Microengineered peripheral nerve-on-a-chip for preclinical physiological testing
Microengineered peripheral nerve-on-a-chip for preclinical physiological testing
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DOI：
10.1039/c4lc01513d
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发表时间：
2015-01-01
期刊：
影响因子：
6.1
通讯作者：
Moore, Michael J.
中科院分区：
文献类型：
--
作者：
Huval, Renee M.;Miller, Oliver H.;Moore, Michael J.
使用先进的体外测试是开发适用于改善新型药物化合物的高消耗率的预测性细胞测定的有力工具。具有模拟临床神经复合动作电位（CAP）和神经纤维密度（NFD）测试的生理测量的神经组织的微尺度器官型模型可以更好地预测临床结果，从而实现更具成本效益的方法来选择具有更高后期成功机会的有前景的先导化合物。然而，神经系统的结构、生理学和周围的细胞外基质很难在体外模拟。使用双水凝胶结构和外植体从大鼠胚胎背根神经节，本研究描述了一种在体外的方法，使用空间控制的，微工程的感觉神经纤维束的细胞内和细胞外的记录的电生理记录。具体而言，这些三维神经文化表现出结构和功能的特点，密切模仿那些在体内发现的传入感觉外周纤维。我们的双水凝胶系统在空间上将生长限制在类似神经纤维束的几何形状，从而允许高密度的平行束状神经生长。也许更重要的是，通过我们的先进模型，类似于临床相关测试标准的输出，包括CAP和NFD的测量是可能的。此外，3D水凝胶构建体允许在掺入的细胞类型、几何构造和电操作中的灵活性，为系统培养、扰动和仿生神经生长的测试提供可行的测定，用于需要生理学相关读数的机械研究。
The use of advanced in vitro testing is a powerful tool to develop predictive cellular assays suitable for improving the high attrition rates of novel pharmaceutical compounds. A microscale, organotypic model of nerve tissue with physiological measures that mimic clinical nerve compound action potential (CAP) and nerve fiber density (NFD) tests may be more predictive of clinical outcomes, enabling a more cost-effective approach for selecting promising lead compounds with higher chances of late-stage success. However, the neurological architecture, physiology, and surrounding extracellular matrix are hard to mimic in vitro. Using a dual hydrogel construct and explants from rat embryonic dorsal root ganglia, the present study describes an in vitro method for electrophysiological recording of intra- and extra-cellular recordings using a spatially-controlled, microengineered sensory neural fiber tract. Specifically, these 3D neural cultures exhibit both structural and functional characteristics that closely mimic those of afferent sensory peripheral fibers found in vivo. Our dual hydrogel system spatially confines growth to geometries resembling nerve fiber tracts, allowing for a high density of parallel, fasciculated neural growth. Perhaps more importantly, outputs resembling clinically relevant test criteria, including the measurement of CAP and NFD are possible through our advanced model. Moreover, the 3D hydrogel constructs allow flexibility in incorporated cell type, geometric fabrication, and electrical manipulation, providing a viable assay for systematic culture, perturbation, and testing of biomimetic neural growth for mechanistic studies necessitating physiologicallyrelevant readouts.