A novel approach for phyto-synthesis of silver nanoparticles using floral extract of Punica Granatum for potent anti-cancerous and antibacterial capabilities

A novel approach for phyto-synthesis of silver nanoparticles using floral extract of Punica Granatum for potent anti-cancerous and antibacterial capabilities | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report A novel approach for phyto-synthesis of silver nanoparticles using floral extract of Punica Granatum for potent anti-cancerous and antibacterial capabilities Sumit K Shrivastava, Asita Kulshreshtha, Rajesh K. Gangwar, Shikha Srivastava, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5054888/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 16 Oct, 2025 Read the published version in Discover Nano → Version 1 posted 14 You are reading this latest preprint version Abstract Synthesis of safest and non-toxic nanoparticles for biomedical applications in various fields of science & Technology remained a vast challenge for researchers. This work aims to create, analyze, and examine the antibacterial effects of silver nanoparticles that are synthesized utilizing the floral extract of Punica Granatum as both a stabilizing and reducing agent. Silver nitrate (AgNO3) solution was reduced using Punica Granatum flower extract which led to the synthesis of stable silver nanoparticles (PG-AgNPs) solution. The PG-AgNPs were examined and characterized using UV-Vis, FT-IR, FE-SEM, XRD, and EDX methodologies. Besides this anti-bacterial & anti-cancerous property were also studied. The results of FE-SEM analysis revealed that majority of the phytosynthesized PG-AgNPs had spherical shape and were evenly dispersed with a diameter of nearly 27.33 nm. EDX results revealed the relative presence of different elements, biomolecules, and capping agents. Additionally, synthesised silver nanoparticles had anti-cancer potential, with an IC 50 of ~ 13 µg. E. coli and S. aureus were tested for silver nanoparticle antimicrobial efficacy. Phytosynthesized PG-AgNPs against Escherichia coli and Staphylococcus aureus were very effective. A new chemotherapeutic medication or innovative supplement to treat various forms of human cancer may be developed after the comprehensive clinical trial using PG-AgNPs containing Punica Granatum flower aqueous extract. Silver Nanoparticles (Ag NPs) Punica Granatum Flower Green Synthesis Eco-friendly Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Nanotechnology heavily depends on metal nanoparticles that are synthesised via phytosynthesis. Silver and gold nanoparticles are becoming more popular due to their versatility and wide range of applications. [1–5] Researchers have created novel synthesis techniques for affordable, non-toxic, and eco-friendly metal nanoparticles. Phyto-synthesis is a well-known example. Nanotechnology has many scientific and technological uses. The famous aphorism "Necessity is the mother of invention" argues that necessity drives every new idea. As metal nanoparticle demand rises, researchers are always looking for new, easy production techniques. Researchers are employing herbs, fruits, and plants to greenly synthesise metal nanoparticles. This methodology encourages green metal nanoparticle manufacturing without hazardous ingredients. Green or phytosynthesis is a simple, non-toxic, eco-friendly synthesis process. Because many bio molecules work as reducing and capping agents, phyto-synthesis processes are cost-effective and versatile. [6–10] Due to their unique features and beneficial medicinal uses, silver and gold nanoparticles are continually drawing researchers. The reducing agent for many green metal nanoparticle manufacturing processes is plants, bacteria, and fungus. [11–20] According to earlier research, metallic nanoparticles phytosynthesized by ethno medicinal plants rich in antioxidant compounds have increased anti-bacterial and anti-cancer activities. Many cancer physicians utilise chemotherapy, immunotherapy, and radiation. Because chemotherapy is harmful, phytosynthesized metallic nanoparticles are needed to develop an effective chemotherapy medication. [21–30] An interdisciplinary and cross-sectoral area uses nanotechnology in medical diagnostics, food, medicine, biotechnology, environment, energy, chemistry, and physics. Nanotechnology is also applied in the field of biotechnology. Nanoscience and nanotechnology's multidisciplinary nature yields current diagnostics tools. [31] The European, American, and Japanese National Nanotechnology Project is prioritised in several domains. To comprehend and manage unparalleled basic components of physical things, nanotechnology and nanoscience may transfer materials precisely. These advances may impact vaccination and computer design. Nanotechnology involves creating metal nanoparticle-based medications. These uses in biology, health, and medicine have made nanoparticles popular today. Structured, they are 100 nanometers. They improve illness diagnosis and therapy extensively. Nanoparticles size allows them to pass the blood-brain barrier. Despite their modest size, nanoparticles can treat brain tumours. Nanoparticles are designed by managing the particle size, surface features, and release of a specific drug at a certain time and place. This is accomplished via the process of nanoparticle design. Disease diagnostics, tissue engineering scaffolds, and targeted medicine delivery employ nanoparticles. [32–34] Metallic nanoparticle applications research has become popular worldwide. The uses of metal nanoparticles are primarily influenced by elements such as particle size, dispersion, architecture, chemical assembly, colloidal stability, and biocompatibility. Metallic nanoparticles with medicinal herbs work well against cancer. Recent years have seen metal nanoparticles carrying herbs heal many malignancies. Recently, nanoparticle technology has advanced the manufacture of various medications, which might help cure and diagnose many illnesses, including cancer. Cancer’s high death rate causes psychological and economic difficulties. A major determinant in cancer incidence is lifestyle. The primary factors contributing to the development of cancer include environmental contaminants, carcinogens, mutagens, bacterial and viral infections, and genetic susceptibility. A combination of surgical intervention, radiation therapy, and chemotherapy is used to battle and manage cancer, with the specific treatment approach determined by the kind and stage of cancer as well as the overall health condition of the patient. Although there is a potential for negative consequences in advanced malignancies, the majority of these approaches are unsuccessful, underscoring the need for innovative cancer therapies and scientific investigation. Manufacturing materials, devices, and systems efficiently using nanometre-scale matter control and novel features and phenomena is nanotechnology. Nanoparticles are atomic or molecule assemblies with diameters under 100 nm, exhibiting unique physicochemical features relative to their bulk. Nanotechnology helped create a novel cancer approach. We've long investigated metallic nanoparticles for cancer therapy. Natural metal oxides are abundant, making nanoparticle processing and manufacturing one of the cheapest methods. Metallic nanoparticles are a new form of commonly utilised mineral particle that researchers have evaluated owing to their appropriate physical and chemical features and higher adsorption power than other metallic nanoparticle compositions. Metallic nanoparticles are biocompatible and nontoxic when used as medical fillers, cosmetics, and pharmaceutical carriers. Metallic nanoparticles possess exceptional chemical stability, a low dielectric constant, catalytic activity, the capacity to absorb infrared and ultraviolet light, and notably, antibacterial properties. The therapeutic and anticancer capabilities of these compounds might potentially enhance cancer therapy. Gangwar et al. developed a green technique for improving bioavailability using curcumin-conjugated silica nanoparticles [35]. According to Shrivastava et al., Plumeria Pudica Plant sections can synthesise silver nanoparticles. They phytosynthesized silver nanoparticles from Plumeria Pudica flowers [36]. They also phytosynthesised silver nanoparticles from Plumeria Pudica leaves and found anti-cancerous action [37]. Huang et al. used the aqueous extract of Fumaria officinalis to synthesise nickel nanoparticles, which were subsequently identified as a groundbreaking chemotherapeutic agent for the management of ovarian cancer. [38] Zhang et al. used Leucus aspera to phytosynthesize AgNPs and observed significant efficacy in inhibiting alveolar carcinoma, demonstrating promising anticancer