Department of Advanced Materials, College of Life Science and Nano Technology, Hannam University, Yuseong-gu, Daejeon 305-811, South Korea.
J Biomed Mater Res A. 2013 Apr;101(4):942-53. doi: 10.1002/jbm.a.34393. Epub 2012 Sep 8.
The purpose of this study was to generate tissue-engineered bone using human periosteal-derived osteoblasts (PO) and polydioxanone/pluronic F127 (PDO/pluronic F127) scaffold with preseeded human periosteal-derived CD146 positive endothelial-like cells (PE). PE were purified from the periosteal cell population by cell sorting. One of the important factors to consider in generating tissue-engineered bone using osteoprecursor and endothelial cells and a specific scaffold is whether the function of osteoprecursor and endothelial cells can be maintained in originally different culture medium conditions. After human PE were preseeded into PDO/pluronic F127 scaffold and cultured in endothelial cell basal medium-2 for 7 days, human PO were seeded into the PDO/pluronic F127 scaffold with PE, and then, this cell-scaffold construct was cultured in endothelial cell basal medium-2 with osteogenic induction factors, including ascorbic acid, dexamethasone, and β-glycerophosphate, for a further 7 days. Then, this 2-week cultured construct was grafted into the mandibular defect of miniature pig. Twelve weeks after implantation, the animal was sacrificed. Clinical examination revealed that newly formed bone was seen more clearly in the defect with human PO and PDO/pluronic F127 scaffold with preseeded human PE. The experimental results suggest that tissue-engineered bone formation using human PO and PDO/pluronic F127 scaffold with preseeded human PE can be used to restore skeletal integrity to various bony defects when used in clinics.
本研究旨在利用人骨膜源性成骨细胞(PO)和聚二氧六环酮/泊洛沙姆 F127(PDO/ 泊洛沙姆 F127)支架,以及预先接种人骨膜源性 CD146 阳性内皮样细胞(PE)来生成组织工程骨。PE 通过细胞分选从骨膜细胞群体中纯化出来。在使用成骨前体细胞和内皮细胞以及特定支架生成组织工程骨时,需要考虑的一个重要因素是,成骨前体细胞和内皮细胞的功能是否可以在原本不同的培养条件下得以维持。将人 PE 预先接种到 PDO/ 泊洛沙姆 F127 支架中,并在内皮细胞基础培养基-2 中培养 7 天后,将人 PO 接种到含有 PE 的 PDO/ 泊洛沙姆 F127 支架中,然后将这个细胞-支架构建体在含有成骨诱导因子(包括抗坏血酸、地塞米松和 β-甘油磷酸)的内皮细胞基础培养基-2 中再培养 7 天。然后,将这个培养了 2 周的构建体移植到小型猪的下颌骨缺损处。植入 12 周后,处死动物。临床检查显示,在预先接种人 PE 的人 PO 和 PDO/ 泊洛沙姆 F127 支架的缺损处,可见到更多的新形成骨。实验结果表明,当用于临床时,利用人 PO 和 PDO/ 泊洛沙姆 F127 支架预先接种人 PE 可以用于恢复各种骨缺损的骨骼完整性。