Dilemmas in the reliable estimation of the in-vitro cell viability in magnetic nanoparticle engineering: which tests and what protocols?
Dilemmas in the reliable estimation of the in-vitro cell viability in magnetic nanoparticle engineering: which tests and what protocols?
复制标题
磁性纳米颗粒工程中可靠估计体外细胞活力的困境：哪些测试和哪些协议？
DOI：
10.1186/1556-276x-7-77
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发表时间：
2012-01-16
影响因子：
--
通讯作者：
Cuschieri A
中科院分区：
文献类型：
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作者：
Hoskins C;Wang L;Cheng WP;Cuschieri A
由铁氧化物制成的磁性纳米颗粒在生物医学中有着广泛的应用。充分了解MNPs与哺乳动物细胞之间的相互作用是其应用的关键问题。在这项研究中，将聚乙亚胺和聚乙二醇包覆在MNPs上，以提供它们的表面电荷和细胞毒性的细微差异，并通过三种标准的细胞存活率分析方法：3-(4，5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium[MTS]，CellTiter-Blue和CellTiter-Glo(Promega，Southampton，UK)对SH-SY5Y和RAW 264.7细胞进行数据验证。与台盼蓝人工计数相比，MTS和滴度蓝检测似乎始终高估了生存能力。滴度-Glo也经历了轻微的高估。我们假设检测系统和纳米颗粒之间发生了相互作用，导致了不正确的细胞活性评估。为了进一步了解纳米粒子对这些细胞的细胞毒性作用，我们对这些细胞的活性氧产生、脂质过氧化和细胞膜完整性进行了研究。聚乙二醇化后，MNP-PEI-PEI具有较低的表面正电荷，与MNP-PEI相比具有更好的生物相容性，不仅表现为更高的细胞存活率，而且显著减少了氧化应激和细胞膜的损伤。这些发现突显了在纳米结构的体外表征中选择化验方法和解剖不同细胞反应的重要性。
Magnetic nanoparticles [MNPs] made from iron oxides have many applications in biomedicine. Full understanding of the interactions between MNPs and mammalian cells is a critical issue for their applications. In this study, MNPs were coated with poly(ethylenimine) [MNP-PEI] and poly(ethylene glycol) [MNP-PEI-PEG] to provide a subtle difference in their surface charge and their cytotoxicity which were analysed by three standard cell viability assays: 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium [MTS], CellTiter-Blue and CellTiter-Glo (Promega, Southampton, UK) in SH-SY5Y and RAW 264.7 cells The data were validated by traditional trypan blue exclusion. In comparison to trypan blue manual counting, the MTS and Titer-Blue assays appeared to have consistently overestimated the viability. The Titer-Glo also experienced a small overestimation. We hypothesise that interactions were occurring between the assay systems and the nanoparticles, resulting in incorrect cell viability evaluation. To further understand the cytotoxic effect of the nanoparticles on these cells, reactive oxygen species production, lipid peroxidation and cell membrane integrity were investigated. After pegylation, the MNP-PEI-PEG possessed a lower positive surface charge and exhibited much improved biocompatibility compared to MNP-PEI, as demonstrated not only by a higher cell viability, but also by a markedly reduced oxidative stress and cell membrane damage. These findings highlight the importance of assay selection and of dissection of different cellular responses in in-vitro characterisation of nanostructures.