Highlights
What are the main findings?
- Colloidal form of silver is biologically more reactive than the ionic form.
- Silver nanoparticles (AgNPs) can strongly bind to the spikes of the SARS-CoV-2 virus and prevent host cell attachment.
- Anti-viral activity of the AgNPs mainly depends on their size and structural features.
What is the implication of the main finding?
- AgNPs could be used to tackle the variants of SARS-CoV-2 virus effectively.
- Increased toxicity of AgNPs at the higher concentrations is a point of concern to be addressed.
Abstract
COVID-19 is an epizootic and life-threatening outbreak affecting millions of people globally. Coronavirus variants have emerged in different locations since their origin. Silver and its compounds, including silver nanoparticles (AgNPs), have been used in the medical field for a long period, especially in surgical treatments. The anti-microbial and anti-viral properties of silver are well documented. These properties depend on the size of the particles, concentration, precursor, method of preparation, and the presence of other benefiting compounds. Several experiments were conducted by researchers worldwide to prove the anti-bacterial and anti-viral properties of silver (Ag) and AgNPs, emphasizing that silver can be introduced to multiple organs in the human body and exhibit the expected antiviral characteristics. In this review article, use of silver nanoparticles to fight the COVID-19 pandemic according with the current information is discussed. The mechanisms involving antiviral activity and toxicity are discussed in detail. This article concludes that strong binding of AgNPs with SARS-CoV-2 virus prevents binding with the host cell, leading to the death of the virus. However, increased cytotoxic effect of the silver compounds at higher concentrations is a matter of concern.
1. Introduction
Public concerns regarding the emergence of infectious diseases are on the rise. Chronic respiratory diseases such as asthma and acute respiratory distress syndrome are long-term diseases characterized by the destruction of neutrophils or cycles of infection. The reports from 2010 show that, in India, acute respiratory diseases account for 15.9% of the total death rate in children under the age of 5 [1]. In a global scenario, 1.4 million infants (20% of mortality rate) were found to be the victims of acute lower respiratory infection (ALRI) [1,2]. In recent years, several variants of coronavirus have been imputed for the rapidly increasing severe respiratory diseases, with Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) observed in 2002 and Middle East Respiratory Syndrome coronavirus (MERS CoV) observed in 2012 leading the table. The new version of SARS coronavirus, i.e., SARS-CoV-2, turned out to be more contagious and engendered a severe respiratory disease. The coronavirus disease (COVID-19) has deeply affected the world with high mortality rates [3]. As per the reports of the World Health Organization (WHO), this COVID-19 pandemic has caused more than 6.3 million deaths in the world (as of September 2022) [4] and has emerged as the most significant global health crisis after the Spanish flu pandemic of 1918 [5]. As stated by the World Health Organization (WHO), the outbreak of SARS-CoV-2 might have been initiated between mid-November and early December 2019. SARS-CoV-2 is the ninth documented coronavirus to torment human beings. Although bats, pangolins, and civets function as hosts for other coronaviruses, epidemiological and epizootic studies have revealed that there is no clear evidence of transmission of SARS-CoV-2 by the human–animal interface. SARS-CoV-2 is found in frozen food, packaging, and cold storage, indicating that the viruses’ transmission could occur between various surfaces [6].
The main reason for the transmission of coronaviruses is respiratory droplets [6,7,8]. Excrements from the infected animals and untreated wastewater are also proven to spread the infection [9]. Some studies [10,11] have demonstrated that blood plasma of SARS-CoV-2 infected people could also transmit the disease. However, these findings are not substantial [12]. Studies have also shown that the indiscretion of touching contaminated surfaces before contacting the mouth and other associated organs is the main cause of the mass transmission of viruses in a confined region [13]. Surface contamination raises concerns regarding transmission through food contamination, bioaccumulation of viruses in shellfish, and consumption of gobbets without proper cooking, even though transmission via food and water are not yet proven [14]. In an experiment conducted on a group of students, unintended touching of the nose and other facial organs was found to be an average of 23 times in an hour, which signifies the compulsory use of masks and standard clothing as a preventative measure to stop the spread of the coronavirus [15]. The lifetime of coronavirus was found to be unalike on different surfaces and under different conditions. Coronavirus was found to survive with an average of 672 h (28 days) at a temperature around 4 °C, while at higher temperatures (viz., 30 to 40 °C), the lifetime decreases to 2–9 h [16]. It was seen that, on steel surfaces, the coronavirus can survive for a period of about 120 h (5 days), whereas on aluminum surfaces, it can last for only about 2–8 h [17,18,19]. On the other materials, such as wood, paper, and glass, the coronavirus was found to survive for an average of 96 h [16]. Surfaces/materials made of metals, alloys, and metal oxides are highly effective in deactivating the coronavirus [20,21,22]. Especially, pure nanoparticles of copper [23,24,25,26], zinc [27,28,29], magnesium [30,31], and silver [32,33,34] and the corresponding oxides have proven to be highly effective against coronavirus. This article focuses on the antiviral and antimicrobial applications of silver ions and silver nanoparticles and their potential utility in tackling the life-threatening SARS-CoV-2 virus and its variants. Analysis of the impact of silver-based materials in the various facets of COVID-19 treatment is described. The different synthetic methods of AgNPs and their advantages and disadvantages are also discussed. Furthermore, toxicity profile of the AgNPs is assessed.
