Nano-silver is used in an increasing number of products. Some of the applications have resulted in the concern of governments and the public, since little is known about the potential risks of nano-silver. In this review, an inventory is made to identify knowledge gaps that have to be filled before risks for both man and the environment can be assessed as reliable as for ‘non-nanosized’ chemicals. It is hypothesized that the toxic effects of nano-silver are due to a combination of the specific properties of silver nanoparticles and the generation of ions from them. The main topic for future research is validation of our ‘0-hypothesis’ that toxic effects of nano-silver are proportional to the activity of free silver ions released by the nanoparticles. Furthermore, it must be determined whether – or to what extent – nano-silver particles will enter the body. The outcomes of these tests will determine the requirements for further toxicity testing.

Nano-silver – a review of available data and knowledge gaps in human and environmental risk assessment

In the present scenario, pharmaceutical and biomedical sectors are facing the challenges of continuous increase in the multidrug-resistant (MDR) human pathogenic microbes. Re-emergence of MDR microbes is facilitated by drug and/or antibiotic resistance, which is acquired way of microbes for their survival and multiplication in uncomfortable environments. MDR bacterial infections lead to significant increase in mortality, morbidity and cost of prolonged treatments. Therefore, development, modification or searching the antimicrobial compounds having bactericidal potential against MDR bacteria is a priority area of research. Silver in the form of various compounds and bhasmas have been used in Ayurveda to treat several bacterial infections since time immemorial. As several pathogenic bacteria are developing antibiotic resistance, silver nanoparticles are the new hope to treat them. This review discusses the bactericidal potential of silver nanoparticles against the MDR bacteria. This multiactional nanoweapon can be used for the treatment and prevention of drug-resistant microbes.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2672.2012.05253.x

Silver nanoparticles: the powerful nanoweapon against multidrug-resistant bacteria

Chronic wound pathogenic biofilms are host-pathogen environments that colonize and exist as a cohabitation of many bacterial species. These bacterial populations cooperate to promote their own survival and the chronic nature of the infection. Few studies have performed extensive surveys of the bacterial populations that occur within different types of chronic wound biofilms. The use of 3 separate16S-based molecular amplifications followed by pyrosequencing, shotgun Sanger sequencing, and denaturing gradient gel electrophoresis were utilized to survey the major populations of bacteria that occur in the pathogenic biofilms of three types of chronic wound types: diabetic foot ulcers (D), venous leg ulcers (V), and pressure ulcers (P). There are specific major populations of bacteria that were evident in the biofilms of all chronic wound types, including Staphylococcus, Pseudomonas, Peptoniphilus, Enterobacter, Stenotrophomonas, Finegoldia, and Serratia spp. Each of the wound types reveals marked differences in bacterial populations, such as pressure ulcers in which 62% of the populations were identified as obligate anaerobes. There were also populations of bacteria that were identified but not recognized as wound pathogens, such as Abiotrophia para-adiacens and Rhodopseudomonas spp. Results of molecular analyses were also compared to those obtained using traditional culture-based diagnostics. Only in one wound type did culture methods correctly identify the primary bacterial population indicating the need for improved diagnostic methods. If clinicians can gain a better understanding of the wound's microbiota, it will give them a greater understanding of the wound's ecology and will allow them to better manage healing of the wound improving the prognosis of patients. This research highlights the necessity to begin evaluating, studying, and treating chronic wound pathogenic biofilms as multi-species entities in order to improve the outcomes of patients. This survey will also foster the pioneering and development of new molecular diagnostic tools, which can be used to identify the community compositions of chronic wound pathogenic biofilms and other medical biofilm infections.

/pdf/survey-of-bacterial-diversity-in-chronic-wounds-using-4r1b1c70h8.pdf

Survey of bacterial diversity in chronic wounds using Pyrosequencing, DGGE, and full ribosome shotgun sequencing

Nanoparticles (NPs) can potentially cause adverse effects on organ, tissue, cellular, subcellular, and protein levels due to their unusual physicochemical properties (e.g., small size, high surface area to volume ratio, chemical composition, crystallinity, electronic properties, surface structure reactivity and functional groups, inorganic or organic coatings, solubility, shape, and aggregation behavior). Metal NPs, in particular, have received increasing interest due to their widespread medical, consumer, industrial, and military applications. However, as particle size decreases, some metal-based NPs are showing increased toxicity, even if the same material is relatively inert in its bulk form (e.g., Ag, Au, and Cu). NPs also interact with proteins and enzymes within mammalian cells and they can interfere with the antioxidant defense mechanism leading to reactive oxygen species generation, the initiation of an inflammatory response and perturbation and destruction of the mitochondria causing apoptosis or necrosis. As a result, there are many challenges to overcome before we can determine if the benefits outweigh the risks associated with NPs. WIREs Nanomed Nanobiotechnol 2010 2 544–568

For further resources related to this article, please visit the WIREs website

The view presented in this manuscript does not necessarily reflect those of Food and Drug Administration.

/pdf/metal-based-nanoparticles-and-their-toxicity-assessment-3a512ilb5p.pdf

Metal-based nanoparticles and their toxicity assessment.

