Department of Mechanical and Materials Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588-0526, USA.
Acta Biomater. 2013 Jan;9(1):4592-8. doi: 10.1016/j.actbio.2012.08.039. Epub 2012 Aug 30.
Creating physical-biochemical superposed microenvironments optimal for stimulating neurite outgrowth would be beneficial for neuronal regenerative medicine. We investigated potential co-regulatory effects of cell micropatterning and retinoic acid (RA) soluble factor on neuronal cell morphology and neurite outgrowth. Human neuroblastoma (SH-SY5Y) cell patterning sensitivity could be enhanced by poly-L-lysine-g-polyethylene glycol cell-repellent back-filling, enabling cell confinement in lanes as narrow as 5 μm. Cells patterned on narrow (5 and 10 μm) lanes showed preferred nucleus orientation following the patterning direction. These cells also showed high nucleus aspect ratio but constrained nucleus spreading. On the other hand, cells on wide (20 μm and above) lanes showed random nucleus orientation and cell and nucleus sizes similar to those on unpatterned controls. All these changes were generally maintained with or without RA. Confining cells on narrow (5 and 10 μm) lanes, even without RA, significantly enhanced neurite extension relative to unpatterned control, which was further stimulated by RA. Interestingly, cell patterning on 5 and 10 μm lanes without RA produced longer neurites relative to the RA treatment alone case. Our data on the potential interplay between microscale physical cell confinement and RA-soluble stimulation may provide a new, integrative insight on how to trigger neurite/axon formation for neuronal regenerative medicine.
创建有利于刺激神经突生长的物理-生化超叠加微环境将有益于神经再生医学。我们研究了细胞微图案和维甲酸（RA）可溶性因子对神经元细胞形态和神经突生长的潜在协同调节作用。聚-L-赖氨酸-g-聚乙二醇细胞排斥性背填充可增强人神经母细胞瘤（SH-SY5Y）细胞的图案化敏感性，从而使细胞能够局限在 5μm 宽的狭缝中。在狭窄（5 和 10μm）狭缝中图案化的细胞在图案化方向上显示出优先的细胞核取向。这些细胞还表现出高核纵横比，但限制了核的扩散。另一方面，在宽（20μm 及以上）狭缝上的细胞显示出随机的细胞核取向以及与未图案化对照相似的细胞和细胞核大小。所有这些变化通常在有或没有 RA 的情况下都能维持。将细胞限制在狭窄（5 和 10μm）狭缝中，即使没有 RA，也能显著增强相对于未图案化对照的神经突延伸，而 RA 进一步刺激了这一过程。有趣的是，即使没有 RA，细胞在 5μm 和 10μm 狭缝上的图案化也会产生比单独用 RA 处理更长的神经突。我们关于微尺度物理细胞限制和 RA 可溶性刺激之间潜在相互作用的数据可能为如何触发神经突/轴突形成提供新的、综合的见解，以用于神经再生医学。
