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用于制备应用于神经组织工程的纳米结构混合支架的马尾神经衍生细胞外基质。

Cauda equina-derived extracellular matrix for fabrication of nanostructured hybrid scaffolds applied to neural tissue engineering.

作者信息

Wen Xiaoxiao, Wang Yu, Guo Zhiyuan, Meng Haoye, Huang Jingxiang, Zhang Li, Zhao Bin, Zhao Qing, Zheng Yudong, Peng Jiang

机构信息

1 School of Materials Science and Engineering, University of Science and Technology Beijing , Beijing, P.R. China .

出版信息

Tissue Eng Part A. 2015 Mar;21(5-6):1095-105. doi: 10.1089/ten.TEA.2014.0173. Epub 2014 Dec 16.

Abstract

Extracellular matrix (ECM) components have become important candidate materials for use as neural scaffolds for neural tissue engineering. In the current study, we prepared cauda equina-derived ECM materials for the production of scaffolds. Natural porcine cauda equina was decellularized using Triton X-100 and sodium deoxycholate, shattered physically, and made into a suspension by differential centrifugation. The decellularization procedure resulted in the removal of >94% of the nuclear material and preserved the extracellular collagen and sulfated glycosaminoglycan. Immunofluorescent staining confirmed the presence of collagen type I, laminin, and fibronectin in the ECM. The cauda equine-derived ECM was blended with poly(l-lactide-co-glycolide) (PLGA) to fabricate nanostructured scaffolds using electrospinning. The incorporation of the ECM increased the hydrophilicity of the scaffolds. Fourier transform infrared spectroscopy and multiphoton-induced autofluorescence images showed the presence of the ECM in the scaffolds. ECM/PLGA scaffolds were beneficial for the survival of Schwann cells compared with scaffolds consisting of PLGA alone, and the aligned fibers could regulate cell morphologic features by modulating cellular orientation. Axons in the dorsal root ganglia explants extended to a greater extent along ECM/PLGA compared with PLGA-alone fibers. The cauda equina ECM might be a promising material for forming scaffolds for use in neural tissue engineering.

摘要

细胞外基质(ECM)成分已成为用于神经组织工程的神经支架的重要候选材料。在本研究中,我们制备了马尾来源的ECM材料用于支架生产。使用 Triton X-100 和脱氧胆酸钠对天然猪马尾进行脱细胞处理,物理粉碎,并通过差速离心制成悬浮液。脱细胞过程导致去除了>94%的核物质,并保留了细胞外胶原蛋白和硫酸化糖胺聚糖。免疫荧光染色证实ECM中存在I型胶原蛋白、层粘连蛋白和纤连蛋白。将马尾来源的ECM与聚(l-丙交酯-共-乙交酯)(PLGA)混合,采用静电纺丝法制备纳米结构支架。ECM的加入增加了支架的亲水性。傅里叶变换红外光谱和多光子诱导自发荧光图像显示支架中存在ECM。与仅由PLGA组成的支架相比,ECM/PLGA支架有利于雪旺细胞的存活,且排列的纤维可通过调节细胞取向来调控细胞形态特征。与仅PLGA纤维相比,背根神经节外植体中的轴突在ECM/PLGA上延伸的程度更大。马尾ECM可能是一种用于形成神经组织工程支架的有前途的材料。

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