Discovery of high antibacterial and antitumor effects against multi-drug resistant clinically isolated bacteria and MCF-7 and AGS cell lines by biosynthesized silver nanoparticles using Oxalis corniculata extract European Journal of Chemistry 14 (2) (2023) 202-210 European Journal of Chemistry ISSN 2153-2249 (Print) / ISSN 2153-2257 (Online) – Copyright © 2023 The Authors – Atlanta Publishing House LLC – Printed in the USA. This work is published and licensed by Atlanta Publishing House LLC – CC BY NC – Some Rights Reserved. https://dx.doi.org/10.5155/eurjchem.14.2.202-210.2406 European Journal of Chemistry View Journal Online View Article Online Discovery of high antibacterial and antitumor effects against multi-drug resistant clinically isolated bacteria and MCF-7 and AGS cell lines by biosynthesized silver nanoparticles using Oxalis corniculata extract Mohammad Ali Ebrahimzadeh 1, Seyedeh Roya Alizadeh 1 and Zahra Hashemi 2,* 1 Department of Medicinal Chemistry, School of Pharmacy and Pharmaceutical Sciences Research Center, Mazandaran University of Medical Sciences, Sari, Iran 2 Department of Medicinal Chemistry, Faculty of Pharmacy, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran * Corresponding author at: Department of Medicinal Chemistry, Faculty of Pharmacy, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran. e-mail: hngmhashemi@gmail.com (Z. Hashemi). 10.5155/eurjchem.14.2.202-210.2406 Received: 25 December 2023 Received in revised form: 29 January 2023 Accepted: 21 February 2023 Published online: 30 June 2023 Printed: 30 June 2023 The green technique is a unique way to produce functional nanoparticles. We examined the green synthesis of Ag nanoparticles (O-AgNPs) by the extract of Oxalis corniculata. Green- synthesized O-AgNPs were accomplished by monitoring critical factors such as concentration, pH, reaction time, and temperature. Several analytical techniques, including scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction analysis, and UV-Vis spectroscopy, were applied to characterize O-AgNPs. The SEM analysis showed O-AgNPs with a spherical shape and an average size of 33.57 nm. The XRD pattern indicated the face-centered cubic (fcc) structure of the prepared O-AgNPs. The anticancer activity of the synthesized O-AgNPs was investigated in MCF-7 (breast) and AGS (gastric) cell lines, indicating high anticancer effects against selected cell lines. The growth of all selected bacteria containing Gram+ and Gram- was inhibited by O-AgNPs. O-AgNPs showed greater inhibition in comparison to conventional antibiotics. As a result, our green synthesized AgNPs using plant extracts exhibited anticancer and antibacterial activities. Green synthesis Characterization Oxalis corniculate Anticancer activity Silver nanoparticles Antibacterial activity Cite this: Eur. J. Chem. 2023, 14(2), 202-210 Journal website: www.eurjchem.com 1. Introduction Nanotechnology is a scientific breakthrough that uses molecules at the level of atoms or the production of materials and devices with inimitable properties [1]. Nanomaterials were considered due to their high specific surface area and high reactivity [2,3]. In terms of shape, size, and crystal structure, nanoparticles significantly affect their catalytic, optical, and electronic properties [1]. The site-specific activity of the nanoparticles makes a safe dose in administration that induces the efficacy of the drug and reduces unwanted effects [4]. Nanoparticles have shown useful applications and may also have dangerous side effects on humans, aquatic organisms, and plants [5]. Cancer is a rapid and abnormal cell division [6-8] that is the second most current cause of death worldwide, killing more than eight million people per year. Cancer is predicted to improve by more than 50% in the future decades [9]. Current treatments are limited by side effects, drug resistance, and solubility [1]. The use of nanomaterials is a new strategy for cancer therapy that can be used as pharmaceutical carriers to increase drug efficacy [10,11]. Nanotechnology can lead to significant advances in the detection, diagnosis, and treatment of cancer [7]. Fabrication of nanomaterials can be achieved using various physical, chemical, and biogenic synthesis techniques. Synthesis using physical and chemical methods may be created using toxic high radiation and reductants that affect both humans and other living organisms. However, the green synthesis method is a single step for low-cost and environmentally friendly synthesis of nanoparticles. The synt- hesized nanoparticles exhibited antifungal, anti-insecticidal, and antimicrobial activity. Due to their characteristics, all metallic nanoparticles have paved the way for nanoscale researchhers to advance medical research [1]. Green synthesis of Ag nanoparticles (AgNPs) was performed using various plant extracts such as Feijoa sellowiana [12], Crataegus pentagyna [13], Crataegus microphylla [14], Convolvulus fruticosus [15], Ferula persica [16], Stachys inflate [17], Scrophularia striata [18], Allium paradoxum [19], Allium sativum [20], Cleistanthus collinus [21], Piper longum fruit [22], Aloe vera [23], Melia azedarach [24], Pleurotus florida [25] and Dregea volubilis [26]. Effective anticancer, antibacterial, and antioxidant activities have been reported from biosynthesized Ag nanoparticles [14,22-24,27]. Various mechanisms have been proposed for the anticancer effects of AgNPs, including induction of apoptosis, cell cycle