INTRODUCTION:

Nanotechnology is an advanced field that allows for the precise manipulation and examination of structures and devices ranging from 1 to 100 nanometers in size, with remarkable accuracy and efficiency^1,55.

Nanomaterials are not simple molecules and their surface effects differ from those of micromaterials or bulk materials for three key reasons. Firstly, nanomaterials that are dispersed have a very large surface area and high particle number per mass unit.

Secondly, there is an increase in the percentage of atoms present on the surface of nanomaterials. And thirdly, there is a decrease in the number of direct neighbours of atoms present on the surface of nanomaterials².

Silver nanoparticles (AgNPs) are widely used in various fields such as medicine, food, healthcare, consumer goods, and industry⁵⁶, due to their unique physical and chemical properties. AgNPs have many applications, including as antibacterial agents in industrial, household, and healthcare products. They are also used in consumer goods, medical device coatings, optical sensors, cosmetics, pharmaceuticals, and the food industry. In healthcare, AgNPs are used for diagnostics, orthopaedics, and drug delivery and have potential as anticancer agents. Notably, they have been observed to augment the efficacy of anticancer drugs, enhancing their tumour-killing effects³.

There are various methods to produce nanoparticles, but the most common way is through chemical processes. However, some of these chemical methods require the use of toxic compounds, which can be harmful. Noble metal nanoparticles such as gold, silver, and platinum are commonly used in situations where people may come into contact with them. Alternatively, there are biological techniques that use microorganisms to synthesise nanoparticles⁴.

CLASSIFICATION OF NANOPARTICLES:

Nanoparticles fall into three main categories based on their composition: organic, carbon-based, and inorganic².

Organic based NPs:

This group comprises nanoparticles made from proteins, carbohydrates, lipids, polymers, or similar organic compounds. Examples include dendrimers, liposomes, micelles, and protein complexes like ferritin². Generally, these nanoparticles exhibit characteristics such as non-toxicity, biodegradability, and, in cases like liposomes, they may have a hollow core. They are also sensitive to thermal and electromagnetic radiation, such as heat and light⁵.

Carbon based NPs:

Nanoparticles that are made entirely of carbon are called carbon-based nanoparticles⁵. Some well-known examples of this type of nanoparticles are fullerenes, carbon black NPs, and carbon quantum dots. Fullerenes are special carbon molecules that have a symmetrical closed-cage structure. For example, C₆₀ fullerenes consist of 60 carbon atoms that are arranged like a soccer ball. Moreover, there are other types of fullerenes, such as C₇₀ and C₅₄₀, that have also been discovered².

Inorganic Based NPS:

Inorganic nanoparticles are particles that do not contain carbon. Metal and metal oxide-based nanoparticles fall under the category of inorganic nanoparticles. Metal nanoparticles are derived solely from metal precursors and can take the form of monometallic, bimetallic, or polymetallic structures. Their unique optical and electrical properties stem from their localised surface plasmon resonance characteristics. Additionally, specific metal nanoparticles exhibit distinct thermal, magnetic, and biological attributes².

TYPES OF SYNTHESIS:

Two main approaches exist for synthesising nanoparticles: bottom-up and top-down.

1. Bottom-up:

The reverse approach of creating nanoparticles from simpler substances is known as the building-up approach. This can be achieved using techniques such as sedimentation and reduction, biochemical synthesis, sol-gel, green synthesis, and spinning. A recent example is the solvent-exchange method used by Needham et al. to produce limit-sized low-density lipoprotein (LDL) nanoparticles for medical cancer drug delivery. In this process, nucleation serves as the bottom-up approach followed by growth, constituting the upward progression. These LDL nanoparticles were obtained without the use of phospholipids and exhibited high hydrophobicity, which is a critical feature for drug delivery applications⁶.

