Dai Xingliang, Tian Xuefeng, Gu Shengcai, Yang Yafei, Li Huaixu, Gao Peng, Lan Qing, Cheng Hongwei
Department of Neurosurgery, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, P. R. China.
Department of Neurosurgery, the Second Affiliated Hospital of Soochow University, Suzhou, 215004, P. R. China.
Int J Bioprint. 2022 Dec 30;9(2):659. doi: 10.18063/ijb.v9i2.659. eCollection 2023.
The present study aimed to combine extrusion-based three-dimensional (3D) bioprinting and polymer nanofiber electrospinning technology to fabricate tissue-like structures with neurosecretory function . Using neurosecretory cells as cell resources, sodium alginate/gelatin/fibrinogen as matrix, polylactic acid/gelatin electrospun nanofibers as diaphragm, and neurosecretory cells-loaded 3D hydrogel scaffolds were bioprinted and then covered with electrospun nanofibers layer-by-layer. The morphology was observed by scanning electron microscopy and transmission electron microscopy (TEM), and the mechanical characteristics and cytotoxicity of the hybrid biofabricated scaffold structure were evaluated. The 3D-bioprinted tissue activity, including cell death and proliferation, was verified. Western blotting and ELISA experiments were used to confirm the cell phenotype and secretory function, while animal transplantation experiments confirmed the histocompatibility, inflammatory reaction, and tissue remodeling ability of the heterozygous tissue structures. Neurosecretory structures with 3D structures were successfully prepared by hybrid biofabrication . The mechanical strength of the composite biofabricated structures was significantly higher than that of the hydrogel system ( < 0.05). The survival rate of PC12 cells in the 3D-bioprinted model was 92.849 ± 2.995%. Hematoxylin and eosin-stained pathological sections showed that the cells grew in clumps, and there was no significant difference in the expression of MAP2 and tubulin-β between 3D organoids and PC12 cells. The results of ELISA showed that the PC12 cells in 3D structures retained the ability to continuously secrete noradrenaline and met-enkephalin, and the secretory vesicles around and within the cells could be observed by TEM. In transplantation, PC12 cells gathered and grew in clusters, maintained high activity, neovascularization, and tissue remodeling in 3D structures. The neurosecretory structures were biofabricated by 3D bioprinting and nanofiber electrospinning , which had high activity and neurosecretory function. transplantation of neurosecretory structures showed active proliferation of cells and potential for tissue remodeling. Our research provides a new method for biological manufacture of neurosecretory structures , which maintains neurosecretory function and lays the foundation for the clinical application of neuroendocrine tissues.
本研究旨在将基于挤压的三维(3D)生物打印与聚合物纳米纤维静电纺丝技术相结合,以制造具有神经分泌功能的组织样结构。以神经分泌细胞为细胞资源,海藻酸钠/明胶/纤维蛋白原为基质,聚乳酸/明胶静电纺纳米纤维为隔膜,对负载神经分泌细胞的3D水凝胶支架进行生物打印,然后逐层覆盖静电纺纳米纤维。通过扫描电子显微镜和透射电子显微镜(TEM)观察形态,并评估混合生物制造支架结构的力学特性和细胞毒性。验证了3D生物打印组织的活性,包括细胞死亡和增殖。采用蛋白质免疫印迹法和酶联免疫吸附测定(ELISA)实验确认细胞表型和分泌功能,而动物移植实验则证实了杂合组织结构的组织相容性、炎症反应和组织重塑能力。通过混合生物制造成功制备了具有3D结构的神经分泌结构。复合生物制造结构的机械强度显著高于水凝胶系统(<0.05)。3D生物打印模型中PC12细胞的存活率为92.849±2.995%。苏木精-伊红染色病理切片显示细胞成团生长,3D类器官与PC12细胞之间微管相关蛋白2(MAP2)和微管蛋白-β的表达无显著差异。ELISA结果显示,3D结构中的PC12细胞保留了持续分泌去甲肾上腺素和甲硫氨酸脑啡肽的能力,并且通过TEM可以观察到细胞周围和内部的分泌囊泡。在移植中,PC12细胞在3D结构中聚集并成簇生长,保持高活性、新生血管形成和组织重塑。通过3D生物打印和纳米纤维静电纺丝制造的神经分泌结构具有高活性和神经分泌功能。神经分泌结构的移植显示细胞活跃增殖和组织重塑潜力。我们的研究为神经分泌结构的生物制造提供了一种新方法,该方法维持神经分泌功能,为神经内分泌组织的临床应用奠定了基础。