Pereira Inês, Lopez-Martinez Maria J, Villasante Aranzazu, Introna Clelia, Tornero Daniel, Canals Josep M, Samitier Josep
Nanobioengineering Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology (BIST), Barcelona, Spain.
Department of Electronic and Biomedical Engineering, University of Barcelona, Barcelona, Spain.
Front Bioeng Biotechnol. 2023 Mar 3;11:1110547. doi: 10.3389/fbioe.2023.1110547. eCollection 2023.
Three-dimensional (3D) bioprinting is a promising technique for the development of neuronal models because it controls the deposition of materials and cells. Finding a biomaterial that supports neural differentiation while ensuring compatibility with the technique of 3D bioprinting of a self-standing construct is a challenge. In this study, gelatin methacryloyl (GelMA), methacrylated alginate (AlgMA), and hyaluronic acid (HA) were examined by exploiting their biocompatibility and tunable mechanical properties to resemble the extracellular matrix (ECM) and to create a suitable material for printing neural progenitor cells (NPCs), supporting their long-term differentiation. NPCs were printed and differentiated for up to 15 days, and cell viability and neuronal differentiation markers were assessed throughout the culture. This composite biomaterial presented the desired physical properties to mimic the ECM of the brain with high water intake, low stiffness, and slow degradation while allowing the printing of defined structures. The viability rates were maintained at approximately 80% at all time points. However, the levels of -III tubulin marker increased over time, demonstrating the compatibility of this biomaterial with neuronal cell culture and differentiation. Furthermore, these cells showed increased maturation with corresponding functional properties, which was also demonstrated by the formation of a neuronal network that was observed by recording spontaneous activity Ca imaging.
三维（3D）生物打印是一种很有前景的技术，可用于开发神经元模型，因为它能控制材料和细胞的沉积。找到一种既能支持神经分化，又能确保与自立式结构的3D生物打印技术兼容的生物材料是一项挑战。在本研究中，通过利用甲基丙烯酰化明胶（GelMA）、甲基丙烯酰化海藻酸盐（AlgMA）和透明质酸（HA）的生物相容性和可调机械性能，使其类似于细胞外基质（ECM），并创建一种适合打印神经祖细胞（NPC）的材料，以支持其长期分化。将NPC打印并分化长达15天，并在整个培养过程中评估细胞活力和神经元分化标志物。这种复合生物材料呈现出所需的物理特性，能够模拟大脑的ECM，具有高吸水性、低硬度和缓慢降解的特点，同时允许打印特定结构。在所有时间点，存活率均维持在约80%。然而，β-III微管蛋白标志物的水平随时间增加，表明这种生物材料与神经元细胞培养和分化具有兼容性。此外，这些细胞表现出成熟度增加以及相应的功能特性，通过记录自发活动的钙成像观察到的神经元网络形成也证明了这一点。
