Jiang Ziyun, Zhou Linhong, Xiao Miao, Ma Sancheng, Cheng Guosheng
Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, China.
Institute for Cardiovascular Science and Department of Cardiovascular Surgery of the First Affiliated Hospital, Medical College, Soochow University, Suzhou, China.
Cells Tissues Organs. 2024;213(4):316-325. doi: 10.1159/000534255. Epub 2023 Oct 9.
An early substantial loss of basal forebrain cholinergic neurons (BFCNs) is a common property of Alzheimer's disease and the degeneration of functional BFCNs is related to learning and memory deficits. As a biocompatible and conductive scaffold for growth of neural stem cells, three-dimensional graphene foam (3D-GF) supports applications in tissue engineering and regenerative medicine. Although its effects on differentiation have been demonstrated, the effect of 3D-GF scaffold on the generation of BFCNs still remains unknown.
In this study, we used 3D-GF as a culture substrate for neural progenitor cells (NPCs) and demonstrated that this scaffold material promotes the differentiation of BFCNs while maintaining excellent cell viability and proliferation.
Immunofluorescence analysis, real-time polymerase chain reaction, Western blotting, and ELISA revealed that the proportion of BFCNs at 21 days of differentiation reached approximately 30.5% on 3D-GF compared with TCPS group that only presented 9.7%. Furthermore, a cell adhesion study suggested that 3D-GF scaffold enhances the expression of adhesion proteins including vinculin, integrin, and N-cadherin. These findings indicate that 3D-GF scaffold materials are preferable candidates for the differentiation of BFCNs from NPCs.
These results suggest new opportunities for the application of 3D-GF scaffold as a neural scaffold for cholinergic neurons therapies based on NPCs.
基底前脑胆碱能神经元（BFCNs）早期大量丧失是阿尔茨海默病的一个常见特征，功能性BFCNs的退化与学习和记忆缺陷有关。作为神经干细胞生长的生物相容性和导电性支架，三维石墨烯泡沫（3D-GF）支持在组织工程和再生医学中的应用。尽管其对分化的影响已得到证实，但3D-GF支架对BFCNs生成的影响仍然未知。
在本研究中，我们使用3D-GF作为神经祖细胞（NPCs）的培养底物，并证明这种支架材料在保持优异细胞活力和增殖的同时促进BFCNs的分化。
免疫荧光分析、实时聚合酶链反应、蛋白质印迹法和酶联免疫吸附测定显示，与仅呈现9.7%的传统细胞培养聚苯乙烯（TCPS）组相比，在3D-GF上分化21天时BFCNs的比例达到约30.5%。此外，细胞黏附研究表明，3D-GF支架增强了包括纽蛋白、整合素和N-钙黏蛋白在内的黏附蛋白的表达。这些发现表明，3D-GF支架材料是NPCs分化为BFCNs的优选候选材料。
这些结果为3D-GF支架作为基于NPCs的胆碱能神经元治疗的神经支架的应用提供了新的机会。
