Golubev Alexander A, Prilepskii Artur Y, Dykman Lev A, Khlebtsov Nikolai G, Bogatyrev Vladimir A
*Laboratory of Immunochemistry, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov 410049, Russian Federation.
Laboratory of Nanobiotechnology, Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov 410049, Russian Federation.
Toxicol Sci. 2016 May;151(1):115-25. doi: 10.1093/toxsci/kfw023. Epub 2016 Feb 10.
A diagnostic test system was developed to determine the toxicity of nanomaterials to the saltwater microalga Dunaliella salina through evaluation of cell death and changes in the culture growth rate at various toxicant concentrations, providing LC50 and other toxicological metrics. The viability of cells was shown to decrease with decreasing chlorophyll absorption of red light by damaged cells. This correlation was confirmed by independent fluorescence microscopic measurements of live and dead cells in the population. Two standard colorless pollutants, hydrogen peroxide and formaldehyde, were used to validate the colorimetric method. The method's performance is exemplified with three Ag-containing preparations (Ag nitrate, Ag proteinate, and 20-nm Ag nanoparticles) and with cetyltrimethylammonium bromide (CTAB) mixed with colloidal 15-nm Au and 20-nm Ag nanoparticles. The toxicity of the Ag-containing preparations to D. salina decreased in the order Ag nitrate ≥ Ag proteinate ≫ colloidal Ag. The toxicity of colloidal Au-CTAB mixtures was found to depend mostly on the content of free CTAB. The toxicity of colloidal Ag increased substantially in the presence of CTAB. The results suggest that our D. salina-based colorimetric test system can be used for simple and rapid preliminary screening of the toxicity of different nanomaterials.
开发了一种诊断测试系统,通过评估不同毒物浓度下的细胞死亡和培养物生长速率变化,来确定纳米材料对盐水微藻杜氏盐藻的毒性,从而提供半数致死浓度(LC50)和其他毒理学指标。细胞活力随着受损细胞对红光叶绿素吸收的减少而降低。通过对群体中活细胞和死细胞进行独立的荧光显微镜测量,证实了这种相关性。使用两种标准无色污染物过氧化氢和甲醛来验证比色法。用三种含银制剂(硝酸银、蛋白银和20纳米银纳米颗粒)以及与胶体15纳米金和20纳米银纳米颗粒混合的十六烷基三甲基溴化铵(CTAB)举例说明了该方法的性能。含银制剂对杜氏盐藻的毒性按硝酸银≥蛋白银≫胶体银的顺序降低。发现胶体金-CTAB混合物的毒性主要取决于游离CTAB的含量。在CTAB存在下,胶体银的毒性大幅增加。结果表明,我们基于杜氏盐藻的比色测试系统可用于简单快速地初步筛选不同纳米材料的毒性。