action. [39] Approaches to the production of nanomaterials that are natural, non-hazardous economically viable, and environmentally friendly have gained prominence over physicochemical ones. There are many approaches to make nanomaterials from metal ion salts biochemically. Use an ecologically friendly solvent solution with stabilising and reducing components to synthesise nanoparticles 'green'. Nanoparticle biogenesis results from lowering metal ions of biomolecules. This method produces huge numbers of well-defined nanoparticles without contamination and reduces the environmental effect of biological synthesis. All types of plants, as well as microbes, underwater algae, plant-derived substances, tissue, and fruits, are used in the production of nanomaterials. Beginning in the 1900s, biomaterials have been able to decrease metal ions. In the last half-century, there has been an increase in curiosity in lowering agents, which has been driven by a dearth of understanding about the mechanisms involved in their production. It has been revealed by researchers that nanoparticles made from biological components possess anticancer capabilities that are unique to them. Recent developments in medicine have made use of metallic nanoparticles. Recent studies have shown that some nanoparticles possess therapeutic properties and may serve as a viable alternative to physicochemical different metal-supported nanoparticles, as well as antibacterial and anticancer medications. Pomegranate is said to have originated in Iran. The Punica granatum, or pomegranate tree, grows to five to eight metres in Iran, the northern Indian Himalayas, China, the US, and the Mediterranean regions. PG is a key Iranian plant because of its versatility. It grows in dry and semiarid regions nationwide. More than seven hundred Punica granatum (PG) varieties in Iran's Saveh and Yazd cities share size, colour, flavour, ripening duration, and disease resistance. [40] The plant's root, juice, peel, leaves, blossoms, bark, and seed of PG contain ingredients that have unique toxicological and medicinal characteristics.[41] The edible berry has a thick crimson skin, a hexagonal shape, and 600 seeds. Each seed has a water-laden pulp (aril) that may be white, deep crimson, or purple and is delicious. White, spongey, astringent pulp encases seeds. Pomegranates are twice mentioned in the Quran as God's magnificent creations and as growing in paradise's gardens. The PG fruit has been used to treat acidosis, dysentery, microbiological infections, diarrhoea, helminth infections, haemorrhage, and respiratory illnesses. PG seeds contain estrone and estradiol. [42–43] Both the dried pericarp and fruit juice are used to treat dental issues, headaches, piles, acne, colitis, menorrhagia, oxyuriasis, and diuretics. Recent scientific researchers have examined traditional PG uses. Many ancient cultures used PG juice, fruit, and extracts for medicine. PG may include phytochemicals with poisonous and pharmacological effects. The mechanisms and pharmacological and toxicological consequences of many of these substances are yet unknown. There is a growing interest in herbal remedies and medicinal plants all over the globe with the purpose of determining whether or not they are effective in preventing and treating chronic illnesses. It is possible to treat a variety of conditions with PG, such as cancers of the prostate, colon, breast, lung, and skin; leukaemia; diabetes; hyperlipidaemia; hypertension; myocardial ischaemia; myocardial perfusion; infections; erectile dysfunction; male infertility; neonatal hypoxia-ischemic brain damage; obesity; and Alzheimer's disease. [44] This research presents a basic phytosynthesis process for the creation of silver nanoparticles from a solution of silver nitrate (AgNO 3 ) which makes use of an extract from the Punica Granatum plant as a reducing agent. The procedure is described in detail in this article. These phytosynthesized silver nanoparticles (PG-AgNPs) were characterised using a wide range of various procedures, and their appearance and structure were examined using multiple methods. After conducting an analysis of the FE-SEM picture, it was found that the silver nanoparticles that were produced using green synthesis had an average size of around 27.33 nanometres of size. There are reasons to be optimistic about the potential of these PG-AgNPs to combat cancer and bacteria, as shown by the findings of study that was carried out on these properties. 2. Methods 2.1 Experimental Protocol/Design 2.1.1 Materials Trypan blue, 0.25% Trypsin-EDTA solution, Dulbecco's Modified Eagle Medium (DMEM), Fetal bovine serum (FBS), Antibiotic-antimycotic (Ab/Am) solution, Fetal bovine serum (FBS), PBS: Phosphate Buffered Saline, 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide tetrazolium salt (MTT), PBS: Phosphate Buffered Saline, Silver Nitrate (AgNO 3 , 99.99%) and other chemicals required for the experiment, Fresh Punica Granatum flowers , commonly known as Anar or pomegranate. 2.1.2 Phytosynthesis of Silver Nano Particles The blooms of Punica Grantum were carefully washed under running water to eliminate any dust or other impurities. The petals of these flowers were cleaned six times with double-distilled water before being let to dry in room temperature air and then chopped into little bits. Boiling the flower petals (~ 60 gramme) in a beaker with 300 mL of double distilled water for 28 minutes was the procedure used. After undergoing six passes through sartorius filter discs (Grade 292), the extract was left to cool. The extract was concentrated to 920 millilitres and kept at 4 degrees Celsius until needed. In 1000 ml of deionized water, 1.6985 gramme of 99.99 percent pure Silver Nitrate (AgNO 3 ) salt (Merck) was dissolved to produce a 10mM aqueous solution of silver ions. As a starting point, 10 millilitres of Punica Granatum L. flower extract (PG-ex) was combined with 100 millilitres of an aqueous AgNO 3 solution (10 mM). Under constant stirring on a magnetic hot plate set at 65°C, the reaction was allowed to proceed for 75 minutes. The synthesis of Silver NPs, or the reduction of Silver ions (Ag + ) to Ag-NPs (Ag 0 ), was indicated by a shift in the colour of the mixed solution from a brilliant yellow to a dark brownish tint within 25 minutes. In order to monitor any further reactions, the solution was stirred at the same temperature for 50 minutes longer. After 50 minutes, there was no more noticeable change in hue, indicating that the reaction had finished. For months, phytosynthesized AgNPs (PG-AgNPs) remained stable and spread evenly. The synthesis technique is shown schematically in Fig. 1.0. 2.2 Characterization & Biomedical Applications A UV-Vis spectroscopy analysis was performed on PG-AgNPs derived from Punica Granatum flower extract (PG-ex) using the Evolution 201 system. As for the morphological and elemental analysis of the phytosynthesized PG-AgNPs, TESCAN MAIA3 with EDX, a FeSEM, was used, while the FTIR of sample was recorded in transmittance mode. Spin coating on a cleaned Silica wafer with a diluted solution of phytosyntesized PG-AgNPs at a speed of 1000 revolutions per minute (RPM) was used to prepare samples for FE-SEM. These samples were subsequently dried in a vacuum oven at a temperature of 60 degrees Celsius. For the purpose of obtaining structural information of the PG-AgNPs, the PANalytical X'Pert pro 9kW X-Ray diffractometer was used. The step size utilised was 0.010, and the radiation utilised was Cu Kα, with a wavelength of 1.54 Å. In order to get the samples ready for XRD analysis, a diluted solution of PG-AgNPs was spin coated five times at a speed of one thousand revolutions per minute. The National Centre for Cell Sciences in Pune, India, was kind enough to provide us with our SiHa cell line, which was produced from cervical squamous cell carcinoma obtained from human patients. The procedure of doing research ultimately resulted in the acquisition of this cell line. Supporting the cells in DMEM was accomplished by the use of a medium that included 10% FBS and 1% antibiotic-antimycotic solution. This action was taken in order to achieve the goal of maintaining the cells in a state of excellent repair. After that, they were put into a CO 2 incubator that was established at 37 degrees Celsius and contained 5% (v/v) CO 2 . There were further exposure experiments in which the cells were reused after being cultured for twenty-four hours. In order to determine whether or not the PG-AgNPs had a cytotoxic effect, the researchers used the SiHa cancer cell line. The reduction test that was developed by Mosmann (1983) was used in order to assess the viability of the cells that received treatment as well as those that served as the control.