Silver Nanoparticles Induced Cell Apoptosis, Membrane Damage of Azotobacter vinelandii and Nitrosomonas europaea via Generation of Reactive Oxygen Species

  • Li Zhang
  • Lingli Wu
  • Yazhu Mi
  • Youbin SiEmail author


Silver nanoparticles (AgNPs) is widely used as an antibacterial agent, but the specific antibacterial mechanism is still conflicting. This study aimed to investigate the size dependent inhibition of AgNPs and the relationship between inhibition and reactive oxygen species (ROS). Azotobactervinelandii and Nitrosomonaseuropaea were exposed to AgNPs with different particles size (10 nm and 50 nm). The ROS production was measured and the results showed that the generation of ROS related to the particle size and concentrations of AgNPs. At 10 mg/L of 10 nm Ag particles, the apoptosis rate of A. vinelandii and N. europaea were 20.23% and 1.87% respectively. Additionally, the necrosis rate of A. vinelandii and N. europaea reached to 15.20% and 42.20% respectively. Furthermore, transmission electron microscopy images also indicated that AgNPs caused severely bacterial cell membrane damage. Together these data suggested that the toxicity of AgNPs depends on its particle size and overproduction of ROS.


Silver nanoparticles Reactive oxygen species Azotobactervinelandii Nitrosomonaseuropaea Apoptosis 



This work was supported by the National Natural Science Foundation of China [Grant No. 41430752].

Compliance with Ethical Standards

Conflict of interest

We declare that we have no conflict of interest.

Research Involving Human Participants and/or Animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

128_2019_2622_MOESM1_ESM.doc (36 kb)
Supplementary file1 (DOC 35 kb)


