Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology, Beijing, China; National Clinical Research Center for Oral Diseases, Beijing, China; National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing, China; Beijing Key Laboratory of Digital Stomatology, Beijing, China.
National Clinical Research Center for Oral Diseases, Beijing, China; National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing, China; Beijing Key Laboratory of Digital Stomatology, Beijing, China; Center of Digital Dentistry, Peking University School and Hospital of Stomatology, Beijing, China.
Mater Sci Eng C Mater Biol Appl. 2021 Oct;129:112393. doi: 10.1016/j.msec.2021.112393. Epub 2021 Aug 25.
Peripheral nerve injury can cause various degrees of damage to the morphological structure and physiological function of the peripheral nerve. At present, compared with "gold standard" autologous nerve transplantation, tissue engineering has certain potential for regeneration and growth; however, achieving oriented guidance is still a challenge. In this study, we used 3D bioprinting to construct a nerve scaffold of RSC96 cells wrapped in sodium alginate/gelatin methacrylate (GelMA)/bacterial nanocellulose (BNC) hydrogel. The 5% sodium alginate+5% GelMA+0.3% BNC group had the thinnest lines among all groups after printing, indicating that the inherent shape of the scaffold could be maintained after adding BNC. Physical and chemical property testing (Fourier transform infrared, rheometer, conductivity, and compression modulus) showed that the 5% alginate+5% GelMA+0.3% BNC group had better mechanical and rheological properties. Live/dead cell staining showed that no mass cell death was observed on days 1, 3, 5, and 7 after printing. In the 5% alginate+5% GelMA group, the cells grew and formed linear connections in the scaffold. This phenomenon was more obvious in the 5% alginate+5% GelMA+0.3% BNC group. In the 5% alginate+5% GelMA+0.3% BNC group, S-100β immunofluorescence staining and cytoskeleton staining showed oriented growth. Polymerase chain reaction (PCR) array results showed that mRNA levels of related neurofactors ASCL1, POU3F3, NEUROG1, DLL1, NOTCH1 and ERBB2 in the 5%GelMA+0.3%BNC group were higher than those of other groups. Four weeks after implantation in nude mice, RSC96 cells grew and proliferated well, blood vessels grew, and S-100β immunofluorescence was positive. These results indicate that a 3D-bioprinted sodium alginate/GelMA/BNC composite scaffold can improve cell-oriented growth, adhesion and the expression of related factors. This 3D-bioprinted composite scaffold has good biocompatibility and is expected to become a new type of scaffold material in the field of neural tissue engineering.
周围神经损伤可导致周围神经的形态结构和生理功能发生不同程度的损伤。目前，与“金标准”自体神经移植相比，组织工程在再生和生长方面具有一定的潜力；然而，实现定向引导仍然是一个挑战。在本研究中，我们使用 3D 生物打印技术构建了一种 RSC96 细胞包裹在海藻酸钠/甲基丙烯酰化明胶（GelMA）/细菌纳米纤维素（BNC）水凝胶中的神经支架。在打印后，所有组中 5%海藻酸钠+5%GelMA+0.3%BNC 组的线最细，表明添加 BNC 后支架的固有形状可以保持。理化性能测试（傅里叶变换红外、流变仪、电导率和压缩模量）表明，5%海藻酸钠+5%GelMA+0.3%BNC 组具有更好的机械和流变性能。活/死细胞染色显示，打印后第 1、3、5 和 7 天，没有观察到大量细胞死亡。在 5%海藻酸钠+5%GelMA 组中，细胞在支架中生长并形成线性连接。在 5%海藻酸钠+5%GelMA+0.3%BNC 组中，这种现象更为明显。在 5%海藻酸钠+5%GelMA+0.3%BNC 组中，S-100β免疫荧光染色和细胞骨架染色显示出定向生长。聚合酶链反应（PCR）阵列结果显示，5%GelMA+0.3%BNC 组中相关神经因子 ASCL1、POU3F3、NEUROG1、DLL1、NOTCH1 和 ERBB2 的 mRNA 水平高于其他组。裸鼠体内植入 4 周后，RSC96 细胞生长增殖良好，血管生长，S-100β 免疫荧光阳性。这些结果表明，3D 生物打印的海藻酸钠/GelMA/BNC 复合支架可以改善细胞的定向生长、黏附和相关因子的表达。这种 3D 生物打印的复合支架具有良好的生物相容性，有望成为神经组织工程领域的新型支架材料。