[45] For this particular objective, the tetrazolium dye known as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used. This dye was sourced from Himedia in Pennsylvania, United States of America. The yellow 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide salt (MTT) turns into the purple formazan crystals that are employed in the MTT experiment. This transformation is caused by the mitochondrial translocation protein (MTT), which is responsible for the transformation. According to Parveen et al. ( 2023 ), the function and health of the mitochondria are considered to be a measure of the vitality of the cell. [46] Inoculation was carried out for a period of twenty-four hours after the seeding of cells in 96-well plates at a density of one million cells per well. After that, the cells were subjected to different concentrations of PG-ex and PG-AgNPs, ranging from 5 to 30 microgrammes, and then they were permitted to incubate at a temperature of 37 0 degrees Celsius in an incubator with a CO 2 content of 5% for a period of twenty-four hours. After being exposed for twenty-four hours, ten microlitres of MTT solution with a concentration of five milligrammes per millilitre was added to each well. The wells were then heated in a carbon dioxide incubator at 37 0 degrees Celsius for the subsequent four hours. Following the incubation process, 100 microlitres of DMSO was applied in order to dissolve the formazan crystals. On the other hand, the culture medium was discarded. Once the cells had been incubated for fifteen minutes, the optical density (OD) was measured at 540 nm using a microplate reader (BioTek, Epoch 2, USA). This was done in order to evaluate the vitality of the cells. The statistical analysis was carried out with the assistance of GraphPad Prism. When calculating the vitality of cells, the formula that was utilised was as follows: $$\:\mathbf{C}\mathbf{e}\mathbf{l}\mathbf{l}\:\mathbf{v}\mathbf{i}\mathbf{a}\mathbf{b}\mathbf{i}\mathbf{l}\mathbf{i}\mathbf{t}\mathbf{y}\left(\mathbf{\%}\right)=\:\left[\frac{\left\{\left(\mathbf{C}\mathbf{o}\mathbf{n}\mathbf{t}\mathbf{r}\mathbf{o}\mathbf{l}\:\mathbf{a}\mathbf{b}\mathbf{s}\mathbf{o}\mathbf{r}\mathbf{b}\mathbf{a}\mathbf{n}\mathbf{c}\mathbf{e}\right)\:-\:\left(\mathbf{t}\mathbf{e}\mathbf{s}\mathbf{t}\:\mathbf{a}\mathbf{b}\mathbf{s}\mathbf{o}\mathbf{r}\mathbf{b}\mathbf{a}\mathbf{n}\mathbf{c}\mathbf{e}\right)\right\}}{\left(\mathbf{c}\mathbf{o}\mathbf{n}\mathbf{t}\mathbf{r}\mathbf{o}\mathbf{l}\:\mathbf{a}\mathbf{b}\mathbf{s}\mathbf{o}\mathbf{r}\mathbf{b}\mathbf{a}\mathbf{n}\mathbf{c}\mathbf{e}\right)}\right]\:\times\:100$$ Two harmful microorganisms, namely E. coli and S. aureus, were subjected to antimicrobial activity testing on silver nanoparticles in order to assess the efficiency of these particles against these bacteria. It is well recognised that silver nanoparticles have the ability to function as antimicrobial agents against bacteria that cause an infection. The traditional agar well diffusion technique was used in order to validate the effectiveness of silver nanoparticles as antibacterial agents. These nanoparticles were produced by using the flower of Punica Granatum, which was a green synthesis process. To ascertain the zones of inhibition (ZOI) against every kind of bacteria, the cultures were put in an incubator and heated to 37 degrees Celsius for a period of twenty-four hours. This was done in order to discover the ZOI. 3. Result and discussions Figure 2.0 (a) displays the UV-Vis Spectra of PG-ex, revealing a prominent absorption peak at around 312 nm. This peak is likely caused by the existence of functional groups within the extract. Figure 2.0 (b) displays the UV-Vis Spectra of PG-AgNPs, showing a prominent absorption peak at around 453 nm. This peak is likely caused by the presence of functional molecules in the extract. The presence of a distinctive peak in the spectrum of silver nanoparticles is attributed to surface plasmon resonance. This was also apparent from the change in hue of the mixture, transitioning from a translucent state to a golden shade and eventually to a dark brown colour towards the conclusion of the synthesis process. The observed shift in hue occurred due to the stimulation of the surface plasmon resonance band of unbound electrons in silver nanoparticles. The presence of a plasmon peak is contingent upon the specific form and size of the nanoparticles that have been synthesised. Figure 2.0 (c) depicts The samples were analysed using FTIR spectroscopy in transmittance mode, and the resulting plot can be shown in Fig. 2.0 (c). The FTIR spectra show prominent peaks at 3449.43 cm − 1, corresponding to intermolecular hydrogen bonding O-H stretching vibrations of secondary amines, and at 2086.54 cm − 1, representing C ≡ C stretching. The peaks seen at 1640.82 cm − 1 are likely due to the C = O stretching of tertiary amide. Furthermore, the peak at 689.68 cm − 1 corresponded to the stretching vibrations of C-Cl bonds and the vibrations of the halide group. The synthesis of PG-AgNPs also entails the use of several bio-compounds as capping agents throughout the synthesis procedure. Figure 2.0 (d) displays the X-ray diffraction (XRD) spectra of the silver nanoparticles that were produced. The XRD pattern showed distinct peaks at positions 38.43, 44.64, 64.84, and 77.75, which match the Face Centred Cubic (FCC) structure of silver nanoparticles (NPs). The peaks seen at 2θ values of 38.43, 44.64, 64.84, and 77.75 correspond to the (111), (200), (220), and (311) planes, respectively, of the face-centered cubic (FCC) structure of silver. These values are consistent with the data provided in the JCPDS File No. 04-0783. Thus, it was determined that the silver nanoparticles produced using PG flower extract had a polycrystalline structure. The lattice parameter was determined and measured to be 4.06203 Å, which closely matches the standard value of 4.0865 Å (JCPDS File No. 04-0783). The particles' crystallite domain size was determined using the Debye Scherrer formula and was found to be around 17.81 nm.[47] In collaboration with EDX, we have selected a specific region for the aim of conducting elemental mapping, as seen in Fig. 2(g). The elemental silver nanoparticles were seen to have uniform growth and distribution inside the nanoparticle dispersion, as confirmed by the EDX analysis of the Ag nanoparticle dispersion. Table 1 provides a comprehensive overview of the parameters and data, Wells 4 and 5 are specifically designated as the positive and negative controls, respectively. The negative control group, consisting of distilled water, exhibited no inhibitory zones. Conversely, the positive group, which consisted of Gentamicin, exhibited inhibitory zones of 25 mm for E. coli and 24 mm for S. aureus. Table 1 Antibacterial activity study : Naoparticle Zone of Inhibition against certain bacteria. S. No. Sample Zone of Inhibition in mm S. aureus E.coli 1 50µg/ml NP Nil Nil 2 100µg/ml NP 10 11 3 150µg/ml NP 12 13 4 (-) control Nil Nil 5 (+) control 24 25 The antibacterial properties of the substance were examined in bacterial cultures using the disc diffusion method. In order to determine the efficacy of the PG-AgNPs as an antibacterial agent, they were subjected to testing against both Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). The antibacterial capabilities of silver nanoparticles were evaluated against Escherichia coli and Staphylococcus aureus, which are two pathogenic microorganisms. Wells 1–50µg/ml NP, 2–100µg/ml NP, 3–150µg/ml NP, 4–distilled water negative control, and 5–gentamicin positive control are indicated in the image above. Evaluating Cytotoxic Activity (MTT Assay) revealed that compounds considerably reduced cell viability of SiHa cells from cervical cancer. Various quantities of silver nanoparticles were used to test the cytotoxicity of substances by calculating the % cell viability. As the dosage concentrations increased, the findings showed that cancer cell viability dropped dramatically compared to untreated cells (control cells). The IC50 values for PG-ex and PG-AgNPs were determined to be around 25µg and 13µg, respectively, as shown in Fig. 1 . In order to suppress cells by 50% in vitro, this concentration is required. 