Department of Molecular Pharmacology, Physiology and Biotechnology, Center for Biomedical Engineering, Brown University, Providence, RI, USA.
Acta Biomater. 2013 Jul;9(7):7158-68. doi: 10.1016/j.actbio.2013.03.032. Epub 2013 Apr 1.
The path created by aligned Schwann cells (SCs) after nerve injury underlies peripheral nerve regeneration. We developed geometric bioinspired substrates to extract key information needed for axon guidance by deconstructing the topographical cues presented by SCs. We have previously reported materials that directly replicate SC topography with micro- and nanoscale resolution, but a detailed explanation of the means of directed axon extension on SC topography has not yet been described. Here, using neurite tracing and time-lapse microscopy, we analyzed the SC features that influence axon guidance. Novel poly(dimethylsiloxane) materials, fabricated via photolithography, incorporated bioinspired topographical components with the shapes and sizes of aligned SCs, namely somas and processes, where the lengths of the processes were varied but the soma geometry and dimensions were kept constant. Rat dorsal root ganglia neurites aligned to all materials presenting bioinspired topography after 5days in culture and aligned to bioinspired materials presenting soma and process features after only 17h in culture. The key findings of this study were: neurite response to underlying bioinspired topographical features was time dependent, with neurites aligned most strongly to materials presenting combinations of soma and process features at 5days, with higher than average density of either process or soma features, but at 17h they aligned more strongly to materials presenting average densities of soma and process features and to materials presenting process features only. These studies elucidate the influence of SC topography on axon guidance in a time-dependent setting and have implications for the optimization of nerve regeneration strategies.
在神经损伤后,排列整齐的施万细胞(Schwann cells,SCs)所形成的轨迹是周围神经再生的基础。我们开发了几何形状仿生基底,通过解构 SC 提供的地形线索,提取出轴突导向所需的关键信息。我们之前已经报道了可以直接以微纳尺度复制 SC 形貌的材料,但尚未详细解释 SC 形貌对轴突导向的影响机制。在这里,我们使用神经突追踪和延时显微镜分析了影响轴突导向的 SC 特征。通过光刻技术制造的新型聚二甲基硅氧烷(poly(dimethylsiloxane),PDMS)材料,整合了具有与排列整齐的 SC 形貌形状和尺寸相匹配的仿生拓扑结构组件,即胞体和突起,其中突起的长度不同,但胞体的形状和尺寸保持不变。在培养 5 天后,大鼠背根神经节(dorsal root ganglia,DRG)神经突会与所有呈现仿生形貌的材料对齐,而在培养 17 小时后,仅与呈现胞体和突起特征的仿生材料对齐。这项研究的主要发现包括:神经突对底层仿生形貌特征的反应是时间依赖性的,在培养 5 天时,神经突与呈现胞体和突起特征的材料结合时对齐最强,具有高于平均密度的突起或胞体特征,但在培养 17 小时时,它们与呈现平均密度的胞体和突起特征的材料以及仅呈现突起特征的材料对齐更强。这些研究阐明了 SC 形貌在时间依赖性条件下对轴突导向的影响,并对优化神经再生策略具有重要意义。