School of Physics, Dublin Institute of Technology, Kevin Street, Dublin 8, Ireland.
Analyst. 2017 Oct 9;142(20):3848-3856. doi: 10.1039/c7an00969k.
Although consumer exposure to nanomaterials is ever increasing, with potential increased applications in areas such as drug and/or gene delivery, contrast agents and diagnosis, the determination of the cyto- and geno-toxic effects of nanomaterials on human health and the environment still remains challenging. Although many techniques have been established and adapted to determine the cytotoxicity and genotoxicity of nano-sized materials, these techniques remain limited by the number of assays required, total cost, and use of labels and they struggle to explain the underlying interaction mechanisms. In this study, Raman microspectroscopy is employed as an in vitro label-free, high content screening technique to observe toxicological changes within the cell in a multi-parametric fashion. The evolution of spectral markers as a function of time and applied dose has been used to elucidate the mechanism of action of polyamidoamine (PAMAM) dendrimers associated with cytotoxicity and their impact on nuclear biochemistry. PAMAM dendrimers are chosen as a model nanomaterial due to their widely studied cytotoxic and genotoxic properties and commercial availability. Point spectra were acquired from the cytoplasm to monitor the cascade of toxic events occurring in the cytoplasm upon nanoparticle exposure, whereas the spectra acquired from the nucleus and the nucleolus were used to explore PAMAM-nuclear material interaction as well as genotoxic responses.
尽管消费者接触纳米材料的情况日益增多，纳米材料在药物和/或基因传递、对比剂和诊断等领域的潜在应用也在增加，但纳米材料对人类健康和环境的细胞毒性和遗传毒性的确定仍然具有挑战性。尽管已经建立并改编了许多技术来确定纳米材料的细胞毒性和遗传毒性，但这些技术仍然受到所需检测数量、总成本以及标签使用的限制，并且难以解释潜在的相互作用机制。在本研究中，拉曼显微镜被用作一种非标记的、高通量的体外筛选技术，以多参数的方式观察细胞内的毒理学变化。通过随时间和施加剂量的变化来研究光谱标记物的演变，以阐明与细胞毒性相关的聚酰胺-胺（PAMAM）树枝状聚合物的作用机制及其对核生物化学的影响。选择 PAMAM 树枝状聚合物作为模型纳米材料，是因为它们具有广泛研究的细胞毒性和遗传毒性特性以及商业可用性。从细胞质中获取点光谱，以监测纳米颗粒暴露后细胞质中发生的一系列毒性事件，而从细胞核和核仁中获取的光谱则用于探索 PAMAM-核材料相互作用以及遗传毒性反应。
