The biological synthesis, characterization, and therapeutic utility of Fusarium oxysporum silver nanoparticles

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Mohamed Sikkander, Khadeeja Yasmeen, Mohamed Haseeb. This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4649729/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The use of fungi in the safe, economical, and ecologically benign synthesis of silver nanoparticles is emerging as a major field in nanotechnology. The fungus Fusarium oxysporum is used in the current study to investigate the biological synthesis of silver nanoparticles. Since putrefying banana fruit, the assessment fungus that was derivative after PDA was inaccessible. Proceeding the foundation of morphologic traits, Fusarium oxysporum was acknowledged. The mechanism of silver nanoparticle making by the fungus Fusarium oxysporum was considered. The situation remained originate that as soon as exposed to silver ions, Fusarium oxysporum harvests silver nanoparticles. When the produced nanoparticles were examined using UV-Vis spectroscopy, the peak of the spectra was found to be at 420 nm. Silver nanoparticles were subjected to a TEM-based morphological analysis, which revealed that the particles are spherical in shape and have a diameter of between 50 and 100 nm. The TEM analysis of the fungus's response to the silver ion suggests that the protein may be in charge of stabilizing the silver nanoparticles. A large-scale biosynthesis process for "microbial nanotechnology" would benefit greatly from the speedy synthesis of silver nanoparticles. Fusarium oxysporum silver nanoparticles Characterization of nanoparticles Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Nanotechnology is a fascinating new manufacturing technique that works with atoms. It will make it possible to produce a large variety of goods at low cost and with no pollution. It will lead to the development of nanomachines, also referred to as nanodevices [ 1 ]. Nanotechnology has developed rapidly over the past ten years and is now applied in a wide range of technical domains. In particular, the most productive fields will benefit from the use of nanoparticles and other engineered nonmaterial [ 2 ]. Nanoparticles (NP) can enter the body through a variety of "ports of entry" and accumulate preferentially in specific subcellular structures, but the potential harmful effects of NP on humans and the environment are still largely unknown. Because they can behave like "traditional" ultrafine particles, manufactured nanoscale materials have the potential to pass through cell membranes and have a variety of adaptive or harmful effects, including genotoxic ones. Nanotechnology is the engineering of tiny machines that could build things inside of personal nano factories (PNs) from the ground up. It uses instruments and methods that are currently available to create highly sophisticated, fully functional products. The famous speech by Richard P. Feynman is titled There's Plenty of Room at the Bottom. In the end, mechanochemistry will allow nanotechnology to control matter at the nanoscale scale. Novel features that distinguish nanoparticles from the surrounding material typically emerge at a critical length scale of less than 100 nm [ 3 ]. A particle having one or more dimensions of the order of 100nm or less. It was making a major contribution to several contemporary trends concerning the improvement of human life. Nanotechnology is being applied in many fields, which has created a great demand for human resources [ 4 – 7 ]. Applications of nanotechnology based on DNA, RNA, and proteins are referred to as bimolecular nanotechnology; applications in medicine, such as illness diagnosis and treatment, are included in nanomedical technology [ 8 ]. Numerous physical and chemical methods have been employed thus far in the synthesis of the AgNPs [ 9 ]. However, attention has been drawn to the toxicity of the chemical agents used in the synthesis of AgNPs. It is vital to develop a green strategy for AgNP production that avoids using materials that are hazardous to the environment or public health. Compared to traditional synthetic methods, biological systems present a novel idea for the synthesis of nanomaterials [ 10 – 19 ]. It has been shown that a wide variety of microorganisms, including fungi and bacteria, are capable of synthesizing inorganic materials both inside and outside of cells. They could therefore be employed as ecologically friendly nanofactories. Placed in a concentrated solution of silver nitrate, the periplasmic space of Pseudoman stutzeri AG259, isolated from silver mines, produced silver nanoparticles with a distinct morphology and well-defined size. Ag bioreduction by Bacillus licheniformis has also been documented [ 20 ]. In general, atoms in clusters with sizes between 1 and 100 nm are referred to as nanoparticles (NP). A metal's properties are known to be influenced by its size, shape, composition, crystallinity, and structure. An important metal with many uses, such as infection prevention, medical diagnosis, and electronic catalysis, is silver nanoparticles. For example, AgNPs may be useful substrates for Surface Enhanced Raman Scattering (SERS) to study single molecules and catalysts for the oxidation of methanol to formaldehyde. AgNPs have been used to expedite wound healing due to their well-known antimicrobial and anti-inflammatory properties. According to a recent demonstration of an advancement in the biological synthesis approach, AgNPs' shape could be changed from nanospheres to nano prisms by controlling the growth kinetics of the silver-resistant bacteria Morganella psychrotolerans [ 21 ]. The same research team also demonstrated that all members of the genus Morganella were capable of producing extracellular silver nanoparticles, a finding related to the organisms' defense mechanisms against silver. It has been demonstrated that fungi secrete far more bioactive substances than bacteria, making them a better choice for the large-scale synthesis of nanoparticles [ 22 ]. However, hazardous chemicals are not required for enzymatic catalysis when creating nanoparticles via biosynthesis, and the final products have higher catalytic reactivity. There are several advantages to using biological organisms in the synthesis of metal nanoparticles, such as lower toxicity, less pollution, high monodispersed, low cost, and ease of use. Silver nanoparticles, which are produced by a variety of microorganisms, from prokaryotes to eukaryotes, have effective antimicrobial activity against E. Coli, V. cholera, P. aeruginosa, Candida spp., and powdery mildew. Apart from microbes, plant leaves and their extracts were also utilized as beneficial components in the creation of silver nanoparticles [ 23 – 25 ]. Moreover, the extracellular biosynthesis of fungi has the potential to greatly simplify downstream processing compared to that of bacteria. Fusarium oxysporum was used to demonstrate the cell-associated biosynthesis of silver, and the resulting particles ranged in size from 5 to 15 nm and were essentially quasi-spherical. This is a fascinating illustration of fungal biosynthesis. Sastry et al. report that the fungi Verticillium spp. and Fusarium oxysporum formed the corresponding metallic nanoparticles rather quickly after being exposed to gold and silver ions. There have also been several published studies on the biosynthesis of AgNPs using fungi like Fusarium acuminatum and fellutanum. Despite these encouraging results, it is still unclear where exactly fungi get their silver nanoparticles from and what exact mechanism underlies their ability to synthesize them. It has previously been documented that a variety of active substances secreted by fungi were important capping and reducing agents in the reaction [ 26 – 30 ]. Therefore, it was essential to look into novel fungal strains for AgNP synthesis based on biodiversity. More importantly, it may also facilitate understanding of the molecular mechanism underlying AgNP biosynthesis. With the use of nanocrystalline silver dressings, creams, and gels, bacterial infections in chronic wounds have been effectively reduced—just one of the many fields in which silver nanoparticles have proven instrumental. Silver nanoparticles with polyvinyl nanofiber also have good antibacterial qualities when applied as a wound dressing. Reports state that the silver nanoparticles showed enhanced cosmetic appearance, less scarring, and enhanced wound healing capacity when tested on an animal model. Medical devices impregnated with silver, such as surgical masks and implantable devices, exhibit significant antimicrobial efficacy. Environmentally acceptable antimicrobial nano paint can be produced []. Inorganic composite-based preservatives are found in a wide range of products. However, the mechanisms for using microorganisms to create metal nanoparticles are not fully understood due to the complexity of biological reactions [ 31 – 40 ]. This process of fungus-based nanoparticle formation can be understood as a nitrate reductase-based reduction process. In the presence of the metal salt solution, the fungus's secreted nitrate reductase assisted in the reduction process, lowering it from Ag⁺ to Ag⁰. One novel biological method for producing silver nanoparticles with Verticillium spp. included a two-step mechanism. The first step is to capture silver ions at the surface of the fungal cells. In the second step, the cell's enzymes reduced the silver ions. Thus, our objective is to biologically synthesize silver nanoparticles using fungus, which will subsequently be examined under a TEM and UV-Vis [ 41 – 45 ]. Materials and Methods Materials PDA (Potato Dextrose Agar), PDB (Potato Dextrose nutrient broth) and silver nitrate. All the experiments were performed by using Double distilled water. Fungus used for screening For the synthesis of silver nanoparticles two fungal species were used i.e. Aspergillus Niger, Fusarium oxysporum. The a bove fungus species were cultured and maintained in PDA (Potato Dextrose Agar) medium. Isolation of test fungus Aspergillus niger Soil sample were collected from an area of carpenter shop. The soil sample were taken from a depth of 5-10cm and kept in plastic bags until drying was performed immediately. After sampling in the laboratory. The soil samples were air dried at room