首页> 外文期刊>Acta Oto-Laryngologica >Transplantation of neural stem cells overexpressing glia-derived neurotrophic factor promotes facial nerve regeneration
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Transplantation of neural stem cells overexpressing glia-derived neurotrophic factor promotes facial nerve regeneration

机译:过度表达神经胶质细胞源性神经营养因子的神经干细胞移植促进面神经再生

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Conclusion. Combining neurotrophic factor support and neural stem cell (NSC) transplantation may improve regeneration of the peripheral nervous system. Objectives. We constructed a biodegradable nerve conduit (NC) filled with NSCs overexpressing glia-derived neurotrophic factor (GDNF), which is known to protect facial motoneurons, and tested the effect of this NC on facial nerve regeneration. Materials and methods. Primary cultured NSCs were transduced with a lentiviral vector encoding enhanced green fluorescent protein (EGFP) and GDNF. GDNF expression was confirmed by Western blotting and ELISA. Sprague Dawley (SD) rats were subjected to right facial nerve transection, and polyglycolic/ polyglycolic acid (PLGA) NCs filled with NSCs-GDNF were used to bridge the nerve gap (n=24). In vivo GDNF expression was confirmed by real-time PCR. NCs containing NSCs, transgenic Schwann cells (SCs-GDNF), or empty NCs served as controls (n=24 per group). Facial nerve regeneration was assessed 2-12 weeks after surgery, by electro-physiological testing, immunohistochemical staining, and morphometric analysis of axons. Results. NSCs exhibited sustained and robust GDNF expression in culture and following implantation. Nerve action potential amplitude, axonal area, and axonal number were significantly greater in the NSCs-GDNF group than in the NSCs or empty NC groups. Axonal area and number were also greater in the NSCs-GDNF group than the SCs-GDNF group, although this was not statistically significant. The enhanced regeneration observed in the NSCs-GDNF group was accompanied by increased labeling for S100, NF, and betaIII tubulin.
机译:结论。结合神经营养因子支持和神经干细胞(NSC)移植可以改善周围神经系统的再生。目标。我们构建了一个可生物降解的神经导管(NC),其中充满了过表达神经胶质细胞衍生的神经营养因子(GDNF)的NSC,已知该NSC保护面部运动神经元,并测试了该NC对面部神经再生的影响。材料和方法。用编码增强的绿色荧光蛋白(EGFP)和GDNF的慢病毒载体转导原代培养的NSC。通过蛋白质印迹和ELISA确认GDNF表达。对Sprague Dawley(SD)大鼠进行右面部神经横断,并用充满NSCs-GDNF的聚乙醇酸/聚乙醇酸(PLGA)NC桥接神经间隙(n = 24)。通过实时PCR证实了体内GDNF表达。包含NSC,转基因雪旺细胞(SCs-GDNF)或空NC的NC用作对照(每组n = 24)。术后2-12周,通过电生理测试,免疫组织化学染色和轴突形态分析评估面部神经再生。结果。 NSC在培养物中和植入后表现出持续且稳定的GDNF表达。 NSCs-GDNF组的神经动作电位振幅,轴突面积和轴突数目显着大于NSCs或空NC组。 NSCs-GDNF组的轴突面积和数量也比SCs-GDNF组的大,尽管这在统计学上并不显着。在NSCs-GDNF组中观察到增强的再生,同时伴随着S100,NF和betaIII微管蛋白的标记增加。

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