Beruflich Dokumente
Kultur Dokumente
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
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.
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JAC vol.54 no.6 q The British Society for Antimicrobial Chemotherapy 2004; all rights reserved.
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.
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Silver nanoparticle impregnation and infection
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
F. Furno et al.
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)
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Silver nanoparticle impregnation and infection
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
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