2. Top-down:

This method involves breaking down larger molecules into smaller units, which are then transformed into nanoparticles. Methods such as grinding, chemical vapor deposition (CVD), physical vapor deposition (PVD), and other decomposition methods come under this category. For instance, a top-down method can be used to synthesize colloidal carbon spherical particles with precisely controlled sizes. The synthesis process involves the continuous chemical adsorption of polyoxometalates (POM) onto the carbon interfacial surface⁶.

METHOD OF PREPARATION:

1. Physical method preparation:

Using a "top-down" methodology, the physical method employs crushing, impacting, disrupting, degrading, cutting, cryogrinding, grinding, processing, and homogenization to break down bulk material into fine particulate matter. Physical methods such as refining, high-energy ball milling (HEBM), ball milling, grinding, and homogenization techniques like high process homogenization (HPH) and medium-pressure homogenization (UHPH) fragment microparticles during the milling process. While these physical techniques offer versatility in producing nanoparticles with controlled larger size, diameter, and volume, they may introduce surface defects, contamination, and entail high costs and time consumption. Although achieving nanoparticles of uniform size is feasible, the equipment setup is prohibitively expensive, with maintenance costs also posing significant challenges. These limitations have spurred interest in exploring biological sources and methods for nanoparticle manufacturing, particularly for space travel applications, given their efficiency and cost-effectiveness⁷.

2. Chemical method preparation:

Chemical reduction with a range of organic and inorganic reducing agents is the main technique used to produce silver nanoparticles. Aqueous and non-aqueous solutions are used to employ these agents, which include ascorbate, Sodium Borohydride (NaBH₄), Sodium citrate, elemental hydrogen, Tollens reagent, N, N-dimethylformamide (DMF), and Poly ethylene glycol block polymers. These agents cause silver ions (Ag⁺) to reduce to metallic silver (Ag0), which starts the process of forming metallic colloidal silver particles. These particles start as oligomeric clusters that eventually aggregate. To maintain the stability of the dispersed nanoparticles during synthesis, protective agents are essential. These agents, typically surfactants with functional groups like thiols, alcohols and acids, adsorb onto nanoparticle surfaces, preventing their agglomeration and preserving their surface properties⁸.

3. Biological method preparation:

Biological synthesis involves creating nanoparticles using plant extracts, bacteria, and fungi.

Phytonanotechnology is a rapidly growing field that shows promise for synthesising nanoparticles in an environmentally friendly, straightforward, and cost-effective manner. One of the main advantages of this technology is that it allows for scalability and the use of water as a universal solvent for reducing agents, making it a biocompatible solution. Different parts of plants such as roots, stems, seeds, and leaves can be used for nanoparticle synthesis. However, the exact mechanism involved in this process is not yet fully understood. It has been observed that organic acids, proteins, and secondary metabolites such as alkaloids, flavonoids, polysaccharides, and heterocyclic compounds play a crucial role in synthesising various types of nanoparticles.

By acting as "nanofactories," microorganisms offer a viable and economical alternative to hazardous chemicals and high energy requirements in the synthesis of nanoparticles. This approach is also environmentally friendly. Microorganisms can accumulate and detoxify heavy metals due to the presence of various reductase enzymes, which are essential in the reduction of metal salts into nanoparticles. The ability of yeast, fungi, and bacteria to synthesise nanoparticles has been the subject of much research in recent years. In the synthesis of nanoparticles, proteins, reducing cofactors, metal-resistant genes, enzymes, and organic materials all play a major role as capping agents⁹.

CHARACTERIZATION:

1. UV visible Spectroscopy:

UV-Vis spectroscopy is vital for understanding the formation of silver nanoparticles during the initial synthesis stage.

UV-Vis absorption spectroscopy is a valuable technique for characterising absorbance bands and band gaps of nanoparticles, especially those made of noble metals. This is particularly true for diffuse reflectance spectroscopy (DRS), which enables the examination of nanoparticles optical properties. Noble metals are highly coloured and show absorptions that are caused by oscillations of surface plasmons. Light waves that are restricted to the surface as a result of their interaction with free electrons in the metal are known as surface plasmons. It is noteworthy that the spectrum of metal nanoparticles lacks the surface plasmon resonance (SPR) band, which is specific to these particles¹⁰.