4. Conclusions It has been established that the floral extract of Punica Granatum may be used as a reducing agent in order to construct a noval synthesis method for the phytosynthesis of silver nano size particles. This was accomplished by looking at the flowers of the plant. Not only is the method of synthesis simple and inexpensive, but it is also environmentally friendly. Methods such as UV-Vis Spectroscopy, FE-SEM, XRD, and FTIR were used in order to carry out an examination of the structural and morphological properties of the silver nanoparticles that were synthesised. It was found that the silver nanoparticles that were made have a spherical form, a smooth surface, and are distributed equally throughout the material. This was determined via the processes of manufacturing. It was established that the average size of the nanoparticles that were synthesised was about 27.33 nanometers, and the FE-SEM image was utilised to create an estimate of this size. In the study, it was discovered that the phytosynthesized PG-AgNPs displayed antibacterial activity against two harmful bacteria, namely Staphylococcus aureus and Escherichia coli. The capacity of phytosynthesized silver nanoparticles (PG-AgNPs) to limit the growth of SiHa cells derived from cervical cancer has been shown to have considerable favourable outcomes in terms of their anticancer efficacy. When compared to cells that were not exposed to the nanoparticles, the viability of cancer cells was significantly reduced when quantities of silver nanoparticles were present. This is the reason why this is the case. Additional silver nanoparticles that have been synthesised have the potential to be useful for applications such as the delivery of medicine, bioimaging, and the treatment of bacterial infections. Declarations CRediT authorship contribution statement Sumit Kumar Shrivastava : Conceptualization, Formal Analysis, Writing – original draft, Writing – review & Editing, Validation, Visualization, Methodology, Investigation, Data Curation, Resources. Asita Kulshreshtha: Formal Analysis, Writing – review & Editing, Supervision. Shikha Srivastava: Validation, Writing – review & Editing. Rajesh K. Gangwar: Formal Analysis, Methodology, Writing – review & Editing, Supervision. Saurabh Kumar: Validation, Resources. Monisha Banerjee: Validation, Resources Writing – review & Editing, Dhirendra K. Chaudhary: Formal Analysis, Methodology, Writing – review & Editing, Supervision, Dimple Kumari: Formal Analysis, Writing – review & Editing. Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Data Availability The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Plant guidelines The collection of the plants used in the study complies with local or national guidelines with no need for further affirmation. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Acknowledgements We would like to express our gratitude to Amity University Lucknow and the Council of Science and Technology Uttar Pradesh Lucknow for providing us with the chance and further assistance. In addition, we would like to express our gratitude to Professor B. C. Yadav and Abhishek Prakash Tiwari of BBAU Lucknow for providing their assistance. We would also want to express our gratitude to ACMS and IIT Kanpur for providing us with FE-SEM and XRD capabilities. For the cell culture facility at the Molecular and Human Genetics Laboratory, Department of Zoology, University of Lucknow, Lucknow, the authors would like to express their gratitude to the Centre of Excellence Programme, Higher Education, Government of Uttar Pradesh, India. Both SK and SP would like to express their gratitude to the Indian Council of Medical Research (ICMR) and the Central Services of Research (CSIR) in New Delhi for providing them with research scholarships. References Rudrappa M, Rudayni HA, Assiri RA, Bepari A, Basavarajappa DS, Nagaraja SK, Chakraborty B, Swamy PS, Agadi SN, Niazi SK, Nayaka S (2022) Plumeria alba-Mediated Green Synthesis of Silver Nanoparticles Exhibits Antimicrobial Effect and Anti-Oncogenic Activity against Glioblastoma U118 MG Cancer Cell Line. Nanomaterials. 12(3):493, https://doi.org/10.3390/nano12030493 . 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Springer, Dordrecht. https://doi.org/10.1007/978-94-007-5653-3_10 Borges MGSSA, Lima LDO, Veras ACMF, Ferreira RC, Alves DDN, Medeiros IAD, Magnani M, Sobral MV, Castro RDD, Tintino SR, Oliveira-Tintino CDDM, Coutinho HDM, Guerra FQS, Barros DBD, Oliveira MBMD (2024) Chem. Biodiversity 21:e202301982. https://doi.org/10.1002/cbdv.202301982 Parveen S, Masood S, Kumar S, Banerjee M (2023) Phytonanomedicine: A therapeutic approach for cervical cancer. OpenNano 13:100178. https://doi.org/10.1016/j.onano.2023.100178 Muniz FTL, Miranda MAR, Santos CMD, Sasaki JM (2016) Acta Crystallogr., Sect. A: Found. Crystallogr. 72 (3):385–390. https://doi.org/10.1107/S205327331600365X . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 16 Oct, 2025 Read the published version in Discover Nano → Version 1 posted Editorial decision: Revision requested 01 Oct, 2024 Reviews received at journal 30 Sep, 2024 Reviews received at journal 28 Sep, 2024 Reviews received at journal 28 Sep, 2024 Reviewers agreed at journal 26 Sep, 2024 Reviewers agreed at journal 25 Sep, 2024 Reviewers agreed at journal 24 Sep, 2024 Reviewers agreed at journal 24 Sep, 2024 Reviewers agreed at journal 24 Sep, 2024 Reviewers agreed at journal 23 Sep, 2024 Reviewers invited by journal 23 Sep, 2024 Editor assigned by journal 18 Sep, 2024 Submission checks completed at journal 18 Sep, 2024 First submitted to journal 08 Sep, 2024 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. 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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-5054888","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":361198495,"identity":"ae3eadff-c3c7-46fc-809c-112121b50852","order_by":0,"name":"Sumit K Shrivastava","email":"","orcid":"","institution":"Amity University U.P","correspondingAuthor":false,"prefix":"","firstName":"Sumit","middleName":"K","lastName":"Shrivastava","suffix":""},{"id":361198496,"identity":"14d31182-bbec-4a2a-93c9-ded2a25c4ae7","order_by":1,"name":"Asita Kulshreshtha","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBUlEQVRIiWNgGAWjYDACHgbGA4wNQIYEAxsDQwUDiGSAkbi0MEC1MAOVnQEqZiNJC2MbTDUeLfw9ZwwO/txxT05+dv+xBz/n2UXzyfcYMHwoO4xTi8TZHoPDvGeKjQ3uHGY37N2WnNvGxmPAOOMcbi0M53kMDjO2JSRukEhmk+DdxgzWwszbhluLPFDLwZ9tCfXzZySzSf6dUw/R8hePFgOgww7wtiUkMNxIZpPmbTgM0cKIR4vhmWMFh4FaDDfcSDaTljl2HKglreBgz7l0nFrkziRvfAh0mLz8jMRnkm9qqnPnNx/e+OBHmTVu72MBHAYHSFIPBOwPSNUxCkbBKBgFwxsAAABVVzp9anLBAAAAAElFTkSuQmCC","orcid":"","institution":"Amity University U.P","correspondingAuthor":true,"prefix":"","firstName":"Asita","middleName":"","lastName":"Kulshreshtha","suffix":""},{"id":361198497,"identity":"9f7a3264-0502-4e24-b4f3-c1b2f23282de","order_by":2,"name":"Rajesh K. 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(b). UV-Vis spectra of PG-AgNPs. (c). FTIR Spectra of PG-AgNPs. (d). XRD spectra of PGNPs.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/557f4db87058e081f9dc78d4.png"},{"id":68263088,"identity":"80e1eb0b-9c0b-40f2-867d-438cf1b70a5d","added_by":"auto","created_at":"2024-11-05 12:19:53","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":229651,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a-d):\u003c/strong\u003e \u003cem\u003eThe FE-SEM images of phytosynthesized PG-AgNPs by using Punica Granatum floral extract\u003c/em\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/f217d8f92096d29f0b062388.png"},{"id":68264206,"identity":"b296f336-b3dc-484f-80f0-6af390b65c3e","added_by":"auto","created_at":"2024-11-05 12:27:53","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":181227,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a-c) :\u003c/strong\u003e \u003cem\u003eDistribution of particle size in EDX Spectra of the nanoparticle\u003c/em\u003e \u003cem\u003edispersion\u003c/em\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/6ef9aa65d1d6330ab33c5ff6.png"},{"id":68264205,"identity":"15bdb63c-2489-450c-9418-92aa6d4210ae","added_by":"auto","created_at":"2024-11-05 12:27:53","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":486396,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a-b) :\u003c/strong\u003e \u003cem\u003eNanoparticle antibacterial activity against (a)- E. coli and (b)- S. aureus.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/e6d42487c70d3415c5a2e73d.png"},{"id":68263084,"identity":"16037c0b-947f-4cc7-b620-eaff391adcc1","added_by":"auto","created_at":"2024-11-05 12:19:53","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":103162,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eCell viability assay in SiHa cells after treatment with different concentration of PG-ex and PG-AgNPs.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/94979b4232609e061d4b7cfe.png"},{"id":93955768,"identity":"592ee496-2453-4426-9753-5d4a4b013fdd","added_by":"auto","created_at":"2025-10-20 16:00:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2394892,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5054888/v1/1d2743d9-54b2-479a-b158-65bb361750ce.