temperature at 27⁰C for a week and grind it using a mortar pestle. Then soil sample were sieved with 0.5mm sieve to remove larger particles such as stone and plant debris in order to obtain a consistent soil particle size for isolation using the soil dilution technique. Isolation of test fungus Fusarium oxysporum The test fungus was isolated from decayed banana fruit in PDA (potato, dextrose, agar) and incubated at 28⁰C for a week. Individual fungal colonies were picked and further purified by sub culturing on PDA media. Identification of fungus The fungus was identified by cultural (mycelia, colony color, shape and size) and microscopic characteristics (macro and micro conidia and chlamydospores) by using Siefert’s key and Leslie’s Laboratory manual [ 46 – 50 ]. Maintenance of cultures For maintaining the culture using appropriate medium. Fungus cultures were incubating in to the PDA plates. The plates were maintained at room temperature at 27⁰C for week for further use. Synthesis of Silver nanoparticles Production of biomass To prepare the biomass for biosynthesis, the fungus culture obtained were inoculate in liquid broth for growth containing potato, dextrose and nutrient broth. The culture flasks were incubated on room temperature at 27⁰C. The biomass was harvested after 120 hours of growth. Sieving it through a plastic sieve followed by extensive washing with sterile double distilled water to remove any medium components from the biomass. Synthesis of Silver nanoparticles Typically, 10 gm of biomass (wet weight) were brought in to contact with 100 ml sterile doubled distilled water for 48 hours at 27⁰C in an Erlenmeyer flask and agitated 150 rpm. After incubation the cell filtrate was filtered by Whatman filter paper no. 1. After filtration the observed pH of the cell filtrate was 7.2. In to 80 ml of filtrate, a carefully weighed quantity of silver nitrate was added to the Erlenmeyer flask and incubated at room temperature in dark. Control containing cell free filtrate without silver nitrate was run simultaneously as standard with the experimental flask. Silver nanoparticles were concentrated by centrifugation of the reaction mixture at 11, 000 rpm. Cell free filtrate incubated with silver nitrate get change in color, was visually observed over a period of time. Silver nanoparticles (AgNPs) synthesized using biological methods, such as the fungus Fusarium oxysporum, have garnered significant attention for their medicinal properties. This biogenic synthesis method is considered eco-friendly and cost-effective compared to chemical and physical methods [ 51 – 54 ]. Antimicrobial Activity Silver nanoparticles exhibit broad-spectrum antimicrobial properties. Silver nanoparticles (AgNPs) are renowned for their broad-spectrum antimicrobial properties, which make them effective against a wide variety of microorganisms, including bacteria, fungi, and viruses. These properties have significant implications for medical applications, such as wound dressings, coatings for medical devices, and as components of disinfectants [ 55 – 60 ]. Mechanisms of Antimicrobial Action Disruption of Cell Membranes: Attachment and Penetration AgNPs can attach to the microbial cell membrane and penetrate it, leading to increased membrane permeability and eventual cell lysis. Structural Damage This interaction can cause structural changes in the membrane, including the formation of pores, which disrupts the integrity of the cell membrane and leads to cell death. Generation of Reactive Oxygen Species (ROS): Oxidative Stress AgNPs can induce the production of ROS, such as hydrogen peroxide, superoxide anions, and hydroxyl radicals. These ROS can damage cellular components, including lipids, proteins, and DNA, leading to cell death. Mitochondrial Damage The oxidative stress caused by ROS can also affect the mitochondria, impairing their function and contributing to cell death. Interaction with Cellular Proteins: Enzyme Inhibition AgNPs can interact with thiol groups in proteins and enzymes, leading to their inactivation. This inhibition can disrupt essential cellular processes, such as metabolism and replication. Protein Denaturation By binding to proteins, AgNPs can cause their denaturation, further disrupting cellular function and viability. DNA Interaction: DNA Damage AgNPs can enter the cell and interact with DNA, causing structural damage and affecting replication and transcription processes. This damage can result in mutations and cell death. Inhibition of Replication By binding to DNA, AgNPs can inhibit the replication process, preventing the cell from proliferating [ 61 – 70 ]. Antimicrobial Spectrum Bacterial Infections: Gram-positive Bacteria AgNPs are effective against Gram-positive bacteria, such as Staphylococcus aureus and Streptococcus pneumoniae. They can penetrate the thick peptidoglycan layer and disrupt cellular functions. Gram-negative Bacteria AgNPs also show efficacy against Gram-negative bacteria, such as Escherichia coli and Pseudomonas aeruginosa. Their action is facilitated by the thin peptidoglycan layer and the presence of an outer membrane. Fungal Infections: AgNPs exhibit antifungal properties against various fungal species, including Candida albicans and Aspergillus niger. They can disrupt fungal cell membranes and inhibit fungal growth. Viral Infections: Viral Inhibition AgNPs can inhibit viral replication and prevent viruses from entering host cells. They have shown efficacy against viruses such as HIV, hepatitis B, and influenza. Viral Deactivation AgNPs can bind to viral particles, deactivating them and preventing them from infecting host cells [ 71 – 74 ]. Anti-inflammatory Properties Silver nanoparticles have shown potential in reducing inflammation. Silver nanoparticles (AgNPs) have shown significant potential in reducing inflammation, a beneficial property that can be leveraged in various medical applications [ 75 – 80 ]. Mechanisms of Anti-inflammatory Action Inhibition of Pro-inflammatory Cytokines: AgNPs can reduce the production of pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). These cytokines are key mediators of the inflammatory response, and their reduction helps to alleviate inflammation. Modulation of Immune Cells: AgNPs can influence the activity of various immune cells, including macrophages, neutrophils, and lymphocytes, which play critical roles in the inflammatory process. By modulating these cells' activities, AgNPs help in reducing inflammation. Inhibition of NF-κB Pathway: The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway is a significant signaling pathway involved in the regulation of inflammation. AgNPs can inhibit the activation of the NF-κB pathway, thereby reducing the expression of inflammatory genes. Reduction of Reactive Oxygen Species (ROS): AgNPs can decrease the levels of ROS, which are chemically reactive molecules that contribute to inflammation by activating various inflammatory pathways. By reducing ROS levels, AgNPs can mitigate oxidative stress and associated inflammation. Promotion of Anti-inflammatory Cytokines: AgNPs can enhance the production of anti-inflammatory cytokines, such as interleukin-10 (IL-10). These cytokines help resolve inflammation and promote tissue repair and healing [ 81 – 89 ]. Anticancer Activity Research indicates that silver nanoparticles (AgNPs) possess significant anticancer properties. These properties arise from their ability to induce apoptosis, generate reactive oxygen species (ROS), and disrupt critical cellular functions specific to cancer cells. Mechanisms of Anticancer Action Induction of Apoptosis: Mitochondrial Pathway AgNPs can disrupt the mitochondrial membrane potential, leading to the release of cytochrome c and activation of caspases, which are enzymes crucial for apoptosis (programmed cell death). Death Receptor Pathway AgNPs can activate death receptors on the cell surface, triggering the extrinsic pathway of apoptosis. Generation of Reactive Oxygen Species (ROS): Silver nanoparticles can induce the production of ROS within cancer cells. High levels of ROS cause oxidative stress, damaging cellular components such as DNA, proteins, and lipids, leading to apoptosis or necrosis. DNA Damage: AgNPs can interact directly with the DNA of cancer cells, causing structural damage and interfering with replication and transcription processes, leading to cell cycle arrest and apoptosis. Inhibition of Cell Proliferation: AgNPs can inhibit the proliferation of cancer cells by interfering with the cell cycle, causing cell cycle arrest at various phases (G0/G1, S, G2/M), which prevents the cells from dividing and growing. Disruption of Cellular Functions: AgNPs can disrupt essential cellular functions specific to cancer cells, including signal transduction pathways, protein synthesis, and metabolic processes, leading to cell death. Anti-angiogenic Effects: AgNPs can inhibit angiogenesis, the formation of new blood vessels from existing ones, which is crucial for tumor growth and metastasis. By inhibiting angiogenesis, AgNPs can starve the tumor of nutrients and oxygen, slowing its growth and spread [ 90 – 103 ]. Wound Healing Silver nanoparticles have been extensively used in wound dressings due to their antimicrobial and anti-inflammatory properties. The antimicrobial properties of AgNPs help prevent infections in wounds, which is crucial for proper healing and reducing complications such as sepsis. By reducing inflammation, AgNPs help in alleviating pain, swelling, and redness at the wound site, creating a more conducive environment for healing. AgNPs have been shown to promote the proliferation of keratinocytes and fibroblasts, which are essential for tissue regeneration and wound closure. Studies have demonstrated that wound dressings containing AgNPs can accelerate the healing process, resulting in faster wound closure and reduced healing times. These dressings are impregnated with silver nanoparticles, providing sustained antimicrobial activity. They are available in various forms, such as gauzes, foams, and hydrocolloids. These dressings have a coating of silver nanoparticles on their surface, offering direct antimicrobial contact with the wound bed. These dressings combine silver nanoparticles with other materials, such as hydrogels or alginates, to enhance their antimicrobial and moisture-retentive properties. AgNPs are particularly