Applied Microbiology and Biotechnology

, Volume 91, Issue 4, pp 1149–1157 | Cite as

The influence of nanoscopically thin silver films on bacterial viability and attachment

  • Elena P. Ivanova
  • Jafar Hasan
  • Vi Khanh Truong
  • James Y. Wang
  • Massimo Raveggi
  • Christopher Fluke
  • Russell J. Crawford
Applied Microbial and Cell Physiology


The physicochemical and bactericidal properties of thin silver films have been analysed. Silver films of 3 and 150 nm thicknesses were fabricated using a magnetron sputtering thin-film deposition system. X-ray photoelectron and energy dispersive X-ray spectroscopy and atomic force microscopy analyses confirmed that the resulting surfaces were homogeneous, and that silver was the most abundant element present on both surfaces, being 45 and 53 at.% on the 3- and 150-nm films, respectively. Inductively coupled plasma time of flight mass spectroscopy (ICP-TOF-MS) was used to measure the concentration of silver ions released from these films. Concentrations of 0.9 and 5.2 ppb were detected for the 3- and 150-nm films, respectively. The surface wettability of the films remained nearly identical for both film thicknesses, displaying a static water contact angle of 95°, while the surface free energy of the 150-nm film was found to be slightly greater than that of the 3-nm film, being 28.8 and 23.9 mN m−1, respectively. The two silver film thicknesses exhibited statistically significant differences in surface topographic profiles on the nanoscopic scale, with R a, R q and R max values of 1.4, 1.8 and 15.4 nm for the 3-nm film and 0.8, 1.2 and 10.7 nm for the 150-nm film over a 5 × 5 μm scanning area. Confocal scanning laser microscopy and scanning electron microscopy revealed that the bactericidal activity of the 3-nm silver film was not significant, whereas the nanoscopically smoother 150-nm silver film exhibited appreciable bactericidal activity towards Pseudomonas aeruginosa ATCC 9027 cells and Staphylococcus aureus CIP 65.8 cells, obtaining up to 75% and 27% sterilisation effect, respectively.


Nanoscopically thin silver coating Film Bactericidal activity S. aureus P. aeruginosa 



This study was supported in part by the Advanced Manufacturing Co-operative Research Centre.

Supplementary material

253_2011_3195_Fig4_ESM.gif (176 kb)
Fig. S1a

SEM, XPS and EDX spectra of silver coatings of 3 and 150 nm thicknesses. Shows typical SEM images of the silver films. Peaks in the EDX spectra indicating silica are due to the detection of the substrate beneath the silver coatings (GIF 176 kb)

253_2011_3195_MOESM1_ESM.tif (394 kb)
High-resolution image (TIFF 393 kb)
253_2011_3195_Fig5_ESM.gif (237 kb)
Fig. S1b

SEM, XPS and EDX spectra of silver coatings of 3 and 150 nm thicknesses. Shows high-resolution XPS spectra of Ag 3d and O 1s on 3 nm (left) and 150 nm (right). Peaks in the EDX spectra indicating silica are due to the detection of the substrate beneath the silver coatings (GIF 236 kb)

253_2011_3195_MOESM2_ESM.tif (200 kb)
High-resolution image (TIFF 200 kb)
253_2011_3195_Fig6_ESM.gif (258 kb)
Fig. S1c

SEM, XPS and EDX spectra of silver coatings of 3 and 150 nm thicknesses. Shows typical distribution maps (above) showing uniform distribution of the silver over the coating area (white dots) and EDX spectra (below). Peaks in the EDX spectra indicating silica are due to the detection of the substrate beneath the silver coatings (GIF 257 kb)

253_2011_3195_MOESM3_ESM.tif (418 kb)
High-resolution image (TIFF 417 kb)
253_2011_3195_Fig7_ESM.gif (47 kb)
Fig. S2

Silver ion migration from silver films into PBS medium at regular intervals over 18 h as quantified by ICP-TOF-MS (GIF 46 kb)

