Background aims

Spinal cord injury (SCI) is one of the most complex and destructive diseases of the nervous system, which can lead to permanent loss of tactile perception. But existing treatment methods have limited effects. To establish a novel method that may be therapeutic in repairing the injured spinal cord, gene-modified dental pulp stem cells (DPSCs) were injected in situ.

Methods

Adenovirus carrying osteopontin (OPN), Insulin-like growth factor 1 (IGF-1) and cailiary-derived neurotrophic factor (CNTF) (Ad-OIC) was constructed. After modified with Ad-OIC, supernatant of DPSC were co-cultured with HT-22 cells and the effect of DPSC-OIC on the HT-22 cells were evaluated via Cell Counting Kit-8 (CCK-8) assay, Real-Time polymerase chain reaction (PCR) analysis, laser confocal microscopy and fluorescence activating cell sorter (FACS). DPSC-OIC were injected in the lesion area of injured spinal cord and the survival time of transplanted cells were measured by bioluminescence imaging system. The recovery of the injured spinal cord was evaluated by behavioral score, radiological evaluation and immunopathological analysis.

Results

DPSC-OIC could enhance the proliferation and axon growth of HT-22 cells, and protect HT-22 cells from H₂O₂ induced apoptosis. The transplanted DPSC-Null or DPSC-OIC could survive for more than two weeks in local injection site. DPSC-OIC treatment could increase Basso-Mouse Scale (BMS) scores, improve Magnetic Resonance Imaging (MRI) manifestation and promote bladder function recovery. Less apoptotic neurons and more proliferative cells were found in the lesion area of DPSC-OIC treated spinal cord. Nestin⁺ cells and neural stem cell marker (Sox2) were both up-regulated after DPSC-OIC treatment. Additionally, inhibitory extracellular matrix proteoglycan Neural/Glial Antigen 2 (NG2) was down-regulated and axon growth promotive factor fibronectin was up-regulated after both DPSC-Null (DPSCs infected with Ad-Null) and DPSC-OIC treatments.

Conclusions

DPSC-OIC could be a novel effective method for treating SCI.

中文翻译：

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OPN、IGF-1 和 CNTF 共过表达增强 DPSC 对脊髓损伤的治疗作用

背景目标

脊髓损伤（SCI）是神经系统最复杂和最具破坏性的疾病之一，可导致触觉永久丧失。但现有的治疗方法效果有限。为了建立一种可用于修复受损脊髓的新方法，原位注射了基因修饰的牙髓干细胞（DPSC）。

方法

构建了携带骨桥蛋白（OPN）、胰岛素样生长因子1（IGF-1）和睫状源性神经营养因子（CNTF）（Ad-OIC）的腺病毒。经过 Ad-OIC 修饰后，DPSC 上清液与 HT-22 细胞共培养，并通过 Cell Counting Kit-8 (CCK-8) 检测实时评估 DPSC-OIC 对 HT-22 细胞的影响聚合酶链反应 (PCR) 分析、激光共聚焦显微镜和荧光激活细胞分选仪 (FACS)。将DPSC-OIC注射到损伤脊髓的病变区域，并通过生物发光成像系统测量移植细胞的存活时间。通过行为评分、放射学评估和免疫病理学分析来评估受伤脊髓的恢复情况。

结果

DPSC-OIC可以增强HT-22细胞的增殖和轴突生长，并保护HT-22细胞免受H ₂ O ₂诱导的细胞凋亡。移植的DPSC-Null或DPSC-OIC在局部注射部位可存活两周以上。DPSC-OIC治疗可以提高巴索小鼠量表（BMS）评分，改善磁共振成像（MRI）表现并促进膀胱功能恢复。DPSC-OIC处理的脊髓损伤区发现凋亡神经元较少，增殖细胞较多。DPSC-OIC 处理后， Nestin ⁺细胞和神经干细胞标记物 (Sox2) 均上调。此外，在 DPSC-Null（感染 Ad-Null 的 DPSC）和 DPSC-OIC 治疗后，抑制性细胞外基质蛋白聚糖神经/胶质抗原 2 (NG2) 下调，轴突生长促进因子纤连蛋白上调。

结论

DPSC-OIC 可能是治疗 SCI 的一种新的有效方法。