beneficial in the treatment of chronic wounds, such as diabetic ulcers, pressure ulcers, and venous leg ulcers, where infection and inflammation are major concerns. Silver nanoparticle-based dressings are extensively used in the management of burn wounds to prevent infections and promote healing. AgNP dressings are used post-surgery to reduce the risk of infections and aid in faster recovery. The potential cytotoxic effects of AgNPs on human cells need to be evaluated to ensure their safe use. Studies suggest that the concentration and size of nanoparticles play a crucial role in their safety profile. Although rare, some individuals may experience allergic reactions to silver. Monitoring and addressing such reactions are important in clinical settings. The potential for microorganisms to develop resistance to silver should be monitored to ensure the long-term efficacy of silver-based dressings. Silver nanoparticles have proven to be highly effective in wound dressings due to their potent antimicrobial and anti-inflammatory properties. They help prevent infections, reduce inflammation, and promote faster healing, making them invaluable in the treatment of various types of wounds, including chronic, burn, and surgical wounds. While their benefits are well-documented, ongoing research is essential to optimize their use and ensure their safety and efficacy in clinical applications [ 104 – 105 ]. Antiviral Coatings: AgNPs can be incorporated into coatings for medical devices, surfaces, and personal protective equipment (PPE) to reduce the transmission of viruses in healthcare settings and public spaces. Topical Formulations: AgNPs can be formulated into gels, creams, or ointments for topical application to prevent or treat viral infections, particularly in the case of skin or mucosal infections. Pharmaceutical Formulations: AgNPs can be included in pharmaceutical formulations, such as oral tablets, nasal sprays, or injectable solutions, to deliver antiviral agents directly to the site of infection. Preventive Measures: AgNPs can be used in the development of preventive measures, such as antiviral masks and air filters, to reduce the spread of respiratory viruses. Safety and Efficacy Considerations Cytotoxicity : While AgNPs have demonstrated antiviral activity, their potential cytotoxic effects on human cells need to be carefully evaluated. Optimal dosing and delivery methods must be determined to minimize toxicity. Resistance Development: The potential for viruses to develop resistance to silver should be monitored. Combining AgNPs with other antiviral agents may help reduce the risk of resistance. Regulatory Approval: Comprehensive studies and clinical trials are required to gain regulatory approval for the use of AgNPs in antiviral applications. Ensuring their safety, efficacy, and quality is crucial for their successful implementation [ 105 – 112 ]. Anti-biofilm Activity Silver nanoparticles can disrupt biofilms, which are structured communities of microorganisms that are resistant to conventional antibiotics. Silver nanoparticles (AgNPs) can disrupt biofilms, which are structured communities of microorganisms that exhibit high resistance to conventional antibiotics. Biofilms are a significant problem in medical and industrial contexts because they protect microorganisms from environmental stresses, including antibiotic treatment, making infections difficult to eradicate. Mechanisms of Biofilm Disruption Penetration of Biofilm Matrix : AgNPs can penetrate the extracellular polymeric substance (EPS) matrix of biofilms, which is a protective layer composed of proteins, polysaccharides, and nucleic acids. This penetration allows AgNPs to reach and affect the microorganisms embedded within the biofilm. Disruption of Biofilm Structure: AgNPs can disrupt the structural integrity of biofilms by degrading the EPS matrix. This disruption can be due to the generation of reactive oxygen species (ROS) or direct interactions between AgNPs and matrix components, leading to the breakdown of the biofilm's physical structure. Antimicrobial Activity Against Biofilm-Embedded Cells: AgNPs exhibit strong antimicrobial properties against both planktonic (free-floating) and biofilm-embedded microorganisms. By interacting with bacterial cell membranes and internal structures, AgNPs can kill or inhibit the growth of biofilm-associated cells. Inhibition of Biofilm Formation: AgNPs can prevent the initial adhesion and aggregation of microorganisms on surfaces, thereby inhibiting the formation of biofilms. This is particularly important in preventing biofilm-related infections and contamination in medical devices and industrial systems. Synergistic Effects with Antibiotics: AgNPs can enhance the efficacy of conventional antibiotics against biofilms. They can disrupt the biofilm matrix and increase the permeability of microbial cells, allowing antibiotics to penetrate and act more effectively. Wound Dressings: AgNPs are used in wound dressings to prevent biofilm formation and promote wound healing. These dressings help manage chronic wounds and burns, where biofilms often complicate the healing process. Medical Device Coatings: Coating medical devices, such as catheters, implants, and prosthetics, with AgNPs can prevent biofilm formation and reduce the risk of device-associated infections. This is particularly important for indwelling devices that are prone to biofilm-related complications. Dental Applications: AgNPs can be incorporated into dental materials, such as composite resins, adhesives, and sealants, to prevent biofilm formation and dental plaque, thereby reducing the incidence of dental caries and periodontal diseases [ 113 – 120 ]. Characterization of silver nanoparticles UV-Visible spectroscopy The reaction mixture was subjected to UV-Vis Spectrophotometric Measurements (Model UV-1601 PC). According to this technique many molecules absorb ultraviolet or visible light. The percentage of transmittance light radiations determines when light of certain frequency passed through the samples. This spectrophotometer analyses records the intensity of absorbance or optical density (O.D) as a function of wavelength. Absorption is directly proportional to the concentration of the absorbing species (Beer’s law) [ 121 ]. Transmission Electron Microscope Analysis This study was undertaken to know the morphology and particle size distribution of silver nanoparticles. In TEM there is an electron source at the top of the microscope, one-meter-long column is attached for vacuum, allows following down the electron. Electron gun, Electron lens, specimen and image forming system are different components of the microscope used for imaging. It has resolving power of 1nm and provide 2D image of the sample. TEM micrographs of the sample were taken using the JEOL JSM 1oocx instrument [ 122 ]. Results and Discussion Fungus used for Screening : Different fungus Aspergillus niger and Fusarium oxysporum were isolated from different sources in Fig. 1 & Fig. 2 . Culture was maintained on PDA (Potato, Dextrose and Nutrient Agar) plate, and then transferred to PDB (Potato, Dextrose and Nutrient broth) through inoculation and incubate at room temperature. This medium is prepared for synthesis of silver nanoparticles. After adding chemical salt (AgNO₃) the color of the culture broth of Fusarium oxysporum were changed from yellow to brown. Synthesis of silver nanoparticles Two different fungal species was used for biological synthesis of silver nanoparticles and then Fusarium oxysporum was found to be capable of synthesizing silver nanoparticles. After reduction for 2 days, Culture filtrate color changed from yellow to brown. Formation of brown is due to the Surface Plasmon Resonance property of silver nanoparticles. Surface Plasmon Resonance Aqueous silver nitrate ions were reduced during exposure to the Fusarium oxysporum cell filtrate. The color of the reaction mixture changed from yellow to brown indicates the formation of silver nanoparticles in Fig. 3 . Due to excitation of surface Plasmon vibration in metal nanoparticles, silver nanoparticles exhibit yellowish brown color in water [ 123 ]. Control shows no color changes (yellow) with aqueous silver nitrate solution when incubated at same condition. Silver nanoparticles showed dark brown color solution after 24 hours of incubation Formations of silver nanoparticles were characterized by UV-Visible spectroscopy and this technique has proved to be very useful for the analysis of nanoparticles. UV-Vis analysis Stability of synthesized nanoparticles was monitored regularly for about three months. It was observed that the nanoparticles solution was extremely stable at room temperature. This indicated that the nanoparticles were well dispersed in the solution without aggregation. Figure 4 , shows that strong surface Plasmon Resonance centered at 420nm, which indicates the formation of silver nanoparticles. Transmission electron microscopy studies- TEM analyzed the silver nanoparticles coated on carbon coated copper TEM grid in Fig. 5 . This micrograph showed that they are well-disperse and size ranging from 50-100nm. The morphology of nanoparticles is essentially spherical. Conclusions Silver nanoparticles synthesized from Fusarium oxysporum possess a range of medicinal properties, including antimicrobial, anti-inflammatory, anticancer, wound healing, antiviral, and anti-biofilm activities. These properties make them promising candidates for various medical applications, such as in antimicrobial coatings, drug delivery systems, wound dressings, and therapeutic agents. Further research and clinical trials are necessary to fully understand their mechanisms and to develop safe and effective medical products based on these nanoparticles. On combining all optimized conditions, ecofriendly and inexpensive method has been developed for the rapid and large-scale synthesis of Silver Nanoparticles. In this current work nanoparticles synthesized biologically using fungus Fusarium Oxysporum , which is a pure green chemistry as well as completely toxic free compared to chemical synthesis methods. The Surface Plasmon Resonance (SPR) property of synthesized nanoparticles was studied by UV-Vis spectroscopy and the peak of the spectra was found to be at 420nm. The morphological study of silver nanoparticles using TEM suggests that the nanoparticles