Bactericidal Activity of a Silver-Coated Nylon Fiber Hydrogel Wound Dressing

S.
1 Krupa ,

LB-45
*Corresponding author Joseph B. Laudano (jlaudano@alliqua.com) Vice President, Medical Affairs, Alliqua, Inc. 850 3rd Avenue, New York, New York 10022
Presenting

J.

2 Laudano *,

J.

3 Smiell ,

S.

4 Snyder ,

P.

5 1 Forman AquaMed

Technologies Inc.,

2Alliqua

Inc.,

3JMS

Clinical LLC,

4Advanced

Clinical Perspectives LLC,

5Center

for Wound Healing Inc.

author

abstract
Introduction
Nonadherent silver-containing hydrogel wound dressings have demonstrated a benefit in reducing the incidence of infection and, therefore, in speeding the healing of a variety of common wounds, including partial and full thickness dermal ulcers, leg ulcers, superficial wounds, abrasions, first and second degree burns, donor sites, and over-debrided and grafted partial thickness wounds.

Objective
To assess the bactericidal properties of single layers of a silver-coated nylon fiber hydrogel wound dressing that has been shown to provide silver ions at concentrations well above 1 ppm (the minimum associated with therapeutic activity).1,9

Table. Log Reduction in Various Organisms Organism MRSA (ATCC 33591) VRE (ATCC 51575) Escherichia coli (ATCC 8739) Pseudomonas aeruginosa (ATCC 9027) Enterococcus faecalis (ATCC 29212) Staphylococcus aureus (ATCC 6538) Klebsiella pneumoniae (ATCC 4352) Log10 Reduction 5 4 4 4 3 4 4 Time (h) 12 24 24 12 12 12 12 % Reduction >99.99 99.99 99.99 99.99 99.66 99.99 99.99

Figure 5. P. aeruginosa (ATCC 9027)

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Discussion
As tested, the silver-coated nylon fiber hydrogel wound dressing reduced the bioburden of MRSA, VRE, E. coli, P. aeruginosa, E. faecalis, S. aureus, and K. pneumoniae. The silver ion hydrogel dressing produced reductions in bacterial bioburden that met the definition of therapeutic activity for all species tested, such that within 12 or 24 hours, contamination was reduced by >99%.

Organism Count (CFU/mL)

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Methods
Bacterial-time kill studies were performed by North American Science Associates (NAMSA) (Irvine, CA). Single-layer samples of the silver-ion dressing were exposed to common pathogens associated with wound infections, as follows: 1 g dressing swatches for MRSA and VRE studies 1 g dressing swatches for E. coli, P. aeruginosa, and E. faecalis studies 0.750.01 g dressing swatches for S. aureus and K. pneumoniae studies Swatches were inoculated with the following organisms and incubated using the following test methods: AATCC Test Method 100: The silver ion hydrogel dressing and a no-silver ion hydrogel dressing (negative control) were inoculated with MRSA (target 107 CFU/mL). ASTM E2149: The silver ion hydrogel dressing and negative control were inoculated with VRE, E. coli, P. aeruginosa, and E. faecalis (target 107 CFU/mL). Dow Corning Corporate Test Method 0923: The silver ion hydrogel dressing and negative control were inoculated with S. aureus and K. pneumoniae (target 105 CFU/mL). Colony-forming units (CFU) were counted at 0 and 12 hours for all tests and at 10 minutes and 3, 8 and/or 24 hours for some tests.

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Negative control Silver-containing hydrogel dressing

The silver ion hydrogel dressing reduced the bioburden of antibiotic-resistant bacteria with similar rapidity as normal bacteria.
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Methods

Organism Count (CFU/mL)

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Organism Count (CFU/mL)

The bactericidal properties of single layers of silver-coated nylon fiber hydrogel wound dressing* (size based on test method) were assessed (NAMSA, Irvine, CA) against bacterial pathogens commonly associated with wound infections, including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VRE), and Pseudomonas aeruginosa. This dressing has been shown to provide silver ion at concentrations well above 1 ppm, which is the minimum associated with therapeutic activity. Use of the wound dressing resulted in a 99.7% reduction in E. faecalis in 12 hours, a >99% reduction in VRE in 24 h, a >99% reduction in E. coli, P. aeruginosa, and Klebsiella pneumoniae in 12 hours; and an 86.7% reduction in MRSA in 8 hours. At 12 h, the silver-coated nylon fiber hydrogel wound dressing resulted in a >99.99% reduction in MRSA compared to no reduction with a similar hydrogel dressing with no silver coating. In some cases, measurable reductions in pathogen counts compared to controls were seen in 3 hours. All results were greater than the accepted standard for bactericidal activity of 3 log10 reduction in a given organism. These data indicate that this silver-coated nylon fiber hydrogel wound dressing can provide rapid and sustained efficacy in managing the occurrence of wound contamination.
*SilverSeal Hydrogel Dressing (Alliqua Inc., New York, NY) Hydress Dressing (Alliqua Biomedical Inc.)

