Mrakovcic Maria, Meindl Claudia, Roblegg Eva, Fröhlich Eleonore
Center for Medical Research, Medical University of Graz, Stiftingtalstr. 24, Graz, Austria.
Institute of Pharmaceutical Sciences, Department of Pharmaceutical Technology, Karl-Franzens-University of Graz, Helmholtzstr. 46, Graz, Austria.
Toxicol Res (Camb). 2014 Mar 1;3(2):86-97. doi: 10.1039/C3TX50112D.
Nanoparticles (NPs) are increasingly used in industrial, health and consumer products. In addition to the intended effects, NPs may also cause cell damage. Typical cytotoxicity assays assess short-term effects in adherent cells but do not evaluate longer exposure times and do not focus on cells in suspension. Since NPs are not removed easily from the organism, non-biodegradable NPs may persist in the systemic circulation and affect monocyte function at low concentrations. To mimic this situation, THP-1 monocytes were exposed to low concentrations of plain polystyrene particles (PPP) in different sizes for short (24 h) and prolonged (16 d) time periods. CELLine CL350, a small two-chamber bioreactor, and sub-culturing in flasks were compared regarding prolonged cytotoxicity testing. Uptake rates of the particles, cytotoxicity screening assays, and interleukin secretion were used for the identification of adverse effects. After 24 h, 50 μg ml 20 nm PPP did not affect cellular viability and interleukin secretion, while at higher concentrations the cytotoxicity of PPP (20 nm-500 nm) was correlated to surface area. After 16 d of exposure at 50 μg ml 20 nm PPP, the decrease in cell number and the increase in interleukins were significant. 200 nm PPP, by contrast, caused only minimal effects. Due to lower reproducibility, CELLine proved to be less suitable for the assessment as compared to sub-culturing in flasks. After prolonged exposure, silica Aerosil OX50 particles also were more cytotoxic towards THP-1 monocytes. The data suggest that prolonged exposure to NPs leads to cytotoxicity at low doses and that induction of cell death may be involved in the observed pro-inflammatory action of NPs.
纳米颗粒(NPs)越来越多地应用于工业、健康和消费品中。除了预期的效果外,纳米颗粒还可能导致细胞损伤。典型的细胞毒性试验评估贴壁细胞的短期效应,但不评估更长的暴露时间,也不关注悬浮细胞。由于纳米颗粒不易从生物体中清除,不可生物降解的纳米颗粒可能会在体循环中持续存在,并在低浓度下影响单核细胞功能。为模拟这种情况,将THP-1单核细胞暴露于不同大小的低浓度普通聚苯乙烯颗粒(PPP)中,暴露时间分别为短时间(24小时)和长时间(16天)。比较了小型双室生物反应器CELLine CL350和在培养瓶中传代培养在延长细胞毒性测试方面的情况。颗粒摄取率、细胞毒性筛选试验和白细胞介素分泌用于确定不良反应。24小时后,50μg/ml 20nm的PPP不影响细胞活力和白细胞介素分泌,而在较高浓度下,PPP(20nm - 500nm)的细胞毒性与表面积相关。在50μg/ml 20nm PPP暴露16天后,细胞数量减少和白细胞介素增加显著。相比之下,200nm的PPP仅产生最小影响。由于重现性较低,与在培养瓶中传代培养相比,CELLine被证明不太适合用于评估。长时间暴露后,二氧化硅Aerosil OX50颗粒对THP-1单核细胞的细胞毒性也更大。数据表明,长时间暴露于纳米颗粒会导致低剂量细胞毒性,并且细胞死亡的诱导可能参与了纳米颗粒观察到的促炎作用。