Department of Neurosurgery, Hainan Cancer Hospital, Hainan, China.
Department of Neurobiology, Harbin Medical University, Heilongjiang, China.
Int J Neurosci. 2024 Aug;134(8):889-898. doi: 10.1080/00207454.2022.2155155. Epub 2022 Dec 16.
This study aimed to produce an acellular spinal cord scaffold-bone marrow stromal cell (ASCS-BMSC) complex in which the growth of BMSCs transplanted into the spinal cord of rats could be simulated , facilitating the observation and evaluation of the growth of BMSCs on the ASCS for the first time.
Freeze-thaw, chemical extraction and mechanical shaking approaches were used to remove the cellular components and prepare a rat ASCS containing only the extracellular matrix (ECM) structure from the rat spinal cord. BMSCs were embedded into ASCSs and freeze-dried agarose scaffolds (FASs), and cell migration and proliferation were observed fluorescence microscopy and the MTT assay.
Compared with the normal rat spinal cord, the ASCS had no cell structure and retained ECM components such as type IV collagen, fibronectin and laminin, showing a three-dimensional network structure with good voids. The growth and proliferation of BMSCs on the ASCS was good, as shown by the MTT assay. Scanning electron microscopy showed that BMSCs covered 65% of the ASCS surface, and the mitochondria of BMSCs were developed and adhered to collagen fibres, as demonstrated by transmission electron microscopy. HE staining showed that BMSCs could grow inside the ASCS, and immunohistochemical staining showed that BMSCs still expressed CD44 and CD90 on the ASCS and had stem cell characteristics.
The results of the experiment indicate that the ASCS has the ability to improve cell adhesion and proliferation. Thus, the ASCS-BMSC combination may be used to treat spinal cord injury.
本研究旨在构建去细胞脊髓支架-骨髓基质细胞(ASCS-BMSC)复合物,模拟移植到大鼠脊髓内的 BMSCs 的生长,首次便于观察和评估 BMSCs 在 ASCS 上的生长情况。
采用冻融、化学提取和机械搅拌法去除细胞成分,从大鼠脊髓中制备仅含细胞外基质(ECM)结构的大鼠 ASCS。将 BMSCs 包埋于 ASCS 与琼脂糖冻干支架(FAS)中,荧光显微镜和 MTT 检测观察细胞迁移和增殖情况。
与正常大鼠脊髓相比,ASCS 无细胞结构,保留了 IV 型胶原、纤维连接蛋白和层粘连蛋白等 ECM 成分,呈三维网络结构,具有良好的空隙。MTT 检测显示 BMSCs 在 ASCS 上生长和增殖良好。扫描电镜显示 BMSCs 覆盖 ASCS 表面的 65%,透射电镜显示 BMSCs 的线粒体发达,黏附于胶原纤维。HE 染色显示 BMSCs 可在 ASCS 内生长,免疫组织化学染色显示 BMSCs 在 ASCS 上仍表达 CD44 和 CD90,具有干细胞特性。
实验结果表明 ASCS 具有促进细胞黏附和增殖的能力。因此,ASCS-BMSC 复合物可能用于治疗脊髓损伤。