2. Transmission electron microscopy (TEM):

The use of transmission electron microscopy (TEM) in the characterization of nanoparticles is unquestionably essential. It creates micrographs of nanoscale materials with remarkable lateral spatial resolution by focusing an electron beam onto a thin sample, typically less than 200 nm thick. Additionally, by concentrating and containing the electron beam and examining the resulting electron diffraction pattern, TEM enables the study of the crystalline structure of particular microscopic regions within crystalline materials. This method makes it easier to examine the specific particle level characteristics of shape, size, and crystal structure. Even though it can provide a nanometer-level visual inspection of individual particles, the entire procedure—sample preparation, measurement, and analysis—can be very labor-intensive. In addition, TEM requires highly skilled workers to function properly and is associated with high acquisition and maintenance costs¹¹.

3. X-Ray Diffraction:

A common technique for analyzing atomic spacings and crystalline structures is X-ray diffraction. This technique provides information on phases, topologies, desired textures, and other structural characteristics like average grain size, crystallinity, strain, and lattice parameters. The constructive interference of a coherent X-ray beam reflected at particular angles from each pair of crystal planes within a material produces the patterns seen in X-ray diffraction. The arrangement of atoms within the crystal lattice determines the intensity of the peaks in the diffraction pattern.

An X-ray tube, a sample stage, and an X-ray detecting sensor are the three main parts of an X-ray diffractometer. Initially, a cathode ray tube emits electrons through a heated filament, which are then accelerated by voltage towards the sample target. Diffractograms are the term used to describe the output of the XRD instrument. The y-axis shows the intensity, and the x-axis shows the scanning angle. The diffractogram's peaks' width and form reveal details about antiphase boundaries, stacked faults, and crystallite size. X-ray diffraction is widely used in thin-film observations, texture analysis, and stress-strain observations in addition to crystalline structure and phase analysis¹².

4. Scanning electron microscopy:

Visualising sample surfaces is made possible by the scanning electron microscope (SEM), which picks up secondary electrons released when an electron beam interacts with the sample. Surface sensitivity and shallow beam penetration are the results of SEM's use of lower beam energies than TEM. As a result, SEM can be used to analyse the morphology of "thick" samples (>100 nm), something that TEM cannot do. In comparison to TEM, SEM is more user-friendly, quicker, and less expensive to maintain, but it also offers moderate resolution (>2–3 nm) and a lower chance of sample damage. High-resolution imaging with SEM usually requires conductive substrates, and nonconductive samples might require a thin metallic film coating. When analysing SEM images, it's critical to take sample preparation into account as it affects the size and surface structure of non-conductive nanoparticles. Furthermore, surface structure information is provided by SEM, and sample structure throughout its volume is revealed by TEM¹¹.

5. Fourier Transform Infrared (FT-IR) Spectroscopy:

The identification of chemical residues on the surface of silver nanoparticles (Ag NPs) as well as the roles that chemicals and metabolites play in the reduction and capping of Ag NPs are made possible by Fourier transform infrared spectroscopy (FTIR). This method is based on the interaction of molecular bonding with infrared electromagnetic radiation, which produces observable stretching and bending vibrations that are usually located in the 4000-400 cm^-1 region. Fastness, economy, non-destructiveness (especially with the attenuated total reflectance accessory), and excellent repeatability are the reasons why FTIR is highly regarded. It mostly offers qualitative insights, though, and has a limited sensitivity to nanoscale analysis. The role of functional groups such as amide (-CO-NH2), carbonyl (-CO), and hydroxyl (-OH) in mediating the reduction, capping, and stabilization of Ag NPs is highlighted by FTIR spectral analysis¹³.

