ABSTRACT
The hemolytic activity assay is a versatile tool for fast primary toxicity studies. This work presents a systematic study of the hemolytic properties of ArgovitTM 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.
ABSTRACT
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.
ABSTRACT
Silver nanoparticles (AgNPs) are the most used nanomaterials worldwide due to their excellent antibacterial, antiviral, and antitumor activities, among others. However, there is scarce information regarding their genotoxic potential measured using human peripheral blood lymphocytes. In this work, we present the cytotoxic and genotoxic behavior of two commercially available poly(vinylpyrrolidone)-coated silver nanoparticle (PVP-AgNPs) formulations that can be identified as noncytotoxic and nongenotoxic by just evaluating micronuclei (MNi) induction and the mitotic index, but present enormous differences when other parameters such as cytostasis, apoptosis, necrosis, and nuclear damage (nuclear buds (NBUDs) and nucleoplasmic bridges (NPBs)) are analyzed. The results show that Argovit (35 nm PVP-AgNPs) and nanoComposix (50 nm PVP-AgNPs), at concentrations from 0.012 to 12 µg/mL, produce no changes in the nuclear division index (NDI) or micronuclei (MNi) frequency compared with the values found on control cultures of human blood peripheral lymphocytes from a healthy donor. Still, 50 nm PVP-AgNPs significantly decrease the replication index and significantly increase cytostasis, apoptosis, necrosis, and the frequencies of nuclear buds (NBUDs) and nucleoplasmic bridges (NPBs). These results provide evidence that the cytokinesis-block micronucleus (CBMN) assay using human lymphocytes and evaluating the eight parameters provided by the technique is a sensitive, fast, accurate, and inexpensive detection tool to support or discard AgNPs or other nanomaterials, which is worthwhile for continued testing of their effectiveness and toxicity for biomedical applications. In addition, it provides very important information about the role played by the [coating agent]/[metal] ratio in the design of nanomaterials that could reduce adverse effects as much as possible while retaining their therapeutic capabilities.
