Zn2+交联的海藻酸钠三维支架的理化特性及其与脊髓神经干/祖细胞的生物相容性研究

周军;唐志坚;吴雅奇;刘胜文;王煜

神经损伤与功能重建 ›› 2022, Vol. 17 ›› Issue (10) : 567-570.

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神经损伤与功能重建 ›› 2022, Vol. 17 ›› Issue (10) : 567-570.
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Zn2+交联的海藻酸钠三维支架的理化特性及其与脊髓神经干/祖细胞的生物相容性研究

  • 周军,唐志坚,吴雅奇,刘胜文,王煜
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The Physicochemical Properties of Alginate Hydrogel 3D Scaffold Crosslinked by Zn2+ and It’s Biocompatibility with Spinal Cord Neural Stem/Progenitor Cells

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摘要

目的:明确Zn2+ 交联的海藻酸钠水凝胶(AH)三维支架的理化特性、超微结构、管道结构的稳定性;评 价AH与脊髓神经干/祖细胞(NSPCs)的生物相容性。方法:采用Zn2+ 与海藻酸钠交联制备AH三维支架;光 镜和扫描电镜评估其微管道结构;光谱分析仪测定盐酸洗涤后水凝胶内部残留Zn2+ 含量;分离和培养胎鼠 (E14)脊髓NSPCs;细胞培养环境下用磷酸盐缓冲液孵育AH三维支架18周,光镜下观察内部管道变化情况, 测量重量变化;脊髓NSPCs与AH三维支架共培养,MTT法测定细胞存活率,免疫荧光染色检测NSPCs的 分化。结果:AH三维支架管道直径为(93.26±6.24)μm,管道密度为(9.72±2.37)/mm2 ,管道孔隙率为(66.22± 8.97)%;经过盐酸洗涤8次后,AH三维支架内Zn2+ 浓度为0.08 nmoL/mg;脊髓NSPCs在AH三维支架表面的 存活率>90%,且可以分化为神经元、星形胶质细胞和少突胶质细胞。结论:AH三维支架仅含有极低浓度 的Zn2+ ,且具有稳定的拓扑结构,与脊髓NSPCs有良好的生物相容性。

Abstract

To clarify the physicochemical properties, ultrastructure, stability of alginate hydrogel (AH) crosslinked by Zn2+ and assess the biocompatibility of AH 3D scaffold with spinal cord neural stem/progenitor cells (NSPCs). Methods: AH 3D scaffolds were fabricated by crosslinking of alginate with Zn2+ . Then the scaffolds were viewed under the optical microscope and scanning electron microscope to view general and ultrastructure respectively. Optical spectrum analyzer was used to test the concentration of Zn2+ remained in the scaffold after being washed by HCl solution. After incubation in phosphate buffer saline for 18 weeks, the AH scaffolds were weighed and checked under the optical microscope for capillary structure. MTT method was used to quantify the survival rate of NSPCs co-cultured with AH for different days. The differentiation of NSPCs on AH 3D scaffolds was determined by immunofluorescence. Results: The capillary diameter of AH 3D scaffolds was (93.26±6.24) μm, while the porosity of capillaries was (7.83±1.58)% and the density was (68.41±6.51)/mm2 . The concentration of Zn2+ within the scaffold was 0.08 nmoL/mg after 8-time washing by hydrochloric acid. The spinal cord NSPCs could survive (>90%) well and differentiate into neurons, astrocytes and oligodendrocytes on the AH 3D scaffolds. Conclusion: AH 3D scaffolds have a stable topological structure with few Zn2 + left and show great biocompatibility with spinal cord NSPCs.

关键词

海藻酸钠水凝胶 / 三维支架 / 神经干/祖细胞 / 生物相容性

Key words

alginate hydrogel

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周军;唐志坚;吴雅奇;刘胜文;王煜. Zn2+交联的海藻酸钠三维支架的理化特性及其与脊髓神经干/祖细胞的生物相容性研究[J]. 神经损伤与功能重建. 2022, 17(10): 567-570
The Physicochemical Properties of Alginate Hydrogel 3D Scaffold Crosslinked by Zn2+ and It’s Biocompatibility with Spinal Cord Neural Stem/Progenitor Cells[J]. Neural Injury and Functional Reconstruction. 2022, 17(10): 567-570

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