Treatment of infection and inflammation associated with COVID-19, multi-drug resistant pneumonia and fungal sinusitis by nebulizing a nanosilver solution.

Solutions containing Ag0 nanoclusters, Ag+1, and higher oxidation state silver, generated from nanocrystalline silver dressings, were anti-inflammatory against porcine skin inflammation. The dressings have clinically-demonstrated broad-spectrum antimicrobial activity, suggesting application of nanosilver solutions in treating pulmonary infection. Nanosilver solutions were tested for antimicrobial efficacy; against HSV-1 and SARS-CoV-2; and nebulized in rats with acute pneumonia. Patients with pneumonia (ventilated), fungal sinusitis, burns plus COVID-19, and two non-hospitalized patients with COVID-19 received nebulized nanosilver solution. Nanosilver solutions demonstrated pH-dependent antimicrobial efficacy; reduced infection and inflammation without evidence of lung toxicity in the rat model; and inactivated HSV-1 and SARS-CoV-2. Pneumonia patients had rapidly reduced pulmonary symptoms, recovering pre-illness respiratory function. Fungal sinusitis-related inflammation decreased immediately with infection clearance within 21 days. Non-hospitalized patients with COVID-19 experienced rapid symptom remission. Nanosilver solutions, due to anti-inflammatory, antiviral, and antimicrobial activity, may be effective for treating respiratory inflammation and infections caused by viruses and/or microbes.

Incorporating gold into nanocrystalline silver dressings reduces grain boundary size and maintains suitable antimicrobial properties.

Nanocrystalline silver dressings are widely known to be potent antimicrobial and anti-inflammatory agents and have long been used to treat topical wounds. Gold is known to be a strong anti-inflammatory agent and has been used in the treatment of rheumatoid arthritis for >70 years. The purpose of this work was to study the effect of incorporating gold into nanocrystalline silver dressings from antimicrobial and anti-inflammatory perspectives. Gold and silver dressing alloys were created by direct current magnetron sputtering and compared with pure silver nanocrystalline dressings using conventional biological (log reduction and corrected zone of inhibition) and physical (X-ray diffraction, X-ray photoelectron spectroscopy, energy-dispersive X-ray spectroscopy, atomic absorption spectroscopy, atomic force microscopy and scanning electron microscopy) characterisation techniques. While the gold/silver dressings were slightly weaker antimicrobials than the pure silver nanocrystalline structures, the addition of gold to the nanostructure reduces the minimum crystallite size from 17 to 4 nm. This difference increases the number of grain boundary atoms from 12% to 40% which could augment the anti-inflammatory properties of the dressings. The formation of gold oxide (Au2O3) was thought to be responsible for the observed decrease in crystallite size.

Atomic force microscopy: a nanoscopic view of microbial cell surfaces.

The atomic force microscope (AFM) is a powerful instrument for microbiological investigation. It has evolved from an imaging tool used to investigate microbial surfaces at high resolution in their physiological environment into a lab-on-a-tip device, which allows more quantitative analysis of biological samples (from molecules to cells) in aqueous liquids. Atomic force microscopy provides information about the nanoscale architecture of microbes and about the localization and interactions of their individual constituents. Microbial interactions play essential roles in biology, medicine, ecology, biotechnology, food science and contribute to phenomena as varied as bacterial infections, biofilm formation, and bacterial adhesion to surfaces. In this review, we focus on recent developments offered by the rapid advances in AFM imaging and force spectroscopy with emphasizes on microbial research.

Special considerations in wound bed preparation 2011: an update©.

This article builds and expands upon the concept of wound bed preparation introduced by Sibbald et al in 2000 as a holistic approach to wound diagnosis and treatment of the cause and patient-centered concerns such as pain management, optimizing the components of local wound care: Debridement, Infection and persistent Inflammation, along with Moisture balance before Edge effect for healable but stalled chronic wounds.

Evaluating antimicrobial efficacy of new commercially available silver dressings.