253_2011_3195_MOESM4_ESM.tif (38 kb)
High-resolution image (TIFF 37 kb)
253_2011_3195_Fig8_ESM.gif (2.1 mb)
Fig. S3

AFM surface roughness analysis showing two dimensional AFM images and corresponding surface profiles of 3 nm (left) and 150 nm (right) silver coatings on approximately 10 × 10 μm scanned areas and three-dimensional visualisation of the silver coating surfaces of 3 nm (left bottom) and 150 nm (right bottom) (GIF 2200 kb)

253_2011_3195_MOESM5_ESM.tif (4.9 mb)
High-resolution image (TIFF 4969 kb)
253_2011_3195_Fig9_ESM.jpg (181 kb)
Fig. S4

AFM analysis of height distribution on the 3- and 150-nm silver film surfaces. Highlight in yellow areas indicates the characteristics height for each type of the surfaces (JPEG 181 kb)

253_2011_3195_MOESM6_ESM.pdf (6 mb)
AFM surface roughness analysis of silver coatings of 3 nm and 150 nm thickness. Two dimensional AFM images and cross section surface profiles of 3 nm (left) and 150 nm (right) silver coatings on approximately 5 μm × 5 μm scanned areas and three-dimensional visualization of the silver coating surfaces of 3 nm (left bottom) and 150 nm (right bottom). 3D images were produced using an Innova atomic force microscope (Veeco) by exporting raw data files to Avizo data processing software (v6.2, Visual Sciences Group). Readers using version 8.0 or higher of Acrobat Reader can enable interactive, three-dimensional (3-d) views of representative 1.5 μm x 1.5 μm subsections of the data by clicking on the figure panels. Once enabled, 3-d mode allows the reader to rotate and zoom the view using the computer mouse. While all of the datasets are presented with the same vertical scale, the colour scale is unique to each sample (PDF 6 mb)


