Bellák Tamás, Fekécs Zoltán, Török Dénes, Kristóf Rebeka, Esezoobo Omoikhoje, Marton Annamária, Vizler Csaba, Nógrádi Antal, Pajer Krisztián
Department of Anatomy, Histology and Embryology, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, Hungary.
Present address: Galway University Hospitals, Galway, Ireland.
Stem Cell Res Ther. 2025 May 13;16(1):237. doi: 10.1186/s13287-025-04283-9.
Avulsion injury results in motoneuron death due to the increased cytotoxicity developing after the injury. We have earlier shown that intraspinally grafted immortalized NE-4C neuroectodermal stem cells derived from 9-day old mouse forebrain vesicles produced a secretome, which induced decreased microglia/macrophage reaction, and promoted the neuroprotection and regeneration following avulsion injury. Here we intended to prove the motoneuron rescuing effect of intravenously grafted NE-4C stem cells and reveal the mechanism of action used by the grafted cells.
In our experimental model the left lumbar 4 (L4) ventral root of the spinal cord was avulsed and then reimplanted into the L4 spinal segment. Treated animals received various doses of NE-4C stem cells intravenously and the survival and regeneration of the affected motoneurons was checked by morphological and functional analysis. The molecular changes within the treated cord were followed by the ELISA Proteome Profiler rat cytokine array and qPCR analysis. To mimic the effect of stem cells fucoidan treatment (a specific selectin inhibitor, 50 and 100 mg/kg bw) was applied for two weeks intraperitoneally.
High doses of intravenous stem cell treatment (4 × 10 and 1 × 10 cells) induced the reinnervation of the reimplanted ventral root by surviving injured motoneurons (up to 38% of the total L4 pool). Proteome Profiler analysis showed that systemic stem cell treatment downregulated the level of L-selectin, that promotes leukocyte rolling on vascular endothelium. Both systemic stem cell and fucoidan treatment reduced macrophage and microglial densities in the affected spinal segment and administration of fucoidan downregulated inflammatory cytokine and inflammasome levels along with improved morphological and functional reinnervation.
Blocking L-selectin, similarly to systemic NE-4C stem cell treatment decreases the neuroinflammation in the injured spinal cord segment after ventral root avulsion and induces significant motoneuron survival and functional reinnervation of the denervated hind limb muscles.
撕脱伤会导致运动神经元死亡，这是由于损伤后细胞毒性增加所致。我们之前已经表明，脊髓内移植源自9日龄小鼠前脑泡的永生化NE-4C神经外胚层干细胞会产生一种分泌组，该分泌组可诱导小胶质细胞/巨噬细胞反应减弱，并促进撕脱伤后的神经保护和再生。在此，我们旨在证明静脉内移植NE-4C干细胞对运动神经元的拯救作用，并揭示移植细胞所使用的作用机制。
在我们的实验模型中，脊髓左腰4（L4）腹侧神经根被撕脱，然后重新植入L4脊髓节段。接受治疗的动物静脉内给予不同剂量的NE-4C干细胞，并通过形态学和功能分析检查受影响运动神经元的存活和再生情况。通过ELISA蛋白质组分析大鼠细胞因子阵列和qPCR分析追踪治疗脊髓内的分子变化。为模拟干细胞的作用，腹腔内应用岩藻依聚糖治疗（一种特异性选择素抑制剂，50和100mg/kg体重）两周。
高剂量静脉内干细胞治疗（4×10和1×10个细胞）诱导存活的损伤运动神经元对重新植入的腹侧神经根进行再支配（高达L4总池的38%）。蛋白质组分析表明，全身干细胞治疗下调了L-选择素的水平，L-选择素可促进白细胞在血管内皮上滚动。全身干细胞和岩藻依聚糖治疗均降低了受影响脊髓节段中的巨噬细胞和小胶质细胞密度，并且岩藻依聚糖的给药下调了炎性细胞因子和炎性小体水平，同时改善了形态学和功能再支配。
与全身NE-4C干细胞治疗类似，阻断L-选择素可降低腹侧神经根撕脱后损伤脊髓节段中的神经炎症，并诱导显著的运动神经元存活以及失神经后肢肌肉的功能再支配。
