Covalent introduction of bioactive molecules is one of main strategies to significantly enhance the biological activities of bone repair materials. In this study, three most-commonly used chemical groups were respectively introduced on graphene (GP), followed by covalent binding with bone morphogenetic protein-2 (BMP-2) -derived peptides, ensuring that the same molar mass of peptides was bound to different functionalized GP (f-GP). Then the same amount of composites composed of different f-GP and peptides were respectively compounded with poly (lactic-co-glycolic acid) to fabricate 3D scaffolds. In vivo study demonstrated that the scaffolds containing ammonized GP covalently bound with the peptides through amide binding could reach best efficiency of promoting ectopic bone regeneration and repairing calvarial defect probably because the most positive charges on the peptide chain and surface of the ammonized GP could absorb more specific proteins in vivo and have better interactions with them, thereby differentiating most inducible cells into osteogenic cells. Our results indicate that the performances of scaffolds containing covalently bound bioactive molecules can be controlled by the covalent binding mode, and that our prepared scaffold containing ammonized GP covalently bound with the BMP-2-derived peptides through amide binding possess inspiring potential applicable prospects for bone tissue regeneration and engineering.

共价引入生物活性分子是显着增强骨修复材料生物活性的主要策略之一。在这项研究中，三个最常用的化学基团分别被引入到石墨烯（GP）上，然后与骨形态发生蛋白-2（BMP-2）衍生的肽共价结合，确保相同摩尔质量的肽结合到不同功能化的 GP (f-GP)。然后将等量的由不同f-GP和肽组成的复合物分别与聚（乳酸-乙醇酸共聚物）复合以制造3D支架。体内研究表明，含有通过酰胺结合与肽共价结合的氨化 GP 的支架可以达到促进异位骨再生和修复颅骨缺损的最佳效率，这可能是因为氨化 GP 的肽链和表面上的大部分正电荷可以吸附更多特异性蛋白质体内并与它们有更好的相互作用，从而将大多数可诱导细胞分化为成骨细胞。我们的研究结果表明，共价结合模式可以控制含有共价结合生物活性分子的支架的性能，并且我们制备的含有通过酰胺结合与 BMP-2 衍生肽共价结合的氨化 GP 的支架具有鼓舞人心的潜在应用前景组织再生和工程。