Potent antiviral effect of silver nanoparticles on SARS-CoV-2

Sundararaj S Jeremiah, Kei Miyakawa, Takeshi Morita, Yutaro Yamaoka, Akihide Ryo, Sundararaj S Jeremiah, Kei Miyakawa, Takeshi Morita, Yutaro Yamaoka, Akihide Ryo

Abstract

The pandemic of COVID-19 is spreading unchecked due to the lack of effective antiviral measures. Silver nanoparticles (AgNP) have been studied to possess antiviral properties and are presumed to inhibit SARS-CoV-2. Due to the need for an effective agent against SARS-CoV-2, we evaluated the antiviral effect of AgNPs. We evaluated a plethora of AgNPs of different sizes and concentration and observed that particles of diameter around 10 nm were effective in inhibiting extracellular SARS-CoV-2 at concentrations ranging between 1 and 10 ppm while cytotoxic effect was observed at concentrations of 20 ppm and above. Luciferase-based pseudovirus entry assay revealed that AgNPs potently inhibited viral entry step via disrupting viral integrity. These results indicate that AgNPs are highly potent microbicides against SARS-CoV-2 but should be used with caution due to their cytotoxic effects and their potential to derange environmental ecosystems when improperly disposed.

Keywords: COVID-19; Colloidal silver; SARS-CoV-2; Silver nanoparticles.

Conflict of interest statement

Declaration of competing interest The authors have no conflicts of interest directly relevant to the content of this article. Y.Y. is a current employee of Kanto Chemical Co., Inc.

Copyright © 2020 Elsevier Inc. All rights reserved.

Figures

Fig. 1
Fig. 1
Cytotoxicity of colloidal Silver on mammalian cells 1A: Cytotoxicity exhibited by serial concentrations of colloidal silver on VeroE6/TMPRSS2 cells. 1B: Cytotoxicity exhibited by serial concentrations of colloidal silver on Calu-3 cells.
Fig. 2
Fig. 2
Concentration and dose dependent antiviral effect of naked Silver nanoparticles on SARS-CoV-2.2A: Colloidal Silver rescues VeroE6/TMPRSS2 cells from SARS-CoV-2 mediated cell death in a concentration dependent manner. Error bars obtained from triplicate testing. p value ≤ 0.005 (∗∗∗). 2B: Concentration dependent inhibition of SARS-CoV-2 replication in Calu-3 cells by colloidal Silver. Error bars obtained from triplicate testing. p value ≤ 0.001 (∗∗∗). 2C: Silver nanoparticles exhibit size-dependent antiviral action against SARS-CoV-2 in Vero/TMPRSS2 cells. Error bars obtained from triplicate testing. p value ≤ 0.005 (∗∗∗). 2D: Size-dependent viral inhibition of SARS-CoV-2 by Silver nanoparticles in Calu-3 cells. Error bars obtained from triplicate testing. p value ≤ 0.001 (∗∗∗).
Fig. 3
Fig. 3
Silver nanoparticles effectively inhibit extracellular SARS-CoV-2.3A: Schematic representation of virus pre-treatment assay (top panel), cell post-treatment assay (central panel) and cell pre-treatment assay (bottom panel). 3B: Performance of PVP coated 10 nm Silver nanoparticles in the three study designs with respect to rescue of cells from SARS-CoV-2 infection. Error bars obtained from triplicate testing. p value ≤ 0.005 (∗∗∗). 2C: Performance of PVP coated 10 nm Silver nanoparticles in the three study designs with respect to reduction of SARS-CoV-2 replication. Error bars obtained from triplicate testing. p value ≤ 0.001 (∗∗∗).
Fig. 4
Fig. 4
Characteristics of PVP coated 10 nm Silver nanoparticles in SARS-CoV-2 infection. 4A: Immunofluorescence imaging comparing the effect of 10 nm and 100 nm Silver nanoparticles against SARS-CoV-2 infection in VeroE6/TMPRSS2 cells. Cell nuclei (blue) and SARS-CoV-2 nucleocapsid protein in cytoplasm (red). NC - Negative control. 4B: PVP coated 10 nm Silver nanoparticles protect VeroE6/TMPRSS2 cells from SARS-CoV-2 infection mediated cell death. Crystal violet staining reveals partial protection with visible plaques (red arrowheads) and complete protection with absence of plaques (black arrowheads). 4C: Pseudovirus entry assay. PVP coated 10 nm Silver nanoparticles inhibit entry of pseudovirus in VeroE6/TMPRSS2 cells. NC - Negative control, nAb - Neutralizing antibody. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

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