Lindner Karina, Ströbele Michael, Schlick Sandra, Webering Sina, Jenckel André, Kopf Johannes, Danov Olga, Sewald Katherina, Buj Christian, Creutzenberg Otto, Tillmann Thomas, Pohlmann Gerhard, Ernst Heinrich, Ziemann Christina, Hüttmann Gereon, Heine Holger, Bockhorn Henning, Hansen Tanja, König Peter, Fehrenbach Heinz
Institut für Anatomie, Zentrum für medizinische Struktur- und Zellbiologie, Universität zu Lübeck (UzL), Airway Research Center North (ARCN), German Center for Lung Research (DZL), Ratzeburger Allee 160, 23562, Lübeck, Germany.
Karlsruher Institut für Technologie, Engler-Bunte-Institut, Bereich Verbrennungstechnik, Karlsruhe, Germany.
Part Fibre Toxicol. 2017 Mar 21;14(1):8. doi: 10.1186/s12989-017-0189-1.
Carbon black nanoparticles (CBNP) are mainly composed of carbon, with a small amount of other elements (including hydrogen and oxygen). The toxicity of CBNP has been attributed to their large surface area, and through adsorbing intrinsically toxic substances, such as polycyclic aromatic hydrocarbons (PAH). It is not clear whether a PAH surface coating changes the toxicological properties of CBNP by influencing their physicochemical properties, through the specific toxicity of the surface-bound PAH, or by a combination of both.
Printex90 (P90) was used as CBNP; the comparators were P90 coated with either benzo[a]pyrene (BaP) or 9-nitroanthracene (9NA), and soot from acetylene combustion that bears various PAHs on the surface (AS-PAH). Oxidative stress and IL-8/KC mRNA expression were determined in A549 and bronchial epithelial cells (16HBE14o-, Calu-3), mouse intrapulmonary airways and tracheal epithelial cells. Overall toxicity was tested in a rat inhalation study according to Organization for Economic Co-operation and Development (OECD) criteria. Effects on cytochrome monooxygenase (Cyp) mRNA expression, cell viability and mucociliary clearance were determined in acute exposure models using explanted murine trachea.
All particles had similar primary particle size, shape, hydrodynamic diameter and ζ-potential. All PAH-containing particles had a comparable specific surface area that was approximately one third that of P90. AS-PAH contained a mixture of PAH with expected higher toxicity than BaP or 9NA. PAH-coating reduced some effects of P90 such as IL-8 mRNA expression and oxidative stress in A549 cells, granulocyte influx in the in vivo OECD experiment, and agglomeration of P90 and mucus release in the murine trachea ex vivo. Furthermore, P90-BaP decreased particle transport speed compared to P90 at 10 μg/ml. In contrast, PAH-coating induced IL-8 mRNA expression in bronchial epithelial cell lines, and Cyp mRNA expression and apoptosis in tracheal epithelial cells. In line with the higher toxicity compared to P90-BaP and P90-9NA, AS-PAH had the strongest biological effects both ex vivo and in vivo.
Our results demonstrate that the biological effect of CBNP is determined by a combination of specific surface area and surface-bound PAH, and varies in different target cells.
炭黑纳米颗粒（CBNP）主要由碳组成，含有少量其他元素（包括氢和氧）。CBNP的毒性归因于其较大的表面积，以及通过吸附本质上有毒的物质，如多环芳烃（PAH）。目前尚不清楚PAH表面涂层是否通过影响CBNP的物理化学性质、表面结合PAH的特定毒性或两者的结合来改变CBNP的毒理学特性。
使用Printex90（P90）作为CBNP；比较物为涂有苯并[a]芘（BaP）或9-硝基蒽（9NA）的P90，以及表面带有各种PAH的乙炔燃烧烟灰（AS-PAH）。在A549细胞和支气管上皮细胞（16HBE14o-、Calu-3）、小鼠肺内气道和气管上皮细胞中测定氧化应激和IL-8/KC mRNA表达。根据经济合作与发展组织（OECD）标准，在大鼠吸入研究中测试总体毒性。在使用离体小鼠气管的急性暴露模型中测定对细胞色素单加氧酶（Cyp）mRNA表达、细胞活力和黏液纤毛清除的影响。
所有颗粒具有相似的初级粒径、形状、流体动力学直径和ζ电位。所有含PAH的颗粒具有相当的比表面积，约为P90的三分之一。AS-PAH含有PAH混合物，其毒性预计高于BaP或9NA。PAH涂层降低了P90的一些影响，如A549细胞中的IL-8 mRNA表达和氧化应激、体内OECD实验中的粒细胞流入，以及离体小鼠气管中P90的聚集和黏液释放。此外，与10μg/ml的P90相比，P90-BaP降低了颗粒运输速度。相反，PAH涂层诱导支气管上皮细胞系中的IL-8 mRNA表达，以及气管上皮细胞中的Cyp mRNA表达和凋亡。与P90-BaP和P90-9NA相比毒性更高，AS-PAH在离体和体内均具有最强的生物学效应。
我们的结果表明，CBNP的生物学效应由比表面积和表面结合的PAH共同决定，并且在不同靶细胞中有所不同。