Nanoparticles possess exceptional physical and chemical properties which led to rapid commercialisation. Silver nanoparticles (Ag-np) are among the most commercialised nanoparticles due to their antimicrobial potential. Ag-np based cosmetics, therapeutic agents and household products are in wide use, which raised a public concern regarding their safety associated with human and environmental use. No safety regulations are in practice for the use of these nanomaterials. The interactions of nanomaterials with cells, uptake mechanisms, distribution, excretion, toxicological endpoints and mechanism of action remain unanswered. Normal human lung fibroblasts (IMR-90) and human glioblastoma cells (U251) were exposed to different doses of Ag-nps in vitro. Uptake of Ag-nps occurred mainly through endocytosis (clathrin mediated process and macropinocytosis), accompanied by a time dependent increase in exocytosis rate. The electron micrographs revealed a uniform intracellular distribution of Ag-np both in cytoplasm and nucleus. Ag-np treated cells exhibited chromosome instability and mitotic arrest in human cells. There was efficient recovery from arrest in normal human fibroblasts whereas the cancer cells ceased to proliferate. Toxicity of Ag-np is mediated through intracellular calcium (Ca2+) transients along with significant alterations in cell morphology and spreading and surface ruffling. Down regulation of major actin binding protein, filamin was observed after Ag-np exposure. Ag-np induced stress resulted in the up regulation of metallothionein and heme oxygenase -1 genes. Here, we demonstrate that uptake of Ag-np occurs mainly through clathrin mediated endocytosis and macropinocytosis. Our results suggest that cancer cells are susceptible to damage with lack of recovery from Ag-np-induced stress. Ag-np is found to be acting through intracellular calcium transients and chromosomal aberrations, either directly or through activation of catabolic enzymes. The signalling cascades are believed to play key roles in cytoskeleton deformations and ultimately to inhibit cell proliferation.

/pdf/anti-proliferative-activity-of-silver-nanoparticles-4q46hctxox.pdf

Anti-proliferative activity of silver nanoparticles

Antimicrobial dressings such as those containing silver are now being used widely to control wound bioburden, and tests to demonstrate their efficacy predominantly involve in vitro models using free-living or planktonic bacteria. In this present study a wide range of antibiotic-sensitive and resistant bacteria were tested in their quasi-sessile state using a standard agar assay and a second method used a poloxamer gel (true biofilm state - poloxamer encourages microorganisms to exhibit a more clinically relevant biofilm phenotype) technique. The antimicrobial activity of two silver dressings, a silver-containing Hydrofiber (SCH) dressing and a nanocrystalline silver-containing dressing (NCS), were evaluated on a variety of microorganisms, using a zone-of-inhibition (ZOI) test. When grown on agar (presenting a quasi-sessile state of each organism), the antibiotic-susceptible microorganisms were generally more susceptible to the SCH dressing compared with the NCS. ZOIs associated with SCH dressing ranged between 5.7 and 17.5 mm; those for the NCS against the same group of organisms ranged between 1.9 and 8.6 mm. When grown on poloxamer gel, (presenting the biofilm state of each organism) the same group of microorganisms were less susceptible to both dressings. The SCH dressing was most effective against strains of Pseudomonas aeruginosa, Candida albicans and Staphylococcus aureus (ZOI range: 2.6-6 mm); the NCS was most effective against strains of Klebsiella pneumoniae, Enterococcus faecalis and Escherichia coli (i.e. ZOI range: 1-2.8 mm). Similarly to the antibiotic-susceptible microorganisms, nine of ten antibiotic-resistant bacterial strains when grown on agar were more susceptible to the SCH dressing compared with the NCS. Although the microorganisms tested were universally less susceptible to the silver dressings when in their biofilm state, in the majority of cases, the SCH dressing demonstrated greater biofilm-inhibiting activity than the NCS.

Antimicrobial activity of silver‐containing dressings on wound microorganisms using an in vitro biofilm model

Wound infections, due to biofilms, are a constant problem because of their recalcitrant nature towards antibiotics. Appropriate antibiotic selection for the treatment of these biofilm infections is important. The traditional in vitro disc diffusion method for antibiotic selection uses bacterial cultures grown on agar plates. However, the form of bacterial growth on agar is not representative of how bacteria grow in wounds and other tissue sites as here bacteria grow naturally in a biofilm. The aim of this research was to test a more appropriate method for testing antimicrobial efficacy on biofilms and compare with the standard methods used for antibiotic sensitivity testing. Outer Membrane Protein analysis was performed on E.coli, Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis and Acinetobacter juni when grown on Mueller Hinton agar ('quasi-biofilm state') and 30% Poloxamer hydrogel ('true- biofilm state). Susceptibility to antibiotics on 28 clinical isolates was determined using the modified Kirby Bauer disc diffusion method, on agar and 30% Poloxamer. Similar outer membrane proteins [OMPs] were identified in bacteria grown in a biofilm state and on a 30% poloxamer hydrogel, which were very different to the OMPs identified in bacteria grown on Mueller-Hinton agar and broth. There was a significant difference between the means of the clearance zones around the antibiotic discs on standard agar and poloxamer gels [P 0.05]. The findings of this experiment suggest that poloxamer gel could be used as an appropriate medium on which to conduct biofilm antibiotic susceptibility tests as it enables bacteria to be grown in a state representative of the infected surface from which the culture was taken.

/pdf/evaluating-antibiotics-for-use-in-medicine-using-a-poloxamer-1nx4wka7sa.pdf

Jayne Dolman

Papers

Antimicrobial activity of silver‐containing dressings on wound microorganisms using an in vitro biofilm model

Evaluating antibiotics for use in medicine using a poloxamer biofilm model

Susceptibility of silver resistant bacteria to silver containing wound dressings