Prevention and treatment of bacterial colonised/infected wounds are critical. Many commercially available silver dressings claim broad-spectrum bactericidal activity over days and are indicated for serious conditions including burns and ulcers. However, there is no peer-reviewed literature available for many newer dressings. This study compared the activity of some of these dressings. Six silver-containing dressings were compared using log reduction, silver release and corrected zone of inhibition assays. Only the nanocrystalline silver dressing was bactericidal against Staphylococcus aureus, and the only other dressing that produced any log reduction was a silver collagen matrix dressing. These two dressings and a silver alginate dressing produced zones of inhibition, although the collagen matrix and alginate dressings had decreasing zone sizes over time, and the latter liquefied after five transfers. The remaining dressings (two ionic silver foam dressings and a silver sulphate dressing) did not produce zones of inhibition. For the foam, alginate and collagen matrix dressings, antimicrobial activity was related to silver release. The silver sulphate dressing released large quantities of silver, but only through the dressing edges, as the wound-contacting surface appeared to be hydrophobic. The results of this study emphasise the importance of confirming product claims regarding silver dressing efficacy.

Does nanocrystalline silver have a transferable effect?

This study examined the mechanism of nanocrystalline silver antiinflammatory activity, and tested nanocrystalline silver for systemic antiinflammatory effects. Secondary ion mass spectroscopy of skin treated directly with nanocrystalline silver for 24 hours showed that at skin surfaces there were significant deposits at weights corresponding to Ag, AgO, AgCl, AgNO(3), Ag(2)O, and silver clusters Ag(2-6), but silver penetration was minimal. To test for translocation of the effect, a porcine contact dermatitis model in which wounds were induced on one side of the back and then treated with nanocrystalline silver on the opposite side of the back was used. Visual and histological data showed improvement relative to animals treated with saline only. Significantly increased induction of apoptosis in the inflammatory cells present in the dermis was observed with remote nanocrystalline silver treatments. In addition, immunohistochemical analysis showed decreased levels of proinflammatory cytokines tumor necrosis factor-alpha and interleukin-8, and increased levels of antiinflammatory cytokine interleukin-4, epidermal growth factor, keratinocyte growth factor, and keratinocyte growth factor-2. Thus, the antiinflammatory effects of nanocrystalline silver appear to be induced by interactions with cells in the top layers of the skin, which then release biological signals resulting in widespread antiinflammatory activity.

Anti-inflammatory activity of nanocrystalline silver-derived solutions in porcine contact dermatitis.

BACKGROUND: Nanocrystalline silver dressings have anti-inflammatory activity, unlike solutions containing Ag+ only, which may be due to dissolution of multiple silver species. These dressings can only be used to treat surfaces. Thus, silver-containing solutions with nanocrystalline silver properties could be valuable for treating hard-to-dress surfaces and inflammatory conditions of the lungs and bowels. This study tested nanocrystalline silver-derived solutions for anti-inflammatory activity. METHODS: Inflammation was induced on porcine backs using dinitrochlorobenzene. Negative and positive controls were treated with distilled water. Experimental groups were treated with solutions generated by dissolving nanocrystalline silver in distilled water adjusted to starting pHs of 4 (using CO2), 5.6 (as is), 7, and 9 (using Ca(OH)2). Solution samples were analyzed for total silver. Daily imaging, biopsying, erythema and oedema scoring, and treatments were performed for three days. Biopsies were processed for histology, immunohistochemistry (for IL-4, IL-8, IL-10, TNF-alpha, EGF, KGF, KGF-2, and apoptotic cells), and zymography (MMP-2 and -9). One-way ANOVAs with Tukey-Kramer post tests were used for statistical analyses. RESULTS: Animals treated with pH 7 and 9 solutions showed clear visual improvements. pH 9 solutions resulted in the most significant reductions in erythema and oedema scores. pH 4 and 7 solutions also reduced oedema scores. Histologically, all treatment groups demonstrated enhanced re-epithelialisation, with decreased inflammation. At 24 h, pMMP-2 expression was significantly lowered with pH 5.6 and 9 treatments, as was aMMP-2 expression with pH 9 treatments. In general, treatment with silver-containing solutions resulted in decreased TNF-alpha and IL-8 expression, with increased IL-4, EGF, KGF, and KGF-2 expression. At 24 h, apoptotic cells were detected mostly in the dermis with pH 4 and 9 treatments, nowhere with pH 5.6, and in both the epidermis and dermis with pH 7. Solution anti-inflammatory activity did not correlate with total silver content, as pH 4 solutions contained significantly more silver than all others. CONCLUSIONS: Nanocrystalline silver-derived solutions appear to have anti-inflammatory/pro-healing activity, particularly with a starting pH of 9. Solutions generated differently may have varying concentrations of different silver species, only some of which are anti-inflammatory. Nanocrystalline silver-derived solutions show promise for a variety of anti-inflammatory treatment applications.

