Zhang Xue, Guo Wen-Guang, Cui Helen, Liu Huan-Ye, Zhang Yang, Müller Werner E G, Cui Fu-Zhai
Department of Orthodontics, School of Stomatology, China Medical University, Shenyang, 110001, China.
Beijing Allgens Medical Science & Technology Company, Beijing, 100085, China.
J Tissue Eng Regen Med. 2016 Feb;10(2):99-107. doi: 10.1002/term.1705. Epub 2013 Jan 31.
Enhancement of osteogenic capacity was achieved in a mineralized collagen composite, nano-hydroxyapatite/collagen (nHAC), by loading with synthetic peptides derived from BMP-2 residues 32-48 (P17-BMP-2). Rabbit marrow stromal cells (MSCs) were used in vitro to study cell biocompatibility, attachment and differentiation on the mineralized collagen composite by a cell counting kit, scanning electron microscopy (SEM) and real-time reversed transcriptase-polymerase chain reaction analysis (RT-PCR). Optimal peptide dosage (1.0 µg/mL) was obtained by RT-PCR analysis in vitro. In addition, the relative expression level of OPN and OCN was significantly upregulated on P17-BMP-2/nHAC compared with nHAC. In vitro results of P17-BMP-2 release kinetics demonstrated that nHAC released P17-BMP-2 in a controlled and sustained manner. In the rabbit mandibular box-shaped bone defect model, osteogenic capacity of three groups (nHAC, P17-BMP-2/nHAC, rhBMP-2/nHAC) was evaluated. Compared to the nHAC group, bone repair responses in both P17-BMP-2/nHAC and rhBMP-2/nHAC group implants were significantly improved based on histological analysis. The osteogenic response of the P17-BMP-2/nHAC group was similar to that of the rhBMP-2/nHAC group.
通过负载源自骨形态发生蛋白-2(BMP-2)第32 - 48位残基的合成肽(P17 - BMP - 2),在矿化胶原复合材料纳米羟基磷灰石/胶原(nHAC)中实现了成骨能力的增强。体外使用兔骨髓间充质干细胞(MSCs),通过细胞计数试剂盒、扫描电子显微镜(SEM)和实时逆转录聚合酶链反应分析(RT-PCR)来研究细胞在矿化胶原复合材料上的生物相容性、黏附及分化情况。通过体外RT-PCR分析获得了最佳肽剂量(1.0 µg/mL)。此外,与nHAC相比,P17 - BMP - 2/nHAC上骨桥蛋白(OPN)和骨钙素(OCN)的相对表达水平显著上调。P17 - BMP - 2释放动力学的体外结果表明,nHAC以可控且持续的方式释放P17 - BMP - 2。在兔下颌盒状骨缺损模型中,评估了三组(nHAC、P17 - BMP - 2/nHAC、重组人骨形态发生蛋白-2/nHAC)的成骨能力。基于组织学分析,与nHAC组相比,P17 - BMP - 2/nHAC组和重组人骨形态发生蛋白-2/nHAC组植入物的骨修复反应均显著改善。P17 - BMP - 2/nHAC组的成骨反应与重组人骨形态发生蛋白-2/nHAC组相似。