RADA16-I is an ion-complementary self-assembled peptide with a regular folded secondary conformation and can be assembled into an ordered nanostructure. Dentonin is an extracellular matrix phosphate glycoprotein functional peptide motif-containing RGD and SGDG motifs. In this experiment, we propose to combine RAD and Dentonin to form a functionalized self-assembled peptide RAD/Dentonin hydrogel scaffold. Furthermore, we expect that the RAD with the addition of functional motif Dentonin can promote pulp regeneration. The study analyzed the physicochemical properties of RAD/Dentonin through circular dichroism, morphology scanning, and rheology. Besides, we examined the scaffold’s biocompatibility by immunofluorescent staining, CCK-8 method, Live/Dead fluorescent staining, and 3D reconstruction. Finally, we applied ALP activity assay, RT-qPCR, and Alizarin red S staining to detect the effect of RAD/Dentonin on the odontogenic differentiation of human dental pulp stem cells (hDPSCs). The results showed that RAD/Dentonin spontaneously assembles into a hydrogel with a β-sheet-based nanofiber network structure. In vitro, RAD/Dentonin has superior biocompatibility and enhances adhesive proliferation, migration, odontogenic differentiation, and mineralization deposition of hDPSCs. In conclusion, the novel self-assembled peptide RAD/Dentonin is a new scaffold material suitable for cell culture and has promising applications as a scaffold for endodontic tissue engineering.

RADA16-I是一种离子互补的自组装肽，具有规则折叠的二级构象，可以组装成有序的纳米结构。Dentonin 是一种细胞外基质磷酸糖蛋白功能肽基序，含有 RGD 和 SGDG 基序。在本实验中，我们建议将 RAD 和 Dentonin 结合形成功能化的自组装肽 RAD/Dentonin 水凝胶支架。此外，我们预计添加功能性基序 Dentonin 的 RAD 可以促进牙髓再生。该研究通过圆二色性、形态扫描和流变学分析了 RAD/Dentonin 的物理化学性质。此外，我们通过免疫荧光染色、CCK-8方法、活/死荧光染色和3D重建检查了支架的生物相容性。最后，我们应用了 ALP 活性测定、RT-qPCR、和茜素红 S 染色检测 RAD/Dentonin 对人牙髓干细胞 (hDPSCs) 成牙本质分化的影响。结果表明，RAD/Dentonin 自发组装成水凝胶，具有基于β-折叠的纳米纤维网络结构。在体外，RAD/Dentonin 具有优异的生物相容性，并增强 hDPSCs 的粘附增殖、迁移、牙源性分化和矿化沉积。总之，新型自组装肽RAD/Dentonin是一种适用于细胞培养的新型支架材料，作为牙髓组织工程支架具有广阔的应用前景。