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"A novel approach for phyto-synthesis of silver nanoparticles using floral extract of Punica Granatum for potent anti-cancerous and antibacterial capabilities","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eNanotechnology heavily depends on metal nanoparticles that are synthesised via phytosynthesis. Silver and gold nanoparticles are becoming more popular due to their versatility and wide range of applications. [1\u0026ndash;5] Researchers have created novel synthesis techniques for affordable, non-toxic, and eco-friendly metal nanoparticles. Phyto-synthesis is a well-known example. Nanotechnology has many scientific and technological uses. The famous aphorism \"Necessity is the mother of invention\" argues that necessity drives every new idea. As metal nanoparticle demand rises, researchers are always looking for new, easy production techniques. Researchers are employing herbs, fruits, and plants to greenly synthesise metal nanoparticles. This methodology encourages green metal nanoparticle manufacturing without hazardous ingredients. Green or phytosynthesis is a simple, non-toxic, eco-friendly synthesis process. Because many bio molecules work as reducing and capping agents, phyto-synthesis processes are cost-effective and versatile. [6\u0026ndash;10]\u003c/p\u003e \u003cp\u003eDue to their unique features and beneficial medicinal uses, silver and gold nanoparticles are continually drawing researchers. The reducing agent for many green metal nanoparticle manufacturing processes is plants, bacteria, and fungus. [11\u0026ndash;20]\u003c/p\u003e \u003cp\u003e According to earlier research, metallic nanoparticles phytosynthesized by ethno medicinal plants rich in antioxidant compounds have increased anti-bacterial and anti-cancer activities. Many cancer physicians utilise chemotherapy, immunotherapy, and radiation. Because chemotherapy is harmful, phytosynthesized metallic nanoparticles are needed to develop an effective chemotherapy medication. [21\u0026ndash;30]\u003c/p\u003e \u003cp\u003eAn interdisciplinary and cross-sectoral area uses nanotechnology in medical diagnostics, food, medicine, biotechnology, environment, energy, chemistry, and physics. Nanotechnology is also applied in the field of biotechnology. Nanoscience and nanotechnology's multidisciplinary nature yields current diagnostics tools. [31]\u003c/p\u003e \u003cp\u003eThe European, American, and Japanese National Nanotechnology Project is prioritised in several domains. To comprehend and manage unparalleled basic components of physical things, nanotechnology and nanoscience may transfer materials precisely. These advances may impact vaccination and computer design. Nanotechnology involves creating metal nanoparticle-based medications. These uses in biology, health, and medicine have made nanoparticles popular today. Structured, they are 100 nanometers. They improve illness diagnosis and therapy extensively. Nanoparticles size allows them to pass the blood-brain barrier. Despite their modest size, nanoparticles can treat brain tumours. Nanoparticles are designed by managing the particle size, surface features, and release of a specific drug at a certain time and place. This is accomplished via the process of nanoparticle design. Disease diagnostics, tissue engineering scaffolds, and targeted medicine delivery employ nanoparticles. [32\u0026ndash;34]\u003c/p\u003e \u003cp\u003eMetallic nanoparticle applications research has become popular worldwide. The uses of metal nanoparticles are primarily influenced by elements such as particle size, dispersion, architecture, chemical assembly, colloidal stability, and biocompatibility. Metallic nanoparticles with medicinal herbs work well against cancer. Recent years have seen metal nanoparticles carrying herbs heal many malignancies. Recently, nanoparticle technology has advanced the manufacture of various medications, which might help cure and diagnose many illnesses, including cancer. Cancer\u0026rsquo;s high death rate causes psychological and economic difficulties. A major determinant in cancer incidence is lifestyle. The primary factors contributing to the development of cancer include environmental contaminants, carcinogens, mutagens, bacterial and viral infections, and genetic susceptibility.\u003c/p\u003e \u003cp\u003eA combination of surgical intervention, radiation therapy, and chemotherapy is used to battle and manage cancer, with the specific treatment approach determined by the kind and stage of cancer as well as the overall health condition of the patient. Although there is a potential for negative consequences in advanced malignancies, the majority of these approaches are unsuccessful, underscoring the need for innovative cancer therapies and scientific investigation. Manufacturing materials, devices, and systems efficiently using nanometre-scale matter control and novel features and phenomena is nanotechnology. Nanoparticles are atomic or molecule assemblies with diameters under 100 nm, exhibiting unique physicochemical features relative to their bulk. Nanotechnology helped create a novel cancer approach.\u003c/p\u003e \u003cp\u003eWe've long investigated metallic nanoparticles for cancer therapy. Natural metal oxides are abundant, making nanoparticle processing and manufacturing one of the cheapest methods. Metallic nanoparticles are a new form of commonly utilised mineral particle that researchers have evaluated owing to their appropriate physical and chemical features and higher adsorption power than other metallic nanoparticle compositions. Metallic nanoparticles are biocompatible and nontoxic when used as medical fillers, cosmetics, and pharmaceutical carriers. Metallic nanoparticles possess exceptional chemical stability, a low dielectric constant, catalytic activity, the capacity to absorb infrared and ultraviolet light, and notably, antibacterial properties. The therapeutic and anticancer capabilities of these compounds might potentially enhance cancer therapy.\u003c/p\u003e \u003cp\u003eGangwar et al. developed a green technique for improving bioavailability using curcumin-conjugated silica nanoparticles [35]. According to Shrivastava et al., Plumeria Pudica Plant sections can synthesise silver nanoparticles. They phytosynthesized silver nanoparticles from Plumeria Pudica flowers [36]. They also phytosynthesised silver nanoparticles from Plumeria Pudica leaves and found anti-cancerous action [37]. Huang et al. used the aqueous extract of Fumaria officinalis to synthesise nickel nanoparticles, which were subsequently identified as a groundbreaking chemotherapeutic agent for the management of ovarian cancer. [38] Zhang et al. used Leucus aspera to phytosynthesize AgNPs and observed significant efficacy in inhibiting alveolar carcinoma, demonstrating promising anticancer action. [39]\u003c/p\u003e \u003cp\u003eApproaches to the production of nanomaterials that are natural, non-hazardous economically viable, and environmentally friendly have gained prominence over physicochemical ones. There are many approaches to make nanomaterials from metal ion salts biochemically. Use an ecologically friendly solvent solution with stabilising and reducing components to synthesise nanoparticles 'green'. Nanoparticle biogenesis results from lowering metal ions of biomolecules. This method produces huge numbers of well-defined nanoparticles without contamination and reduces the environmental effect of biological synthesis. All types of plants, as well as microbes, underwater algae, plant-derived substances, tissue, and fruits, are used in the production of nanomaterials. Beginning in the 1900s, biomaterials have been able to decrease metal ions. In the last half-century, there has been an increase in curiosity in lowering agents, which has been driven by a dearth of understanding about the mechanisms involved in their production. It has been revealed by researchers that nanoparticles made from biological components possess anticancer capabilities that are unique to them. Recent developments in medicine have made use of metallic nanoparticles. Recent studies have shown that some nanoparticles possess therapeutic properties and may serve as a viable alternative to physicochemical different metal-supported nanoparticles, as well as antibacterial and anticancer medications.\u003c/p\u003e \u003cp\u003ePomegranate is said to have originated in Iran. The Punica granatum, or pomegranate tree, grows to five to eight metres in Iran, the northern Indian Himalayas, China, the US, and the Mediterranean regions. PG is a key Iranian plant because of its versatility. It grows in dry and semiarid regions nationwide. More than seven hundred Punica granatum (PG) varieties in Iran's Saveh and Yazd cities share size, colour, flavour, ripening duration, and disease resistance. [40] The plant's root, juice, peel, leaves, blossoms, bark, and seed of PG contain ingredients that have unique toxicological and medicinal characteristics.