NT-3/BMSCs-导电性三维神经网络支架复合体移植联合督脉电针修复脊髓损伤的研究

The effective regeneration of neuronal cells and axons play an important key to the repaired of spinal cord injury (SCI). SCI can cause complex pathophysiological changes. And the local microenvironment is not conducive to the nerve regeneration microenvironment. These factors etermine that a single strategy intervention is unable to meet the effective repair for structure and function. The development of genetic engineering and neural tissue engineering technology, as well as electro acupuncture therapy,provides new ideas and means for the research of SCI repair. In this research, due to the conductivity of single wall carbon nanotube (SWCNT), we modify the hydrogel prepared in our before research with the SWCNT, synthesizing hydrogel scaffolds with electrical conductivity. Using genetic engineering techniques, the gene of neurotrophic factor-3 (NT-3) is import into the bone marrow mesenchymal stem cells (BMSCs) to prepare NT-3/ BMSCs which can long-term and stably express the NT-3. Then NT-3/ BMSCs are planted in the conductive hydrogel scaffolds, and construct NT-3/ BMSCs-conductive ydrogel complexes in vitro. And the complexes are implanted into the parts with SCI. After that, the electro-acupuncture therapy is ntroduced and to study the effect of the complexes on the repair of the structure and function of SCI under the electro-acupuncture therapy. This research not only provides new ideas and strategies for the study of SCI, but also provides experimental basis for future clinical treatment of SCI.

脊髓损伤（SCI）后修复的关键在于神经元细胞和轴突的有效再生。SCI复杂的病理生理变化和不利于神经元有效再生的局部微环境，决定了单一策略的干预手段，远远无法满足神经元细胞和轴突的有效再生，以基因工程技术和神经组织工程技术为基础，结合中医针灸疗法，为研究SCI修复提供了新的思路和手段。本课题利用基因工程技术将神经营养因子-3（NT-3）基因导入骨髓间充质干细胞（BMSCs）中，制备长期稳定表达NT-3的NT-3/BMSCs，将其接种于导电性三维神经网络支架中，在体外构建NT-3/BMSCs-导电性三维神经网络支架复合体，并将复合体植入SCI部位后，联合督脉电针疗法，研究该复合体在督脉电针疗法的作用下对SCI的修复作用。本项目为研究SCI修复提供新的思路和策略，为将来临床治疗SCI提供实验依据。

脊髓损伤后修复的关键在于神经元细胞和轴突的有效再生。本课题紧紧围绕脊髓损伤后神经环路的有效重建这一科学问题，主要完成了以下研究内容：将神经营养因子-3（NT-3）基因导入骨髓间充质干细胞（BMSCs）中，合成导电性三维神经网络支架并检测其生物学特性，构建NT-3/BMSCs-导电性三维神经网络支架复合体；探究 NT-3/BMSCs-导电性三维神经网络支架复合体移植SCI处后，联合督脉电针，检测移植的外源性干细胞与宿主脊髓神经环路整合及脊髓修复情况。研究结果表明：NT-3基因修饰的骨髓间充质干细胞能够稳定的表达NT-3，导电性三维神经网络支架的构建，有利于移植干细胞的归巢、生长、分化；在督脉电针的联合治疗下，能够提高脊髓损伤组织内NT-3的表达水平，促进部分移植干细胞源性神经样细胞与宿主脊髓神经元形成更多接触；导电微环境能够促进脊髓损伤区域新生血管的形成，修复受损区域的电传导性能，从而改善脊髓损伤大鼠的瘫痪肢体自主运动功能。项目执行期间，申请人以第1作者或通讯作者，正式发表标注项目资助的SCI论文5篇，其中影响因子＞10的2篇，申报发明专利3项，获得授权实用新型专利1项，申请人获评湖南省青年骨干教师。本项研究从导电三维神经网络支架构建、干细胞基因修饰、督脉经穴电针、移植的外源性干细胞与宿主脊髓神经环路的重建修复脊髓损伤这一主干线，进一步阐明了中医在应用针灸治疗脊髓损伤时首选督脉穴的机理，为将来应用干细胞联合中医药治疗脊髓损伤提供新的理论和策略。