Patntirapong Somying, Janvikul Wanida, Theerathanagorn Tharinee, Singhatanadgit Weerachai
1 Thammasat University, Pathumthani, Thailand.
2 National Metal and Materials Technology Center, Pathumthani, Thailand.
J Biomater Appl. 2017 Jan;31(6):859-870. doi: 10.1177/0885328216684374.
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.
骨替代物是治疗骨异常的一种治疗方法。支架主要作为骨传导元件。为了促进更好的生物学性能，将短胶原肽固定在水解聚丁二酸丁二醇酯/β-磷酸三钙（HPBSu/TCP）支架上。通过超临界CO₂技术制备PBSu/TCP（80:20）支架，用0.6 M NaOH水解，并与标记有或没有红色荧光的短胶原肽偶联。通过扫描电子显微镜和微计算机断层扫描对支架的表面形态和多孔结构进行表征。将人间充质干细胞培养在支架上，并通过碱性磷酸酶活性、茜素红染色和逆转录-聚合酶链反应在体外检测其成骨分化和生物矿化。成功制备了PBSu/TCP和HPBSu/TCP支架。扫描电子显微镜和微计算机断层扫描结果表明，孔隙率分布在整个支架中，孔径范围为250-900 µm。荧光显微镜显示标记的短胶原肽保留在支架上。间充质干细胞在该材料上粘附并良好生长。在成骨诱导下，培养在固定有短胶原肽的支架上的细胞比培养在对照支架上的细胞显著产生更多的碱性磷酸酶活性和阳性矿化。目前的结果表明，固定有短胶原肽的HPBSu/TCP支架增强了干细胞来源的成骨细胞的骨诱导和生物矿化，可能是通过刺激碱性磷酸酶活性。这表明成骨肽固定材料在骨组织工程中用于修复骨缺损具有潜在用途。