[41] The edible berry has a thick crimson skin, a hexagonal shape, and 600 seeds. Each seed has a water-laden pulp (aril) that may be white, deep crimson, or purple and is delicious. White, spongey, astringent pulp encases seeds. Pomegranates are twice mentioned in the Quran as God's magnificent creations and as growing in paradise's gardens. The PG fruit has been used to treat acidosis, dysentery, microbiological infections, diarrhoea, helminth infections, haemorrhage, and respiratory illnesses. PG seeds contain estrone and estradiol. [42\u0026ndash;43] Both the dried pericarp and fruit juice are used to treat dental issues, headaches, piles, acne, colitis, menorrhagia, oxyuriasis, and diuretics. Recent scientific researchers have examined traditional PG uses. Many ancient cultures used PG juice, fruit, and extracts for medicine. PG may include phytochemicals with poisonous and pharmacological effects. The mechanisms and pharmacological and toxicological consequences of many of these substances are yet unknown. There is a growing interest in herbal remedies and medicinal plants all over the globe with the purpose of determining whether or not they are effective in preventing and treating chronic illnesses. It is possible to treat a variety of conditions with PG, such as cancers of the prostate, colon, breast, lung, and skin; leukaemia; diabetes; hyperlipidaemia; hypertension; myocardial ischaemia; myocardial perfusion; infections; erectile dysfunction; male infertility; neonatal hypoxia-ischemic brain damage; obesity; and Alzheimer's disease. [44]\u003c/p\u003e \u003cp\u003eThis research presents a basic phytosynthesis process for the creation of silver nanoparticles from a solution of silver nitrate (AgNO\u003csub\u003e3\u003c/sub\u003e) which makes use of an extract from the Punica Granatum plant as a reducing agent. The procedure is described in detail in this article. These phytosynthesized silver nanoparticles (PG-AgNPs) were characterised using a wide range of various procedures, and their appearance and structure were examined using multiple methods. After conducting an analysis of the FE-SEM picture, it was found that the silver nanoparticles that were produced using green synthesis had an average size of around 27.33 nanometres of size. There are reasons to be optimistic about the potential of these PG-AgNPs to combat cancer and bacteria, as shown by the findings of study that was carried out on these properties.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"2. Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Experimental Protocol/Design\u003c/h2\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003e2.1.1 Materials\u003c/h2\u003e \u003cp\u003eTrypan blue, 0.25% Trypsin-EDTA solution, Dulbecco's Modified Eagle Medium (DMEM), Fetal bovine serum (FBS), Antibiotic-antimycotic (Ab/Am) solution, Fetal bovine serum (FBS), PBS: Phosphate Buffered Saline, 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide tetrazolium salt (MTT), PBS: Phosphate Buffered Saline, Silver Nitrate (AgNO\u003csub\u003e3\u003c/sub\u003e, 99.99%) and other chemicals required for the experiment, Fresh \u003cem\u003ePunica Granatum flowers\u003c/em\u003e, commonly known as Anar or pomegranate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.1.2 Phytosynthesis of Silver Nano Particles\u003c/h2\u003e \u003cp\u003eThe blooms of Punica Grantum were carefully washed under running water to eliminate any dust or other impurities. The petals of these flowers were cleaned six times with double-distilled water before being let to dry in room temperature air and then chopped into little bits. Boiling the flower petals (~\u0026thinsp;60 gramme) in a beaker with 300 mL of double distilled water for 28 minutes was the procedure used. After undergoing six passes through sartorius filter discs (Grade 292), the extract was left to cool. The extract was concentrated to 920 millilitres and kept at 4 degrees Celsius until needed. In 1000 ml of deionized water, 1.6985 gramme of 99.99 percent pure Silver Nitrate (AgNO\u003csub\u003e3\u003c/sub\u003e) salt (Merck) was dissolved to produce a 10mM aqueous solution of silver ions. As a starting point, 10 millilitres of Punica Granatum L. flower extract (PG-ex) was combined with 100 millilitres of an aqueous AgNO\u003csub\u003e3\u003c/sub\u003e solution (10 mM). Under constant stirring on a magnetic hot plate set at 65\u0026deg;C, the reaction was allowed to proceed for 75 minutes. The synthesis of Silver NPs, or the reduction of Silver ions (Ag\u003csup\u003e+\u003c/sup\u003e) to Ag-NPs (Ag\u003csup\u003e0\u003c/sup\u003e), was indicated by a shift in the colour of the mixed solution from a brilliant yellow to a dark brownish tint within 25 minutes. In order to monitor any further reactions, the solution was stirred at the same temperature for 50 minutes longer. After 50 minutes, there was no more noticeable change in hue, indicating that the reaction had finished. For months, phytosynthesized AgNPs (PG-AgNPs) remained stable and spread evenly. The synthesis technique is shown schematically in Fig.\u0026nbsp;1.0.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Characterization \u0026amp; Biomedical Applications\u003c/h2\u003e \u003cp\u003eA UV-Vis spectroscopy analysis was performed on PG-AgNPs derived from Punica Granatum flower extract (PG-ex) using the Evolution 201 system. As for the morphological and elemental analysis of the phytosynthesized PG-AgNPs, TESCAN MAIA3 with EDX, a FeSEM, was used, while the FTIR of sample was recorded in transmittance mode. Spin coating on a cleaned Silica wafer with a diluted solution of phytosyntesized PG-AgNPs at a speed of 1000 revolutions per minute (RPM) was used to prepare samples for FE-SEM. These samples were subsequently dried in a vacuum oven at a temperature of 60 degrees Celsius. For the purpose of obtaining structural information of the PG-AgNPs, the PANalytical X'Pert pro 9kW X-Ray diffractometer was used. The step size utilised was 0.010, and the radiation utilised was Cu Kα, with a wavelength of 1.54 \u0026Aring;. In order to get the samples ready for XRD analysis, a diluted solution of PG-AgNPs was spin coated five times at a speed of one thousand revolutions per minute.\u003c/p\u003e \u003cp\u003eThe National Centre for Cell Sciences in Pune, India, was kind enough to provide us with our SiHa cell line, which was produced from cervical squamous cell carcinoma obtained from human patients. The procedure of doing research ultimately resulted in the acquisition of this cell line. Supporting the cells in DMEM was accomplished by the use of a medium that included 10% FBS and 1% antibiotic-antimycotic solution. This action was taken in order to achieve the goal of maintaining the cells in a state of excellent repair. After that, they were put into a CO\u003csub\u003e2\u003c/sub\u003e incubator that was established at 37 degrees Celsius and contained 5% (v/v) CO\u003csub\u003e2\u003c/sub\u003e. There were further exposure experiments in which the cells were reused after being cultured for twenty-four hours.\u003c/p\u003e \u003cp\u003eIn order to determine whether or not the PG-AgNPs had a cytotoxic effect, the researchers used the SiHa cancer cell line. The reduction test that was developed by Mosmann (1983) was used in order to assess the viability of the cells that received treatment as well as those that served as the control.