are spherical in shape with a diameter around 50-100nm.However, development of simple and eco-friendly synthetic route would help promoting further interest in the synthesis and application of metallic nanoparticles. In this respect, nature has provided exciting possibilities of utilizing biological systems for this purpose. This comes from the fact that micro-organisms while interacting with metal ions have shown to reduce the ions into metallic particles. Thus, fungi have shown ability to reduce metal ions to form metallic nanoparticles. Silver nanoparticles exhibit significant antiviral properties against various viruses, including HIV, hepatitis B, and influenza. Their mechanisms of action include direct viral inactivation, inhibition of viral entry and replication, generation of ROS, and modulation of the host immune response. The potential applications of AgNPs in antiviral coatings, topical formulations, pharmaceutical products, and preventive measures highlight their versatility and promise as antiviral agents. However, further research and clinical trials are essential to ensure their safety, efficacy, and regulatory compliance for widespread use in antiviral therapy and prevention. Silver nanoparticles have demonstrated a strong ability to disrupt biofilms, making them valuable in both medical and industrial applications. Their mechanisms of action include penetration and disruption of the biofilm matrix, direct antimicrobial activity against biofilm-embedded cells, and inhibition of biofilm formation. These properties enable their use in wound dressings, medical device coatings, dental materials, water treatment systems, food processing, and industrial equipment. However, ensuring the safe and effective use of AgNPs requires thorough evaluation of their toxicity, regulatory approval, and environmental impact. Declarations Funding: The authors received no financial support for the research, authorship, and/or publication of this article. Author Contribution: Dr Mohamed Haseeb has provided this paper problem statement. Dr.A. Mohamed Sikkander & Dr. Khadeeja Yasmeen have done, consolidated and framed this research data appropriately. Conflict of Interest: The authors declare that there are no conflicts of interest. Acknowledgement: We would like to acknowledge and give our warmest thanks to Dr.P. Manisankar , Former Vice Chancellor, Bharathidasan University, Trichy, and Tamil Nadu, INDIA who made this work possible. His guidance and advice carried us through all the stages of writing our paper. We would also like to thank coauthors for letting our defense be an enjoyable moment, and for your brilliant comments and suggestions. Finally, we would like to thank God, for letting us through all the difficulties. We have experienced your guidance day by day. Data Availability Statement: This article does not qualify for data sharing because no datasets were created or examined for this investigation. References Malik, S.; Muhammad, K.; Waheed, Y. Nanotechnology: A Revolution in Modern Industry. Molecules 2023, 28, 661. https://doi.org/10.3390/molecules28020661 Malik S, Muhammad K, Waheed Y. 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International Journal of Molecular Sciences. 2016; 17(9):1534. https://doi.org/10.3390/ijms17091534 Gudikandula K, Vadapally P, Charya MAS. Biogenic synthesis of silver nanoparticles from white rot fungi: Their characterization and antibacterial studies. OpenNano. 2017;2:64–78. doi: 10.1016/j.onano.2017.07.002 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4649729","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":326874682,"identity":"360cdbcd-f101-4601-bc5e-4b7a57d66d9c","order_by":0,"name":"A. Mohamed Sikkander","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYHACxgcSBjZy/OwNQLaBBVFamA0sKtKMJXsOgLRIEKWFTaLizOFEgxsJIA4RWvinHX8gcbMtLUFy5vOrG34USDDwt3cn4NUicTvHwHBmm00ev3RO2c0eoMMkzpzdgN+a2zkMyZJtacWSs3PSbvAAtRhI5OLXIn87/cHhv22HEzfcPJN28w8xWgxuJxg2SAC9v+EG+7HbRNlieDvHmEECHMg5bLdlDCR4CPpF7nb68x+QqDz+7OabP0BGey8B7yMAjwGYJFY5CLA/IEX1KBgFo2AUjCAAAFy8TIIU0dG0AAAAAElFTkSuQmCC","orcid":"","institution":"Velammal Engineering College","correspondingAuthor":true,"prefix":"","firstName":"A.","middleName":"Mohamed","lastName":"Sikkander","suffix":""},{"id":326874686,"identity":"0eeaac58-38bb-4b5d-8e56-bbb36cb008d7","order_by":1,"name":"Khadeeja Yasmeen","email":"","orcid":"","institution":"North East Frontier Technical University","correspondingAuthor":false,"prefix":"","firstName":"Khadeeja","middleName":"","lastName":"Yasmeen","suffix":""},{"id":326874687,"identity":"f3f7bc37-686e-4ac7-8571-7a6443212c46","order_by":2,"name":"Mohamed Haseeb.","email":"","orcid":"","institution":"Bezmiâlem Vakıf Üniversitesi","correspondingAuthor":false,"prefix":"","firstName":"Mohamed","middleName":"","lastName":"Haseeb.","suffix":""}],"badges":[],"createdAt":"2024-06-27 15:56:58","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4649729/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4649729/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":61288641,"identity":"ae7a8ade-2ef2-48c0-a739-b05cb734e02b","added_by":"auto","created_at":"2024-07-29 06:53:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":357087,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eAspergillus niger Fungus in PDA medium\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/aa0436018cbbb452adb18408.png"},{"id":61287679,"identity":"8db5d481-3549-44f0-972e-9760da820e7e","added_by":"auto","created_at":"2024-07-29 06:45:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":336213,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFusarium oxysporum Fungus in PDA medium\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/168a5d883aff582cd220bfab.png"},{"id":61288640,"identity":"2c551880-8b93-4e27-9f47-8213c6bc701a","added_by":"auto","created_at":"2024-07-29 06:53:50","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":273496,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSynthesis of silver nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/eb8ad89a38d10e0fadb94659.png"},{"id":61287676,"identity":"7f17ce3c-e378-48ca-a624-4ae775ea5588","added_by":"auto","created_at":"2024-07-29 06:45:50","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":57880,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eUV-visible absorbance spectra obtained for silver nanoparticles synthesized by \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eFusarium oxysporum\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/08f73b33c3350833635392c1.png"},{"id":61287680,"identity":"21c6ef39-be95-4661-8cf1-38d6a41a997e","added_by":"auto","created_at":"2024-07-29 06:45:51","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":320184,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTEM image of Silver Nanoparticles\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/d0b45037f15597f281df543a.png"},{"id":62430017,"identity":"af211d7b-effa-4ea3-9c3d-cb5d5a7b55a9","added_by":"auto","created_at":"2024-08-14 06:27:03","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2887262,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4649729/v1/8b10afec-7842-4ae2-a830-e6d6fc991710.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"The biological synthesis, characterization, and therapeutic utility of Fusarium oxysporum silver nanoparticles","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNanotechnology is a fascinating new manufacturing technique that works with atoms. It will make it possible to produce a large variety of goods at low cost and with no pollution. It will lead to the development of nanomachines, also referred to as nanodevices [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Nanotechnology has developed rapidly over the past ten years and is now applied in a wide range of technical domains. In particular, the most productive fields will benefit from the use of nanoparticles and other engineered nonmaterial [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Nanoparticles (NP) can enter the body through a variety of \"ports of entry\" and accumulate preferentially in specific subcellular structures, but the potential harmful effects of NP on humans and the environment are still largely unknown. Because they can behave like \"traditional\" ultrafine particles, manufactured nanoscale materials have the potential to pass through cell membranes and have a variety of adaptive or harmful effects, including genotoxic ones. Nanotechnology is the engineering of tiny machines that could build things inside of personal nano factories (PNs) from the ground up. It uses instruments and methods that are currently available to create highly sophisticated, fully functional products. The famous speech by Richard P. Feynman is titled There's Plenty of Room at the Bottom. In the end, mechanochemistry will allow nanotechnology to control matter at the nanoscale scale. Novel features that distinguish nanoparticles from the surrounding material typically emerge at a critical length scale of less than 100 nm [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. A particle having one or more dimensions of the order of 100nm or less.