The kinetics of thermal instability in nanocrystalline silver and the effect of heat treatment on the antibacterial activity of nanocrystalline silver dressings.

The kinetics of nanocrystalline silver dressing heat treatment was investigated via isothermal heat treatments at 90 degrees C, 100 degrees C, and 110 degrees C lasting 2-50h. Bactericidal efficacy of the dressings was measured via log reductions, while bacteriostatic longevity was determined via plate-to-plate transfer corrected zones of inhibition. Morphological evolution of the dressing was studied by X-ray diffraction, scanning electron microscopy, and X-ray photoelectron spectroscopy, while changes in heat flow were measured by differential scanning calorimetry. Increasing temperature increased the rate at which dressing bactericidal activity and bacteriostatic longevity decreased. Once changes in dressing properties began, they occurred nonlinearly with time. The earliest biological, chemical, and physical indicators of altered dressing properties were loss of bacteriostatic longevity, silver-oxygen bonds, and fine features, respectively. An early change in heat flow appeared to be responsible for these indicators, while a later change corresponded to rapid grain growth occurring after a critical crystallite size (approximately 30 nm) was reached. The grain growth exponent was determined to be 2.8 for temperatures of 100-110 degrees C, with an activation energy of 177 kJ/mol, suggesting that normal grain growth occurred, with volume and/or grain boundary diffusion as the dominant forms of diffusion. The thermal instability of nanocrystalline silver should be accounted for during production, storage, and use of dressings. The properties required for nanosilver antimicrobial efficacy demonstrated in this study, as well as its thermal instability, should be taken into consideration for the development of nanosilver products in the future.

Anti-inflammatory activity of nanocrystalline silver in a porcine contact dermatitis model.

The anti-inflammatory activity of nanocrystalline silver was examined using a porcine model of contact dermatitis. Inflammation was induced with dinitrochlorobenzene and then treated daily with nanocrystalline silver dressings, 0.5% silver nitrate, or saline. Erythema, edema, and histological data showed that nanocrystalline silver-treated pigs had near-normal skin after 72 hours, while other treatment groups remained inflamed. The decreased inflammation in the nanocrystalline silver-treated group was associated with increased inflammatory cell apoptosis, a decreased expression of proinflammatory cytokines, and decreased gelatinase activity. Silver nitrate treatments induced apoptosis in all cell types, including keratinocytes, resulting in delayed wound healing. These results demonstrate that nanocrystalline silver had a direct anti-inflammatory effect in the porcine contact dermatitis model that improved the overall outcome of the healing process. These data offer support that a species of silver (e.g., Ag(0)) that is uniquely associated with nanocrystalline silver may be responsible for the anti-inflammatory activity and improvement in healing.

The effect of treating infected skin grafts with Acticoat on immune cells.

A study was conducted to determine the effect of Acticoat placed on an infected skin graft on parameters of immunity. Two partial thickness wounds (2 cm x 4 cm) were created on the dorsal midline of Hartley guinea pigs (n=28). Wounds were covered with autologous skin graft and maintained either aseptically (Noninoculated, n=8), inoculated with Staphylococcus aureus (Surgery-Inoculated, n=8) with or without Acticoat bandage (Surgery-Inoculated-Acticoat, n=6). Five days later, splenocytes and blood were collected to estimate natural killer cell (NK) cytotoxicity, proliferative response to T and B cell mitogens and neutrophil oxidative burst. Animals that did not undergo surgery were included as a nonsurgery control group. [(3)H]-thymidine incorporation in response to a variety of T and B cell mitogens was significantly lower for all groups undergoing surgery compared to the nonsurgery control group (p<0.0001) and no additional effect was observed on this immune measure by applying the Acticoat bandage. The Surgery-Inoculated-Acticoat group exhibited greater NK cytotoxic activity (as assessed as the ability to lyse K562 tumor cells) compared to the Surgery-Inoculated group (p<0.006). The Surgery-Inoculated-Acticoat group had higher neutrophil oxidative burst at 5 min post stimulation, but was not different from controls after 15 min. In conclusion, the application of an Acticoat bandage to an inoculated surgery wound did not alter the low cell-mediated immune response that followed surgery, but appeared to increase parameters (NK cytotoxic activity and neutrophil function) of innate immunity.

