Original Article

Antiviral activity of biosynthesized copper nanoparticle by Juglans regia green husk aqueous extract and Iron nanoparticle: molecular docking and in-vitro studies


Background and Objectives: The interaction between nanoparticles (NPs) and viruses is attracting interest because of the antiviral potential of NPs. This study aims to investigate the antiviral potential of NPs against Herpes simplex virus types 1 (HSV-1).
Materials and Methods: Molecular docking studies were conducted by Molegro virtual docker software. An extract of Juglans regia green husk was utilized to biosynthesize copper-oxide nanoparticles (CuNPs). The cytotoxicity of NPs was evaluated by MTT assay. Different treatment assays were conducted. Another assay was designed to employ the concentration of 300 μg/ml of CuNPs, which is the highest concentration that did not precipitate. Finally, chemically synthesized Iron oxide nanoparticles (FeNPs) were utilized to adsorb CuNPs. The antiviral effect of FeNPs was investigated, separately.
Results: Docking results confirmed that NPs could interact with the HSV-1 glycoproteins and prevent viral entry. MTT assay results illustrated that the minimum non-toxic concentration (MNTD) of CuNPs is 100 μg/ml which did not exhibit antiviral properties. Employing a noncytotoxic concentration of FeNPs (300 mg/ml) in combination with cytotoxic concentration of CuNPs (300 μg /ml), eliminated the cytotoxicity effects of CuNPs. Exposure of the virus with the combination of CuNPs and FeNPs resulted in 4.5 log10 TCID50 reductions in HSV-1. While treating HSV-1 with only FeNPs reduced the titer of virus by 3.25 log10 TCID50.
Conclusion: The results highlight that combination of CuNPs and FeNPs have antiviral activity against HSV-1. Moreover, FeNPs demonstrated antiviral properties against HSV-1 separately.

