Development of 3D culture scaffolds for directional neuronal growth using 2-photon lithography
Development of 3D culture scaffolds for directional neuronal growth using 2-photon lithography
复制标题
DOI：
10.1016/j.msec.2021.112502
复制
发表时间：
2021-10-25
影响因子：
7.9
通讯作者：
Terenzio, Marco
中科院分区：
文献类型：
--
作者：
Agrawal, Lokesh;Saidani, Menouer;Terenzio, Marco
由于恢复受损组织功能的复杂性，移植技术的传统应用（应用于神经系统的严重创伤性损伤）在临床上取得的成功有限。神经组织工程旨在部署模仿细胞外基质生理特性的支架，以促进轴突的伸长和受损神经的修复。然而，制造具有精确控制的厚度、纹理、孔隙率、排列以及所需的机械强度和有效临床应用所需的功能的理想支架在技术上仍然具有挑战性。我们利用最先进的 2 光子光刻技术来制造高度有序且生物相容的 3D 纳米网格结构，以增强神经元定向生长。首先，我们通过在其表面成功培养初级感觉和运动神经元来表征所述支架的物理和化学性质，并证明其生物相容性。有趣的是，轴突沿着纤维延伸，与纳米网格的图案高度对齐，这与在平板玻璃或聚合物表面上观察到的缺乏方向性相反，并且可以在支架的不同层之间以 3D 方式生长。观察到的轴突生长模式对于治疗外周和脊髓损伤期间发生的创伤性神经损伤是非常理想的。因此，我们的研究结果提供了概念证明，并探索了部署对齐的纤维 3D 支架/植入物以实现轴突定向生长的可能性，并且可用于设计旨在恢复和修复丢失的神经元连接的支架。
Conventional applications of transplant technology, applied to severe traumatic injuries of the nervous system, have met limited success in the clinics due to the complexity of restoring function to the damaged tissue. Neural tissue engineering aims to deploy scaffolds mimicking the physiological properties of the extracellular matrix to facilitate the elongation of axons and the repair of damaged nerves. However, the fabrication of ideal scaffolds with precisely controlled thickness, texture, porosity, alignment, and with the required mechanical strength, features needed for effective clinical applications, remains technically challenging. We took advantage of stateof-the-art 2-photon photolithography to fabricate highly ordered and biocompatible 3D nanogrid structures to enhance neuronal directional growth. First, we characterized the physical and chemical properties and proved the biocompatibility of said scaffolds by successfully culturing primary sensory and motor neurons on their surface. Interestingly, axons extended along the fibers with a high degree of alignment to the pattern of the nanogrid, as opposed to the lack of directionality observed on flat glass or polymeric surfaces, and could grow in 3D between different layers of the scaffold. The axonal growth pattern observed is highly desirable for the treatment of traumatic nerve damage occurring during peripheral and spinal cord injuries. Thus, our findings provide a proof of concept and explore the possibility of deploying aligned fibrous 3D scaffold/implants for the directed growth of axons, and could be used in the design of scaffolds targeted towards the restoration and repair of lost neuronal connections.