  1. Ahmed B, Hashmi A, Khan MS, Musarrat J (2018) ROS mediated destruction of cell membrane, growth and biofilms of human bacterial pathogens by stable metallic AgNPs functionalized from bell pepper extract and quercetin. Adv Powder Technol 29:1601–1616CrossRefGoogle Scholar
  2. Bao H, Yu X, Xu C, Li X, Li Z, Wei D, Liu Y (2015) New toxicity mechanism of silver nanoparticles: promoting apoptosis and inhibiting proliferation. PLoS ONE 10:e0122535CrossRefGoogle Scholar
  3. Barcinska E, Wierzbicka J, Zauszkiewicz-Pawlak A, Jacewicz D, Dabrowska A, Inkielewicz-Stepniak I (2018) Role of oxidative and nitro-oxidative damage in silver nanoparticles cytotoxic effect against human pancreatic ductal adenocarcinoma cells. Oxid Med Cell Longev 2018:8251961CrossRefGoogle Scholar
  4. Beer C, Foldbjerg R, Hayashi Y, Sutherland DS, Autrup H (2012) Toxicity of silver nanoparticles: nanoparticle or silver ion? Toxicol Lett 208:286–292CrossRefGoogle Scholar
  5. Burdusel AC, Gherasim O, Grumezescu AM, Mogoanta L, Ficai A, Andronescu E (2018) Biomedical applications of silver nanoparticles: an up-to-date overview. Nanomaterials 8:681CrossRefGoogle Scholar
  6. Chen Q, Li T, Gui M, Liu S, Zheng M, Ni J (2017) Effects of ZnO nanoparticles on aerobic denitrification by strain Pseudomonas stutzeri PCN-1. Bioresour Technol 239:21–27CrossRefGoogle Scholar
  7. Choi O, Hu Z (2008) Size dependent and reactive oxygen species related nanosilver toxicity to nitrifying bacteria. Environ Sci Technol 42:4583–4588CrossRefGoogle Scholar
  8. Choi Y, Kim HA, Kim KW, Lee BT (2018) Comparative toxicity of silver nanoparticles and silver ions to Escherichia coli. J Environ Sci 66:50–60CrossRefGoogle Scholar
  9. Dasgupta N, Ramalingam C (2016) Silver nanoparticle antimicrobial activity explained by membrane rupture and reactive oxygen generation. Environ Chem Lett 14:477–485CrossRefGoogle Scholar
  10. Duran N, Duran M, de Jesus MB, Seabra AB, Favaro WJ, Nakazato G (2016) Silver nanoparticles: a new view on mechanistic aspects on antimicrobial activity. Nanomedicine 12:789–799CrossRefGoogle Scholar
  11. Dwyer DJ, Camacho DM, Kohanski MA, Callura JM, Collins JJ (2012) Antibiotic-induced bacterial cell death exhibits physiological and biochemical hallmarks of apoptosis. Mol Cell 46:561–572CrossRefGoogle Scholar
  12. Fabrega J, Luoma SN, Tyler CR, Galloway TS, Lead JR (2011) Silver nanoparticles: behaviour and effects in the aquatic environment. Environ Int 37:517–531CrossRefGoogle Scholar
  13. Fakai MI, Abd Malek SN, Karsani SA (2019) Induction of apoptosis by chalepin through phosphatidylserine externalisations and DNA fragmentation in breast cancer cells (MCF7). Life Sci 220:186–193CrossRefGoogle Scholar
  14. Franci G, Falanga A, Galdiero S, Palomba L, Rai M, Morelli G, Galdiero M (2015) Silver nanoparticles as potential antibacterial agents. Molecules 20:8856–8874CrossRefGoogle Scholar
  15. Gliga AR, Skoglund S, Wallinder IO, Fadeel B, Karlsson HL (2014) Size-dependent cytotoxicity of silver nanoparticles in human lung cells: the role of cellular uptake, agglomeration and Ag release. Part Fibre Toxicol 11:11CrossRefGoogle Scholar
  16. Gruen AY, App CB, Breidenbach A, Meier J, Metreveli G, Schaumann GE, Manz W (2018) Effects of low dose silver nanoparticle treatment on the structure and community composition of bacterial freshwater biofilms. PLoS ONE 13:e0199132CrossRefGoogle Scholar
  17. Hakansson AP, Roche-Hakansson H, Mossberg AK, Svanborg C (2011) Apoptosis-like death in bacteria induced by HAMLET, a human milk lipid-protein complex. PLoS ONE 6:e17717CrossRefGoogle Scholar
  18. Huang CC, Aronstam RS, Chen DR, Huang YW (2010) Oxidative stress, calcium homeostasis, and altered gene expression in human lung epithelial cells exposed to ZnO nanoparticles. Toxicol In Vitro 24:45–55CrossRefGoogle Scholar
  19. Kim S-H et al (2014) Silver nanoparticles induce apoptotic cell death in cultured cerebral cortical neurons. Mol Cell Toxicol 10:173–179CrossRefGoogle Scholar
  20. Li WR, Xie XB, Shi QS, Zeng HY, Ou-Yang YS, Chen YB (2010) Antibacterial activity and mechanism of silver nanoparticles on Escherichia coli. Appl Microbiol Biotechnol 85:1115–1122CrossRefGoogle Scholar
  21. Li J, Tang M, Xue Y (2019) Review of the effects of silver nanoparticle exposure on gut bacteria. J Appl Toxicol 39:27–37CrossRefGoogle Scholar
  22. Long YM, Hu LG, Yan XT, Zhao XC, Zhou QF, Cai Y, Jiang GB (2017) Surface ligand controls silver ion release of nanosilver and its antibacterial activity against Escherichia coli. Int J Nanomed 12:3193–3206CrossRefGoogle Scholar
  23. Mao BH, Chen ZY, Wang YJ, Yan SJ (2018) Silver nanoparticles have lethal and sublethal adverse effects on development and longevity by inducing ROS-mediated stress responses. Sci Rep 8:2445CrossRefGoogle Scholar
  24. Petrov V, Hille J, Mueller-Roeber B, Gechev TS (2015) ROS-mediated abiotic stress-induced programmed cell death in plants. Front Plant Sci 6:69CrossRefGoogle Scholar
  25. Quinteros MA et al (2018) Biosynthesized silver nanoparticles: decoding their mechanism of action in Staphylococcus aureus and Escherichia coli. Int J Biochem Cell Biol 104:87–93CrossRefGoogle Scholar
  26. Riaz Ahmed KB, Nagy AM, Brown RP, Zhang Q, Malghan SG, Goering PL (2017) Silver nanoparticles: significance of physicochemical properties and assay interference on the interpretation of in vitro cytotoxicity studies. Toxicol In Vitro 38:179–192CrossRefGoogle Scholar
  27. Schaumann GE et al (2015) Understanding the fate and biological effects of Ag- and TiO2-nanoparticles in the environment: The quest for advanced analytics and interdisciplinary concepts. Sci Total Environ 535:3–19CrossRefGoogle Scholar
  28. Shi T, Sun X, He QY (2018) Cytotoxicity of silver nanoparticles against bacteria and tumor cells. Curr Protein Pept Sci 19:525–536CrossRefGoogle Scholar
  29. Tang S, Zheng J (2018) Antibacterial activity of silver nanoparticles: structural effects. Adv Healthc Mater 7:e1701503CrossRefGoogle Scholar
  30. Wang YW, Tang H, Wu D, Liu D, Liu Y, Cao A, Wang H (2016) Enhanced bactericidal toxicity of silver nanoparticles by the antibiotic gentamicin. Environ Sci Nano 3:788–798CrossRefGoogle Scholar
  31. Wang E, Huang Y, Du Q, Sun Y (2017a) Silver nanoparticle induced toxicity to human sperm by increasing ROS (reactive oxygen species) production and DNA damage. Environ Toxicol Pharmacol 52:193–199CrossRefGoogle Scholar
  32. Wang G et al (2017b) Antibacterial effects of titanium embedded with silver nanoparticles based on electron-transfer-induced reactive oxygen species. Biomaterials 124:25–34CrossRefGoogle Scholar
  33. Wang J, Shu K, Zhang L, Si Y (2017c) Effects of silver nanoparticles on soil microbial communities and bacterial nitrification in suburban vegetable soils. Pedosphere 27:482–490CrossRefGoogle Scholar
  34. Yan X, He B, Liu L, Qu G, Shi J, Hu L, Jiang G (2018) Antibacterial mechanism of silver nanoparticles in Pseudomonas aeruginosa: proteomics approach. Metallomics 10:557–564CrossRefGoogle Scholar
  35. Yang Y, Wang J, Xiu Z, Alvarez PJ (2013) Impacts of silver nanoparticles on cellular and transcriptional activity of nitrogen-cycling bacteria. Environ Toxicol Chem 32:1488–1494Google Scholar
  36. Zapór L (2016) Effects of silver nanoparticles of different sizes on cytotoxicity and oxygen metabolism disorders in both reproductive and respiratory system cells. Arch Environ Protect 42:32–47CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Anhui Province Key Laboratory of Farmland Ecological Conservation and Pollution Prevention, School of Resources and EnvironmentAnhui Agricultural UniversityHefeiChina

Personalised recommendations