1. Gaikwad S, Ingle A, Gade A, Rai M, Falanga A, Incoronato N, et al. Antiviral activity of mycosynthesized silver nanoparticles against herpes simplex virus and human parainfluenza virus type 3. Int J Nanomedicine 2013; 8: 4303-4314.
2. Halder A, Das S, Ojha D, Chattopadhyay D, Mukherjee A. Highly monodispersed gold nanoparticles synthesis and inhibition of herpes simplex virus infections. Mater Sci Eng C Mater Biol Appl 2018; 89: 413-421.
3. Reske A, Pollara G, Krummenacher C, Chain BM, Katz DR. Understanding HSV-1 entry glycoproteins. Rev Med Virol 2007; 17: 205-515.
4. Trigilio J, Antoine TE, Paulowicz I, Mishra YK, Adelung R, Shukla D. Tin oxide nanowires suppress herpes simplex virus-1 entry and cell-to-cell membrane fusion. PLoS One 2012; 7(10): e48147.
5. Duarte LF, Farías MA, Álvarez DM, Bueno SM, Riedel CA, González PA. Herpes simplex virus type 1 infection of the central nervous system: insights into proposed interrelationships with neurodegenerative disorders. Front Cell Neurosci 2019; 13: 46.
6. Ohtsu Y, Susaki Y, Noguchi K. Absorption, distribution, metabolism, and excretion of the novel Helicase-Primase inhibitor, Amenamevir (ASP2151), in Rodents. Eur J Drug Metab Pharmacokinet 2018; 43: 693-706.
7. Pires de Mello CP, Bloom DC, Paixão IC. Herpes simplex virus type-1: replication, latency, reactivation and its antiviral targets. Antivir Ther 2016; 21: 277-286.
8. Hassan H, Bello RO, Adam SK, Alias E, Meor Mohd Affandi MMR, Shamsuddin AF, et al. Acyclovir-loaded solid lipid nanoparticles: optimization, characterization and evaluation of its Pharmacokinetic profile. Nanomaterials (Basel) 2020; 10: 1785.
9. Rai M, Deshmukh SD, Ingle AP, Gupta IR, Galdiero M, Galdiero S. Metal nanoparticles: The protective nanoshield against virus infection. Crit Rev Microbiol 2016; 42: 46-56.
10. Tavakoli A, Hashemzadeh MS. Inhibition of herpes simplex virus type 1 by copper oxide nanoparticles. J Virol Methods 2020; 275: 113688.
11. Jeyaraj M, Gurunathan S, Qasim M, Kang M-H, Kim J-H. A comprehensive review on the synthesis, characterization, and biomedical application of platinum nanoparticles. Nanomaterials (Basel) 2019; 9: 1719.
12. Azizi S, Namvar F, Mahdavi M, Ahmad MB, Mohamad R. Biosynthesis of silver nanoparticles using brown marine Macroalga, Sargassum Muticum aqueous extract. Materials (Basel) 2013; 6: 5942-5950.
13. Tavakoli A, Ataei-Pirkooh A, Mm Sadeghi G, Bokharaei-Salim F, Sahrapour P, Kiani SJ, et al. Polyethylene glycol-coated zinc oxide nanoparticle: an efficient nanoweapon to fight against herpes simplex virus type 1. Nanomedicine (Lond) 2018; 13: 2675-2690.
14. Ayadi Hassan S, Ghadam P, Abdi Ali A. One step green synthesis of Cu nanoparticles by the aqueous extract of Juglans regia green husk: assessing its physicochemical, environmental and biological activities.
Bioprocess Biosyst Eng 2022; 45: 605-618.
15. Ayadi Hassan S, Gorji V, Ghadam P. "The efficient magnetic separation of the four biogenic nanoparticles from aqueous media by the unmodified iron oxide nanoparticles". Int J Environ Sci Technol 2021; 18: 3883-3894.
16. Singh J, Kumar V, Kim K-H, Rawat M. Biogenic synthesis of copper oxide nanoparticles using plant extract and its prodigious potential for photocatalytic degradation of dyes. Environ Res 2019; 177: 108569.
17. Baram-Pinto D, Shukla S, Gedanken A, Sarid R. Inhibition of HSV-1 attachment, entry, and cell-to-cell spread by functionalized multivalent gold nanoparticles. Small 2010; 6: 1044-1050.
18. Vincent M, Duval RE, Hartemann P, Engels-Deutsch M. Contact killing and antimicrobial properties of copper. J Appl Microbiol 2018; 124: 1032-1046.
19. Aallaei M, Molaakbari E, Mostafavi P, Salarizadeh N, Maleksah RE, Afzali D. Investigation of Cu metal nanoparticles with different morphologies to inhibit SARS-CoV-2 main protease and spike glycoprotein using Molecular Docking and Dynamics Simulation. J Mol Struct 2022; 1253: 132301.
20. El-Sheekh MM, Shabaan MT, Hassan L, Morsi HH. Antiviral activity of algae biosynthesized silver and gold nanoparticles against Herps Simplex (HSV-1) virus in vitro using cell-line culture technique. Int J Environ Health Res 2022; 32: 616-627.
21. Sivaraj R, Rahman PK, Rajiv P, Salam HA, Venckatesh R. Biogenic copper oxide nanoparticles synthesis using Tabernaemontana divaricate leaf extract and its antibacterial activity against urinary tract pathogen. Spectrochim Acta A Mol Biomol Spectrosc 2014; 133: 178-181.
22. Abo-Zeid Y, Ismail NSM, McLean GR, Hamdy NM. A molecular docking study repurposes FDA approved iron oxide nanoparticles to treat and control COVID-19 infection. Eur J Pharm Sci 2020; 153: 105465.
23. Szymańska E, Orłowski P, Winnicka K, Tomaszewska E, Bąska P, Celichowski G, et al. Multifunctional Tannic Acid/Silver Nanoparticle-Based Mucoadhesive Hydrogel for Improved Local Treatment of HSV Infection: in vitro and in vivo Studies. Int J Mol Sci 2018; 19: 387.
24. Haggag EG, Elshamy AM, Rabeh MA, Gabr NM, Salem M, Youssif KA, et al. Antiviral potential of green synthesized silver nanoparticles of Lampranthus coccineus and Malephora lutea. Int J Nanomedicine 2019; 14: 6217-6229.
25. Fujimori Y, Sato T, Hayata T, Nagao T, Nakayama M, Nakayama T, et al. Novel antiviral characteristics of nanosized copper(I) iodide particles showing inactivation activity against 2009 pandemic H1N1 influenza virus. Appl Environ Microbiol 2012; 78: 951-955.
26. Shionoiri N, Sato T, Fujimori Y, Nakayama T, Nemoto M, Matsunaga T, et al. Investigation of the antiviral properties of copper iodide nanoparticles against feline calicivirus. J Biosci Bioeng 2012; 113: 580-586.
27. Choudhary S, Kumar R, Dalal U, Tomar S, Reddy SN. Green synthesis of nanometal impregnated biomass - antiviral potential. Mater Sci Eng C Mater Biol Appl 2020; 112: 110934.
28. Mohamed HEA, Afridi S, Khalil AT, Ali M, Zohra T, Salman M, et al. Bio-redox potential of Hyphaene thebaica in bio-fabrication of ultrafine maghemite phase iron oxide nanoparticles (Fe2O3 NPs) for therapeutic applications. Mater Sci Eng C Mater Biol Appl 2020; 112: 110890.
IssueVol 15 No 1 (2023) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v15i1.11930
Antiviral; Herpes simplex virus types 1; Copper-oxide; Iron oxide; Nanoparticles

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Ahmadi M, Elikaei A, Ghadam P. Antiviral activity of biosynthesized copper nanoparticle by Juglans regia green husk aqueous extract and Iron nanoparticle: molecular docking and in-vitro studies. Iran J Microbiol. 2023;15(1):138-148.