  1. Agarwal A, Weis TL, Schurr MJ, Faith NG, Czuprynski CJ, McAnulty JF, Murphy CJ, Abbott NL (2010) Surfaces modified with nanometer-thick silver-impregnated polymeric films that kill bacteria but support growth of mammalian cells. Biomater 31:680–690CrossRefGoogle Scholar
  2. AshaRani PV, Hande MP, Valiyaveettil S (2009) Anti-proliferative activity of silver nanoparticles. BMC Cell Biol 10:65–71CrossRefGoogle Scholar
  3. Barnes DG, Fluke CJ (2008) Incorporating interactive three-dimensional graphics in astronomy research papers. New Astron 13:599–605CrossRefGoogle Scholar
  4. Barnes DG, Fluke CJ, Bourke PD, Parry OT (2006) An advanced, three-dimensional plotting library for astronomy. Publications of the Astronomical Society of Australia 23:82–93CrossRefGoogle Scholar
  5. Berger TJ, Spadaro JA, Chapin SE, Becker RO (1976) Electrically generated silver ions: quantitative effects on bacterial and mammalian cells. Antimicrob Agents Chemother 9:357–358Google Scholar
  6. Bosetti M, Massè A, Tobin E, Cannas M (2002) Silver coated materials for external fixation devices: in vitro biocompatibility and genotoxicity. Biomater 23:887–892CrossRefGoogle Scholar
  7. Brutel De La Riviere A, Dossche KME, Birnbaum DE, Hacker R (2000) First clinical experience with a mechanical valve with silver coating. J Heart Valve Dis 9:123–130Google Scholar
  8. Cai K, Müller M, Bossert J, Rechtenbach A, Jandt KD (2005) Surface structure and composition of flat titanium thin films as a function of film thickness and evaporation rate. Appl Surf Sci 250:252–267CrossRefGoogle Scholar
  9. Chen W, Liu Y, Courtney HS, Bettenga M, Agrawal CM, Bumgardner JD, Ong JL (2006) In vitro anti-bacterial and biological properties of magnetron co-sputtered silver-containing hydroxyapatite coating. Biomater 27:5512–5517CrossRefGoogle Scholar
  10. Crawford R, Koopal LK, Ralston J (1987) Contact angles on particles and plates. Colloids Surf 27:57–64Google Scholar
  11. Donlan RM (2001) Biofilms and device-associated infections. Emerg Infect Dis 7:277–281CrossRefGoogle Scholar
  12. Dowling DP, Donnelly K, McConnell ML, Eloy R, Arnaud MN (2001) Deposition of anti-bacterial silver coatings on polymeric substrates. Thin Solid Films 398–399:602–606CrossRefGoogle Scholar
  13. Ewald A, Glückermann SK, Thull R, Gbureck U (2006) Antimicrobial titanium/silver PVD coatings on titanium. Biomed Eng Online 5:22CrossRefGoogle Scholar
  14. Gadelmawla ES, Koura MM, Maksoud TMA, Elewa IM, Soliman HH (2002) Roughness parameters. J Mater Process Technol 123:133–145CrossRefGoogle Scholar
  15. Gibbins B, Warner L (2005) The role of antimicrobial silver nanotechnology. Med Device Diagn Ind pp. 27–33Google Scholar
  16. Gosheger G, Hardes J, Ahrens H, Streitburger A, Buerger H, Erren M, Gunsel A, Kemper FH, Winkelmann W, Von Eiff C (2004) Silver-coated megaendoprostheses in a rabbit model—an analysis of the infection rate and toxicological side effects. Biomater 25:5547–5556CrossRefGoogle Scholar
  17. Groessner-Schreiber B, Hannig M, Dück A, Griepentrog M, Wenderoth DF (2004) Do different implant surfaces exposed in the oral cavity of humans show different biofilm compositions and activities? Eur J Oral Sci 112:516–522CrossRefGoogle Scholar
  18. Guy DW, Crawford RJ, Mainwaring DE (1996) The wetting behaviour of several organic liquids in water on coal surfaces. Fuel 75:238–242CrossRefGoogle Scholar
  19. Hachem RY, Wright KC, Zermeno A, Bodey GP, Raad II (2003) Evaluation of the silver iontophoretic catheter in an animal model. Biomater 24:3619–3622CrossRefGoogle Scholar
  20. Hardes J, Streitburger A, Ahrens H, Nusselt T, Gebert C, Winkelmann W, Battmann A, Gosheger G (2007) The influence of elementary silver versus titanium on osteoblasts behaviour in vitro using human osteosarcoma cell lines. Sarcoma. doi: 10.1155/2007/26539 Google Scholar
