Babaloo Hamideh, Barati Shirin, Haghir Hossein, Gholami Arman Abroumand, Moharreri Payam, Fallahnezhad Somaye, Asl Elmira Roshani, Noorzehi Golaleh, Tahmasebi Fatemeh
Biotechnology Research Center, International Campus, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Department of Anatomy, Saveh University of Medical Sciences, Saveh, Iran.
Neuroscience. 2025 Sep 13;583:53-62. doi: 10.1016/j.neuroscience.2025.07.044. Epub 2025 Jul 29.
Spinal cord injury (SCI) is a neurodegenerative disease involving inflammation, microgliosis, and astrocytosis. This study examined how polyurethane/multi-walled carbon nanotube (PU/MWCNT) nanofiber scaffolds containing hesperidin nanoparticles (NPs) and adipose-derived mesenchymal stem cells (AMSCs) affect the phenotype of microglia and astrocytes in an SCI model. Thirty-six male rats were divided into six groups: SCI, sham, scaffold, scaffold with AMSCs, scaffold with NPs, and scaffold with AMSCs and NPs. Motor performance in the hind limbs was evaluated using the Basso, Beattie, and Bresnahan (BBB) behavioral test. Tissue sections were prepared and stained with Luxol Fast Blue (LFB) to examine the white matter; Nissl staining was performed to examine neurons. The populations of microglia and astrocytes were assessed using immunofluorescence with Iba-1 and GFAP antibodies, respectively. The expression levels of the A1 (pro-inflammatory) astrocyte gene (C3), the M1 (pro-inflammatory) microglia gene (iNOS), and inflammatory factors, such as IL-1β and TNF-α, were assessed using qRT-PCR. All data were analyzed using SPSS and GraphPad Prism software. Functional evaluation revealed significant behavioral improvement in the group containing a scaffold, AMSCs, and NPs. Nissl and LFB images revealed increased numbers of neurons and remyelination areas in this group, accompanied by downregulation of the IL-1β and TNF-α genes. The M1 and A1 phenotypes significantly decreased in this group. Ultimately, PU/MWCNT nanofiber scaffolds containing hesperidin nanoparticles and AMSCs reduced M1 microglia and A1 astrocytes, as well as inflammatory factors, while increasing neurons and remyelination in the SCI model.
脊髓损伤（SCI）是一种涉及炎症、小胶质细胞增生和星形胶质细胞增生的神经退行性疾病。本研究考察了含有橙皮苷纳米颗粒（NPs）和脂肪来源间充质干细胞（AMSCs）的聚氨酯/多壁碳纳米管（PU/MWCNT）纳米纤维支架如何影响SCI模型中小胶质细胞和星形胶质细胞的表型。将36只雄性大鼠分为六组：SCI组、假手术组、支架组、含AMSCs的支架组、含NPs的支架组以及含AMSCs和NPs的支架组。使用Basso、Beattie和Bresnahan（BBB）行为测试评估后肢的运动性能。制备组织切片并用Luxol Fast Blue（LFB）染色以检查白质；进行尼氏染色以检查神经元。分别使用Iba-1和GFAP抗体通过免疫荧光评估小胶质细胞和星形胶质细胞的数量。使用qRT-PCR评估A1（促炎）星形胶质细胞基因（C3）、M1（促炎）小胶质细胞基因（iNOS）以及炎症因子如IL-1β和TNF-α的表达水平。所有数据均使用SPSS和GraphPad Prism软件进行分析。功能评估显示，含有支架、AMSCs和NPs的组在行为上有显著改善。尼氏和LFB图像显示该组中神经元数量和髓鞘再生区域增加，同时IL-1β和TNF-α基因下调。该组中M1和A1表型显著减少。最终，含有橙皮苷纳米颗粒和AMSCs的PU/MWCNT纳米纤维支架在SCI模型中减少了M1小胶质细胞和A1星形胶质细胞以及炎症因子，同时增加了神经元数量和髓鞘再生。
