Axonal Tract Reconstruction Using a Tissue-Engineered Nigrostriatal Pathway in a Rat Model of Parkinson's Disease.
Axonal Tract Reconstruction Using a Tissue-Engineered Nigrostriatal Pathway in a Rat Model of Parkinson's Disease.
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DOI：
10.3390/ijms232213985
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发表时间：
2022-11-12
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
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作者：
帕金森病（PD）影响1-2%的65岁以上的人，在进行性疾病过程中导致显著的发病率。经典的PD运动缺陷是由黑质多巴胺能神经元的变性引起的，导致其调节纹状体输出的长距离轴突投射的丧失。虽然当代治疗暂时缓解了这种断开的症状，但没有能够取代黑质纹状体通路的方法。我们应用微组织工程技术来创建一个活的、可植入的组织工程化黑质纹状体通路（TE-NSP），该通路模拟了天然通路的结构和功能。TE-NSP包括在水凝胶微柱的管腔内延伸长的成束轴突束的多巴胺能神经元的离散群体。从选择性表达多巴胺能神经元中的绿色荧光蛋白的转基因大鼠的腹侧中脑分离神经元，随后进行荧光激活细胞分选以富集群体至60%纯度。腔细胞外基质和生长因子的变化，以优化细胞结构和神经突长度，而免疫细胞化学和快速扫描循环伏安法（FSCV）显示，TE-NSP轴突释放多巴胺和整合与纹状体神经元在体外。最后，在单侧6-羟基多巴胺SNpc损伤的大鼠PD模型中植入TE-NSP以跨越黑质纹状体通路。免疫组织化学和FSCV证实，移植的TE-NSP存活，维持其轴突束投射，将多巴胺能神经突延伸到宿主组织中，并在纹状体中释放多巴胺。这项工作证明了TE-NSPs可以重建黑质纹状体通路的概念，为未来评估这种策略的潜在功能益处和长期持久性的研究提供了动力。这种通路重建策略可能最终取代失去的神经结构，并减轻PD患者运动症状的原因。
Parkinson’s disease (PD) affects 1–2% of people over 65, causing significant morbidity across a progressive disease course. The classic PD motor deficits are caused by the degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNpc), resulting in the loss of their long-distance axonal projections that modulate striatal output. While contemporary treatments temporarily alleviate symptoms of this disconnection, there is no approach able to replace the nigrostriatal pathway. We applied microtissue engineering techniques to create a living, implantable tissue-engineered nigrostriatal pathway (TE-NSP) that mimics the architecture and function of the native pathway. TE-NSPs comprise a discrete population of dopaminergic neurons extending long, bundled axonal tracts within the lumen of hydrogel micro-columns. Neurons were isolated from the ventral mesencephalon of transgenic rats selectively expressing the green fluorescent protein in dopaminergic neurons with subsequent fluorescent-activated cell sorting to enrich a population to 60% purity. The lumen extracellular matrix and growth factors were varied to optimize cytoarchitecture and neurite length, while immunocytochemistry and fast-scan cyclic voltammetry (FSCV) revealed that TE-NSP axons released dopamine and integrated with striatal neurons in vitro. Finally, TE-NSPs were implanted to span the nigrostriatal pathway in a rat PD model with a unilateral 6-hydroxydopamine SNpc lesion. Immunohistochemistry and FSCV established that transplanted TE-NSPs survived, maintained their axonal tract projections, extended dopaminergic neurites into host tissue, and released dopamine in the striatum. This work showed proof of concept that TE-NSPs can reconstruct the nigrostriatal pathway, providing motivation for future studies evaluating potential functional benefits and long-term durability of this strategy. This pathway reconstruction strategy may ultimately replace lost neuroarchitecture and alleviate the cause of motor symptoms for PD patients.
影响因子：
3.3
作者：
Espana, R. A.;Roberts, D. C. S.;Jones, S. R.
通讯作者：
Jones, S. R.