  21. Heydorn A, Nielsen AT, Hentzer M, Sternberg C, Givskov M, Ersboll BK, Molin S (2000) Quantification of biofilm structures by the novel computer program comstat. Microbiology 146:2395–2407Google Scholar
  22. Hussain SM, Hess KL, Gearhart JM, Geiss KT, Schlager JJ (2005) In vitro toxicity of nanoparticles in brl 3a rat liver cells. Toxicol In Vitro 19:975–983CrossRefGoogle Scholar
  23. Ivanova EP, Mitik-Dineva N, Wang J, Pham DK, Wright JP, Nicolau DV, Mocanasu RC, Crawford RJ (2008) Staleya guttiformis attachment on poly(tert-butylmethacrylate) polymeric surfaces. Micron 39:1197–1204CrossRefGoogle Scholar
  24. Ivanova EP, Truong VK, Wang JY, Bemdt CC, Jones RT, Yusuf II, Peake I, Schmidt HW, Fluke C, Barnes D, Crawford RJ (2010) Impact of nanoscale roughness of titanium thin film surfaces on bacterial retention. Langmuir 26:1973–1982CrossRefGoogle Scholar
  25. Jeyachandran YL, Karunagaran B, Narayandass SK, Mangalaraj D, Jenkins TE, Martin PJ (2006) Properties of titanium thin films deposited by dc magnetron sputtering. Mat Sci Eng A 431:277–284CrossRefGoogle Scholar
  26. Joyce-Wöhrmann RM, Münstedt H (1999) Determination of the silver ion release from polyurethanes enriched with silver. Infection 27:S46–S48CrossRefGoogle Scholar
  27. Kaiser N (2002) Review of the fundamentals of thin-film growth. Appl Opt 41:3053–3060CrossRefGoogle Scholar
  28. Kelly PJ, Li H, Whitehead KA, Verran J, Arnell RD, Iordanova I (2009) A study of the antimicrobial and tribological properties of tin/ag nanocomposite coatings. Surf Coat Technol 204:1137–1140CrossRefGoogle Scholar
  29. Kim JS, Kuk E, Yu KN, Kim JH, Park SJ, Lee HJ, Kim SH, Park YK, Park YH, Hwang CY, Kim YK, Lee YS, Jeong DH, Cho MH (2007) Antimicrobial effects of silver nanoparticles. Nanomedicine 3:95–101Google Scholar
  30. Kora AJ, Manjusha R, Arunachalam J (2009) Superior bactericidal activity of sds capped silver nanoparticles: synthesis and characterization. Mater Sci Eng C 29:2104–2109CrossRefGoogle Scholar
  31. Kumar R, Münstedt H (2005) Silver ion release from antimicrobial polyamide/silver composites. Biomater 26:2081–2088CrossRefGoogle Scholar
  32. Lamolle SF, Monjo M, Lyngstadaas SP, Ellingsen JE, Haugen HJ (2009) Titanium implant surface modification by cathodic reduction in hydrofluoric acid: surface characterization and in vivo performance. J Biomed Mater Res A 88:581–588Google Scholar
  33. Liedberg H, Lundeberg T (1989) Assessment of silver-coated urinary catheter toxicity by cell culture. Urol Res 17:359–360CrossRefGoogle Scholar
  34. Liu X, Chu PK, Ding C (2004) Surface modification of titanium, titanium alloys, and related materials for biomedical applications. Mater Sci Eng R Rep 47:49–121. doi: 10.1016/j.mser.2004.11.001 CrossRefGoogle Scholar
  35. Marambio-Jones C, Hoek E (2010) A review of the antibacterial effects of silver nanomaterials and potential implications for human health and the environment. J Nanopart Res 12:1531–1551CrossRefGoogle Scholar
  36. Marini M, De Niederhausern S, Iseppi R, Bondi M, Sabia C, Toselli M, Pilati F (2007) Antibacterial activity of plastics coated with silver-doped organic-inorganic hybrid coatings prepared by sol-gel processes. Biomacromolecules 8:1246–1254CrossRefGoogle Scholar
  37. Mitik-Dineva N, Wang J, Mocanasu RC, Stoddart PR, Crawford RJ, Ivanova EP (2008) Impact of nano-topography on bacterial attachment. Biotechnol J 3:536–544CrossRefGoogle Scholar
  38. Mitik-Dineva N, Wang J, Truong VK, Stoddart P, Malherbe F, Crawford RJ, Ivanova EP (2009) Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus attachment patterns on glass surfaces with nanoscale roughness. Curr Microbiol 58:268–273CrossRefGoogle Scholar