APPLICATIONS:

AgNPs find utility across various domains such as thin films^14,57, surface coatings¹⁵, batteries¹⁶, energy harvesting¹⁷, and conductors¹⁸. However, it is their application in clinical settings that has garnered significant attention, owing to the escalating global threat of hazardous diseases and the challenge of multidrug resistance in conventional drug delivery systems. AgNPs are widely recognized for their ability to combat a broad spectrum of infections, despite being economically limited in feasibility.

When AgNPs interact with host/pathogenic cell proteins and enzymes, they cause cell death and demonstrate strong antimicrobial properties. Ag+ ions are released during their antibacterial actions, and this produces reactive oxygen species that interfere with vital protein functions and bacterial growth signalling pathways. Silver ions also covalently bind to bacterial cell surfaces, disrupting cell membranes and inhibiting essential cellular processes. Additionally, Ag+ particles interact with tyrosinaseenzymes and proteins in bacterial membranes and cytoplasm¹⁹, crucial for bacterial respiration and substance transport, further contributing to their antimicrobial efficacy²⁰. Moreover, silver particles are recognized for their effectiveness in preventing wound infections^21,54.

Functionalized AgNPs, along with those combined with antimicrobials, have demonstrated effectiveness. Recent advancements include the incorporation of AgNPs into polymer matrices^22,58, enhancing their reactivity. Numerous studies have explored their diverse biomedical properties, including insecticidal, anti larval, antibiofilm, and anticancer effects²³.

Research design would be aided and additional research into the effectiveness of AgNPs in clinical applications would be encouraged with a comprehensive review that includes tables and schematic representations to help with understanding of the many facets of AgNPs.

1. Drug Delivery:

Nanoparticles containing one or more therapeutic drugs that can bind, scatter, or absorb into polymer matrices are a common component of nanotechnology-based drug delivery systems⁵⁸. The creation of nanodrugs for diagnosis, treatment, and imaging has significantly increased in recent years. These systems are principally focused on improving the oral administration of pharmaceuticals⁵⁹, extending the half-life of injectable drugs, and improving the bioavailability of targeted tissue delivery. Most of the time, nano-drugs are used at lower dosages, which improves their pharmacological effects and lowers risks to health and adverse reactions²⁴.

2. Potential Applications of Nanoparticles:

· Antimicrobial assay:

Silver nanoparticles, or AgNPs, are naturally occurring antimicrobial agents that have the ability to inhibit a variety of microorganisms, including yeasts, Escherichia coli, and Staphylococcus aureus. In the pharmaceutical sector, they provide a number of benefits for the treatment of viral and bacterial illnesses²⁵. Their efficiency against clinical bacteria, such as extended-spectrum beta-lactamase (ESBL) and multidrug-resistant (MDR) bacteria, is especially remarkable²².

AgNPs demonstrate higher antibacterial activities compared to silver itself, primarily due to their wide surface areas and greater surface molecule divisions²⁶. Because of these properties, AgNPs are able to enter bacterial cells and cause cell death. AgNPs damage DNA and interfere with cellular processes when they enter bacterial cells. Additionally, when they attach to proteins that contain sulphur, the bacterial cell wall is broken down and protein synthesis is inhibited²⁷. AgNPs further suppress harmful microbes by encouraging the generation of reactive oxygen species (ROS), such as hydrogen peroxide²⁸.

AgNPs synthesised using the fungus Fusarium Oxysporum have shown antibacterial activity and could be incorporated into textiles to reduce the risk of Staphylococcus aureus infections in hospitals²⁹.

Size, shape, and surface charge are some of the variables that affect AgNPs' antibacterial activities. As an illustration, the most potent antibacterial properties are found in triangular silver nanoplates with large surface area-to-volume ratios and crystal structures³⁰. AgNPs with a positive charge have a greater inhibitory effect on gram-negative bacteria even though they are more resistant than gram-positive bacteria^31,52.