2. Role of Silver (Ag) in Medicine
Metallic nanoparticles/compounds/oxides are widely recommended materials for the medical field (silver, titanium, copper, zinc oxide, and iron oxide) for disinfectants, cosmetics, and personal protective equipment [35]. Silver nanoparticles (AgNPs) possesses special characteristics of targeting viruses and can easily attach to viral surface glycoproteins. Then, AgNPs can enter into virus cells to exhibit virucidal activity by attacking the viral genome [36]. Silver (Ag) is one of the seven metals of antiquity known to ancient humans. Although it was established as a precious metal, its use in human healthcare and medicine can be found in prehistoric data. The term silver has been derived from an Anglo-Saxon word siolfur, which means shiny substance, and its symbol ‘Ag’ was acquired from the Latin word argentum and Sanskrit term argunas [37]. Early applications of silver were in disinfection and storage of water. Figure 1 portrays various applications of silver from prehistoric to the present period.
Figure 1.
Timeline of various applications of silver (Ag).
Silver has been extensively used in the medical field even before the advancements in medicine. Use of silver plates for cranioplasty [38,39], catheters [40], sutures [32,38], and silver wire for replacing fractured bones [41], and silver-coated endotracheal tubes [42] is well known. The anti-microbial property of silver is indirectly exploited in household applications and home appliances. Refrigerators and washing machines made by Samsung electronics use AgNPs, as they possess the features of fumigating and sterilizing apparel and help in the safekeeping of the machines from foreign microbes. The refrigerators manufactured by LG Electronics and Hitachi dishwashers also follow the same principles [32,38].
Various methods, including chemical, physical–biological, and hybrid techniques, can be adopted to manufacture nano- and micro-silver particles, as shown in Figure 2 by either a top-to-bottom or bottom-to-top approach. Physical methods of synthesis include plasma arcing [43], ball milling [44], pulsed laser deposition [45], spray pyrolysis [46], and lithographic techniques [47]. The chemical methods used to produce AgNPs are electro-deposition method, sol–gel process [48], chemical solution, chemical vapor deposition methods [49], hydrolysis co-precipitation, and wet chemical method [50]. Microwaves-assisted (frequency range 300 MHz to 300 GHz) nanoparticle synthesis from a blend of silver nitrate and carboxymethyl chitosan was also proven to be a good method for the efficient synthesis of silver nanoparticles [51]. Both physical and chemical methods produce high radiation and utilize stabilizing chemical agents that are harmful to human beings and the environment. Therefore, the biosynthetic route has gained much attention. AgNPs are manufactured with greater stability and precise dimension by biosynthetic routes, as they are made in a single step. In addition, this method provides an eco-friendly, non-toxic and safe route. Biosynthesis of AgNPs is normally achieved by treating the silver salts with plant extracts or microorganisms such as bacteria or fungi. The bio-reducing components of the plant extract polyphenols or proteins and enzymes of the microorganisms are found to play a key role in this bottom-to-top synthetic approach and serve as stabilizing agents.
Figure 2.
Different methods of synthesizing silver nanoparticles (AgNPs).
Although silver possesses many advantages, its toxic effects cannot be neglected. A high concentration of silver in water bodies can affect the aquatic species and cause ecological imbalance [52,53]. Silver components enter the body through absorption, inhalation, and excretion. Ingestion causes the deposition of silver in the stomach and other intestinal regions, whereas inhalation deposits silver in the respiratory tract, such as the larynx trachea, and lungs [52,53,54,55]. Silver particles can induce alveolar inflammation and disrupt the heart and vascular function. Continuous inhalation of silver particles for more than 60 s can damage the lungs and cause many diseases [56]. Excessive consumption of the silver solution leads to a bluish-gray color on the skin known as Argyria [57]. The same effect that occurs in the eye is called Argyrosis. The use of silver-based creams for the treatment of wounds and burns is found to have severe side effects, such as methemoglobinemia, hypochloremia, hyponatremia, and eschars [58]. Aqueous solutions of silver compounds are known to damage various organs, including the liver and kidney, cause infections in the respiratory tract and colons, and affect platelet, white blood cells (WBCs), and red blood cells (RBCs) concentrations. Although the high antimicrobial efficacy of silver is beneficial, the lack of selectivity toward the particular strains of microbes also makes it dangerous for essential ecological bacteria [59]. Several experimental studies have validated the toxicity profile of the AgNPs. Experiments on the shrimp Penaeus monodon showed that AgNPs exhibit high anti-bacterial properties at lower concentrations along with a high survival rate. However, at higher concentrations, the toxic effect of the AgNPs becomes predominant, leading to the death of the specimen. Similar observations are made in the case of other aquatic animals and plants [60].