[45] For this particular objective, the tetrazolium dye known as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used. This dye was sourced from Himedia in Pennsylvania, United States of America. The yellow 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide salt (MTT) turns into the purple formazan crystals that are employed in the MTT experiment. This transformation is caused by the mitochondrial translocation protein (MTT), which is responsible for the transformation. According to Parveen et al. (\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), the function and health of the mitochondria are considered to be a measure of the vitality of the cell. [46] Inoculation was carried out for a period of twenty-four hours after the seeding of cells in 96-well plates at a density of one million cells per well. After that, the cells were subjected to different concentrations of PG-ex and PG-AgNPs, ranging from 5 to 30 microgrammes, and then they were permitted to incubate at a temperature of 37\u003csup\u003e0\u003c/sup\u003e degrees Celsius in an incubator with a CO\u003csub\u003e2\u003c/sub\u003e content of 5% for a period of twenty-four hours. After being exposed for twenty-four hours, ten microlitres of MTT solution with a concentration of five milligrammes per millilitre was added to each well. The wells were then heated in a carbon dioxide incubator at 37\u003csup\u003e0\u003c/sup\u003e degrees Celsius for the subsequent four hours. Following the incubation process, 100 microlitres of DMSO was applied in order to dissolve the formazan crystals. On the other hand, the culture medium was discarded. Once the cells had been incubated for fifteen minutes, the optical density (OD) was measured at 540 nm using a microplate reader (BioTek, Epoch 2, USA). This was done in order to evaluate the vitality of the cells. The statistical analysis was carried out with the assistance of GraphPad Prism. When calculating the vitality of cells, the formula that was utilised was as follows:\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:\\mathbf{C}\\mathbf{e}\\mathbf{l}\\mathbf{l}\\:\\mathbf{v}\\mathbf{i}\\mathbf{a}\\mathbf{b}\\mathbf{i}\\mathbf{l}\\mathbf{i}\\mathbf{t}\\mathbf{y}\\left(\\mathbf{\\%}\\right)=\\:\\left[\\frac{\\left\\{\\left(\\mathbf{C}\\mathbf{o}\\mathbf{n}\\mathbf{t}\\mathbf{r}\\mathbf{o}\\mathbf{l}\\:\\mathbf{a}\\mathbf{b}\\mathbf{s}\\mathbf{o}\\mathbf{r}\\mathbf{b}\\mathbf{a}\\mathbf{n}\\mathbf{c}\\mathbf{e}\\right)\\:-\\:\\left(\\mathbf{t}\\mathbf{e}\\mathbf{s}\\mathbf{t}\\:\\mathbf{a}\\mathbf{b}\\mathbf{s}\\mathbf{o}\\mathbf{r}\\mathbf{b}\\mathbf{a}\\mathbf{n}\\mathbf{c}\\mathbf{e}\\right)\\right\\}}{\\left(\\mathbf{c}\\mathbf{o}\\mathbf{n}\\mathbf{t}\\mathbf{r}\\mathbf{o}\\mathbf{l}\\:\\mathbf{a}\\mathbf{b}\\mathbf{s}\\mathbf{o}\\mathbf{r}\\mathbf{b}\\mathbf{a}\\mathbf{n}\\mathbf{c}\\mathbf{e}\\right)}\\right]\\:\\times\\:100$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eTwo harmful microorganisms, namely E. coli and S. aureus, were subjected to antimicrobial activity testing on silver nanoparticles in order to assess the efficiency of these particles against these bacteria. It is well recognised that silver nanoparticles have the ability to function as antimicrobial agents against bacteria that cause an infection. The traditional agar well diffusion technique was used in order to validate the effectiveness of silver nanoparticles as antibacterial agents. These nanoparticles were produced by using the flower of Punica Granatum, which was a green synthesis process. To ascertain the zones of inhibition (ZOI) against every kind of bacteria, the cultures were put in an incubator and heated to 37 degrees Celsius for a period of twenty-four hours. This was done in order to discover the ZOI.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"3. Result and discussions","content":"\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2.0\u003c/span\u003e (a) displays the UV-Vis Spectra of PG-ex, revealing a prominent absorption peak at around 312 nm. This peak is likely caused by the existence of functional groups within the extract. Figure \u003cspan class=\"InternalRef\"\u003e2.0\u003c/span\u003e (b) displays the UV-Vis Spectra of PG-AgNPs, showing a prominent absorption peak at around 453 nm. This peak is likely caused by the presence of functional molecules in the extract. The presence of a distinctive peak in the spectrum of silver nanoparticles is attributed to surface plasmon resonance. This was also apparent from the change in hue of the mixture, transitioning from a translucent state to a golden shade and eventually to a dark brown colour towards the conclusion of the synthesis process. The observed shift in hue occurred due to the stimulation of the surface plasmon resonance band of unbound electrons in silver nanoparticles. The presence of a plasmon peak is contingent upon the specific form and size of the nanoparticles that have been synthesised. Figure \u003cspan class=\"InternalRef\"\u003e2.0\u003c/span\u003e (c) depicts The samples were analysed using FTIR spectroscopy in transmittance mode, and the resulting plot can be shown in Fig. \u003cspan class=\"InternalRef\"\u003e2.0\u003c/span\u003e (c). The FTIR spectra show prominent peaks at 3449.43 cm\u0026thinsp;\u0026minus;\u0026thinsp;1, corresponding to intermolecular hydrogen bonding O-H stretching vibrations of secondary amines, and at 2086.54 cm\u0026thinsp;\u0026minus;\u0026thinsp;1, representing C\u0026thinsp;\u0026equiv;\u0026thinsp;C stretching. The peaks seen at 1640.82 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 are likely due to the C\u0026thinsp;=\u0026thinsp;O stretching of tertiary amide. Furthermore, the peak at 689.68 cm\u0026thinsp;\u0026minus;\u0026thinsp;1 corresponded to the stretching vibrations of C-Cl bonds and the vibrations of the halide group. The synthesis of PG-AgNPs also entails the use of several bio-compounds as capping agents throughout the synthesis procedure. Figure \u003cspan class=\"InternalRef\"\u003e2.0\u003c/span\u003e (d) displays the X-ray diffraction (XRD) spectra of the silver nanoparticles that were produced. The XRD pattern showed distinct peaks at positions 38.43, 44.64, 64.84, and 77.75, which match the Face Centred Cubic (FCC) structure of silver nanoparticles (NPs). The peaks seen at 2\u0026theta; values of 38.43, 44.64, 64.84, and 77.75 correspond to the (111), (200), (220), and (311) planes, respectively, of the face-centered cubic (FCC) structure of silver. These values are consistent with the data provided in the JCPDS File No. 04-0783. Thus, it was determined that the silver nanoparticles produced using PG flower extract had a polycrystalline structure. The lattice parameter was determined and measured to be 4.06203 \u0026Aring;, which closely matches the standard value of 4.0865 \u0026Aring; (JCPDS File No. 04-0783). The particles\u0026apos; crystallite domain size was determined using the Debye Scherrer formula and was found to be around 17.81 nm.[47]\u003c/p\u003e\n\u003cp\u003eIn collaboration with EDX, we have selected a specific region for the aim of conducting elemental mapping, as seen in Fig.\u0026nbsp;2(g). The elemental silver nanoparticles were seen to have uniform growth and distribution inside the nanoparticle dispersion, as confirmed by the EDX analysis of the Ag nanoparticle dispersion.\u003c/p\u003e\n\u003cp\u003eTable 1 provides a comprehensive overview of the parameters and data, Wells 4 and 5 are specifically designated as the positive and negative controls, respectively. The negative control group, consisting of distilled water, exhibited no inhibitory zones. Conversely, the positive group, which consisted of Gentamicin, exhibited inhibitory zones of 25 mm for E. coli and 24 mm for S. aureus. \u0026nbsp;\u003c/p\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003e\u003cem\u003eAntibacterial activity study : Naoparticle Zone of Inhibition against certain bacteria.\u003c/em\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eS. No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eZone of Inhibition in mm\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eS. aureus\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eE.coli\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u0026micro;g/ml NP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026micro;g/ml NP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e150\u0026micro;g/ml NP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(-) control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNil\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(+) control\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eThe antibacterial properties of the substance were examined in bacterial cultures using the disc diffusion method. In order to determine the efficacy of the PG-AgNPs as an antibacterial agent, they were subjected to testing against both Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). The antibacterial capabilities of silver nanoparticles were evaluated against Escherichia coli and Staphylococcus aureus, which are two pathogenic microorganisms.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eWells 1\u0026ndash;50\u0026micro;g/ml NP, 2\u0026ndash;100\u0026micro;g/ml NP, 3\u0026ndash;150\u0026micro;g/ml NP, 4\u0026ndash;distilled water negative control, and 5\u0026ndash;gentamicin positive control are indicated in the image above.\u003c/p\u003e\n \u003cp\u003eEvaluating Cytotoxic Activity (MTT Assay) revealed that compounds considerably reduced cell viability of SiHa cells from cervical cancer. Various quantities of silver nanoparticles were used to test the cytotoxicity of substances by calculating the % cell viability. As the dosage concentrations increased, the findings showed that cancer cell viability dropped dramatically compared to untreated cells (control cells). The IC50 values for PG-ex and PG-AgNPs were determined to be around 25\u0026micro;g and 13\u0026micro;g, respectively, as shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. In order to suppress cells by 50% in vitro, this concentration is required.