Department of Health & Environment, Nano Systems, Austrian Institute of Technology (AIT), Donau-City Street 1, 1220 Vienna, Austria.
Lab Chip. 2011 Aug 7;11(15):2551-60. doi: 10.1039/c1lc20256a. Epub 2011 Jun 17.
As nanotechnology moves towards widespread commercialization, new technologies are needed to adequately address the potential health impact of nanoparticles (NPs). Assessing the safety of over 30,000 NPs through animal testing would not only be expensive, but it would also raise a number of ethical considerations. Furthermore, existing in vitro cell-based assays are not sufficient in scope to adequately address the complexity of cell-nanoparticle interactions including NP translocation, accumulation and co-transport of e.g. allergens. In particular, classical optical/fluorescent endpoint detection methods are known to provide irreproducible, inaccurate and unreliable results since these labels can directly react with the highly catalytic surfaces of NP. To bridge this technological gap we have developed a lab-on-a-chip capable of continuously and non-invasively monitoring the collagen production of primary human fibroblast cells (NHDF) using contactless dielectric microsensors. Human dermal fibroblast cells are responsible for the maintenance of soft tissue integrity, are found throughout the human body and their primary function is collagen expression. We show that cellular collagen production can be readily detected and used to assess cellular stress responses to a variety of external stimuli, including exposure to nanoparticles. Results of the study showed a 20% and 95% reduction of collagen production following 4 hour exposure to 10 μg mL(-1) gold and silver nanoparticles (dia.10 nm), respectively. Furthermore a prolonged perfusion of sub-toxic concentrations (0.1 μg mL(-1)) of silver NP reduced NHDF collagen production by 40% after 10 h indicating increased NP take up and accumulation. We demonstrate that the application of microfluidics for the tailored administration of different NP treatments constitutes a powerful new tool to study cell-nanoparticle interactions and nanoparticle accumulation effects in small cell populations.
随着纳米技术向广泛商业化迈进，需要新的技术来充分应对纳米颗粒（NPs）的潜在健康影响。通过动物测试评估超过 30000 种 NPs 的安全性不仅昂贵，而且还会引发许多伦理问题。此外，现有的基于细胞的体外检测方法在范围上不足以充分解决细胞-纳米颗粒相互作用的复杂性，包括 NP 的易位、积累和共转运，例如过敏原。特别是，经典的光学/荧光终点检测方法由于这些标记物可以直接与 NP 的高催化表面反应，因此提供的结果是不可重复的、不准确的和不可靠的。为了弥补这一技术差距，我们开发了一种能够使用非接触式介电微传感器连续和非侵入式监测原代人成纤维细胞（NHDF）胶原蛋白产生的芯片实验室。人真皮成纤维细胞负责维持软组织的完整性，遍布人体，其主要功能是胶原蛋白的表达。我们表明，可以轻松检测到细胞胶原蛋白的产生，并将其用于评估细胞对各种外部刺激（包括暴露于纳米颗粒）的应激反应。研究结果表明，在暴露于 10μgmL（-1）金和银纳米颗粒（直径 10nm）4 小时后，胶原蛋白的产生分别减少了 20%和 95%。此外，在 10 小时后，长时间灌注亚毒性浓度（0.1μgmL（-1））的银 NP 会使 NHDF 胶原蛋白的产生减少 40%，表明 NP 摄取和积累增加。我们证明，微流控技术在不同 NP 处理的定制管理中的应用构成了研究细胞-纳米颗粒相互作用和纳米颗粒积累效应的一种强大的新工具，可用于小细胞群体中。