\u003c/p\u003e \u003cp\u003eIt was making a major contribution to several contemporary trends concerning the improvement of human life. Nanotechnology is being applied in many fields, which has created a great demand for human resources [\u003cspan additionalcitationids=\"CR5 CR6\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Applications of nanotechnology based on DNA, RNA, and proteins are referred to as bimolecular nanotechnology; applications in medicine, such as illness diagnosis and treatment, are included in nanomedical technology [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Numerous physical and chemical methods have been employed thus far in the synthesis of the AgNPs [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. However, attention has been drawn to the toxicity of the chemical agents used in the synthesis of AgNPs. It is vital to develop a green strategy for AgNP production that avoids using materials that are hazardous to the environment or public health. Compared to traditional synthetic methods, biological systems present a novel idea for the synthesis of nanomaterials [\u003cspan additionalcitationids=\"CR11 CR12 CR13 CR14 CR15 CR16 CR17 CR18\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. It has been shown that a wide variety of microorganisms, including fungi and bacteria, are capable of synthesizing inorganic materials both inside and outside of cells. They could therefore be employed as ecologically friendly nanofactories. Placed in a concentrated solution of silver nitrate, the periplasmic space of Pseudoman stutzeri AG259, isolated from silver mines, produced silver nanoparticles with a distinct morphology and well-defined size. Ag bioreduction by Bacillus licheniformis has also been documented [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn general, atoms in clusters with sizes between 1 and 100 nm are referred to as nanoparticles (NP). A metal's properties are known to be influenced by its size, shape, composition, crystallinity, and structure. An important metal with many uses, such as infection prevention, medical diagnosis, and electronic catalysis, is silver nanoparticles. For example, AgNPs may be useful substrates for Surface Enhanced Raman Scattering (SERS) to study single molecules and catalysts for the oxidation of methanol to formaldehyde. AgNPs have been used to expedite wound healing due to their well-known antimicrobial and anti-inflammatory properties. According to a recent demonstration of an advancement in the biological synthesis approach, AgNPs' shape could be changed from nanospheres to nano prisms by controlling the growth kinetics of the silver-resistant bacteria Morganella psychrotolerans [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The same research team also demonstrated that all members of the genus Morganella were capable of producing extracellular silver nanoparticles, a finding related to the organisms' defense mechanisms against silver. It has been demonstrated that fungi secrete far more bioactive substances than bacteria, making them a better choice for the large-scale synthesis of nanoparticles [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHowever, hazardous chemicals are not required for enzymatic catalysis when creating nanoparticles via biosynthesis, and the final products have higher catalytic reactivity. There are several advantages to using biological organisms in the synthesis of metal nanoparticles, such as lower toxicity, less pollution, high monodispersed, low cost, and ease of use. Silver nanoparticles, which are produced by a variety of microorganisms, from prokaryotes to eukaryotes, have effective antimicrobial activity against E. Coli, V. cholera, P. aeruginosa, Candida spp., and powdery mildew. Apart from microbes, plant leaves and their extracts were also utilized as beneficial components in the creation of silver nanoparticles [\u003cspan additionalcitationids=\"CR24\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMoreover, the extracellular biosynthesis of fungi has the potential to greatly simplify downstream processing compared to that of bacteria. Fusarium oxysporum was used to demonstrate the cell-associated biosynthesis of silver, and the resulting particles ranged in size from 5 to 15 nm and were essentially quasi-spherical. This is a fascinating illustration of fungal biosynthesis. Sastry et al. report that the fungi Verticillium spp. and Fusarium oxysporum formed the corresponding metallic nanoparticles rather quickly after being exposed to gold and silver ions. There have also been several published studies on the biosynthesis of AgNPs using fungi like Fusarium acuminatum and fellutanum. Despite these encouraging results, it is still unclear where exactly fungi get their silver nanoparticles from and what exact mechanism underlies their ability to synthesize them. It has previously been documented that a variety of active substances secreted by fungi were important capping and reducing agents in the reaction [\u003cspan additionalcitationids=\"CR27 CR28 CR29\" citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Therefore, it was essential to look into novel fungal strains for AgNP synthesis based on biodiversity. More importantly, it may also facilitate understanding of the molecular mechanism underlying AgNP biosynthesis. With the use of nanocrystalline silver dressings, creams, and gels, bacterial infections in chronic wounds have been effectively reduced\u0026mdash;just one of the many fields in which silver nanoparticles have proven instrumental. Silver nanoparticles with polyvinyl nanofiber also have good antibacterial qualities when applied as a wound dressing. Reports state that the silver nanoparticles showed enhanced cosmetic appearance, less scarring, and enhanced wound healing capacity when tested on an animal model. Medical devices impregnated with silver, such as surgical masks and implantable devices, exhibit significant antimicrobial efficacy. Environmentally acceptable antimicrobial nano paint can be produced []. Inorganic composite-based preservatives are found in a wide range of products.\u003c/p\u003e \u003cp\u003eHowever, the mechanisms for using microorganisms to create metal nanoparticles are not fully understood due to the complexity of biological reactions [\u003cspan additionalcitationids=\"CR32 CR33 CR34 CR35 CR36 CR37 CR38 CR39\" citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. This process of fungus-based nanoparticle formation can be understood as a nitrate reductase-based reduction process. In the presence of the metal salt solution, the fungus's secreted nitrate reductase assisted in the reduction process, lowering it from Ag⁺ to Ag⁰.\u003c/p\u003e \u003cp\u003eOne novel biological method for producing silver nanoparticles with Verticillium spp. included a two-step mechanism. The first step is to capture silver ions at the surface of the fungal cells. In the second step, the cell's enzymes reduced the silver ions. Thus, our objective is to biologically synthesize silver nanoparticles using fungus, which will subsequently be examined under a TEM and UV-Vis [\u003cspan additionalcitationids=\"CR42 CR43 CR44\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMaterials\u003c/h2\u003e \u003cp\u003ePDA (Potato Dextrose Agar), PDB (Potato Dextrose nutrient broth) and silver nitrate. All the experiments were performed by using Double distilled water.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eFungus used for screening\u003c/h2\u003e \u003cp\u003eFor the synthesis of silver nanoparticles two fungal species were used i.e. Aspergillus \u003cem\u003eNiger, Fusarium oxysporum. The a\u003c/em\u003ebove fungus species were cultured and maintained in PDA (Potato Dextrose Agar) medium.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIsolation of test fungus\u003c/b\u003e \u003cb\u003eAspergillus niger\u003c/b\u003e\u003c/p\u003e \u003cp\u003eSoil sample were collected from an area of carpenter shop. The soil sample were taken from a depth of 5-10cm and kept in plastic bags until drying was performed immediately. After sampling in the laboratory. The soil samples were air dried at room temperature at 27⁰C for a week and grind it using a mortar pestle. Then soil sample were sieved with 0.5mm sieve to remove larger particles such as stone and plant debris in order to obtain a consistent soil particle size for isolation using the soil dilution technique.\u003c/p\u003e \u003cp\u003e \u003cb\u003eIsolation of test fungus\u003c/b\u003e \u003cb\u003eFusarium oxysporum\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe test fungus was isolated from decayed banana fruit in PDA (potato, dextrose, agar) and incubated at 28⁰C for a week. Individual fungal colonies were picked and further purified by sub culturing on PDA media.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eIdentification of fungus\u003c/h2\u003e \u003cp\u003eThe fungus was identified by cultural (mycelia, colony color, shape and size) and microscopic characteristics (macro and micro conidia and chlamydospores) by using Siefert\u0026rsquo;s key and Leslie\u0026rsquo;s Laboratory manual [\u003cspan additionalcitationids=\"CR47 CR48 CR49\" citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eMaintenance of cultures\u003c/h2\u003e \u003cp\u003eFor maintaining the culture using appropriate medium. Fungus cultures were incubating in to the PDA plates. The plates were maintained at room temperature at 27⁰C for week for further use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eSynthesis of Silver nanoparticles\u003c/h2\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003eProduction of biomass\u003c/h2\u003e \u003cp\u003eTo prepare the biomass for biosynthesis, the fungus culture obtained were inoculate in liquid broth for growth containing potato, dextrose and nutrient broth. The culture flasks were incubated on room temperature at 27⁰C. The biomass was harvested after 120 hours of growth. Sieving it through a plastic sieve followed by extensive washing with sterile double distilled water to remove any medium components from the biomass.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eSynthesis of Silver nanoparticles\u003c/h2\u003e \u003cp\u003eTypically, 10 gm of biomass (wet weight) were brought in to contact with 100 ml sterile doubled distilled water for 48 hours at 27⁰C in an Erlenmeyer flask and agitated 150 rpm. After incubation the cell filtrate was filtered by Whatman filter paper no. 1. After filtration the observed pH of the cell filtrate was 7.2. In to 80 ml of filtrate, a carefully weighed quantity of silver nitrate was added to the Erlenmeyer flask and incubated at room temperature in dark. Control containing cell free filtrate without silver nitrate was run simultaneously as standard with the experimental flask. Silver nanoparticles were concentrated by centrifugation of the reaction mixture at 11, 000 rpm. Cell free filtrate incubated with silver nitrate get change in color, was visually observed over a period of time. Silver nanoparticles (AgNPs) synthesized using biological methods, such as the fungus Fusarium oxysporum, have garnered significant attention for their medicinal properties. This biogenic synthesis method is considered eco-friendly and cost-effective compared to chemical and physical methods [\u003cspan additionalcitationids=\"CR52 CR53\" citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial Activity\u003c/h2\u003e \u003cp\u003eSilver nanoparticles exhibit broad-spectrum antimicrobial properties. Silver nanoparticles (AgNPs) are renowned for their broad-spectrum antimicrobial properties, which make them effective against a wide variety of microorganisms, including bacteria, fungi, and viruses. These properties have significant implications for medical applications, such as wound dressings, coatings for medical devices, and as components of disinfectants [\u003cspan additionalcitationids=\"CR56 CR57 CR58 CR59\" citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eMechanisms of Antimicrobial Action\u003c/h2\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003eDisruption of Cell Membranes:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eAttachment and Penetration\u003c/strong\u003e \u003cp\u003eAgNPs can attach to the microbial cell membrane and penetrate it, leading to increased membrane permeability and eventual cell lysis.