Hernández-Ferrer Javier, Pérez-Bruzón Rodolfo N, Azanza María J, González Mónica, Del Moral Raquel, Ansón-Casaos Alejandro, de la Fuente Jesús M, Marijuan Pedro C, Martínez M Teresa
Instituto de Carboquímica (CSIC), C/ Miguel Luesma Castán 4, E-50018, Zaragoza, Spain.
J Biomed Mater Res A. 2014 Dec;102(12):4443-54. doi: 10.1002/jbm.a.35110. Epub 2014 Feb 26.
Cultures of primary embryonic rat brain hippocampus neurons with supporting glia cells were carried out on different substrates containing polypyrrole (PPy) and/or single-walled carbon nanotubes (SWCNTs). Neuron adhesion, neurites and dendrites branching elongation, and development of neuron networks on substrates were followed by phase-contrast optical microscopy and quantified to state cell survival and proliferation. Suspensions of as-grown and purified SWCNTs were sprayed on a glass coverslips and PPy/SWCNTs were deposited by potentiodynamic electrochemical deposition. Cell neurotoxicity revealed by neuron death was very high for purified SWCNTs substrates in good agreement with [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) test showing lower viability on SWCNTs containing substrates compared with PPy-substrates and control samples probably due to the metal content and the carboxylic groups introduced during the purification. It is interesting to highlight that neurons grown on PPy-substrates adhere developing neurites and branching dendrites earlier even than on control cultures. On subsequent days the neurons are able to adapt to nanotube substrates developing neuron networks for 14-day cultures with similar patterns of complexity for control, PPy and PPy/SWCNT substrates. PPy/SWCNT substrates show a lower impedance value at frequencies under 1 Hz. We have come to the conclusion that glia cells and PPy added to the culture medium and substrates respectively, improve in some degree nanotube biocompatibility, cell adhesion and hence cell viability.
在含有聚吡咯(PPy)和/或单壁碳纳米管(SWCNT)的不同基质上,对原代胚胎大鼠脑海马神经元与支持性神经胶质细胞进行培养。通过相差光学显微镜观察基质上神经元的黏附、神经突和树突的分支伸长以及神经网络的发育情况,并进行量化以确定细胞的存活和增殖。将生长并纯化后的SWCNT悬浮液喷在玻璃盖玻片上,通过动电位电化学沉积法沉积PPy/SWCNT。纯化后的SWCNT基质导致的神经元死亡所揭示的细胞神经毒性非常高,这与[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐](MTT)试验结果高度一致,该试验表明与PPy基质和对照样品相比,含SWCNT的基质上细胞活力较低,这可能是由于纯化过程中引入的金属含量和羧基所致。值得注意的是,在PPy基质上生长的神经元比在对照培养物上更早地黏附并发育出神经突和分支树突。在随后的几天里,神经元能够适应纳米管基质,在14天的培养过程中形成神经网络,对照、PPy和PPy/SWCNT基质的神经网络复杂性模式相似。PPy/SWCNT基质在1Hz以下频率时显示出较低的阻抗值。我们得出的结论是,分别添加到培养基和基质中的神经胶质细胞和PPy在一定程度上改善了纳米管的生物相容性、细胞黏附,从而提高了细胞活力。