Professional Documents
Culture Documents
Silver Nanoparticles and Polymeric Medical Devices: A New Approach To Prevention of Infection?
Silver Nanoparticles and Polymeric Medical Devices: A New Approach To Prevention of Infection?
DOI: 10.1093/jac/dkh478
Advance Access publication 10 November 2004
JAC
Silver nanoparticles and polymeric medical devices: a new
approach to prevention of infection?
Franck Furno1,2, Kelly S. Morley2, Ben Wong2, Barry L. Sharp3, Polly L. Arnold2, Steven M.
Howdle2, Roger Bayston1*, Paul D. Brown4, Peter D. Winship3 and Helen J. Reid3
1
Biomaterials-Related Infection Group, School of Medical and Surgical Sciences, University of Nottingham,
Nottingham NG7 2UH; 2School of Chemistry, University of Nottingham, Nottingham NG7 2RD; 3Department
of Chemistry, University of Loughborough, Loughborough LE11 3TU; 4School of Mechanical, Materials and
Manufacturing Engineering, University of Nottingham, Nottingham NG7 2RD, UK
Received 19 May 2004; returned 13 July 2004; revised 24 August 2004; accepted 27 September 2004
Objectives: Implantable devices are major risk factors for hospital-acquired infection. Biomaterials
coated with silver oxide or silver alloy have all been used in attempts to reduce infection, in most
cases with controversial or disappointing clinical results. We have developed a completely new
approach using supercritical carbon dioxide to impregnate silicone with nanoparticulate silver metal.
This study aimed to evaluate the impregnated polymer for antimicrobial activity.
Methods: After impregnation the nature of the impregnation was determined by transmission electron
microscopy. Two series of polymer discs were then tested, one washed in deionized water and the
other unwashed. In each series, half of the discs were coated with a plasma protein conditioning film.
The serial plate transfer test was used as a screen for persisting activity. Bacterial adherence to the
polymers and the rate of kill, and effect on planktonic bacteria were measured by chemiluminescence
and viable counts. Release rates of silver ions from the polymers in the presence and absence of
plasma was measured using inductively coupled plasma mass spectrometry (ICP-MS).
Results: Tests for antimicrobial activity under various conditions showed mixed results, explained by
the modes and rates of release of silver ions. While washing removed much of the initial activity there
was continued release of silver ions. Unexpectedly, this was not blocked by conditioning film.
Conclusions: The methodology allows for the first time silver impregnation (as opposed to coating) of
medical polymers and promises to lead to an antimicrobial biomaterial whose activity is not restricted
by increasing antibiotic resistance.
1019
JAC vol.54 no.6 q The British Society for Antimicrobial Chemotherapy 2004; all rights reserved.
Downloaded from https://academic.oup.com/jac/article-abstract/54/6/1019/856134
by guest
on 17 April 2018
F. Furno et al.
Quantitative adherence
Test bacterium
As with the SPTT, four sets of discs were tested. They were
A clinical isolate of Staphylococcus epidermidis (F22) whose adher- immersed in a suspension of S. epidermidis F22 at A490 for 1 h and
ence and biofilm characteristics were known was kept at 208C in after rinsing to remove non-adhered bacteria they were sonicated
cryoprotectant until use. To resuscitate, 20 mL of tryptone soya at 50 Hz/10 min (Ultrawave, Cardiff, UK). Triplicates of 125 mL of
broth (TSB; Oxoid, Basingstoke, UK) was inoculated and incubated the sonicate were added to the wells of chemiluminescence trays
overnight at 378C. One drop of this was then inoculated into 20 mL (Zeptogen Ltd, Middlesex, UK) and read in a luminometer
of fresh TSB and incubated with shaking at 378C for 4 h to ensure (Berthold Technologies GmbH, Bad-Wildbad, Germany) using
expression of adhesins in early – mid log phase.20 The cells were a Lumitech Vialite kit (BioWhittaker Ltd, Berks, UK). Plate viable
then washed in PBS and resuspended in 2% TSB (found by exper- counts were also carried out.
1020
Figure 2. Serial plate transfer test (SPTT) clearly showing a 15 mm diameter zone of growth inhibition (arrow, circled) surrounding the silicone disc with sil-
ver nanoparticles (unwashed) on the third day of the experiment (b), compared with the control (a).
Planktonic killing unwashed. In the case of the unwashed discs, zones of inhibition
were seen for 10 days, decreasing from a mean of 8.5 mm on
Groups of discs as above were immersed in a suspension of S. epi-
day 1 to 3 mm on day 5, showing clear evidence of persisting,
dermidis F22 at A490 0.7– 0.8 for 1, 2, 3, 4 and 5 h at 378C and the
suspensions assayed by chemiluminescence and plate counting. An diffusible activity (Figure 2). No residual bacteria were seen
additional two groups of discs, one washed and the other unwashed, under the discs after removal, but further studies were not car-
were plasma-coated for 1 h before testing. ried out to detect them. This activity remained unchanged in the
presence of a conditioning film, that is in those discs which had
been immersed in plasma before being applied to the plate.
