School of Environmental Science and Engineering, Kochi University of Technology, 185 Miyanokuchi, Tosayamada, Kami City, Kochi, 782-8502, Japan.
Chikami Miltec Inc, 1-6-3 Ohtesuji, Kochi City, Kochi, 780-0842, Japan.
J Phycol. 2019 Feb;55(1):118-133. doi: 10.1111/jpy.12798. Epub 2018 Nov 26.
Nanoparticles have unique properties that make them attractive for use in industrial and medical technology industries but can also be harmful to living organisms, making an understanding of their molecular mechanisms of action essential. We examined the effect of three different sized poly(isobutyl-cyanoacrylate) nanoparticles (iBCA-NPs) on the unicellular green alga Chlamydomonas reinhardtii. We found that exposure to iBCA-NPs immediately caused C. reinhardtii to display abnormal swimming behaviors. Furthermore, after one hour, most of the cells had stopped swimming and 10%-30% of cells were stained with trypan blue, suggesting that these cells had severely impaired plasma membranes. Observation of the cyto-ultrastructure showed that the cell walls had been severely damaged and that many iBCA-NPs were located in the space between the cell wall and plasma membrane, as well as inside the cytosol in some cases. A comparison of three strains of C. reinhardtii with different cell wall conditions further showed that the cell mortality ratio increased more rapidly in the absence of a cell wall. Interestingly, cell mortality over time was essentially identical regardless of iBCA-NP size if the total surface area was the same. Furthermore, direct observation of the trails of iBCA-NPs indicated that the first trigger was their contact with the cell wall, which is most likely accompanied by the inactivation or removal of adsorbed proteins from the cell wall surface. Cell mortality was accompanied by the overproduction of reactive oxygen species, which was detected more readily in cells grown under constant light rather than in the dark.
纳米粒子具有独特的性质,使其在工业和医疗技术行业中具有吸引力,但也可能对生物体造成危害,因此了解其分子作用机制至关重要。我们研究了三种不同大小的聚异丁基氰基丙烯酸酯纳米粒子(iBCA-NPs)对单细胞绿藻莱茵衣藻的影响。我们发现,iBCA-NPs 的暴露立即导致莱茵衣藻表现出异常的游动行为。此外,一小时后,大多数细胞停止游动,10%-30%的细胞被台盼蓝染色,表明这些细胞的质膜严重受损。细胞超微结构观察表明细胞壁严重受损,许多 iBCA-NPs 位于细胞壁和质膜之间的空间,以及细胞质内的某些情况下。对具有不同细胞壁条件的三种莱茵衣藻菌株的比较进一步表明,在没有细胞壁的情况下,细胞死亡率增加得更快。有趣的是,如果总表面积相同,那么随着时间的推移,细胞死亡率基本相同,而与 iBCA-NP 的大小无关。此外,直接观察 iBCA-NP 的轨迹表明,第一个触发因素是它们与细胞壁的接触,这很可能伴随着细胞壁表面吸附蛋白的失活或去除。细胞死亡率伴随着活性氧的过度产生,在持续光照下生长的细胞中比在黑暗中更容易检测到。