Table 1. Silver nanoparticles and their antimicrobial activities³².

Organisms used for AgNPs Synthesis	Pathogenic Bacteria
Fungus Alternaria sp.	Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Serratiamarcescens
Actinobacteria Streptacidiphilusdurhamensis	Pseudomonas aeruginosa, Staphylococcus aureus and Proteus mirabilis
Lysinibacillusvarians	Candida albicans and Candida glabrata
Root extract of Helicteresisora	Bacillus subtilis and Micrococcus luteus
Bacterium Ochrobactrumanhtropi	Salmonella typhi, Salmonella paratyphi, Vibrio cholera and Staphylococcus aureus
Fruit Dimocarpus Longan Lour. Peel	Escherichia coli and Staphylococcus aureus

Information about the antibacterial properties of AgNPs made from different organisms against distinct pathogenic bacteria is probably shown in Table 1 of the text. Overall, AgNPs offer promising potential in combating microbial infections, and their properties can be optimized for specific applications through careful design and synthesis.

· Anti-inflammatory:

The immune system uses inflammation as a reaction to infections and cell damage in the body. Its goals include getting rid of dangerous substances and helping the body's tissues reorganize and restore cell function³³. Different inflammatory disorders can arise as a result of this intricate process of dysregulation³⁴. Keratinocytes produce immune-regulatory substances like cytokines and interleukins during an inflammatory response³⁵. The primary immune organs release cytokines like IL-1 and IL-2, which are potential anti-inflammatory agents, to start the healing process, while the endocrine system secretes some inflammatory mediators like enzymes and antibodies³⁶.

Positive impacts on tissue regeneration and wound healing have been shown for biosynthesized gold and platinum nanoparticles³⁷. Table 2 enumerates certain silver nanoparticles (AgNPs) that exhibit anti-inflammatory properties. Plant-derived AgNPs have been observed to induce cytokine production, potentially because the plants alkaloids or flavonoids function as coating agents and add further pharmacological characteristics³⁴.

Increased nanosilver dosages have been shown to stimulate Th1 cell production and the release of inflammatory cytokines, such as IL-2 and INF-γ, which are essential for cellular immunity³⁸. In human peripheral blood mononuclear cells, AgNPs coated with alkaloids from unripe fruits of Piper nigrum exhibit increased anti-inflammatory activity³⁴. Additionally, AgNPs synthesized from various plants, such as Pteris tripartite³⁹, Acalyphaindica, Garciniamangostana³⁸, Centratherumpunctatum Cass, Rosa damascene⁴⁰, and Abutilon indicum³⁷, have exhibited anti-inflammatory activities.

Table 2. Role of bio-synthesized AgNPs in anti-inflammation³².

Organisms used for AgNPs Synthesis	Constituents as Stabilizing Agents	Production of Antiinflammatory Agents
Terminalia sp. (T. bentazoe, T. bellerica, T. mellueri and Terminaliacatapa) leaves	Polysaccharides, protein, polyphenolic and flavonoid compounds	Reactive oxygen species (ROS)
Leaf extract of PteristripartitaSw	Phenolics, flavonoids, terpenoids, tannins, proteins and glycosides	Histamine, serotonin, and prostaglandins
Plant Leucasaspera (willd.), Abutilon indicum	Terpenoids, alkaloids, flavonoids, phenol, tannins, phytosterols, carbohydrates, aromatics, aldehydes, alkenes, aromatics, alkyl halides, aliphatic amines, amines, alkyl halides	Indomethacin

· Anticancer Activity of Silver Nanoparticles:

With its ability to create new, more invasive, and targeted therapeutic approaches, nanomedicine presents a great deal of promise for the treatment of cancer⁴¹. One particularly intriguing area of research is the use of metallic nanoparticles (NPs) for cancer diagnostics and treatment⁴².

