The in Vitro Effect of Polyvinylpyrrolidone and Citrate Coated Silver Nanoparticles on Erythrocytic Oxidative Damage and Eryptosis

Abstract

Background/Aims: Silver nanoparticles (AgNPs) are increasingly used as antimicrobial agents and drug carriers in various biomedical fields. AgNPs can encounter erythrocytes either directly following intravenous injection, or indirectly via translocation from the site of administration. However, information regarding the pathophysiological effects and possible mechanism of action of AgNPs on the erythrocytes are still inadequately studied. Thus, the aim of our study was to investigate the mechanism underlying the effect of coating and concentration of AgNPs on mouse erythrocytes in vitro. Methods: We studied the interaction of polyvinylpyrrolidone (PVP) and citrate (CT) coated AgNPs (10 nm) at various concentrations (2.5, 10, 40 µg/ml) with mouse erythrocytes in vitro using various techniques including transmission electron microscopy (TEM), hemolysis, and colorimetric measurement of markers of oxidative stress comprising malondialdehyde (MDA), reduced glutathione (GSH), and catalase (CAT). Intracellular calcium (Ca²⁺) was determined using Fura 2AM fluorescence. Annexin V was quantified using ELISA and the caspase 3 was determined both flurometrically and by western blot technique. Results: Following incubation of the erythrocytes with AgNPs, both PVP- and CT- AgNPs induced significant and dose - dependent increase in hemolysis. TEM revealed that both PVP- and CT- AgNPs were taken up by erythrocytes. The erythrocyte susceptibility to lipid peroxidation measured by MDA was significantly increased in both PVP-and CT- AgNPs. The concentration of GSH and CAT activity were significantly decreased by both types of AgNPs. Additionally, PVP- and CT- AgNPs significantly increased intracellular Ca²⁺ in a dose -dependent manner. Likewise, the concentration of the cellular protein annexin V was significantly and dose - dependently enhanced by both types of AgNPs. Furthermore, PVP- and CT- AgNPs induced significant increase in calpain activity in incubated erythrocytes. Conclusion: We conclude that both PVP- and CT- AgNPs causes hemolysis, and are taken up by erythrocytes. Moreover, we demonstrated that AgNPs induces oxidative stress and eryptosis. These findings provide evidence for the potential pathophysiological effect of PVP-and CT- AgNPs on erythrocyte physiology.

Bactericidal and Cytotoxic Properties of Silver Nanoparticles

“…Figure 22d showed the internalized AgNPs in RBCs, leading to membrane injury, lipid peroxidation, and eventual hemolysis. Very recently, Ferdous et al studied the interactions of PVP and citrate coated AgNPs (10 nm) of various concentrations (2.5, 10, 40 µg/mL) with murine RBCs [214]. AgNPs induced significant dose-dependent hemolysis, resulting from cellular uptake of AgNPs and oxidative stress generation.…”

Health Impact of Silver Nanoparticles: A Review of the Biodistribution and Toxicity Following Various Routes of Exposure

“…In vitro cytotoxicity studies are often used to characterize the biological response to AgNPs, and the results of these studies may be used to identify hazards associated with exposure to AgNPs. Some important studies that have shown the toxic effects of AgNPs on different cell lines, including macrophages (RAW 264.7), including bronchial epithelial cells (BEAS-2B), alveolar epithelial cells (A549), hepatocytes (C3A, HepG2), colon cells (Caco2), skin keratinocytes (HaCaT), human epidermal keratinocytes (HEKs), erythrocytes, neuroblastoma cells, embryonic kidney cells (HEK293T), porcine kidney cells (Pk 15), monocytic cells (THP-1), and stem cells [20,[108][109][110][111][112][113][114][115][116][117], are discussed below.…”

“…Studies that used human erythrocytes have investigated the effects of AgNPs on hemolysis, morphology, and their uptake [132,134]. In our recent in vitro study, we assessed the effect of the coating and dose of AgNPs on oxidative damage and eryptosis on mice erythrocytes [111]. Both PVP and CT-AgNPs induced oxidative stress and increased cytosolic calcium, annexin V binding, and calpain activity.…”

“…These discrepancies in results, though not fully understood, could be related to the different cell types used in studies. The various blood biological effects of AgNPs, such as hemolysis, the interference of plasma coagulation, the enhancement of platelet aggregation, and the inhibition of lymphocyte proliferation, are size-, coatingand concentration-dependent [111,137,138]. Lin et al [139] studied the potential toxicity of AgNPs on cardiac electrophysiology.…”

Biological and Catalytic Activities of Green Synthesized Silver Nanoparticles from the Leaf Infusion of Dracocephalum kotschyi Boiss

“…[ 9 ] The characteristic properties of AgNPs, including size, surface area, coating, surface chemistry, and charge of the NPs, play a significant role in their toxicity and hemolytic activity. [ 7,9–11 ]…”