SILVER I, II, III: Chemical Characteristics, Properties, and Antimicrobial Activity

By Carla Spina, Ph.D. Exciton Technologies Inc. - Edmonton, Alberta, Canada

HISTORY For centuries, silver has been used for sanitization and medicinal applications. The use of silver vessels for water storage has been recorded as early as 1000 B.C. as a method to produce potable water. In the 8th century silver was first used for medicinal purposes when the Muslim physician, Avicenna used silver as a blood purifier, for bad breath, and for heart palpitations [1]. In the 17th and 18th centuries silver was used to treat ulcers and in the 19th century dilute silver nitrate solutions were utilized to treat burn victims [2]. Investigations into the mechanism of action of silver, however, have only been recently explored. A major step was taken by Acl and Goetz, who determined that the antimicrobial properties of silver were due to the liberation of ionic silver (I) ions, and not due to the metal itself. In the early 1900s, studies into the biocidal properties of silver fell drastically with the introduction of antibiotics. This downward trend is rapidly changing however, due to the everincreasing concern over antibiotic-resistant bacterial strains. Unlike antibiotics, which have a single mechanism of action in causing bacterial death, the mechanism of silvers antimicrobial activity proceeds along several pathways, which decreases the potential for development of bacterial resistance. In the last two decades, the use of silver in medical applications has been growing where silver is now used as an antimicrobial agent in wound care products, as coatings on medical devices, and in disinfectant formulations. Many products on the market today claim silver as an active antimicrobial agent; however, there is no precise explanation of what the active silver species is. Most current silver products contain silver (I) ions (Ag+) and/ or metallic silver (Ag0). In comparison to the current products on the market, Exciton Technologies Inc. offers a unique, effective, antimicrobial product which contains silver, at low concentrations, in three ionic states. The significance of silver in its different ionic forms, biocompatibility, and its use as an antimicrobial agent will be discussed in detail in this report. TOXICITY In minute concentrations, silver is non-toxic to human cells [3]. The most common deleterious side effect of silver products is argyria, where sub-dermal deposits of silver in micro-vessels under the skin result in an irreversible grey-black coloration of the skin. This discoloration is not physically harmful, yet is cosmetically displeasing. The effect of silver towards lower life forms, however, is more severe. Parts per million (ppm) concentrations of silver, which are non-toxic to humans, are known to be lethal towards bacteria and other lower life forms [4].

This biocidal property of silver and other metals has been termed an oligodynamic effect [5]. From the Greek (oligos = few, and dynamis = power), it is a term used to describe how low metal ion concentrations exert powerful biocidal effects towards lower life forms, yet are non-toxic to humans and other higher life forms. This biocidal effect on lower life forms, such as bacterial or fungi, is dependent on the availability of the metal ions in solution, and their size and charge. This effect is found for several metal species where the relative toxicity of these metals has been described in the Horsfall series [6]: Ag > Hg > Cu > Cd > Cr > Ni > Co > Zn > Fe > Ca In this series, the toxicity of the metals to fungal species is compared, with ionic silver being the most effective biocidal agent. A sub-division of this series may be examined for ionic silver: Ag
+3

> Ag > Ag

+2

In this sub-series, in addition to the customary antimicrobial properties of silver, the oxidative strengths of the silver ions are taken into consideration. The result is equivalent or greater biocidal activity for higher ionic or valence states at a lower silver concentration with proven biocompatibility [7]. ANTIMICROBIAL ACTION Although silver has long been recognized for its broadspectrum antimicrobial activity, the mechanism of antimicrobial action of silver is still under investigation. What is known is that the efficacy of silver is related to the ionic state, solubility, availability, and the structural form of silver in solution [8]. When silver is in the metallic state, Ag0, it has no antimicrobial activity, while ionic silver at concentrations as low as 0.001 0.006 ppm is known to be a potent antimicrobial agent [9]. Ionic silver, specifically Ag+, has been shown in bacteria to uncouple the respiratory chain from oxidative phosphorylation, disrupt the proton-motive force across cellular membranes, and interact with thiols and sulphur complexes resulting in protein misfolding and cell structure abnormalities [3, 10-13]. Silver has also been reported to stimulate the release of potassium from bacteria, and bind to DNA thus inhibiting replication [14-15]. Since the mechanism of action of silver proceeds along several pathways, the potential for development of bacterial resistance is greatly reduced. As a result, there is a keen interest in using silver and silver compounds for medical applications. over >

Although singly ionized silver, Ag+, has been previously investigated for its antimicrobial activity, there has been minimal study on the effect and antimicrobial mechanism of higher valence silver complexes due to the difficulty of stabilizing the higher valence states. However, research completed to date, reveals that lower concentrations of Ag+2 and Ag+3 silver ions, are equally or more active than silver Ag+ in killing bacteria [7, 16-17]. The exact mechanism of this activity is still under investigation. exsaltTM At Exciton Technologies Inc., a unique silver product with excellent antimicrobial properties has been developed. This novel exsaltTM SD7 wound care dressing not only has equivalent antimicrobial properties as competitor products, but also demonstrates nominal pH shift, at a markedly lower silver concentration compared to most competitors, Table 1. In addition, biocompatibility studies reveal no toxic effect of exsaltTM products. Reduced staining of the skin is also observed when exsaltTM dressings are compared to other products on the market. Table 1. Silver content of commercially available silver wound care products in comparison to Exciton Technologies exsalt SD7 product [18-22].
Proprietary Name exsalt SD7 Therabond 3D