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eStructural Damage\u003c/strong\u003e \u003cp\u003eThis interaction can cause structural changes in the membrane, including the formation of pores, which disrupts the integrity of the cell membrane and leads to cell death.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eGeneration of Reactive Oxygen Species (ROS):\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eOxidative Stress\u003c/strong\u003e \u003cp\u003eAgNPs can induce the production of ROS, such as hydrogen peroxide, superoxide anions, and hydroxyl radicals. These ROS can damage cellular components, including lipids, proteins, and DNA, leading to cell death.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eMitochondrial Damage\u003c/strong\u003e \u003cp\u003eThe oxidative stress caused by ROS can also affect the mitochondria, impairing their function and contributing to cell death.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eInteraction with Cellular Proteins:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eEnzyme Inhibition\u003c/strong\u003e \u003cp\u003eAgNPs can interact with thiol groups in proteins and enzymes, leading to their inactivation. This inhibition can disrupt essential cellular processes, such as metabolism and replication.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eProtein Denaturation\u003c/strong\u003e \u003cp\u003eBy binding to proteins, AgNPs can cause their denaturation, further disrupting cellular function and viability.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eDNA Interaction:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eDNA Damage\u003c/strong\u003e \u003cp\u003eAgNPs can enter the cell and interact with DNA, causing structural damage and affecting replication and transcription processes. This damage can result in mutations and cell death.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eInhibition of Replication\u003c/strong\u003e \u003cp\u003eBy binding to DNA, AgNPs can inhibit the replication process, preventing the cell from proliferating [\u003cspan additionalcitationids=\"CR62 CR63 CR64 CR65 CR66 CR67 CR68 CR69\" citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e].\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial Spectrum\u003c/h2\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003eBacterial Infections:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eGram-positive Bacteria\u003c/strong\u003e \u003cp\u003eAgNPs are effective against Gram-positive bacteria, such as Staphylococcus aureus and Streptococcus pneumoniae. They can penetrate the thick peptidoglycan layer and disrupt cellular functions.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eGram-negative Bacteria\u003c/strong\u003e \u003cp\u003eAgNPs also show efficacy against Gram-negative bacteria, such as Escherichia coli and Pseudomonas aeruginosa. Their action is facilitated by the thin peptidoglycan layer and the presence of an outer membrane.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eFungal Infections:\u003c/h2\u003e \u003cp\u003eAgNPs exhibit antifungal properties against various fungal species, including Candida albicans and Aspergillus niger. They can disrupt fungal cell membranes and inhibit fungal growth.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eViral Infections:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eViral Inhibition\u003c/strong\u003e \u003cp\u003eAgNPs can inhibit viral replication and prevent viruses from entering host cells. They have shown efficacy against viruses such as HIV, hepatitis B, and influenza.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eViral Deactivation\u003c/strong\u003e \u003cp\u003eAgNPs can bind to viral particles, deactivating them and preventing them from infecting host cells [\u003cspan additionalcitationids=\"CR72 CR73\" citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e].\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eAnti-inflammatory Properties\u003c/h2\u003e \u003cp\u003eSilver nanoparticles have shown potential in reducing inflammation. Silver nanoparticles (AgNPs) have shown significant potential in reducing inflammation, a beneficial property that can be leveraged in various medical applications [\u003cspan additionalcitationids=\"CR76 CR77 CR78 CR79\" citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e80\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eMechanisms of Anti-inflammatory Action\u003c/h2\u003e \u003cdiv id=\"Sec22\" class=\"Section3\"\u003e \u003ch2\u003eInhibition of Pro-inflammatory Cytokines:\u003c/h2\u003e \u003cp\u003eAgNPs can reduce the production of pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). These cytokines are key mediators of the inflammatory response, and their reduction helps to alleviate inflammation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eModulation of Immune Cells:\u003c/h2\u003e \u003cp\u003eAgNPs can influence the activity of various immune cells, including macrophages, neutrophils, and lymphocytes, which play critical roles in the inflammatory process. By modulating these cells' activities, AgNPs help in reducing inflammation.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eInhibition of NF-κB Pathway:\u003c/h2\u003e \u003cp\u003eThe nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway is a significant signaling pathway involved in the regulation of inflammation. AgNPs can inhibit the activation of the NF-κB pathway, thereby reducing the expression of inflammatory genes.\u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003eReduction of Reactive Oxygen Species (ROS):\u003c/h2\u003e \u003cp\u003eAgNPs can decrease the levels of ROS, which are chemically reactive molecules that contribute to inflammation by activating various inflammatory pathways. By reducing ROS levels, AgNPs can mitigate oxidative stress and associated inflammation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003ePromotion of Anti-inflammatory Cytokines:\u003c/h2\u003e \u003cp\u003eAgNPs can enhance the production of anti-inflammatory cytokines, such as interleukin-10 (IL-10). These cytokines help resolve inflammation and promote tissue repair and healing [\u003cspan additionalcitationids=\"CR82 CR83 CR84 CR85 CR86 CR87 CR88\" citationid=\"CR81\" class=\"CitationRef\"\u003e81\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e89\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e \u003ch2\u003eAnticancer Activity\u003c/h2\u003e \u003cp\u003eResearch indicates that silver nanoparticles (AgNPs) possess significant anticancer properties. These properties arise from their ability to induce apoptosis, generate reactive oxygen species (ROS), and disrupt critical cellular functions specific to cancer cells.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003eMechanisms of Anticancer Action\u003c/h2\u003e \u003cdiv id=\"Sec29\" class=\"Section3\"\u003e \u003ch2\u003eInduction of Apoptosis:\u003c/h2\u003e \u003cp\u003e \u003cstrong\u003eMitochondrial Pathway\u003c/strong\u003e \u003cp\u003eAgNPs can disrupt the mitochondrial membrane potential, leading to the release of cytochrome c and activation of caspases, which are enzymes crucial for apoptosis (programmed cell death).\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eDeath Receptor Pathway\u003c/strong\u003e \u003cp\u003eAgNPs can activate death receptors on the cell surface, triggering the extrinsic pathway of apoptosis.\u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eGeneration of Reactive Oxygen Species (ROS):\u003c/h3\u003e\n\u003cp\u003eSilver nanoparticles can induce the production of ROS within cancer cells. High levels of ROS cause oxidative stress, damaging cellular components such as DNA, proteins, and lipids, leading to apoptosis or necrosis.\u003c/p\u003e \u003cdiv id=\"Sec31\" class=\"Section2\"\u003e \u003ch2\u003eDNA Damage:\u003c/h2\u003e \u003cp\u003eAgNPs can interact directly with the DNA of cancer cells, causing structural damage and interfering with replication and transcription processes, leading to cell cycle arrest and apoptosis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec32\" class=\"Section2\"\u003e \u003ch2\u003eInhibition of Cell Proliferation:\u003c/h2\u003e \u003cp\u003eAgNPs can inhibit the proliferation of cancer cells by interfering with the cell cycle, causing cell cycle arrest at various phases (G0/G1, S, G2/M), which prevents the cells from dividing and growing.\u003c/p\u003e \u003cdiv id=\"Sec33\" class=\"Section3\"\u003e \u003ch2\u003eDisruption of Cellular Functions:\u003c/h2\u003e \u003cp\u003eAgNPs can disrupt essential cellular functions specific to cancer cells, including signal transduction pathways, protein synthesis, and metabolic processes, leading to cell death.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec34\" class=\"Section3\"\u003e \u003ch2\u003eAnti-angiogenic Effects:\u003c/h2\u003e \u003cp\u003eAgNPs can inhibit angiogenesis, the formation of new blood vessels from existing ones, which is crucial for tumor growth and metastasis. By inhibiting angiogenesis, AgNPs can starve the tumor of nutrients and oxygen, slowing its growth and spread [\u003cspan additionalcitationids=\"CR91 CR92 CR93 CR94 CR95 CR96 CR97 CR98 CR99 CR100 CR101 CR102\" citationid=\"CR90\" class=\"CitationRef\"\u003e90\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e103\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eWound Healing\u003c/h3\u003e\n\u003cp\u003eSilver nanoparticles have been extensively used in wound dressings due to their antimicrobial and anti-inflammatory properties. The antimicrobial properties of AgNPs help prevent infections in wounds, which is crucial for proper healing and reducing complications such as sepsis. By reducing inflammation, AgNPs help in alleviating pain, swelling, and redness at the wound site, creating a more conducive environment for healing. AgNPs have been shown to promote the proliferation of keratinocytes and fibroblasts, which are essential for tissue regeneration and wound closure. Studies have demonstrated that wound dressings containing AgNPs can accelerate the healing process, resulting in faster wound closure and reduced healing times.