The use of nanomaterials is becoming increasingly widespread, leading to substantial research focused on nanomedicine. Nevertheless, the lack of complete toxicity profiles limits nanomaterials’ uses, despite their remarkable diagnostic and therapeutic results on in vitro and in vivo models. Silver nanoparticles (AgNPs), particularly Argovit™, have shown microbicidal, virucidal, and antitumoral effects. Among the first-line toxicity tests is the hemolysis assay. Here, the hemolytic effect of Argovit™ AgNPs on erythrocytes from one healthy donor (HDE) and one diabetic donor (DDE) is evaluated by the hemolysis assay against AgNO₃. The results showed that Argovit™, in concentrations ≤24 µg/mL of metallic silver, did not show a hemolytic effect on the HDE or DDE. On the contrary, AgNO₃ at the same concentration of silver ions produces more than 10% hemolysis in both the erythrocyte types. In all the experimental conditions assessed, the DDE was shown to be more prone to hemolysis than the HDE elicited by Ag⁺ ions or AgNPs, but much more evident with Ag⁺ ions. The results show that Argovit™ is the least hemolytic compared with the other twenty-two AgNP formulations previously reported, probably due to the polymer mass used to stabilize the Argovit™ formulation. The results obtained provide relevant information that contributes to obtaining a comprehensive toxicological profile to design safe and effective AgNP formulations.

The present work presents translational research with application of AgNPs targeting the global drug resistance problem. In vivo fieldwork was carried out with 400 breeding farm cows sick with a serous mastitis. Ex vivo results revealed that after cow treatment with Lactobay^TM (a mixture of antibiotic drugs) the susceptibility to 31 antibiotics of S. aureus isolates from cow breast secretion decreased by 25%, while after treatment with Argovit–C^TM silver nanoparticles S. aureus susceptibility increased by 11%. The portion of isolates with an efflux effect leading to elimination of antibiotics from S. aureus after Lactobay-treatment resulted in a 15% increase, while Argovit-C-treatment led to a 17.5% decrease. The obtained results showed that mastitis treatments with Argovit-C^TM AgNPs can partially restore the activity of antibiotics towards S. aureus and shorten the duration of mastitis treatment by 33%.

The hemolytic activity assay is a versatile tool for fast primary toxicity studies. This work presents a systematic study of the hemolytic properties of Argovit^TM silver nanoparticles (AgNPs) extensively studied for biomedical applications. The results revealed an unusual and unexpected bell-shaped hemolysis curve for human healthy and diabetic donor erythrocytes. With the decrease of pH from 7.4 and 6.8 to 5.6, the hemolysis profiles for AgNPs and AgNO₃ changed dramatically. For AgNPs, the bell shape changed to a step shape with a subsequent sharp increase, and for AgNO₃ it changed to a gradual increase. Explanations of these changes based on the aggregation of AgNPs due to the increase of proton concentration were suggested. Hemolysis of diabetic donor erythrocytes was slightly higher than that of healthy donor erythrocytes. The meta-analysis revealed that for only one AgNPs formulation (out of 48), a bell-shaped hemolysis profile was reported, but not discussed. This scarcity of data was explained by the dominant goal of studies consisting in achieving clinically significant hemolysis of 5–10%. Considering that hemolysis profiles may be bell-shaped, it is recommended to avoid extrapolations and to perform measurements in a wide concentration interval in hemolysis assays.

The present work is a continuation of our translational research focusing on the use of silver nanoparticles (AgNPs) to solve the global problem of antibiotic resistance. In vivo fieldwork was done with 300 breeding farm cows with serous mastitis. Ex vivo assays revealed that after cow treatment with the antibiotic drug Spectromast LC^TM, S.dysgalactiae susceptibility to 31 antibiotics dropped by 22.9%, but after treatment with Argovit–C^TM AgNPs, it was raised by 13.1%. This was explained by the fact that the percentage of isolates with an efflux effect after Spectromast LC treatment resulted in an 8% increase, while Argovit-C-treatment caused a 19% decrease. The similarity of these results to our previous results on S. aureus isolates from mastitis cows treated with the antibiotic drug Lactobay and Argovit–C^TM AgNPs was shown. So, mastitis treatments with Argovit-C^TM AgNPs can partially return the activity of antibiotics towards S.dysgalactiae and S. aureus, while, in contrast, treatments with antibiotic drugs such as Spectromast LC and Lactobay enhance bacterial resistance to antibiotics. The results of this work strengthen the hope that in the future the use of AgNPs as efflux pump inhibitors will recover the activity of antibiotics, and thus will preserve the wide spectrum of antibiotics on the market.

The current work is a continuation of our studies focused on the application of nanoparticles of metallic silver (AgNPs) to address the global problem of antibiotic resistance. In vivo, fieldwork was carried out with 200 breeding cows with serous mastitis. Ex vivo analyses showed that after the cow was treated with an antibiotic-containing drug Dienomast^TM, E. coli sensibility to 31 antibiotics decreased by 27.3%, but after treatment with AgNPs, it increased by 21.2%. This could be explained by the 8.9% increase in the portion of isolates showing an efflux effect after Dienomast^TM treatment, while treatment with Argovit-C^TM resulted in a 16.0% drop. We verified the likeness of these results with our previous ones on S. aureus and Str. dysgalactiae isolates from mastitis cows processed with antibiotic-containing medicines and Argovit-C^TM AgNPs. The obtained results contribute to the recent struggle to restore the efficiency of antibiotics and to preserve the wide range of antibiotics on the world market.

The growing resistance of bacteria to antibiotics is one of the main public health problems nowadays. The influence of silver nanoparticle (AgNP) pretreatment of 220 cows with mastitis on the susceptibility of Staphylococcus epidermidis bacteria to 31 antibiotics was studied. The obtained results were compared with the previous results for Escherichia coli, Streptococcus dysgalactiae, and Staphylococcus aureus. For all four bacteria, an increase in susceptibility (9.5–21.2%) to 31 antibiotics after cow treatment with AgNPs was revealed, while after first-line antibiotic drug treatment as expected, the susceptibility decreased (11.3–27.3%). These effects were explained by (1) the increase in the contribution of isolates with efflux effect after antibiotic treatments and its decrease after AgNP treatment and (2) the changes in bacteria adhesion and anti-lysozyme activity after these treatments. The effect of the increasing antibacterial activity of antibiotics after AgNP treatment was the most pronounced in the case of E. coli and was minimal in the case of S. epidermidis. With AgNP treatment, the time of recovery decreased by 26.8–48.4% compared to the time of recovery after treatment with the first-line antibiotic drugs. The AgNP treatment allows for achieving the partial restoration of the activity of antibiotics.