The increased activity occurs in close proximity to the dressing which provides an antimicrobial barrier to infection. The higher valent silver species are then reduced to ionic silver (Ag+) which is equivalent to valance state of silver found in the current silver dressings that are on the market [17-18, 23-26]. ExsaltTM SD7 wound dressings therefore provide effective antimicrobial action with excellent biocompatibility, little irritation or sensitization, and minimal pH shift at a lower effective dose of silver than most monovalent Ag+ products on the market. The benefit of exSALTTM technology in combination with its economical value fills a niche in the wound care market. Although the precise mechanism of antimicrobial activity of silver is still under investigation, exsaltTM SD7 wound dressings demonstrate that there is room for improvement over the currently available wound care materials.

REFERENCES
1) 2) 3) 4) 5) 6) Bhattacarya, R., Mukherjee, P., Adv. Drug. Del. Rev. 2008, (60), 1289-1306 Wadhera, A., Fung, M., Dermatol. Online J., 2005, (11), 12 Emsley, John, The Elements, 3rd ed., Clarendon Press, Oxford, 1998 Jung, Woo. K. et al., Appl. Environ. Microbiol. 2008, 2171-2178 Berger, T.J., Spadaro, J.A., Chapin, S.E., Becker, R.O., Antimicrob. Agents. Chemother., 1976, 9(2), 357-8 Horsfall, J. G. , Principles of Fungicidal Action, Chronica Botanica Co., Waltham, Mass., 1956. Antelman MS 1993 US Patent 5.211.855 Warriner, R., Burrell, R., Adv. Skin. Wound Care., 2005, 18(1), 2-12 Walker, M., Cochrane, C.A., Bowler, Ph.G., Parsons, D., Bradshaw, P., Ostomy/ Wound Management, 2006, 52(1), 42-50 Schreurs, W.J.A., Rosenberg, H., J. Bacteriol., 1982, 152, 7-13 Dibrov, P., Dzioba, J., Gosink, K.K., Hase, C.C., Antimicrob. Agents Chemother., 2002, 46, 2668-2670 Liau, S.Y., Read, D.C., Pugh, W.J., Furr, J.R., Russell, A.D., Appl. Microbiol. 1997, 25, 279-283 Zeiri, I., Bronk, B.V., Shabtai, Y., Eichler, J., Efrima, S., Appl. Spectrosc., 2004, 58, 33-40 Kone, Bruce C., et al. J. Membrane Biol., 1988, 102,(1), 11-19 Silver, S., FEMS Microbiol. Rev. 2003, (27), 341-353 Dellasega, D., Facibeni, A., Di Fonzo, F., Bogana, M., Polissi, A., Conti, C., Ducati, C., Casari, C.S., Bassi, A.L., Bottani, C.E., Nanotechnology, 2008, 19, 475602-475607 Djokic, S.S., Journal of the Electrochemical Society, 2004, (151)6, C659-C364 Thomas, S., World Wide Wounds, Nov 2004 Tredget et al., J. Burn Care Rehab., 1998, 19(6), 531-537 Taylor, P.L., Biomaterials, 2005, 26, 7230-7240 http://www.silverlon.com/silverlon_acticoat.html http://www.bbraun.com/cps/rde/xchg/bbraun-com/hs.xsl/products.html?id =00020741510000000229&prid=PRID00002996 Vansek, P., Electrochemical Series, Handbook of Chemistry and Physics, 90th Ed, 2009, CRC Ring, A.; Goertz, O.; Steinstraesser, L.; Cotaru, C.; Muhr, G.; Steinau, H.; Langer, S., Wounds , 2007, 19(2), 39-45 Pal, S., et al., J. Am. Chem. Soc., ASAP Levanson, W., Spicer, M.D., Coord. Chem. Rev., 1987, 76, 45-120

Silver Content (mg/100cm2) 40 440 237 158 141 85 8.3 5.3 (SSD content 3.75%)

Valence States +1, +2, +3 0, +1 0, +1 0, +1 +1 +1 +1 +1 +1

7) 8) 9) 10) 11) 12) 13) 14) 15) 16)

Silverlon Easy Ag

Acticoat 7 Calgitrol Ag Contreet Foam Aquacel Ag SilvaSorb Urgotul SSD

Excitons patented silver wound care product provides the health care industry with a cost-effective, non-toxic, efficacious silver wound dressing. Designed to provide an effective antimicrobial barrier with a lesser pH shift and minimal staining, exsaltTM wound care dressings contain, in addition to ionic silver (Ag+), the higher valent silver ions, Ag+2 and Ag+3. The higher ionic states of the silver species in exsaltTM result in excellent antimicrobial activity at lower total concentrations of silver [17]. The hypothesized mechanism of action is based on exsaltTM solubility characteristics and the different reduction potentials between Ag0 to Ag+3 as shown by equations 1-3 [23]. Standard Reduction Potentials: Ag+3 + e Ag+2 + eAg+ + eAg+2, Ag+, Ag0, E0 = +1.80 V (1) E0 = +1.980 V (2) E0 = +0.7996 V (3)

17) 18) 19) 20) 21) 22) 23) 24) 25) 26)

PR-0028.0