\u003c/p\u003e \u003cp\u003eThese dressings are impregnated with silver nanoparticles, providing sustained antimicrobial activity. They are available in various forms, such as gauzes, foams, and hydrocolloids. These dressings have a coating of silver nanoparticles on their surface, offering direct antimicrobial contact with the wound bed. These dressings combine silver nanoparticles with other materials, such as hydrogels or alginates, to enhance their antimicrobial and moisture-retentive properties. AgNPs are particularly beneficial in the treatment of chronic wounds, such as diabetic ulcers, pressure ulcers, and venous leg ulcers, where infection and inflammation are major concerns. Silver nanoparticle-based dressings are extensively used in the management of burn wounds to prevent infections and promote healing. AgNP dressings are used post-surgery to reduce the risk of infections and aid in faster recovery.\u003c/p\u003e \u003cp\u003eThe potential cytotoxic effects of AgNPs on human cells need to be evaluated to ensure their safe use. Studies suggest that the concentration and size of nanoparticles play a crucial role in their safety profile. Although rare, some individuals may experience allergic reactions to silver. Monitoring and addressing such reactions are important in clinical settings. The potential for microorganisms to develop resistance to silver should be monitored to ensure the long-term efficacy of silver-based dressings. Silver nanoparticles have proven to be highly effective in wound dressings due to their potent antimicrobial and anti-inflammatory properties. They help prevent infections, reduce inflammation, and promote faster healing, making them invaluable in the treatment of various types of wounds, including chronic, burn, and surgical wounds. While their benefits are well-documented, ongoing research is essential to optimize their use and ensure their safety and efficacy in clinical applications [\u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e104\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e105\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eAntiviral Coatings:\u003c/h3\u003e\n\u003cp\u003eAgNPs can be incorporated into coatings for medical devices, surfaces, and personal protective equipment (PPE) to reduce the transmission of viruses in healthcare settings and public spaces.\u003c/p\u003e \u003cdiv id=\"Sec37\" class=\"Section2\"\u003e \u003ch2\u003eTopical Formulations:\u003c/h2\u003e \u003cp\u003eAgNPs can be formulated into gels, creams, or ointments for topical application to prevent or treat viral infections, particularly in the case of skin or mucosal infections.\u003c/p\u003e \u003cdiv id=\"Sec38\" class=\"Section3\"\u003e \u003ch2\u003ePharmaceutical Formulations:\u003c/h2\u003e \u003cp\u003eAgNPs can be included in pharmaceutical formulations, such as oral tablets, nasal sprays, or injectable solutions, to deliver antiviral agents directly to the site of infection.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec39\" class=\"Section2\"\u003e \u003ch2\u003ePreventive Measures:\u003c/h2\u003e \u003cp\u003eAgNPs can be used in the development of preventive measures, such as antiviral masks and air filters, to reduce the spread of respiratory viruses.\u003c/p\u003e \u003cdiv id=\"Sec40\" class=\"Section3\"\u003e \u003ch2\u003eSafety and Efficacy Considerations\u003c/h2\u003e \u003cp\u003e \u003cb\u003eCytotoxicity\u003c/b\u003e:\u003c/p\u003e \u003cp\u003eWhile AgNPs have demonstrated antiviral activity, their potential cytotoxic effects on human cells need to be carefully evaluated. Optimal dosing and delivery methods must be determined to minimize toxicity.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eResistance Development:\u003c/h3\u003e\n\u003cp\u003eThe potential for viruses to develop resistance to silver should be monitored. Combining AgNPs with other antiviral agents may help reduce the risk of resistance.\u003c/p\u003e\n\u003ch3\u003eRegulatory Approval:\u003c/h3\u003e\n\u003cp\u003eComprehensive studies and clinical trials are required to gain regulatory approval for the use of AgNPs in antiviral applications. Ensuring their safety, efficacy, and quality is crucial for their successful implementation [\u003cspan additionalcitationids=\"CR106 CR107 CR108 CR109 CR110 CR111\" citationid=\"CR105\" class=\"CitationRef\"\u003e105\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e112\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eAnti-biofilm Activity\u003c/h3\u003e\n\u003cp\u003eSilver nanoparticles can disrupt biofilms, which are structured communities of microorganisms that are resistant to conventional antibiotics. Silver nanoparticles (AgNPs) can disrupt biofilms, which are structured communities of microorganisms that exhibit high resistance to conventional antibiotics. Biofilms are a significant problem in medical and industrial contexts because they protect microorganisms from environmental stresses, including antibiotic treatment, making infections difficult to eradicate.\u003c/p\u003e\n\u003ch3\u003eMechanisms of Biofilm Disruption\u003c/h3\u003e\n\u003cp\u003e \u003cb\u003ePenetration of Biofilm Matrix\u003c/b\u003e:\u003c/p\u003e \u003cp\u003eAgNPs can penetrate the extracellular polymeric substance (EPS) matrix of biofilms, which is a protective layer composed of proteins, polysaccharides, and nucleic acids. This penetration allows AgNPs to reach and affect the microorganisms embedded within the biofilm.\u003c/p\u003e\n\u003ch3\u003eDisruption of Biofilm Structure:\u003c/h3\u003e\n\u003cp\u003eAgNPs can disrupt the structural integrity of biofilms by degrading the EPS matrix. This disruption can be due to the generation of reactive oxygen species (ROS) or direct interactions between AgNPs and matrix components, leading to the breakdown of the biofilm's physical structure.\u003c/p\u003e\n\u003ch3\u003eAntimicrobial Activity Against Biofilm-Embedded Cells:\u003c/h3\u003e\n\u003cp\u003eAgNPs exhibit strong antimicrobial properties against both planktonic (free-floating) and biofilm-embedded microorganisms. By interacting with bacterial cell membranes and internal structures, AgNPs can kill or inhibit the growth of biofilm-associated cells.\u003c/p\u003e\n\u003ch3\u003eInhibition of Biofilm Formation:\u003c/h3\u003e\n\u003cp\u003eAgNPs can prevent the initial adhesion and aggregation of microorganisms on surfaces, thereby inhibiting the formation of biofilms. This is particularly important in preventing biofilm-related infections and contamination in medical devices and industrial systems.\u003c/p\u003e\n\u003ch3\u003eSynergistic Effects with Antibiotics:\u003c/h3\u003e\n\u003cp\u003eAgNPs can enhance the efficacy of conventional antibiotics against biofilms. They can disrupt the biofilm matrix and increase the permeability of microbial cells, allowing antibiotics to penetrate and act more effectively.\u003c/p\u003e\n\u003ch3\u003eWound Dressings:\u003c/h3\u003e\n\u003cp\u003eAgNPs are used in wound dressings to prevent biofilm formation and promote wound healing. These dressings help manage chronic wounds and burns, where biofilms often complicate the healing process.\u003c/p\u003e\n\u003ch3\u003eMedical Device Coatings:\u003c/h3\u003e\n\u003cp\u003eCoating medical devices, such as catheters, implants, and prosthetics, with AgNPs can prevent biofilm formation and reduce the risk of device-associated infections. This is particularly important for indwelling devices that are prone to biofilm-related complications.\u003c/p\u003e\n\u003ch3\u003eDental Applications:\u003c/h3\u003e\n\u003cp\u003eAgNPs can be incorporated into dental materials, such as composite resins, adhesives, and sealants, to prevent biofilm formation and dental plaque, thereby reducing the incidence of dental caries and periodontal diseases [\u003cspan additionalcitationids=\"CR114 CR115 CR116 CR117 CR118 CR119\" citationid=\"CR113\" class=\"CitationRef\"\u003e113\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR120\" class=\"CitationRef\"\u003e120\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eCharacterization of silver nanoparticles\u003c/h3\u003e\n\u003cp\u003e \u003cb\u003eUV-Visible spectroscopy\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe reaction mixture was subjected to UV-Vis Spectrophotometric Measurements (Model UV-1601 PC). According to this technique many molecules absorb ultraviolet or visible light. The percentage of transmittance light radiations determines when light of certain frequency passed through the samples. This spectrophotometer analyses records the intensity of absorbance or optical density (O.D) as a function of wavelength. Absorption is directly proportional to the concentration of the absorbing species (Beer\u0026rsquo;s law) [\u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e121\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eTransmission Electron Microscope Analysis\u003c/h3\u003e\n\u003cp\u003eThis study was undertaken to know the morphology and particle size distribution of silver nanoparticles. In TEM there is an electron source at the top of the microscope, one-meter-long column is attached for vacuum, allows following down the electron. Electron gun, Electron lens, specimen and image forming system are different components of the microscope used for imaging. It has resolving power of 1nm and provide 2D image of the sample. TEM micrographs of the sample were taken using the JEOL JSM 1oocx instrument [\u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e122\u003c/span\u003e].\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003e\u003cstrong\u003eFungus used for Screening\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003eDifferent fungus Aspergillus niger and Fusarium oxysporum were isolated from different sources in \u003cstrong\u003eFig.