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003eIt has been established that the floral extract of Punica Granatum may be used as a reducing agent in order to construct a noval synthesis method for the phytosynthesis of silver nano size particles. This was accomplished by looking at the flowers of the plant. Not only is the method of synthesis simple and inexpensive, but it is also environmentally friendly. Methods such as UV-Vis Spectroscopy, FE-SEM, XRD, and FTIR were used in order to carry out an examination of the structural and morphological properties of the silver nanoparticles that were synthesised. It was found that the silver nanoparticles that were made have a spherical form, a smooth surface, and are distributed equally throughout the material. This was determined via the processes of manufacturing. It was established that the average size of the nanoparticles that were synthesised was about 27.33 nanometers, and the FE-SEM image was utilised to create an estimate of this size. In the study, it was discovered that the phytosynthesized PG-AgNPs displayed antibacterial activity against two harmful bacteria, namely Staphylococcus aureus and Escherichia coli. The capacity of phytosynthesized silver nanoparticles (PG-AgNPs) to limit the growth of SiHa cells derived from cervical cancer has been shown to have considerable favourable outcomes in terms of their anticancer efficacy. When compared to cells that were not exposed to the nanoparticles, the viability of cancer cells was significantly reduced when quantities of silver nanoparticles were present. This is the reason why this is the case. Additional silver nanoparticles that have been synthesised have the potential to be useful for applications such as the delivery of medicine, bioimaging, and the treatment of bacterial infections.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCRediT authorship contribution statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSumit Kumar Shrivastava : Conceptualization, Formal Analysis, Writing – original draft, Writing – review \u0026amp; Editing, Validation, Visualization, Methodology, Investigation, Data Curation, Resources. Asita Kulshreshtha: Formal Analysis, Writing – review \u0026amp; Editing, Supervision. Shikha Srivastava: Validation, Writing – review \u0026amp; Editing. Rajesh K. Gangwar: Formal Analysis, Methodology, Writing – review \u0026amp; Editing, Supervision. Saurabh Kumar: Validation, Resources. Monisha Banerjee: Validation, Resources Writing – review \u0026amp; Editing, Dhirendra K. Chaudhary: Formal Analysis, Methodology, Writing – review \u0026amp; Editing, Supervision, Dimple Kumari: Formal Analysis, Writing – review \u0026amp; Editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of competing interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePlant guidelines\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe collection of the plants used in the study complies with local or national guidelines with no need for further affirmation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to express our gratitude to Amity University Lucknow and the Council of Science and Technology Uttar Pradesh Lucknow for providing us with the chance and further assistance. \u0026nbsp;In addition, we would like to express our gratitude to Professor B. C. Yadav and Abhishek Prakash Tiwari of BBAU Lucknow for providing their assistance. We would also want to express our gratitude to ACMS and IIT Kanpur for providing us with FE-SEM and XRD capabilities. For the cell culture facility at the Molecular and Human Genetics Laboratory, Department of Zoology, University of Lucknow, Lucknow, the authors would like to express their gratitude to the Centre of Excellence Programme, Higher Education, Government of Uttar Pradesh, India. 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Crystallogr. 72 (3):385\u0026ndash;390. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1107/S205327331600365X\u003c/span\u003e\u003cspan address=\"10.1107/S205327331600365X\" 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":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Silver Nanoparticles (Ag NPs), Punica Granatum Flower, Green Synthesis, Eco-friendly","lastPublishedDoi":"10.21203/rs.3.rs-5054888/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5054888/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSynthesis of safest and non-toxic nanoparticles for biomedical applications in various fields of science \u0026amp; Technology remained a vast challenge for researchers. This work aims to create, analyze, and examine the antibacterial effects of silver nanoparticles that are synthesized utilizing the floral extract of Punica Granatum as both a stabilizing and reducing agent. Silver nitrate (AgNO3) solution was reduced using Punica Granatum flower extract which led to the synthesis of stable silver nanoparticles (PG-AgNPs) solution. The PG-AgNPs were examined and characterized using UV-Vis, FT-IR, FE-SEM, XRD, and EDX methodologies. Besides this anti-bacterial \u0026amp; anti-cancerous property were also studied. The results of FE-SEM analysis revealed that majority of the phytosynthesized PG-AgNPs had spherical shape and were evenly dispersed with a diameter of nearly 27.33 nm. EDX results revealed the relative presence of different elements, biomolecules, and capping agents. Additionally, synthesised silver nanoparticles had anti-cancer potential, with an IC\u003csub\u003e50\u003c/sub\u003e of ~\u0026thinsp;13 \u0026micro;g. E. coli and S. aureus were tested for silver nanoparticle antimicrobial efficacy. Phytosynthesized PG-AgNPs against Escherichia coli and Staphylococcus aureus were very effective. A new chemotherapeutic medication or innovative supplement to treat various forms of human cancer may be developed after the comprehensive clinical trial using PG-AgNPs containing Punica Granatum flower aqueous extract.\u003c/p\u003e","manuscriptTitle":"A novel approach for phyto-synthesis of silver nanoparticles using floral extract of Punica Granatum for potent anti-cancerous and antibacterial capabilities","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-05 12:19:48","doi":"10.21203/rs.3.rs-5054888/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-10-01T15:13:44+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-30T18:57:41+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-28T14:21:58+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-28T12:24:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"196757368045660817961673887904744480831","date":"2024-09-26T08:15:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"331787657207202014702480833836579908635","date":"2024-09-25T16:30:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"128396625714393299497153131728107295347","date":"2024-09-24T12:12:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"290448549358107554398814865850077467529","date":"2024-09-24T09:53:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"301682731973054004076337120987766743628","date":"2024-09-24T07:04:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"297748855908269906935771502609023970436","date":"2024-09-23T18:51:52+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-09-23T18:48:33+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-09-18T06:59:02+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-09-18T06:17:24+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Nano","date":"2024-09-09T03:11:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"discover-nano","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"narl","sideBox":"Learn more about [Discover Nano](https://www.springer.com/journal/11671)","snPcode":"11671","submissionUrl":"https://submission.nature.com/new-submission/11671/3","title":"Discover Nano","twitterHandle":"@SpringerOpen","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5f9ce577-3bfd-4024-80ae-90d8ea3b84f3","owner":[],"postedDate":"November 5th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-10-20T15:58:55+00:00","versionOfRecord":{"articleIdentity":"rs-5054888","link":"https://doi.org/10.1186/s11671-025-04262-2","journal":{"identity":"discover-nano","isVorOnly":false,"title":"Discover Nano"},"publishedOn":"2025-10-16 15:56:51","publishedOnDateReadable":"October 16th, 2025"},"versionCreatedAt":"2024-11-05 12:19:48","video":"","vorDoi":"10.1186/s11671-025-04262-2","vorDoiUrl":"https://doi.org/10.1186/s11671-025-04262-2","workflowStages":[]},"version":"v1","identity":"rs-5054888","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5054888","identity":"rs-5054888","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}