating the possibility that dual bioactive peptides can improve the performance of this material. Although attempts have been made to immobilize dual short peptides on a biomaterial surface, the optimization, characterization and functional analysis of the peptide immobilization method onto HPBSu/TCP composites have not yet been reported.…”

Cell-binding peptides on the material surface guide stem cell fate of adhesion, proliferation and differentiation

“…The amino (primary amine) groups (or carboxylic acids) of peptides can react with carboxylic acids (or amino (primary amine) groups) on the material surface or biomacromolecules using the N-hydroxysuccinimide (NHS)/ 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) reaction (Fig. 16) 68,[93][94][95]105,109,131,132,[159][160][161][162][163][164]168,[232][233][234][235][236][237][238][239][240] or the NHS/N-hydroxysulfosuccinimide (sulfo-NHS) reaction (Fig. 16).…”