Silver (Ag) is a metal with high electrical and thermal conductivity and has been exploited to manufacture various utensils, jewelry, dental alloys, and explosives for a long time [61]. However, it also has good antimicrobial properties resulting in its ability to fight various infections [62,63,64,65,66]. Pure metallic silver is inert and usually does not interact with human tissues or microorganisms. However, in the presence of aqueous media such as wound fluid moisture, silver can readily form silver ions (i.e., Ag+) and exhibit various bio-activities [33]. The Ag+ form is found to be the active ionic form of silver, while the other possible oxidation states of silver, i.e., Ag2+, Ag3+, are unstable under regular conditions and hence, Ag2+, Ag3+ are generally not considered relevant in biological applications. Silver, being a heavy metal, exhibits oligodynamic effect and is therefore used in minimal quantities in said biological applications [67]. Studies have revealed that Ag+ ions form anionic complexes with the halide ions of the form [AgX2]−, where X could be Cl, Br, etc., and can combine with species containing nitrogen or oxygen donor groups to form stable compounds with improved bioactivity [67]. Silver nanoparticles are also known to form a colloidal solution or composite materials with the suitable polymeric matrix [68,69]. Impregnation, complexation, or blending of silver with suitable materials such as activated charcoal, phosphorous, zirconium lactate, allantoin, etc., are also found to be efficient ways of administration of silver in biological systems [70,71,72]. A trace amount of silver is found in all humans, with about 0.1 µg of silver in the blood itself. A general diet supplies about 40 µg of silver per day; however, about 99 percent of it would be readily excreted from the body. Silver in the blood level can increase to 23 μg/L when exposed to silver either as a medicine or in other forms. This absorption rate can be further increased during inflammation and cell growth [32]. Figure 3 shows the different parts of the human body where deposition of silver can occur. Although the effect of deposition of silver in the human brain is not investigated in detail, its effect on other organs such as the gastrointestinal tract, respiratory tract, skin, and genitals has been well documented [73]. Due to the divergent physical, chemical, and enzymatic barriers they encounter in the different organs and tissues of the body, silver particles can undergo drastic alterations in the structural properties and hence can exhibit unique and specific functional properties. The earlier use of silver particles for medicine has been associated with its significant burn and wound healing applications. Figure 4 shows the action of silver on the wound regions. The wound healing process of bandages containing silver particles is well understood. Diffusion of fluids from the wound triggers the release of silver ions (concentration of around 10–40 ppm) from the dressing material, and the free silver ions are found to be responsible for the observed anti-microbial action [74,75].
Figure 3.
Mode of action of silver ions in the human body. Reprinted from Elsevier [73]. 2022, Karthik V Shankar.
Figure 4.
Images showing the action of silver ions in wound dressing. Reprinted with permission from MDPI [76]. 2022, Karthik V Shankar.
Some studies show that silver and mercury ions hold the prime positions of all metal ions in antimicrobial action when used at concentrations less than 1 ppm in vitro [76]. These ions are absorbed into the bacterial membranes when compounded as silver sulfhydryl, which acts as a cytoplasmic poison. It was found that restrained strains of bacteria show a five-fold lower accumulation of silver as compared to the sensitive stains and produce a lower amount of hydrogen sulfide (about 33% only) [77,78]. The resistance observed in the restrained bacteria can be ascribed to the formation of intracellular protective systems. Figure 5 depicts the microbial action of silver from various devices against bacterial cells. In the sensitive bacterial strains, the silver ions are absorbed by phosphate and other electron-donating receptors on the membrane, which will then diffuse into the cell by endocytic vacuoles and phagocytosis. These silver particles inactivate membrane-related enzymes that distort the bacterial cell envelope, disrupting their ability to absorb nutrients. This also prevents the rejection of metabolic waste from the membrane, leading to pitting, which eventually becomes dangerous to bacteria. The inactivation of enzymes also prevents the replication of RNA and DNA in the cell [78]. The silver free radicals also exhibit excellent antimicrobial activity. Due to the presence of unpaired electrons, free radicals can readily react with several amino acids of cell protein such as arginine and glutamic acid and affect their regular functioning, ultimately leading to the destruction of bacterial cells [64,79,80].
Figure 5.
Image showing the antimicrobial action of AgNPs. Reprinted with permission from MDPI [77]. 2022, Karthik V Shankar.
