Grigor'eva Alina, Saranina Irina, Tikunova Nina, Safonov Alexey, Timoshenko Nikolai, Rebrov Alexey, Ryabchikova Elena
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, Lavrent'eva Av., 8, Novosibirsk 630090, Russia.
Biometals. 2013 Jun;26(3):479-88. doi: 10.1007/s10534-013-9633-3. Epub 2013 May 18.
Silver nanoparticles possess antibacterial effect for various bacteria; however mechanisms of the interaction between Ag-NPs and bacterial cells remain unclear. The aim of our study was to obtain direct evidence of Ag-NPs penetration into cells of Gram-negative bacterium S. typhimurium and Gram-positive bacterium S. aureus, and to study cell responses to Ag-NPs. The Ag-NPs (most 8-10 nm) were obtained by gas-jet method. S. typhimurium (7.81 × 10⁷ CFU), or S. aureus (8.96 × 10⁷ CFU) were treated by Ag-NPs (0.05 mg/l of silver) in orbital shaker at 190 rpm, 37 °C. Bacteria were sampled at 0.5, 1, 1.5, 2, 5 and 23 h of the incubation for transmission electron microscopy of ultrathin sections. The Ag-NPs adsorbed on outer membrane of S. typhimurium and cell wall of S. auereus; penetrated and accumulated in cells without aggregation and damaging of neighboring cytoplasm. In cells of S. aureus Ag-NPs bound with DNA fibers. Cell responses to Ag-NPs differed morphologically in S. typhimurium and S. aureus, and mainly were presented by damage of cell structures. The cytoplasm of S. aureus became amorphous, while S. typhimurium showed lumping and lysis of cytoplasm which led to formation of "empty" cells. Other difference was fast change of cell shape in S. typhimurium, and late deformation of S. aureus cells. The obtained results showed how different could be responses induced by the same NPs in relatively simple prokaryotic cells. Evidently, Ag-NPs directly interact with macromolecular structures of living cells and are exert an active influence on their metabolism.
银纳米颗粒对多种细菌具有抗菌作用;然而,银纳米颗粒与细菌细胞之间相互作用的机制仍不清楚。我们研究的目的是获得银纳米颗粒穿透革兰氏阴性菌鼠伤寒沙门氏菌和革兰氏阳性菌金黄色葡萄球菌细胞的直接证据,并研究细胞对银纳米颗粒的反应。通过气体喷射法制备了大多数为8 - 10纳米的银纳米颗粒。将鼠伤寒沙门氏菌(7.81×10⁷CFU)或金黄色葡萄球菌(8.96×10⁷CFU)在37℃、190转/分钟的振荡培养箱中用银纳米颗粒(0.05毫克/升银)处理。在培养0.5、1、1.5、2、5和23小时时取样细菌,用于超薄切片的透射电子显微镜观察。银纳米颗粒吸附在鼠伤寒沙门氏菌的外膜和金黄色葡萄球菌的细胞壁上;穿透并在细胞内积累,没有聚集和对邻近细胞质的损伤。在金黄色葡萄球菌细胞中,银纳米颗粒与DNA纤维结合。鼠伤寒沙门氏菌和金黄色葡萄球菌对银纳米颗粒的细胞反应在形态上有所不同,主要表现为细胞结构的损伤。金黄色葡萄球菌的细胞质变成无定形,而鼠伤寒沙门氏菌显示细胞质结块和溶解,导致形成“空”细胞。另一个差异是鼠伤寒沙门氏菌细胞形状快速变化,而金黄色葡萄球菌细胞后期变形。所获得的结果表明,相同的纳米颗粒在相对简单的原核细胞中诱导的反应可能有多大差异。显然,银纳米颗粒直接与活细胞的大分子结构相互作用,并对其代谢产生积极影响。