Rate of kill Tests for residual organic complexes that could have accounted
Triplicates of washed and unwashed discs with or without condition- for this were negative. However, when the second set of discs
ing film were immersed as above in a suspension of S. epidermidis were washed after impregnation, all the diffusible inhibitory
F22 at A490 0.7 – 0.8 for 1 h. They were then rinsed and immersed in activity was extinguished and no zones were seen, even on day 1.
2% TSB for 2, 3, 4 and 5 h at 378C and after rinsing to remove non- No inhibition zones were seen with plain unprocessed silicone
adhered bacteria they were sonicated. The sonicates were assayed discs or with those which had undergone supercritical fluid pro-
by chemiluminescence and plate counting. cessing but without silver.
1021
Table 1. Rate of killing of adhered bacteria Table 2. Comparison of the mean Ag isotope concentrations, each
assayed in triplicate by inductively coupled plasma mass
Unwashed Washed spectrometry, in water and plasma extracts
Control
107 109
Time (h) after without with without with without Ag isotope Ag isotope
bacterial adherence CF CF CF CF CF Sample day concentration (ppm) concentration (ppm)
1022
progressive decline in viable bacteria over 5 h, again indicating Council, The European Community and the Wade Charitable
high surface activity. Clearly, if there were residues of silver Trust. S.M.H is a Royal Society Wolfson Research Merit Award
precursor compound, or organic ligands, these might also have holder.
an antimicrobial effect. However, thermogravimetric analyses of
a series of the silicone materials after careful and exhaustive
extraction using supercritical carbon dioxide demonstrated that References
all such residues had been removed in the processing. That the 1. Bayston, R. (1999). Medical problems due to biofilms: clinical
antimicrobial effect was due to silver is supported by the ICP- impact, aetiology, molecular pathogenesis, treatment and prevention.
MS results, which were accumulated over a much longer period In Dental Plaque Revisited (Newman, H. N. & Wilson, M., Eds), pp.
of 5 days. The ICP-MS analyses were carried out on washed 111– 23. BioLine, Cardiff, UK.
discs, yet they clearly showed release of silver over the test 2. Elliott, T. S. J., Faroqui, M. H., Tebbs, S. E. et al. (1995). An
period. Interestingly, this occurred most strikingly in the pre- audit programme for central venous catheter-associated infections.
sence of plasma, reaching a peak at 3 days. This finding should Journal of Hospital Infection 30, 181–91.
be seen in conjunction with the results of tests on discs bearing 3. Arnow, P. M., Quimosing, E. M. & Beach, M. (1993).
a plasma conditioning film: while this appeared to slow the Consequences of intravascular catheter sepsis. Clinical Infectious
Diseases 16, 778–84.
release of silver ions, as shown by comparison of the effect of
4. Baumgartner, J. N. & Cooper, S. L. (1996). Bacterial
30 min and 1 h conditioning films on planktonic killing results adhesion on polyurethane surfaces conditioned with thrombus
(Figure 3), the silver ions were clearly able to penetrate the con- components. Journal of American Society of Artificial Internal
ditioning film. This finding is interesting as silver is known to Organs 42, M476 –9.
have a high avidity for protein, and the presence of a protein 5. Green, R. J., Davies, M. C., Roberts, C. J. et al. (1999).
conditioning film, or any extraneous plasma protein, has pre- Competitive protein adsorption as observed by surface plasmon
viously been assumed to inactivate any silver ions released.22 resonance. Biomaterials 20, 385–91.
The finding has obvious clinical implications. The surfaces of 6. Allison, D. G. & Gilbert, P. (1995). Modification by surface
implanted devices rapidly become coated with glycoproteins association of antimicrobial susceptibility of bacterial populations.
from tissue and plasma, with the possible exception of the inner Journal of Industrial Microbiology 15, 311 –7.
7. Proctor, R. A. & von Humboldt, A. (1998). Bacterial energetics
surface of central venous catheters, and this has been cited as
and antimicrobial resistance. Drug Resistance Updates 1, 227– 35.
one of the main reasons for clinical failure of silver-coated 8. Zimmerli, W., Widmer, A. F., Blatter, M. et al. (1998). Role of
devices.22 However, in the case of impregnation, we have now rifampin for treatment of orthopedic implant-related staphylococcal
demonstrated that there is both a depot effect and a diffusion infections—a randomised controlled trial. Journal of the American
pressure available to ‘push’ the silver ions through the condition- Medical Association 279, 1537– 41.
ing film. In order to do this, there must be enough silver ions 9. Everaert, E. P. J. M., Van de Belt-Gritter, B., Van der Mei, H. C.