ATCC=American Type Culture Collection (Rockville, MD); h=hours; MRSA=methacillin-resistant S. aureus; VRE=vancomycin-resistant E. faecalis

Time-Kill Test Results

Figure 2. MRSA (ATCC 33591)
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Time Points (h)

Different analysis times were used across these studies, and the trend lines were extrapolated between measured time points. Increases in bacterial bioburden were not expected where extrapolation was performed while the inoculums and silver remained in contact. In vivo testing is a more accurate measure of overall effectiveness by measuring antibacterial effectiveness, fluid handling, and physical and chemical properties.4,5

Figure 6. E. faecalis (ATCC 29212)

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Negative control Silver-containing hydrogel dressing

Conclusions
The data from this study indicate that the tested silver-coated nylon fiber hydrogel wound dressing may be capable of rapid and sustained efficacy in managing the occurrence of wound contamination by significant reduction of bioburden from multiple bacterial species.
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Results and Conclusion

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These results support the use of a silver ion hydrogel wound dressing for bacterially contaminated wounds.

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References
Figure 3. Vancomycin-Resistant Enterococci (VRE) E. faecalis (ATCC 51575)
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results
At least a 3 log10 reduction in bacterial CFUs was achieved against all species tested (see Table). These reductions occurred in 12-24 hours (see Figures 2-8). An 86.67% reduction in MRSA was evident in 8 hours. Decreases in bacterial bioburden were continuous during the test period. None of the negative control wound dressings met the definition for therapeutic activity. Minimal reductions occurred at 24 hours in bacterial bioburden in a few negative control samples: E. coli changed from 6.90 x 107 to 4.75 x 107 CFU/mL (0 log reduction) P. aeruginosa changed from 4.00 x 107 to 3.60 x 107 CFU/mL (0 log reduction) E. faecalis changed from 2.75 x 107 to 2.40 x 106 CFU/mL (1 log reduction) No reductions were seen in the other negative control samples. Figure 1. Silver Ion Hydrogel Dressing (12x magnification)

Figure 7. S. aureus (ATCC 6538)

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Negative control Silver-containing hydrogel dressing

1. International consensus. Appropriate use of silver dressings in wounds. An expert working group consensus. London, England: Wounds International, 2012. Available at: www.woundsinternational.com. 2. Lindsay S. Silver White Paper. Everything you ever wanted to know about the use of silver in wound therapy. 2011 (Jan) Systagenix. 3. Chaloupka K, Malam Y, Seifalam AM. Nanosilver as a new generation of nanoproduct in biomedical applications. Trends Biotechnol. 2010;28(11):580-588. 4. Lo SF, Chang CJ, Hu WY, Hayter M, Chang YT. The effectiveness of silver-releasing dressings in the management of non-healing chronic wounds: a meta-analysis. J Clin Nurs. 2009;18(5):716-728. 5. Lara HH, Garza-Trevio EN, Ixtepan-Turrent L, Singh DK. Silver nanoparticles are broadspectrum bactericidal and virucidal compounds. J Nanobiotech. 2011;9:30.

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Introduction
Nonadherent silver-containing hydrogel wound dressings have demonstrated benefits in reducing the incidence of infection and may speed the healing of a variety of common wounds, including partial and full thickness dermal ulcers, leg ulcers, superficial wounds, abrasions, first and second degree burns, donor sites, and over-debrided and grafted partial thickness wounds.1 Silver ions adhere to bacterial cell walls and plasma membranes, causing cell lysis and interference with electron transport, and prevent DNA replication and protein synthesis.2,3 Although studies have shown in vitro cytotoxic effects of silver ions on fibroblasts, the use of silver ion-containing dressings provide an environment that promotes more rapid healing.4-6 An international consensus conference supported the use of silver-containing dressings for 2 weeks for wounds with (or at risk of) high bacterial bioburden, and silver-containing dressings are recommended in some treatment guidelines, especially if the wound is contaminated.1,7,8 The accepted standard for acknowledgement of bactericidal activity of an agent is a 3 log10 reduction of the respective organism.1

Negative control Silver-containing hydrogel dressing

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6. Hiro ME, Pierpont YN, Ko F, et al. Comparative evaluation of silver-containing antimicrobial dressings on in vitro and in vivo processes of wound healing. ePlasty. 2012;12(e48):Epub 2012 Oct 11. 7. National Pressure Ulcer Advisory Panel, European Pressure Ulcer Advisory Panel. Pressure ulcer treatment recommendations. In: Prevention and treatment of pressure ulcers: clinical practice guideline. Washington DC: National Pressure Ulcer Advisory Panel; 2009. p. 51-120. 8. Wound, Ostomy, and Continence Nurses Society (WOCN). Guideline for management of wounds in patients with lower-extremity venous disease. Mount Laurel (NJ): Wound, Ostomy, and Continence Nurses Society (WOCN); 2011 Jun 1. 58 p.(WOCN clinical practice guideline series; no. 4). 9. Fluder A, Laudano J, Smiell J, et al. Silver ion release from a silver fiber hydrogel wound dressing. Abstract submitted to the Symposium on Advanced Wound Care/Wound Healing Society. Poster number LB-27. Denver, Colorado. May 15, 2013.

Figure 4. E. coli (ATCC 8739)

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Figure 8. K. pneumoniae (ATCC 4352)

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The 2013 Spring Symposium on Advanced Wound Care (SAWC) May 15, 2013 Denver, Colorado