Comparison of in vitro disc diffusion and time kill-kinetic assays for the evaluation of antimicrobial wound dressing efficacy.

There is a plethora of new silver-containing dressings on the market today. Various manufacturers attempt to show that their dressings are the most efficacious and therefore should be preferentially employed by health care workers based on the results of their in vitro tests. However, there have been no studies that clearly identify which tests are appropriate for comparison purposes. The purpose of this study was to determine which in vitro test is most appropriate for evaluating the antimicrobial efficacy of silver-containing dressings. This was done by testing seven silver-containing dressings and two non-silver-containing topical agents against 17 clinically relevant microorganisms using zone of inhibition assays and time-kill kinetic assays in complex media. The results for the two assays were then correlated to determine whether the methods generated similar results. It was determined that the two methods do not correlate at all. This is most likely a result of the silver interacting with the media in the zone of inhibition test, thus invalidating the results of this test. We therefore conclude that zone of inhibition data generated for silver-containing dressings is of little value when assessing antimicrobial efficacy and that time-kill assays are of greater use.

Pseudomonas infections in the thermally injured patient.

Pseudomonas aeruginosa, remains a serious cause of infection and septic mortality in burn patients, particularly when nosocomially acquired. A prototypic burn patient who developed serious nosocomially acquired Pseudomonas infection is described as an index case which initiated investigations and measures taken to identify the source of the infection. The effect of changes in wound care to avoid further nosocomial infections was measured to provide data on outcome and cost of care. The bacteriology of Pseudomonas is reviewed to increase the burn care providers understanding of the behaviour of this very common and serious pathogen in the burn care setting, before reviewing the approach to detection of the organism and treatment both medically and surgically. After controlling the nosocomial spread of Pseudomonas in our burn unit, we investigated the morbidity and mortality associated with nosocomial infection with an aminoglycoside resistant Pseudomonas and the associated costs compared to a group of case-matched control patients with similar severity of burn injury, that did not acquire resistant Pseudomonas during hospitalization at our institution. We found a significant increase in the mortality rate in the Pseudomonas group compared to controls. The morbidity in terms of length of stay, ventilator days, number of surgical procedures, and the amount of blood products used were all significantly higher in the Pseudomonas group compared to controls. Costs associated with antibiotic requirements were also significantly higher in the Pseudomonas group. Despite this increased resource consumption necessary to treat Pseudomonas infections, these efforts did not prevent significantly higher mortality rates when compared to control patients who avoided infection with the resistant organism. Thus, in addition to the specific measures required to identify and treat nosocomial Pseudomonas infections in burn patients, prevention of infection through modification of treatment protocols together with continuous infection control measures to afford early identification and eradication of nosocomial Pseudomonas infection are critical for cost-effective, successful burn care.

Early healing events in a porcine model of contaminated wounds: effects of nanocrystalline silver on matrix metalloproteinases, cell apoptosis, and healing.

A porcine model of wound healing was employed to examine the impact of nanocrystalline silver-coated dressings on specific wound healing events. Full-thickness wounds were created on the backs of pigs, contaminated with an experimental inoculum containing Pseudomonas aeruginosa, Fusobacterium sp., and coagulase-negative staphylococci, and covered with dressing products either containing silver or not. Nanocrystalline silver-coated dressings promoted rapid wound healing, particularly during the first several days post-injury. Healing was characterized by rapid development of well vascularized granulation tissue that supported tissue grafting 4 days post-injury, unlike control dressed wounds. The proteolytic environment of wounds treated with nanocrystalline silver was characterized by reduced levels of matrix metalloproteinases. Matrix metalloproteinases have been shown to be present in chronic ulcers at abnormally high levels, as compared with acute wounds, and may contribute to the nonhealing nature of these wounds. Cellular apoptosis occurred at a higher frequency in the nanocrystalline silver-treated wounds than in wounds dressed with other products. The results suggest that nanocrystalline silver may play a role in altering or compressing the inflammatory events in wounds and facilitating the early phases of wound healing. These benefits are associated with reduced local matrix metalloproteinase levels and enhanced cellular apoptosis.