  39. Modak SM, Fox CL Jr (1973) Binding of silver sulfadiazine to the cellular components of Pseudomonas aeruginosa. Biochem Pharmacol 22:2391–2404CrossRefGoogle Scholar
  40. Öner D, McCarthy TJ (2000) Ultrahydrophobic surfaces. Effects of topography length scales on wettability. Langmuir 16:7777–7782CrossRefGoogle Scholar
  41. Park HJ, Kim JY, Kim J, Lee JH, Hahn JS, Gu MB, Yoon J (2009) Silver-ion-mediated reactive oxygen species generation affecting bactericidal activity. Water Res 43:1027–1032CrossRefGoogle Scholar
  42. Puckett SD, Taylor E, Raimondo T, Webster TJ (2010) The relationship between the nanostructure of titanium surfaces and bacterial attachment. Biomaterials 31:706–713CrossRefGoogle Scholar
  43. Samokhvalov A, Nair S, Duin EC, Tatarchuk BJ (2010) Surface characterization of ag/titania adsorbents. Appl Surf Sci 256:3647–3652CrossRefGoogle Scholar
  44. Schierholz JM, Lucas LJ, Rump A, Pulverer G (1998) Efficacy of silver-coated medical devices. J Hosp Infect 40:257–262CrossRefGoogle Scholar
  45. Shao W, Zhao Q (2010) Influence of reducers on nanostructure and surface energy of silver coatings and bacterial adhesion. Surf Coat Technol 204:1288–1294CrossRefGoogle Scholar
  46. Silver S (2003) Bacterial silver resistance: molecular biology and uses and misuses of silver compounds. FEMS Microbiol Rev 27:341–353CrossRefGoogle Scholar
  47. Sreekumari KR, Nandakumar K, Takao K, Kikuchi Y (2003) Silver containing stainless steel as a new outlook to abate bacterial adhesion and microbiologically influenced corrosion. ISIJ Int 43:1799–1806CrossRefGoogle Scholar
  48. Stewart PS, Costerton JW (2001) Antibiotic resistance of bacteria in biofilms. Lancet 358:135–138CrossRefGoogle Scholar
  49. Stobie N, Duffy B, Hinder SJ, McHale P, McCormack DE (2009) Silver doped perfluoropolyether-urethane coatings: antibacterial activity and surface analysis. Colloids Surf B Biointerfaces 72:62–67CrossRefGoogle Scholar
  50. Truong VK, Lapovok R, Estrin YS, Rundell S, Wang JY, Fluke CJ, Crawford RJ, Ivanova EP (2010) The influence of nano-scale surface roughness on bacterial adhesion to ultrafine-grained titanium. Biomater 31:3674–3683CrossRefGoogle Scholar
  51. Van Oss CJ, Good RJ, Chaudhury MK (1988) Additive and nonadditive surface tension components and the interpretation of contact angles. Langmuir 4:884–891CrossRefGoogle Scholar
  52. Vik H (1986) Neuropathia caused by silver absoprtion from arthroplasty cement. Lancet 1:872Google Scholar
  53. Wang YP, Yeh CT (1991) Electron paramagnetic resonance study of the interactions of oxygen with silver/titania. J Chem Soc Faraday Trans 87:345–348CrossRefGoogle Scholar
  54. Wang HB, Wei QF, Wang JY, Hong JH, Zhao XY (2008a) Sputter deposition of nanostructured antibacterial silver on polypropylene non-wovens. Surf Eng 24:70–74CrossRefGoogle Scholar
  55. Wang JY, Ghantasala MK, McLean RJ (2008b) Bias sputtering effect on ultra-thin smco5 films exhibiting large perpendicular coercivity. Thin Solid Films 517:656–660CrossRefGoogle Scholar
  56. Williams RL, Doherty PJ, Vince DG, Grashoff GJ, Williams DF (1989) The biocompatibility of silver. Crit Rev Biocompat 5:221–243Google Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Elena P. Ivanova
    • 1
  • Jafar Hasan
    • 1
  • Vi Khanh Truong
    • 1
  • James Y. Wang
    • 2
  • Massimo Raveggi
    • 3
  • Christopher Fluke
    • 4
  • Russell J. Crawford
    • 1
  1. 1.Faculty of Life and Social SciencesSwinburne University of TechnologyMelbourneAustralia
  2. 2.IRIS, Swinburne University of TechnologyMelbourneAustralia
  3. 3.School of GeosciencesMonash UniversityMelbourneAustralia
  4. 4.Centre for Astrophysics and SupercomputingSwinburne University of TechnologyMelbourneAustralia

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