\u0026nbsp;1\u003c/strong\u003e\u0026amp; \u003cstrong\u003eFig.\u0026nbsp;2\u003c/strong\u003e. Culture was maintained on PDA (Potato, Dextrose and Nutrient Agar) plate, and then transferred to PDB (Potato, Dextrose and Nutrient broth) through inoculation and incubate at room temperature. This medium is prepared for synthesis of silver nanoparticles. After adding chemical salt (AgNO₃) the color of the culture broth of Fusarium oxysporum were changed from yellow to brown.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of silver nanoparticles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwo different fungal species was used for biological synthesis of silver nanoparticles and then \u003cem\u003eFusarium oxysporum\u003c/em\u003e was found to be capable of synthesizing silver nanoparticles. After reduction for 2 days, Culture filtrate color changed from yellow to brown. Formation of brown is due to the Surface Plasmon Resonance property of silver nanoparticles.\u003c/p\u003e\n\u003ch3\u003eSurface Plasmon Resonance\u003c/h3\u003e\n\u003cp\u003eAqueous silver nitrate ions were reduced during exposure to the \u003cem\u003eFusarium oxysporum\u003c/em\u003e cell filtrate. The color of the reaction mixture changed from yellow to brown indicates the formation of silver nanoparticles in \u003cstrong\u003eFig.\u0026nbsp;3\u003c/strong\u003e. Due to excitation of surface Plasmon vibration in metal nanoparticles, silver nanoparticles exhibit yellowish brown color in water [\u003cspan class=\"CitationRef\"\u003e123\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eControl shows no color changes (yellow) with aqueous silver nitrate solution when incubated at same condition. Silver nanoparticles showed dark brown color solution after 24 hours of incubation\u003c/p\u003e\n\u003cp\u003eFormations of silver nanoparticles were characterized by UV-Visible spectroscopy and this technique has proved to be very useful for the analysis of nanoparticles.\u003c/p\u003e\n\u003ch3\u003eUV-Vis analysis\u003c/h3\u003e\n\u003cp\u003eStability of synthesized nanoparticles was monitored regularly for about three months. It was observed that the nanoparticles solution was extremely stable at room temperature. This indicated that the nanoparticles were well dispersed in the solution without aggregation. \u003cstrong\u003eFigure\u0026nbsp;4\u003c/strong\u003e, shows that strong surface Plasmon Resonance centered at 420nm, which indicates the formation of silver nanoparticles.\u003c/p\u003e\n\u003ch3\u003eTransmission electron microscopy studies-\u003c/h3\u003e\n\u003cp\u003eTEM analyzed the silver nanoparticles coated on carbon coated copper TEM grid in \u003cstrong\u003eFig.\u0026nbsp;5\u003c/strong\u003e. This micrograph showed that they are well-disperse and size ranging from 50-100nm. The morphology of nanoparticles is essentially spherical.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eSilver nanoparticles synthesized from Fusarium oxysporum possess a range of medicinal properties, including antimicrobial, anti-inflammatory, anticancer, wound healing, antiviral, and anti-biofilm activities. These properties make them promising candidates for various medical applications, such as in antimicrobial coatings, drug delivery systems, wound dressings, and therapeutic agents. Further research and clinical trials are necessary to fully understand their mechanisms and to develop safe and effective medical products based on these nanoparticles. On combining all optimized conditions, ecofriendly and inexpensive method has been developed for the rapid and large-scale synthesis of Silver Nanoparticles. In this current work nanoparticles synthesized biologically using fungus \u003cem\u003eFusarium Oxysporum\u003c/em\u003e, which is a pure green chemistry as well as completely toxic free compared to chemical synthesis methods. The Surface Plasmon Resonance (SPR) property of synthesized nanoparticles was studied by UV-Vis spectroscopy and the peak of the spectra was found to be at 420nm. The morphological study of silver nanoparticles using TEM suggests that the nanoparticles are spherical in shape with a diameter around 50-100nm.However, development of simple and eco-friendly synthetic route would help promoting further interest in the synthesis and application of metallic nanoparticles. In this respect, nature has provided exciting possibilities of utilizing biological systems for this purpose. This comes from the fact that micro-organisms while interacting with metal ions have shown to reduce the ions into metallic particles. Thus, fungi have shown ability to reduce metal ions to form metallic nanoparticles. Silver nanoparticles exhibit significant antiviral properties against various viruses, including HIV, hepatitis B, and influenza. Their mechanisms of action include direct viral inactivation, inhibition of viral entry and replication, generation of ROS, and modulation of the host immune response. The potential applications of AgNPs in antiviral coatings, topical formulations, pharmaceutical products, and preventive measures highlight their versatility and promise as antiviral agents. However, further research and clinical trials are essential to ensure their safety, efficacy, and regulatory compliance for widespread use in antiviral therapy and prevention. Silver nanoparticles have demonstrated a strong ability to disrupt biofilms, making them valuable in both medical and industrial applications. Their mechanisms of action include penetration and disruption of the biofilm matrix, direct antimicrobial activity against biofilm-embedded cells, and inhibition of biofilm formation. These properties enable their use in wound dressings, medical device coatings, dental materials, water treatment systems, food processing, and industrial equipment. However, ensuring the safe and effective use of AgNPs requires thorough evaluation of their toxicity, regulatory approval, and environmental impact.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e The authors received no financial support for the research, authorship, and/or publication of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contribution:\u003c/strong\u003e \u003cstrong\u003eDr Mohamed Haseeb\u003c/strong\u003e has provided this paper problem statement. \u003cstrong\u003eDr.A. Mohamed Sikkander \u0026amp; Dr. Khadeeja\u003c/strong\u003e Yasmeen have done, consolidated and framed this research data appropriately.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest:\u003c/strong\u003e The authors declare that there are no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgement:\u003c/strong\u003e We would like to acknowledge and give our warmest thanks to \u003cstrong\u003eDr.P. Manisankar\u003c/strong\u003e, Former Vice Chancellor, Bharathidasan University, Trichy, and Tamil Nadu, INDIA who made this work possible. His guidance and advice carried us through all the stages of writing our paper. We would also like to thank coauthors for letting our defense be an enjoyable moment, and for your brilliant comments and suggestions. Finally, we would like to thank God, for letting us through all the difficulties. We have experienced your guidance day by day.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis article does not qualify for data sharing because no datasets were created or examined for this investigation.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eMalik, S.; Muhammad, K.; Waheed, Y. Nanotechnology: A Revolution in Modern Industry. Molecules 2023, 28, 661. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/molecules28020661\u003c/span\u003e\u003cspan address=\"10.3390/molecules28020661\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMalik S, Muhammad K, Waheed Y. Emerging Applications of Nanotechnology in Healthcare and Medicine. 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OpenNano. 2017;2:64\u0026ndash;78. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.onano.2017.07.002\u003c/span\u003e\u003cspan address=\"10.1016/j.onano.2017.07.002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Fusarium oxysporum, silver nanoparticles, Characterization of nanoparticles","lastPublishedDoi":"10.21203/rs.3.rs-4649729/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4649729/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe use of fungi in the safe, economical, and ecologically benign synthesis of silver nanoparticles is emerging as a major field in nanotechnology. The fungus Fusarium oxysporum is used in the current study to investigate the biological synthesis of silver nanoparticles. Since putrefying banana fruit, the assessment fungus that was derivative after PDA was inaccessible. Proceeding the foundation of morphologic traits, Fusarium oxysporum was acknowledged. The mechanism of silver nanoparticle making by the fungus Fusarium oxysporum was considered. The situation remained originate that as soon as exposed to silver ions, Fusarium oxysporum harvests silver nanoparticles. When the produced nanoparticles were examined using UV-Vis spectroscopy, the peak of the spectra was found to be at 420 nm. Silver nanoparticles were subjected to a TEM-based morphological analysis, which revealed that the particles are spherical in shape and have a diameter of between 50 and 100 nm. The TEM analysis of the fungus's response to the silver ion suggests that the protein may be in charge of stabilizing the silver nanoparticles. A large-scale biosynthesis process for \"microbial nanotechnology\" would benefit greatly from the speedy synthesis of silver nanoparticles.\u003c/p\u003e","manuscriptTitle":"The biological synthesis, characterization, and therapeutic utility of Fusarium oxysporum silver nanoparticles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-29 06:45:46","doi":"10.21203/rs.3.rs-4649729/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"778dd0ee-267b-4985-a544-39ea7d07c002","owner":[],"postedDate":"July 29th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-11-07T06:53:17+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-29 06:45:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4649729","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4649729","identity":"rs-4649729","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}