The detailed bactericidal process of ferric oxide nanoparticles on E. coli

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Abstract

While nanoparticles exert bactericidal effects through the generation of reactive oxygen species (ROS), the processes of the internalization of and the direct physical damage caused by iron oxide nanoparticles are not completely clear. We hypothesize that direct physical or mechanical damage of the cell membrane and cytoplasmic integrity by nanoparticles is another major cause of bacterial death besides ROS. The aim of this study is to investigate the process of the internalization of iron oxide nanoparticles, and to evaluate the effect of direct physical or mechanical damage on bacterial cell growth and death. The results demonstrate that iron oxide nanoparticles not only inhibited E. coli cell growth, but also caused bacterial cell death. Iron oxide nanoparticles produced significantly elevated ROS levels in bacteria. Transmission electronic microscopy demonstrated that iron oxide nanoparticles were internalized into and condensed the cytoplasm. Strikingly, we observed that the internalized nanoparticles caused intracellular vacuole formation, instead of simply adsorbing thereon; and formed clusters on the bacterial surface and tore up the outer cell membrane to release cytoplasm. This is the first time that the exact process of the internalization of iron oxide nanoparticles has been observed. We speculate that the intracellular vacuole formation and direct physical or mechanical damage caused by the iron oxide nanoparticles caused the bactericidal effect, along with the effects of ROS.

Figures

  • Figure 1. Culturabilityloss of Escherichia coli induced by iron oxide nanoparticle exposure in PBS. (A) E. coli MG1655 at 107 colony forming units (CFU)/mL were exposed to 0, 0.05, 0.5, 5, or 10 mM iron oxide nanoparticles at pH 7.4 and 37 ◦C for 2 h.The presence of iron oxide nanoparticles significantly reduced the culturability of the bacteria (ANOVA, p < 0.05); significant differences between each concentration of the nanoparticles and the control (0 mM) were found with the Student–Newman–Keuls (SNK) test, * p < 0.05; (B) E. coli MG1655 at 107 CFU/mL was exposed to 0.5 mM iron oxide nanoparticles for different time periods at pH 7.4 and 37 ◦C. With prolonge exposure to iron oxide nanoparticles, bacterial viability was significantly reduc d (ANOVA, p < 0.05); significant differences between the iron oxide anoparticle group and the control (0 mM) at each tim point were tested by the SNK test, * p < 0.05.
  • Figure 2. Bacterial growth curves according to qPCR of TolC and mathematical model fitting. (A) No exposure to iron oxide nanoparticles; Ψ = 0.03567 min−1, R2 = 0.89829; (B) Exposure to 0.05 mM iron oxide nanoparticles; Ψ = 0.03166 min−1, R2 = 0.92372; (C) Exposure to 0.5 mM iron oxide nanoparticles; Ψ = 0.03092 min−1, R2 = 0.96471;(D) Exposure to 5 mM iron oxide nanoparticles; Ψ = 0.02762 min−1, R2 = 0.93038;(E) Exposure to 10 mM iron oxide nanoparticles; Ψ = 0.0263 min−1, R2 = 0.93326.
  • Figure 6. The intracellular level of reactive oxygen species. (A)The bacteria were exposed to 0, 0.05, 0.5, 5, or 10 mM iron oxide nanoparticles at 25 °C for 2 h in Luria-Bertani (LB) broth. Iron oxide nanoparticles significantly increased the fluorescence of H2DCF-DA (ANOVA, p < 0.05); significant differences between each concentration group and the control (0 mM) were found with the Student– Newman–Keuls (SNK) test, * p < 0.05; (B) The bacteria were exposed to 0.5 mM iron oxide nanoparticles for the different times indicated, at 25 °C in LB broth. With prolonged exposure to the iron oxide nanoparticles, the fluorescence of H2DCF-DA significantly increased (ANOVA, p < 0.05); significant differences between each of the iron oxide nanoparticle groups and the control (0 mM) at each time point were found with the SNK test, * p < 0.05.

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Li, Y., Yang, D., Wang, S., Li, C., Xue, B., Yang, L., … Qiu, Z. (2018). The detailed bactericidal process of ferric oxide nanoparticles on E. coli. Molecules, 23(3). https://doi.org/10.3390/molecules23030606

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