Metallic NPs offer several advantages that make them attractive for cancer therapy. First and foremost, their special qualities enable high penetration and target specificity, which makes them useful for both treatments and diagnostics⁴³. Furthermore, peptides, monoclonal antibodies, DNA/RNA, and tumour markers can all be functionalized onto metallic NPs. As a result, highly targeted approaches that can bind to cancer cell surface proteins or receptors with selectivity can be developed⁵³.

Through the application of nanobiotechnology, scientists can create therapeutic agents in nanoparticle form that target cancer cells directly, minimizing harm to healthy tissue and minimizing side effects. Furthermore, the ability to incorporate multiple functionalities into metallic NPs enables multimodal imaging and therapy, providing clinicians with valuable tools for both diagnosis and treatment⁴⁴.

All things considered, using metallic nanoparticles in cancer research is a promising way to create new, more potent anticancer treatments. The potential for substantial improvements in cancer patient outcomes while reducing the side effects of conventional treatments is present for further research and development in this area⁴⁵.

· Insecticidal Activity of AgNPs.:

Diseases and insects are major threats to commercial and food-producing crops in agriculture. Pests are defined as insects that feed on plants and can harm developing leaves, fruits, and vegetables. Chemical pesticides are widely used to control pests, but this practice can have detrimental effects on ecosystems that can affect human health⁴⁶, biodiversity loss, disrupt nitrogen fixation⁴⁷, and destroy habitats⁴⁸.

Researchers have suggested using formulations of nanopesticides to manage crop pests and diseases to address these issues. Nano-pesticides offer several advantages over traditional chemical pesticides, including increased efficacy and reduced environmental impact. In particular, biosynthesized silver and lead nanoparticles are gaining attention due to their availability⁴⁹, safety (non-carcinogenic), and diverse biological activities⁵⁰.

For example, silver nanoparticles (AgNPs) synthesized using extracts from plants like Euphorbia prostrata have shown insecticidal activity against pests such as Sitophilusoryzae, which commonly infests rice, wheat, and maize grains. These AgNPs offer a potential alternative to conventional chemical insecticides for controlling pest populations⁵¹.

Additionally, research on AgNPs and sulfur nanoparticles (S NPs) derived from diverse sources has shown their insecticidal activities against the fruit fly Drosophila melanogaster at different life stages (larval, pupal, and adult). While both types of nanoparticles exhibited insecticidal effects, AgNPs derived from olive and mulberry showed particularly high mortality rates and significantly reduced larval longevity.

Overall, the development and application of nano-pesticides, particularly those synthesized from natural sources, hold promise for sustainable pest management in agriculture. Further research into the efficacy, safety, and environmental impacts of these nano-pesticides is essential to their widespread adoption and integration into agricultural practices.

CONCLUSIONS:

Nanoparticles at the nanoscale exhibit significant positive impacts across diverse fields, including their utilisation in various applications within the food industry. Through modifications at the nanolevel, their properties and functions can be tailored to benefit food-related processes. The food packaging and processing industry has made extensive use of nanotechnology, which has led to improvements in antimicrobial paints, textiles, air purification, wastewater treatment, dentistry, plastic materials, ceramics, food preservation, and viruses and bacteria control. In agriculture, nanoparticles are employed for controlling pests, insects, larvae, and pupae through the development of nanoparticle pesticides (NPPs), nanobiopesticides (NBPs), nanofilters, nanofertilizers, and other innovations.

ACKNOWLEDGMENTS:

The authors are thankful to the Management, Principal of Sarojini Naidu Vanita Pharmacy MahaVidyalaya, Osmania University, Hyderabad, India for providing research facilities.

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Received on 08.04.2024 Revised on 10.12.2024

Accepted on 16.05.2025 Published on 06.10.2025

Available online from October 13, 2025

Asian J. Pharm. Res. 2025; 15(4):403-410.

DOI: 10.52711/2231-5691.2025.00063

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