Liu Yijuan, Fan Lina, Lin Xuemei, Zou Luning, Li Yaoyao, Ge Xinting, Fu Weihao, Zhang Zonghao, Xiao Kuancheng, Lv Hongbing
Fujian Key Laboratory of Oral Diseases, Fujian Medical University, Fuzhou, Fujian, People's Republic of China.
Fujian Provincial Engineering Research Center of Oral Biomaterial, Fujian Medical University, Fuzhou, Fujian, People's Republic of China.
Biomed Mater. 2021 Nov 30;17(1). doi: 10.1088/1748-605X/ac3928.
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-sheet-based nanofiber network structure., 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是一种离子互补自组装肽，具有规则折叠的二级构象，可组装成有序的纳米结构。牙本质涎磷蛋白是一种细胞外基质磷酸糖蛋白，含有功能性肽基序RGD和SGDG基序。在本实验中，我们提议将RADA16-I和牙本质涎磷蛋白结合，形成功能化的自组装肽RAD/牙本质涎磷蛋白水凝胶支架。此外，我们期望添加了功能性基序牙本质涎磷蛋白的RADA16-I能够促进牙髓再生。该研究通过圆二色性、形态扫描和流变学分析了RAD/牙本质涎磷蛋白的物理化学性质。此外，我们通过免疫荧光染色、CCK-8法、活/死荧光染色和三维重建检测了该支架的生物相容性。最后，我们应用碱性磷酸酶活性测定、逆转录定量聚合酶链反应和茜素红S染色来检测RAD/牙本质涎磷蛋白对人牙髓干细胞（hDPSC）成牙分化的影响。结果表明，RAD/牙本质涎磷蛋白可自发组装成具有基于α-片层的纳米纤维网络结构的水凝胶。RAD/牙本质涎磷蛋白具有优异的生物相容性，可增强hDPSC的黏附增殖、迁移、成牙分化和矿化沉积。总之，新型自组装肽RAD/牙本质涎磷蛋白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。
