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壳聚糖支架诱导人牙髓干细胞向神经分化:对脊髓损伤治疗的潜在作用

Chitosan scaffolds induce human dental pulp stem cells to neural differentiation: potential roles for spinal cord injury therapy.

作者信息

Zhang Jinlong, Lu Xiaohui, Feng Guijuan, Gu Zhifeng, Sun Yuyu, Bao Guofeng, Xu Guanhua, Lu Yuanzhou, Chen Jiajia, Xu Lingfeng, Feng Xingmei, Cui Zhiming

机构信息

Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, 226001, China.

Department of Stomatology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, 226001, China.

出版信息

Cell Tissue Res. 2016 Oct;366(1):129-42. doi: 10.1007/s00441-016-2402-1. Epub 2016 May 5.

Abstract

Cell-based transplantation strategies hold great potential for spinal cord injury (SCI) repair. Chitosan scaffolds have therapeutic benefits for spinal cord regeneration. Human dental pulp stem cells (DPSCs) are abundant available stem cells with low immunological incompatibility and can be considered for cell replacement therapy. The purpose of this study is to investigate the role of chitosan scaffolds in the neural differentiation of DPSCs in vitro and to assess the supportive effects of chitosan scaffolds in an animal model of SCI. DPSCs were incubated with chitosan scaffolds. Cell viability and the secretion of neurotrophic factors were analyzed. DPSCs incubated with chitosan scaffolds were treated with neural differentiation medium for 14 days and then neural genes and protein markers were analyzed by Western blot and reverse transcription plus the polymerase chain reaction. Our study revealed a higher cell viability and neural differentiation in the DPSC/chitosan-scaffold group. Compared with the control group, the levels of BDNF, GDNF, b-NGF, and NT-3 were significantly increased in the DPSC/chitosan-scaffold group. The Wnt/β-catenin signaling pathway played a key role in the neural differentiation of DPSCs combined with chitosan scaffolds. Transplantation of DPSCs together with chitosan scaffolds into an SCI rat model resulted in the marked recovery of hind limb locomotor functions. Thus, chitosan scaffolds were non-cytotoxic and provided a conducive and favorable microenvironment for the survival and neural differentiation of DPSCs. Transplantation of DPSCs might therefore be a suitable candidate for treating SCI and other neuronal degenerative diseases.

摘要

基于细胞的移植策略在脊髓损伤(SCI)修复方面具有巨大潜力。壳聚糖支架对脊髓再生具有治疗益处。人牙髓干细胞(DPSCs)是丰富可得的干细胞,免疫不相容性低,可考虑用于细胞替代治疗。本研究的目的是探讨壳聚糖支架在体外对DPSCs神经分化的作用,并评估壳聚糖支架在SCI动物模型中的支持作用。将DPSCs与壳聚糖支架一起培养。分析细胞活力和神经营养因子的分泌。将与壳聚糖支架一起培养的DPSCs用神经分化培养基处理14天,然后通过蛋白质印迹法和逆转录加聚合酶链反应分析神经基因和蛋白质标志物。我们的研究显示,DPSC/壳聚糖支架组具有更高的细胞活力和神经分化。与对照组相比,DPSC/壳聚糖支架组中脑源性神经营养因子(BDNF)、胶质细胞源性神经营养因子(GDNF)、β-神经生长因子(b-NGF)和神经营养因子-3(NT-3)的水平显著升高。Wnt/β-连环蛋白信号通路在DPSCs与壳聚糖支架结合的神经分化中起关键作用。将DPSCs与壳聚糖支架一起移植到SCI大鼠模型中可导致后肢运动功能明显恢复。因此,壳聚糖支架无细胞毒性,为DPSCs的存活和神经分化提供了有利的微环境。因此,DPSCs移植可能是治疗SCI和其他神经元退行性疾病的合适候选方法。

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