Nano-silver or silver nanoparticles (AgNPs) are an important form of silver with physical and chemical properties significantly different from those of normal silver particles [81]. Multiple studies have revealed that silver nanoparticles of the size range 50–100 nm exhibit a high order of anti-microbial properties [82,83]. Due to their smaller size, AgNPs can easily reach inaccessible organs and tissues and exhibit superior disinfection effects. AgNPs are found to be very effective against bacteria, fungi, leishmania, plasmodium, and neoplastic cells. AgNPs can activate the enzymes or interfere with the mitotic and meiotic stages of cell division of cancer cells and prevent their growth in various parts of the body. It is seen that in the presence of an appropriate amount of AgNPs, a vital nutrient present in acute lymphoblastic leukemia would exist in an inactive acidic form rather than in an active neutral form, which averts or slows down the growth and spreading of cancer [84]. Extensive experimental analyses conducted by Xiang et al. [85] demonstrated the potential of AgNPs against various viruses such as H1N1 Influenza A, and H3N2 Influenza. Both in vitro and in vivo results showed a significant decrease in the virus concentration and formation of lesions in the lung tissues of the test animals treated with AgNPs [86]. AgNPs were found to be useful in the treatment of genital herpes. From the detailed in vivo and in vitro studies, it was found that the AgNPs inhibit the growth and spread of the virus by boosting the virus immunological response system, which will then efficiently block the penetration of the virus into the cells and suppress inflammation [87]. Shahverdi et al. [88,89] identified that AgNPs can greatly enhance the inhibitory action of various antibiotics such as penicillin G, amoxicillin, erythromycin, clindamycin, and vancomycin against the bacteria E. coli and S. aureus. Detailed in vitro studies carried out on dengue virus (DEN-2) revealed that the mechanism of action of the prominent antiviral activity exhibited by the AgNPs originates from the effective inhibition of gene expression of the virus.
4. Conclusions
The COVID-19 outbreak that began in 2019 led to the rapid spread of the disease worldwide within a few months, resulting in an exponential death toll in most countries. Numerous variants of SARS-CoV-2 emerged during this period, including alpha, beta, gamma, and omicron, and continued with changes in the DNA sequences and mutations. The omicron variant is the current significant strain spread in various parts of the world. Several metals are identified with continuous research on metallo-medicines, and their effects on tackling diseases are established. Silver has been used in medicinal applications since ancient times. The anti-microbial properties of silver nanoparticles (AgNPs) depend on concentration, method of synthesis, size, presence of ‘helper molecules’, and various other factors. Prominent anti-viral properties of AgNPs could be utilized to tackle current variants as well as future mutants of coronavirus. Evidence collected from various experiments showed the inhibition of spikes of SARS-CoV-2 on ACE receptors when treated with AgNPs. AgNPs can also serve as a suitable drug delivery system for hydroxychloroquine and chloroquine medicines used to treat COVID-19. Administration of AgNPs via nasal or oral routes was found to help in controlling respiratory diseases. Silver-based textiles used for hospital requirements, including PPE kits and masks, are proven to limit the surface contact transmission of viruses to a great extent. Silver-based sanitizers and disinfectants have proven their high efficacy. However, high toxicity and adverse effect of AgNPs are still points of concern and should be addressed systematically. Even though extensive research and analysis are continuously being carried out with several materials for understanding their potential for tackling the pandemic, the development of superior inhibitory agents for the SARS-CoV-2 virus or its variants requires more time and effort.
Author Contributions
Conceptualization, K.V.S.; methodology, P.N.J.A. and B.S. (Bipin Sankar); software, K.V.S.; formal analysis, P.N.J.A., B.S. (Bipin Sankar) and K.V.S.; investigation, P.N.J.A., B.S. (Bipin Sankar), K.V.S. and N.V.K.; resources, P.N.J.A. and B.S. (Bipin Sankar); writing—original draft preparation, P.N.J.A., B.S. (Bipin Sankar) and K.V.S.; writing—review and editing, K.V.S., N.V.K., S.S. and B.S. (Balakrishnan Shankar); visualization, B.S. (Bipin Sankar) and P.N.J.A. supervision, N.V.K., S.S., B.S. (Balakrishnan Shankar) and K.V.S.; project administration, N.V.K., S.S., B.S. (Balakrishnan Shankar) and K.V.S.; funding acquisition, K.V.S., S.S. and N.V.K. All authors have read and agreed to the published version of the manuscript.
Funding
This research was funded by Amrita Vishwa Vidyapeetham, Amritapuri, India.
Institutional Review Board Statement
Not applicable.
Informed Consent Statement
Not applicable.
Data Availability Statement
The data presented in this study are available on request from the corresponding author.
Conflicts of Interest
The authors declare no conflict of interest.
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