Silver has long been used as an antibacterial agent for the treatment of many infectious diseases in humans, since the time before the emergence of antibiotics [22
]. The mechanisms of silver's antibacterial properties were suggested to result from its binding to the bacterial cell wall and cell membrane and from the interaction with the thiol groups of bacterial proteins leading to their subsequent inactivation and loss of biochemical competence, without repair of which the cell will die [23
]. The small size of silver nanoparticles gives improved antibacterial effects due to the increase in their surface area for interaction with the microorganisms [24
], as well as potentially enhanced oxidation-solvation and uptake rates across cell membranes into the cytosol to disrupt intracellular protein thiol groups. Application of silver nanoparticles in textile materials is one of the most interesting applications to improve the quality of the products, including wound dressings and anti-bacterial clothes. Meanwhile, there is also increasing concern on the safety of these nanoproducts. To date there has not been any report on how much consumers may be exposed to silver particles during application. Therefore, artificial sweat was used as a model to study the release of silver from manufacturer-claimed nanosilver shirts as well as our laboratory-prepared nanosilver fabrics.
Firstly, the availability of silver in the silver suspension and in each fabric was characterized. From the product's information, the suspension contains silver chloride and titanium dioxide. We focused on the agglomeration of nano-sized particles in the TEM image where a high EDX peak of Ag and a small peak of Ti were demonstrated (Figure ). SEM analysis demonstrated agglomerated particles similar to that found in the TEM images, and with a similar EDX pattern with peaks of Ag and Ti (Figure ). These evidences support that the silver suspension used in this study contains silver nanoparticles. Agglomeration is the natural phenomenon of colloidal nanoparticles due to their ultrafine size [25
]. In the fabric ashes, silver particles detected from the laboratory-prepared fabric A4 (Figure ) were similar in their size to that from fabric E (Figure ), but smaller than that of their original form in the silver suspension (Figure ). It was likely that the burning process reduced the silver particle size agglomeration. The height of the Ag peak in the EDX graph indicated the amount of silver found in the focused area of SEM image. Besides silver, other compounds were also obtained in EDX peaks, the most prominent of which was titanium (Figures and ).
Titanium is one of the frequently used nanomaterials in consumer products, including cosmetics, sunscreens and textiles [2
]. The composition of the silver solution used in this study also contained titanium dioxide which, in the presence of UV light, has the potential to kill bacteria [26
]. However, some forms of titanium dioxide, such as nitrogen- and carbon-doped titanium dioxide, are also effective under visible light [27
]. Hence, titanium dioxide is usually incorporated into most antibacterial materials where it exerts a synergistic antibacterial effect when combined with silver [28
]. In addition, titanium dioxide can reduce the use of silver, with the advantages of an increased whiteness and a reduced cost of the products.
Since the main purpose of fabric samples is to prevent bacteria, the antibacterial effect of the different fabrics was evaluated using S. aureus and E. coli as model Gram-positive and Gram-negative bacteria, respectively. Most of the fabrics used in this study showed a higher antibacterial activity against S. aureus
than that against E. coli
(Table ). The mechanism underlying this antibacterial activity for each of the fabrics was not investigated, but it is likely to not be solely due to nanosilver particles as those fabrics which had silver levels below the detection levels (fabrics B and C) showed essentially as good an antibacterial activity against S. aureus
as the silver impregnated fabrics. Perhaps, this then is accounted for by the titanium dioxide. However, the differences between the inhibition of S. aureus
and E. coli
growth may result from the difference in the compositions of the bacterial cell walls where the cell wall of Gram-negative, but not Gram-positive, bacteria consists of lipopolysaccharides which may provide a more effective protection against bactericides [30
]. Certainly, further screening of more representatives of each bacterial division is required to support such a notion.
By using the coating procedures, it was likely that a 10 g/L silver suspension was sufficient for preparing antibacterial fabrics. At this treatment concentration, the fabrics induced a 99.83% and 99.93% reduction of proliferation competency in S. aureus
and E. coli
, respectively. However, for commercial fabrics, there was no correlation between the initial level of silver in the fabrics and their antibacterial properties, perhaps due to the compounding action of other components such as titanium dioxide. In this light, it is worth noting that fabrics B and C had no detectable silver but could significantly inhibit S. aureus
proliferation competency. In addition, the antibacterial properties of silver nanoparticles can be varied by their size, shape and surface modifications [3
]. These factors might partly explain the uncorrelated results between amount of silver found in the fabrics and their antibacterial properties.
Artificial sweat has been used frequently for assessing the level of metal release from various materials, such as to study nickel released from earrings, which shows a relationship to dermatitis [32
]. However, the composition of sweat not only varies between individuals, but also within an individual according to their body region, age, season, degree of acclimation, diet, infection status and level of activity [33
]. Therefore, the effect of variations in the artificial sweat composition on the silver leaching levels was evaluated using the four international standard artificial sweat formulations (Table ).
Even though the manufacturers claimed nanosilver products, the data indicated that silver was detected only in three out of six commercial fabrics. One of the possibilities might be due to the very low amount of silver available in the products, less than the lower threshold detection limit (0.26 μg/L) of the instrument. In addition, as the level of silver approaches the detection limit, the less accurate are the measured results. Regardless, for those fabrics with detectable levels of silver, it was observed that silver was released into the artificial sweat after incubation at 37°C for 24 h.
For the laboratory-prepared fabrics, the amount of silver released into the artificial sweat was dependent upon the initial amount of silver coated onto each sample. However, nanosilver fabrics can be prepared by several methods [5
]. In this study, some commercial fabrics had different textures and thus likely methods of incorporating silver into them. For instance, unlike the other commercial fabrics, fabric E was prepared by incorporating silver nanoparticles into the fibers (pers. com. with the manufacture). Variation in the fabric quality might be one of the factors that affected the release of silver into artificial sweat.
The ISO artificial sweat formulation (pH 5.5) showed the lowest detected levels of silver leaching from both laboratory-made and commercially obtained fabrics. This pH is similar to the pH of normal human skin [33
]. In contrast, in the EN artificial sweat formulation (pH 6.5), the largest release of silver from all fabrics was observed. This standard formulation is normally used for determining metal release from jewelry [20
]. As shown in Table , it contained urea, which was not included in the other artificial sweat formulations used in this study. Therefore, the release of silver from the fabrics was also likely to be affected by the pH and formulations of artificial sweat.
Several lines of nanosilver-based textile fabrics are already on the market. Dermal exposure represents an important potential route of exposure for these nanoparticles. Yet, data in relation to the potential human health risk are very rare. The release of nanoparticles from the fabrics under various conditions, such as sweating, repetitive attrition and laundering, are considered essential information. In this study, it is possible that artificial sweat might facilitate transfer of silver-nanoparticle-treated fabric to the skin surface [35
]. However, it remains unclear whether these nanoparticles released from the products at the site of application were absorbed into the body. Close contact may allow nanoparticles to penetrate through compromised skin barrier and gain access to the dermal capillaries [12
]. With regard to this notion, abnormal elevation of blood silver levels, Argyria-like symptoms and hepatotoxicity following the use of nanosilver coated dressings for burns in clinical application has been reported [36
]. Our investigation may have a significant advantage for the evaluation of the human health risk of silver nanoparticles released from textile products. Future advanced research in relation to these aspects is needed to be conducted.