Department of Anatomy, Comparative Medicine Institution, Medical School of Nantong University, Nantong, China.
Department of Emergency, Affiliated Hospital of Nantong University, Nantong, China.
J Biomater Sci Polym Ed. 2022 Nov;33(16):2124-2144. doi: 10.1080/09205063.2022.2102275. Epub 2022 Jul 23.
Spinal cord injury (SCI) leads to severe loss of motor and sensory functions, and the rehabilitation of SCI is a worldwide problem. Tissue-engineered scaffolds offer new hope for SCI patients, while the newly developed materials encountered a challenge in modeling the microenvironment around the lesion site. We constructed a new composite scaffold by mixing decellularized spinal cord extracellular matrix (dECM) with gelatin methacryloyl (GelMA). The dECM, as a natural biological material, retained a large number of proteins and growth factors related to neurogenesis. GelMA was a photopolymerizable material, harbored a polymer network structure, soft texture, certain shape and plenty of water. The viability, proliferation, and differentiation of neural stem cells (NSCs) on the composite scaffold were evaluated by cell count kit-8 (CCK8), Live/Dead assay, phalloidin staining, 5-Ethynyl-2'-deoxyurdine (EdU), immunofluorescence staining and western blot. The Live/Dead assay, phalloidin staining, EdU, and CCK8 assay showed that the composite scaffold had good biocompatibility and provided better support for proliferation of NSCs. Results of immunocytochemistry and western blot showed that the composite scaffolds promoted the specific differentiation of NSCs into neuron cells. Together, this dECM/GelMA composite scaffold can be used as a cell culture coating, the isolated NSCs seeded on the surface of composite scaffold expressed neuronal markers and assumed neuronal morphology. Our work provided a new method that would be widely used in tissue engineering of SCI.
脊髓损伤 (SCI) 导致严重的运动和感觉功能丧失，SCI 的康复是一个全球性问题。组织工程支架为 SCI 患者提供了新的希望，而新开发的材料在模拟病变部位周围的微环境方面遇到了挑战。我们通过将去细胞化脊髓细胞外基质 (dECM) 与明胶甲基丙烯酰 (GelMA) 混合来构建新的复合支架。dECM 作为一种天然生物材料，保留了大量与神经发生相关的蛋白质和生长因子。GelMA 是一种光聚合材料，具有聚合物网络结构、柔软的质地、一定的形状和大量的水。通过细胞计数试剂盒-8 (CCK8)、Live/Dead 测定、鬼笔环肽染色、5-乙炔基-2'-脱氧尿苷 (EdU)、免疫荧光染色和 Western blot 评估神经干细胞 (NSCs) 在复合支架上的活力、增殖和分化。Live/Dead 测定、鬼笔环肽染色、EdU 和 CCK8 测定表明，复合支架具有良好的生物相容性，并为 NSCs 的增殖提供了更好的支持。免疫细胞化学和 Western blot 的结果表明，复合支架促进了 NSCs 向神经元细胞的特异性分化。总之，这种 dECM/GelMA 复合支架可用作细胞培养涂层，分离的 NSCs 接种在复合支架表面表达神经元标志物并呈现神经元形态。我们的工作提供了一种新的方法，将广泛应用于 SCI 的组织工程。