available over a sufficient period to exceed those lost to protein et al. (1998). In vitro and in vivo microbial adhesion and growth on
binding. Indeed, this is one of the objectives of impregnation. argon plasma-treated silicone rubber voice prostheses. Journal of
The precipitate found during the ICP-MS analysis was likely to Materials Science—Materials in Medicine 9, 147–57.
be plasma proteins complexed with released silver, and this was 10. Morris, N. & Stickler, D. (1998). Encrustation of indwelling
urethral catheters by Proteus mirabilis biofilms growing in human urine.
supported by the finding of extremely high silver concentrations
Journal of Hospital Infection 39, 227–34.
in the precipitated material. 11. Lai, K. K. & Fontecchio, S. A. (2002). Use of silver-hydrogel
However, another clinically important advantage of impreg- urinary catheters on the incidence of catheter-associated urinary tract
nation is the protection of both inner and outer surfaces of cath- infections in hospitalized patients. American Journal of Infection
eters against bacterial colonization.23 This has been shown to be Control 30, 221–5.
crucial in clinical trials of efficacy.24 These two advantages of 12. Crabtree, J. H., Burchette, R. J., Siddiqi, R. A. et al. (2003).
impregnation, that is the continued release of silver ions in anti- The efficacy of silver-ion implanted catheters in reducing peritoneal
microbial concentrations even in the presence of a conditioning dialysis-related infections. Peritoneal Dialysis International 23,
film, and the ability to protect both inner and outer surfaces of 368– 74.
catheters, could be expected from the use of this novel method 13. Walder, B., Pittet, D. & Tramer, M. R. (2002). Prevention of
bloodstream infections with central venous catheters treated with anti-
to impregnate polymeric biomaterials with silver. A further
infective agents depends on catheter type and insertion time: evidence
advantage is the known wide antimicrobial spectrum of activity from a meta-analysis. Infection Control and Hospital Epidemiology 23,
of silver. 748– 56.
The data presented here clearly demonstrate that supercritical 14. Riley, D. K., Classen, D. C., Stevens, L. E. et al. (1995). A large
fluid impregnation of silver precursor followed by controlled randomised clinical trial of a silver-impregnated urinary catheter: lack
decomposition and extraction leads to a distribution of silver of efficacy and staphylococcal superinfection. American Journal of
throughout the silicone matrix. The impregnation process also Medicine 98, 349–56.
yields a significant surface coating of silver which is easily 15. Bayston, R., Ashraf, W. & Bhundia, C. (2004). Mode of action of
removed by simple washing. However, underlying this is a sus- an antimicrobial biomaterial for use in hydrocephalus shunts. Journal of
tained release which shows great promise, and provides signifi- Antimicrobial Chemotherapy 53, 778– 82.
16. Govender, S. T., Nathoo, N. & Van Dellen, J. R. (2003).
cant scope for optimization of these release kinetics.
Evaluation of an antibiotic-impregnated shunt system for the treatment
of hydrocephalus. Journal of Neurosurgery 99, 831– 9.
Acknowledgements 17. Webb, P. B., Parsons, A. J., Marr, P. C. et al. (2000). Green
chemistry: dissolving biomolecules and modifying biomedical implants
This study was supported by grants from the Engineering and with supercritical carbon dioxide. Pure and Applied Chemistry 72,
Physical Sciences Research Council, The Medical Research 1347–55.
1023
18. Morley, K. S., Marr, P. C., Webb, P. B. et al. (2002). Clean antimicrobials. Journal of Neurology, Neurosurgery, and Psychiatry 52,
preparation of nanoparticulate metals in porous supports: a supercriti- 605–9.
cal route. Journal of Materials Chemistry 12, 1898– 905. 22. Schierholz, J. M., Lucas, L. J., Rump, A. et al. (1998). Efficacy
19. Morley, K. S., Licence, P., Marr, P. C. et al. (2004). Supercritical of silver-coated medical devices. Journal of Hospital Infection 40,
fluids: a route to palladium-aerogel nanocomposites. Journal of 257–62.
Materials Chemistry 14, 1212–7. 23. Wilcox, M., Kite, P. & Dobbins, B. (1998). Antimicrobial
20. Patti, J. M., Allen, B. L., McGavin, M. J. et al. (1994). intravascular catheters—which surface to coat? Journal of Hospital
MSCRAMM—mediated adherence of microorganisms to host tissues. Infection 40, 322– 3.
Annual Review of Microbiology 48, 585–617. 24. Darouiche, R. O., Raad, I. I., Heard, S. O. et al. (1999). A
21. Bayston, R., Grove, N., Siegel, J. et al. (1989). Prevention of comparison of two antimicrobial-impregnated central venous catheters.
hydrocephalus shunt catheter colonisation in vitro by impregnation with New England Journal of Medicine 340, 1 –8.
1024