Kovach Kyle M, Kumsa Doe W, Srivastava Vishnupriya, Hudak Eric M, Untereker Darrel F, Kelley Shawn C, von Recum Horst A, Capadona Jeffrey R
Department of Biomedical Engineering, Case Western Reserve University, 2071 Martin Luther King Jr. Drive, Wickenden Bldg., Cleveland, OH 44106, USA; Advanced Platform Technology Center, Louis Stokes Cleveland Department of Veterans Affairs Medical Center, 10701 East Blvd, 151W/APT, Cleveland, OH 44106, USA.
Department of Biomedical Engineering, Case Western Reserve University, 2071 Martin Luther King Jr. Drive, Wickenden Bldg., Cleveland, OH 44106, USA.
J Neurosci Methods. 2016 Nov 1;273:1-9. doi: 10.1016/j.jneumeth.2016.07.018. Epub 2016 Jul 30.
It is currently unclear how the platinum (Pt) species released from platinum-containing stimulating electrodes may affect the health of the surrounding tissue. This study develops an effective system to assess the cytotoxicity of any electrode-liberated Pt over a short duration, to screen systems before future in vivo testing.
A platinum electrode was stimulated for two hours under physiologically relevant conditions to induce the liberation of Pt species. The total concentration of liberated Pt species was quantified and the concentration found was used to develop a range of Pt species for our model system comprised of microglia and neuron-like cells.
Under our stimulation conditions (k=2.3, charge density of 57.7μC/cm), Pt was liberated to a concentration of 1ppm. Interestingly, after 24h of Pt exposure, the dose-dependent cytotoxicity plots revealed that cell death became statistically significant at 10ppm for microglia and 20ppm for neuronal cells. However, in neuron-like cell cultures, concentrations above 1ppm resulted in significant neurite loss after 24h.
To our knowledge, there does not exist a simple, in vitro assay system for assessing the cytotoxicity of Pt liberated from stimulating neural electrodes.
This work describes a simple model assay that is designed to be applicable to almost any electrode and stimulation system where the electrode is directly juxtaposed to the neural target. Based on the application, the duration of stimulation and Pt exposure may be varied.
目前尚不清楚含铂刺激电极释放的铂(Pt)物种如何影响周围组织的健康。本研究开发了一种有效的系统,用于在短时间内评估任何电极释放的Pt的细胞毒性,以便在未来的体内测试之前筛选系统。
在生理相关条件下对铂电极进行两小时刺激,以诱导Pt物种的释放。对释放的Pt物种的总浓度进行定量,并将所发现的浓度用于为我们由小胶质细胞和神经元样细胞组成的模型系统开发一系列Pt物种。
在我们的刺激条件下(k = 2.3,电荷密度为57.7μC/cm),Pt释放到浓度为1ppm。有趣的是,在Pt暴露24小时后,剂量依赖性细胞毒性图显示,小胶质细胞在10ppm时细胞死亡具有统计学意义,神经元细胞在20ppm时具有统计学意义。然而,在神经元样细胞培养物中,24小时后浓度高于1ppm会导致明显的神经突损失。
据我们所知,不存在一种简单的体外测定系统来评估从刺激神经电极释放的Pt的细胞毒性。
这项工作描述了一种简单的模型测定方法,该方法设计适用于几乎任何电极与神经靶点直接并置的电极和刺激系统。根据应用情况,刺激持续时间和Pt暴露时间可以变化。