Deng Win-Ping, Yang Chi-Chiang, Yang Liang-Yo, Chen Chun-Wei D, Chen Wei-Hong, Yang Charn-Bing, Chen Yu-Hsin, Lai Wen-Fu T, Renshaw Perry F
Graduate Institute of Biomedical Materials and Engineering, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei, Taiwan.
Department of Neurology, Tungs' Taichung Metroharbor Hospital, 699 Taiwan Blvd. 8 Sec., Taitung, Taiwan.
Spine J. 2014 Oct 1;14(10):2488-99. doi: 10.1016/j.spinee.2014.04.024. Epub 2014 Apr 30.
Recent advanced studies have demonstrated that cytokines and extracellular matrix (ECM) could trigger various types of neural differentiation. However, the efficacy of differentiation and in vivo transplantation has not yet thoroughly been investigated.
To highlight the current understanding of the effects of ECM on neural differentiation of human bone marrow-derived multipotent progenitor cells (MPCs), regarding state-of-art cure for the animal with acute spinal cord injury (SCI), and explore future treatments aimed at neural repair.
A selective overview of the literature pertaining to the neural differentiation of the MSCs and experimental animals aimed at improved repair of SCI.
Extracellular matrix proteins, tenascin-cytotactin (TN-C), tenascin-restrictin (TN-R), and chondroitin sulfate (CS), with the cytokines, nerve growth factor (NGF)/brain-derived neurotrophic factor (BDNF)/retinoic acid (RA) (NBR), were incorporated to induce transdifferentiation of human MPCs. Cells were treated with NBR for 7 days, and then TN-C, TN-R, or CS was added for 2 days. The medium was changed every 2 days. Twenty-four animals were randomly assigned to four groups with six animals in each group: one experimental and three controls. Animals received two (bilateral) injections of vehicle, MPCs, NBR-induced MPCs, or NBR/TN-C-induced MPCs into the lesion sites after SCI. Functional assessment was measured using the Basso, Beattie, and Bresnahan locomotor rating score. Data were analyzed using analysis of variance followed by Student-Newman-Keuls (SNK) post hoc tests.
Results showed that MPCs with the transdifferentiation of human MPCs to neurons were associated with increased messenger-RNA (mRNA) expression of neuronal markers including nestin, microtubule-associated protein (MAP) 2, glial fibrillary acidic protein, βIII tubulin, and NGF. Greater amounts of neuronal morphology appeared in cultures incorporated with TN-C and TN-R than those with CS. The addition of TN-C enhanced mRNA expressions of MAP2, βIII tubulin, and NGF, whereas TN-R did not significantly change. Conversely, CS exposure decreased MAP2, βIII tubulin, and NGF expressions. The TN-C-treated MSCs significantly and functionally repaired SCI-induced rats at Day 42. Present results indicate that ECM components, such as tenascins and CS in addition to cytokines, may play functional roles in regulating neurogenesis by human MPCs.
These findings suggest that the combined use of TN-C, NBR, and human MPCs offers a new feasible method for nerve repair.
近期的前沿研究表明，细胞因子和细胞外基质（ECM）可触发多种类型的神经分化。然而，分化的效果以及体内移植的效果尚未得到充分研究。
强调目前对ECM对人骨髓来源的多能祖细胞（MPCs）神经分化影响的理解，涉及急性脊髓损伤（SCI）动物的最新治疗方法，并探索旨在神经修复的未来治疗方法。
对有关MSCs神经分化和旨在改善SCI修复的实验动物的文献进行选择性综述。
将细胞外基质蛋白腱生蛋白 - 细胞趋触蛋白（TN - C）、腱生蛋白 - 限制素（TN - R）和硫酸软骨素（CS）与细胞因子神经生长因子（NGF）/脑源性神经营养因子（BDNF）/视黄酸（RA）（NBR）结合，诱导人MPCs转分化。细胞先用NBR处理7天，然后加入TN - C、TN - R或CS处理2天。每2天更换一次培养基。24只动物随机分为四组，每组6只：一组为实验组，三组为对照组。SCI后，动物在损伤部位接受双侧注射载体、MPCs、NBR诱导的MPCs或NBR/TN - C诱导的MPCs。使用Basso、Beattie和Bresnahan运动评分量表进行功能评估。数据采用方差分析，随后进行Student - Newman - Keuls（SNK）事后检验进行分析。
结果表明，人MPCs向神经元转分化的MPCs与神经元标志物包括巢蛋白、微管相关蛋白（MAP）2、胶质纤维酸性蛋白、βIII微管蛋白和NGF的信使核糖核酸（mRNA）表达增加有关。与CS组相比，加入TN - C和TN - R的培养物中出现了更多的神经元形态。TN - C的添加增强了MAP2、βIII微管蛋白和NGF的mRNA表达，而TN - R没有显著变化。相反，CS处理降低了MAP2、βIII微管蛋白和NGF的表达。在第42天，TN - C处理的MSCs显著且功能性地修复了SCI诱导的大鼠。目前的结果表明，除细胞因子外，ECM成分如腱生蛋白和CS可能在调节人MPCs的神经发生中发挥功能作用。
这些发现表明，联合使用TN - C、NBR和人MPCs为神经修复提供了一种新的可行方法。
