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嗅鞘细胞种植去细胞化支架促进脊髓损伤大鼠轴突再生。

Olfactory ensheathing cells seeded decellularized scaffold promotes axonal regeneration in spinal cord injury rats.

机构信息

Department of Hand Surgery and Peripheral Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.

Department of Plastic Surgery, The First affiliated hospital of Sun Yat-Sen University, Guangdong, China.

出版信息

J Biomed Mater Res A. 2021 May;109(5):779-787. doi: 10.1002/jbm.a.37066. Epub 2020 Aug 22.

Abstract

Spinal cord decellularized (DC) scaffolds can promote axonal regeneration and restore hindlimb motor function of spinal cord defect rats. However, scarring caused by damage to the astrocytes at the margin of injury can hinder axon regeneration. Olfactory ensheathing cells (OECs) integrate and migrate with astrocytes at the site of spinal cord injury, providing a bridge for axons to penetrate the scars and grow into lesion cores. The purpose of this study was to evaluate whether DC scaffolds carrying OECs could better promote axon growth. For these studies, DC scaffolds were cocultured with primary extracted and purified OECs. Immunofluorescence and electron microscopy were used for verification of cells adhere and growth on the scaffold. Scaffolds with OECs were transplanted into rat spinal cord defects to evaluate axon regeneration and functional recovery of hind limbs. Basso, Beattie, and Bresnahan (BBB) scoring was used to assess motor function recovery, and glial fibrillary acidic protein (GFAP) and NF200-stained tissue sections were used to evaluate axonal regeneration and astrological scar distribution. Our results indicated that spinal cord DC scaffolds have good histocompatibility and spatial structure, and can promote the proliferation of adherent OECs. In animal experiments, scaffolds carrying OECs have better axon regeneration promoting protein expression than the SCI model, and improve the proliferation and distribution of astrocytes at the site of injury. These results proved that the spinal cord DC scaffold with OECs can promote axon regeneration at the site of injury, providing a new basis for clinical application.

摘要

去细胞脊髓支架可促进轴突再生并恢复脊髓缺损大鼠的后肢运动功能。然而,损伤边缘的星形胶质细胞引起的瘢痕形成会阻碍轴突再生。嗅鞘细胞(OEC)在脊髓损伤部位与星形胶质细胞整合并迁移,为轴突穿透瘢痕并生长到病变核心提供了桥梁。本研究旨在评估携带 OEC 的去细胞支架是否能更好地促进轴突生长。为此,将去细胞支架与原代提取和纯化的 OEC 共培养。免疫荧光和电子显微镜用于验证细胞在支架上的黏附和生长。将携带 OEC 的支架移植到大鼠脊髓缺损部位,以评估轴突再生和后肢功能恢复。Basso、Beattie 和 Bresnahan(BBB)评分用于评估运动功能恢复,并用胶质纤维酸性蛋白(GFAP)和 NF200 染色组织切片评估轴突再生和星形胶质瘢痕分布。结果表明,脊髓去细胞支架具有良好的组织相容性和空间结构,可促进黏附 OEC 的增殖。在动物实验中,携带 OEC 的支架具有比 SCI 模型更好的促进轴突再生蛋白表达,并改善损伤部位星形胶质细胞的增殖和分布。这些结果证明,携带 OEC 的脊髓去细胞支架可促进损伤部位的轴突再生,为临床应用提供了新的依据。

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