Bhamidipati Manjari, Fabris Laura
Department of Biomedical Engineering, Rutgers University , 599 Taylor Road, Piscataway, New Jersey 08854, United States.
Bioconjug Chem. 2017 Feb 15;28(2):449-460. doi: 10.1021/acs.bioconjchem.6b00605. Epub 2017 Jan 6.
In recent years, we and others have become interested in evaluating the use of surface-enhanced Raman scattering (SERS) tags for early cancer detection and in designing new approaches to demonstrate the applicability of this spectroscopic technique in the clinic. SERS-based imaging in particular offers ultra sensitivity up to the single molecule, multiplexing capability, and increased photostability and has been shown to outperform fluorescence. However, to employ SERS tags for early cancer detection, it is important to understand their interaction with cells and determine their cytotoxicity. We have been particularly interested for quite some time in determining if and how gold nanostars, which have been demonstrated as outstanding SERS enhancing substrates, can be safely employed in living systems and translated to the clinic. In this study, we carried out a multiparametric in vitro study to look at the cytotoxicity and cellular uptake of gold nanoparticles on human glioblastoma and human dermal fibroblast cell lines. Cytotoxicity was evaluated by incubating cells with three different morphologies of AuNPs, namely nanospheres, nanorods, and nanostars, each having three different surface chemistries (cetyltrimethylammonium bromide (CTAB), poly(ethylene glycol) (PEG), and human serum albumin (HSA)). Our results showed that the surface chemistry of the nanoparticles had predominant effects on cytotoxicity, and the morphology and size of the nanoparticles only slightly affected cell viability. CTAB-coated particles were found to be the most toxic to cells, and PEGylated nanostars were determined to be the least toxic. Caspase-3 assay and LDH assay revealed that cell death occurs via apoptosis for cancerous cells and via necrosis for healthy ones. Cellular uptake studies carried out via TEM showed that the particles retain their shape even at long incubation times, which may be beneficial for in vivo SERS-based disease detection. Overall, this study provides valuable information on gold-nanoparticle-induced cytotoxicity that can be leveraged for the development of safe and effective nanoparticle-based therapeutic and diagnostic systems.
近年来，我们和其他一些人开始关注评估表面增强拉曼散射（SERS）标签在早期癌症检测中的应用，并致力于设计新方法来证明这种光谱技术在临床中的适用性。基于SERS的成像尤其具有高达单分子水平的超灵敏度、多路复用能力以及增强的光稳定性，并且已被证明优于荧光成像。然而，要将SERS标签用于早期癌症检测，了解它们与细胞的相互作用并确定其细胞毒性非常重要。一段时间以来，我们一直特别关注确定作为出色SERS增强基底的金纳米星是否以及如何能够安全地应用于生物系统并转化到临床应用中。在这项研究中，我们进行了一项多参数体外研究，以观察金纳米颗粒对人胶质母细胞瘤和人皮肤成纤维细胞系的细胞毒性和细胞摄取情况。通过将细胞与三种不同形态的金纳米颗粒（即纳米球、纳米棒和纳米星）孵育来评估细胞毒性，每种形态的金纳米颗粒又具有三种不同的表面化学性质（十六烷基三甲基溴化铵（CTAB）、聚乙二醇（PEG）和人血清白蛋白（HSA））。我们的结果表明，纳米颗粒的表面化学性质对细胞毒性具有主要影响，而纳米颗粒的形态和大小对细胞活力仅有轻微影响。发现CTAB包被的颗粒对细胞毒性最大，而聚乙二醇化的纳米星毒性最小。半胱天冬酶 - 3检测和乳酸脱氢酶检测表明，癌细胞的细胞死亡通过凋亡发生，而健康细胞则通过坏死发生。通过透射电子显微镜进行的细胞摄取研究表明，即使在长时间孵育后，颗粒仍保持其形状，这可能有利于基于SERS的体内疾病检测。总体而言，这项研究提供了有关金纳米颗粒诱导的细胞毒性的有价值信息，可用于开发安全有效的基于纳米颗粒的治疗和诊断系统。
