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. 2025 Feb 18;18(1):75.
doi: 10.1186/s13104-025-07143-0.

Antiviral activity of silver nanoparticles against H1N1 influenza virus

Affiliations

Antiviral activity of silver nanoparticles against H1N1 influenza virus

Manya Seyfi et al. BMC Res Notes. .

Abstract

Background: Influenza virus is a significant cause of annual global respiratory disease and death. According to the limited availability of effective drugs and vaccines, innovative antivirals are currently being investigated as possible strategies to contain the spread of infectious agents. Among the various types of nanoparticles, silver nanoparticles (Ag-NPs) have attracted great interest due to their exceptional physicochemical properties. This study aims to investigate the antiviral activity of Ag-NPs against the influenza A virus (IAV)/H1N1.

Methods: The MTT assay was used to determine the possible cytotoxicity of the Ag-NPs. Madin-Darby canine kidney (MDCK) cells were exposed to Ag-NPs extract in conjunction with 100 cell culture infectious dose 50% (CCID50) of virus administered at time intervals during the infection process. The antiviral activity of the extract was evaluated under pre-treatment, post-treatment, and simultaneous assay. Viral titer reduction was assayed using hemagglutination (HA) and CCID50 assays. Viral RNA relative quantification by real-time Polymerase Chain Reaction approach was performed in each experimental condition.

Results: The study yielded significant findings regarding the inhibitory effects of Ag-NPs on the IAV/H1N1. Silver nanoparticles showed dose-dependent inhibition of the virus, with the strongest effect observed when administered simultaneously with the virus which the virus titer exhibited a substantial decrease from 5 Log10 in the control group to 1 Log10 in the initial samples, further reducing to 2 Log10 per milliliter at lower concentrations. Notably, Ag-NPs demonstrated a greater reduction in virus titer during the simultaneous stage, showing a statistically significant difference (P < 0.05) between the control and experimental groups). The reduction in viral titer was also evident in both pre- and post-inoculation stages, although the effects were different.

Conclusion: Silver nanoparticles possess inhibitory effects on the growth of the IAV/H1N1, with a significant reduction in virus titer. These findings suggest the potential of Ag-NPs as effective antiviral agents and highlight opportunities for further research and potential clinical applications in combating IAV (H1N1) infections.

Keywords: Anti-viral; CCID50; Hemagglutination; Influenza virus; MTT; Real-time PCR; Silver nanoparticles; Treatment.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: Ethical approval was obtained from Shiraz University of Medical Sciences (IR.SUMS.REC.1393.S6967). A consent letter was not needed due to this experiment was only cell culture-based and no human subject have participated in this research. Consent to participate: No human subjects have participated in the study. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
This curve illustrates the impact of varying concentrations of silver nanoparticles (Ag-NPs) on cell viability, assessed using the MTT assay. The data clearly demonstrate that at the concentrations tested in our study, cell viability remains high
Fig. 2
Fig. 2
a MDCK cells infected with influenza A virus/H1N1 at an MOI of 0.1 were treated in triplicate with different concentrations (0.005, 0.01, 0.05, 0.1, 0.5 mg/mL) of oseltamivir. Oseltamivir demonstrated a significant inhibitory effect on influenza A virus/H1N1 replication compared to the control group, as indicated by differences in CCID50. b Impact of oseltamivir at different concentrations (0.005, 0.01, 0.05, 0.1, 0.5 mg/mL) on the growth of influenza A/H1N1 virus following virus inoculation (MOI: 0.1) into MDCK cells. The influenza A virus (H1N1) titer decreased from 53 HAU/50 μL in the control group to zero, consistently maintained at zero in subsequent samples, demonstrating oseltamivir's inhibitory effect on virus growth. c MDCK cells infected with influenza A virus/H1N1 at an MOI of 0.1 were treated in triplicate with different concentrations (0.005, 0.01, 0.05, 0.1, 0.5 mg/mL) of oseltamivir. Real-Time PCR analysis revealed higher viral genome copies in the control group compared to the experimental group. Oseltamivir reduced influenza A virus (H1N1) titers at all tested concentrations. Statistical significance was determined at p < 0.05 for all tests
Fig. 3
Fig. 3
a Effect of silver nanoparticles one hour before virus inoculation into cell culture assessed by CCID50 assay. Ag-NPs led to a reduction in virus titers compared to the control group, with the most significant reduction observed at concentrations of 1 and 2 μg/ml. b Impact of Ag-NPs one hour before virus inoculation into cell culture assessed by HA assay. The virus titer, calculated based on HAU/50 μL, decreased significantly compared to the control at the utilized concentrations. c Impact of silver nanoparticles one hour before virus inoculation into cell culture assessed by Real-Time PCR analysis conducted in triplicate. Ag-NPs showed a notable reduction in virus titers. Statistical significance was determined at p < 0.05 for all tests
Fig. 4
Fig. 4
a Effect of silver nanoparticles one-hour post-virus inoculation into cell culture assessed by CCID50 assay. Virus titers decreased from 5 Log10 in the control group to 3.25 Log10 initially and to 4 Log10 per milliliter at lower concentrations. At this stage, Ag-NPs did not significantly reduce virus titers. b Impact of silver nanoparticles one-hour post-virus inoculation into cell culture assessed by HA assay. HA results showed no significant reduction in virus titers following Ag-NPs treatment. Specifically, virus titers changed from 298 to 85 HAU per microliter at initial nanoparticle concentrations and to 213 HAU at final concentrations. c Impact of silver nanoparticles one-hour post-virus inoculation into cell culture assessed by Real-Time PCR analysis conducted in triplicate. Ag-NPs showed minimal impact on virus titers. Statistical significance was determined at p < 0.05 for all tests
Fig. 5
Fig. 5
Effect of silver nanoparticles on H1N1 influenza virus replication across different assays. a CCID50 assay: Ag-NPs significantly reduced virus titers from 5 Log10 in the control group to 1 Log10 in initial samples and further to 2 Log10/ml at lower concentrations (P < 0.05). b HA assay: Virus titers decreased from 298 HA Units/μL in the control to 9.3 HA Units/μL initially upon treatment with Ag-NPs, showing a significant reduction compared to the control (P < 0.05). c Real-time PCR analysis showed that Ag-NPs reduced virus genome levels to undetectable levels initially in the control group and to lower levels at reduced concentrations. The control group exhibited significantly higher virus genome levels compared to the experimental groups (P < 0.05). All experiments were conducted in triplicate

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