Laser Zentrum Hannover e.V., Hollerithallee 8, 30419 Hannover, Germany.
ACS Appl Mater Interfaces. 2013 Feb;5(3):1070-7. doi: 10.1021/am3028487. Epub 2013 Jan 30.
To improve neuronal-electrode interfaces, we analyzed the influence of surface topographies combined with coating on the electrochemistry of platinum and neuronal differentiation of PC-12 cells. Surface structuring on nanoscale was realized by femtosecond laser ablation. Additional coating with laminin (LA), collagen type I (COL) or poly-d-lysine (PDL) did not change the produced topography. We further demonstrated that impedance could be improved in all cases. The pre-requisites of differentiation - viability and attachment - were fulfilled on the topography. Cell attachment of non-differentiated and differentiated cells and their formation of focal adhesion complexes were even enhanced compared to unstructured platinum. However, without the nerve growth factor (NGF) no cellular outgrowth and differentiation were possible. The topography enabled cell elongation and reduced the amount of rounded cells, but less effective than coating. Differentiation was either comparable or increased on the structures when compared with unstructured coatings. For instance, microtubule associated protein (MAP2) was detected most on the topography alone. But a combination of surface structuring and coating had the strongest impact on differentiation: the usage of COL provoked best cell elongation and beta III tubulin expression, PDL best synaptophysin. LA-coating had no noteworthy effect. These findings point out that innovative electronic devices like cochlear implants include two aspects: (a) nanotopography to improve the transmission of electrical signals and neuronal attachment; and (b) an additional coating to stimulate neuronal differentiation.
为了改善神经元-电极界面,我们分析了表面形貌与涂层相结合对铂电化学和 PC-12 细胞神经元分化的影响。利用飞秒激光烧蚀实现了纳米级的表面结构化。用层粘连蛋白 (LA)、胶原蛋白 I (COL) 或聚-d-赖氨酸 (PDL) 进行额外的涂层不会改变所产生的形貌。我们进一步证明,在所有情况下,阻抗都可以得到改善。分化的前提 - 活力和附着 - 在形貌上得到满足。未分化和分化细胞的细胞附着及其焦点粘附复合物的形成甚至比未结构化的铂更增强。然而,没有神经生长因子 (NGF),细胞就不可能生长和分化。形貌使细胞伸长并减少了圆形细胞的数量,但效果不如涂层。与未结构化的涂层相比,在结构上的分化要么相似,要么增加。例如,微管相关蛋白 (MAP2) 仅在形貌上检测到最多。但是,表面结构和涂层的结合对分化有最强的影响:使用 COL 可引起最佳的细胞伸长和 β III 微管蛋白表达,PDL 可引起最佳的突触素表达。LA 涂层没有明显的效果。这些发现指出,像耳蜗植入物这样的创新电子设备包括两个方面:(a) 纳米形貌以改善电信号的传输和神经元附着;和 (b) 额外的涂层以刺激神经元分化。