arrest, alteration in P53 utilization, and regulation of ABSTRACT RESEARCH ARTICLE KEYWORDS https://dx.doi.org/10.5155/eurjchem.14.2.202-210.2406 https://www.eurjchem.com/ https://dx.doi.org/10.5155/eurjchem.14.2.202-210.2406 mailto:hngmhashemi@gmail.com http://www.eurjchem.com/ https://crossmark.crossref.org/dialog/?doi=10.5155/eurjchem.14.2.202-210.2406&domain=pdf&date_stamp=2023-06-30 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 203 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 cytokine genes [4]. In this study, Oxalis corniculata extract (O. corniculata) extracts with phenol and flavonoids as useful bioactive compounds were used as stabilizing and capping agents to synthesize Ag nanoparticles from Ag ions. O. corniculata Linn. belongs to the Oxalidaceae family and is found in Asia, Europe, America, and Africa. These plants have essential phytochemical compounds for normal human health [28], including β-carotene, Vitamin C, and niacin. The plant also contains different phytomolecules, such as flavonoids, alka- loids, tannins, steroids, polyphenols, glycosidic compounds, lipids, and volatile oil. Leave infusion is used to remove warts and corneal opacities. It works for fever, cold, cough, diarrhea, dysentery, skin problems, urinary tract infections, and sprains [29]. Antiscorbutic, refrigerant, anti-inflammatory, diaphoretic, hepatoprotective [30], antiseptic, antidiabetic, diuretic activi- ties [28], anticancer [31], antimicrobial and antifungal activities [32,33] have been reported in different studies for O. corniculata [34]. This plant is also used to improve cuts, anemia, dyspepsia, piles, dementia, and convulsions [29,35]. The extract exhibited antioxidant and radical scavenging activity [34]. The present study disclosed the biosynthesis route for silver nanoparticles using O. corniculata extract and investigated their anticancer and antimicrobial activities. The green synthesized nanoparticles were characterized using different analytical methods such as UV/vis spectroscopy, energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay exhibited that the prepared AgNPs possess anticancer activities against two cancer cell lines, MCF-7 (breast cancer) and AGS (human gastric carcinoma). In addition, the characterized O-AgNPs were effective antibacterial agents against ATCC strains and resistant clinically isolated bacteria. 2. Experimental 2.1. Preparation of O. corniculata extract The fresh plant was collected in September 2021 from the city of Lahijan in Gilan Province, in the north of Iran. The methanolic extract was obtained by percolating 20 g of powdered O. corniculata aerial parts for 24 h at room tempe- rature (3×1). The extract was filtered using Whatman filter paper (No. 1) and then concentrated at 35 ° C using a rotary evaporator and freeze-dried. 2.2. Biosynthesis of O-AgNPs Silver nitrate (AgNO3) was provided by Fluka Company. Other reagents were used in analytical grade. In the procedure, 0.1 g of crude extract was diluted in 100 mL of deionised water to make it 1 mg/mL, and 15 mL of extract solution at pH = 10 was mixed with 15 mL of 1 mM of aqueous AgNO3 solution, heated at 65 °C with constant stirring for 30 min, and AgNPs were gradually prepared. The same reactions were carried out with various concentrations of AgNO3 at various temperatures and pH levels at various times during the reaction to determine the best conditions. 2.3. Characterization of O-AgNPs and instrumentation The UV-Vis spectrum was recorded at 420 nm for Ag nanoparticles using a UV-Vis spectrophotometer (T80+, China.). The size and morphology of biosynthesized AgNPs were studied by SEM analysis (SCAN BRNO-Mira3 LMU, Germany), and XRD patterns were documented on (Philips PW 1800) using Cu Kα (λ = 1.5406 Å, 40 kV, 40 mA) radiation. 2.4. Antibacterial activity 2.4.1. Minimal inhibitory concentration (MIC) of synthesized O-AgNPs The antibacterial potential of silver nanoparticles was experienced at concentrations of 10 to 200 µg/mL against Pseudomonas aeruginosa (P. aeruginosa, ATCC 27853), Klebsiella pneumonia (K. pneumonia, ATCC 700603), Acinetobacter baumannii (A. baumannii, ATCC 29606), Proteus mirabilis (P. mirabilis, ATCC 25933), Staphylococcus aureus (S. aureus, ATCC 27853), Enterococcus faecalis (E. faecalis, ATCC 29213) and Escherichia coli bacteria (E. coli, ATCC 25922) bacteria. In the process, 100 μL of Mueller Hinton broth culture medium was added to all wells in the microplate, and the appropriate concentrations of the antibiotics examined were obtained. Subsequently, 100 μL of dilute bacterial suspension was moved to all wells; MIC results were studied after 24 hours of incubation. The MIC was stated to be the first well to show no growth. The positive control (bacterial culture medium without extracts) and the negative control (non-bacterial culture medium) were applied for this study. For ATCC strains, ciprofloxacin was utilised as a reference chemical for anti- bacterial activity. Several clinical centers provided seven bacterial pathogens and were known for their biochemical ways. The influence of synthesized O-AgNPs was studied on these resistant bacteria S. aureus, A. baumannii, and P. aeruginosa (the presence of the genes encoding the enzymes of carbapenemase OXA-24 and OXA-23 is confirmed), P. mirabilis (the presence of the genes encoding OXA-23 is positive and NMD, VIM, PhosA3 is negative), peripheral E. faecalis (positive for ermB, TetM, TetL, VanA, VanB), E. coli (positive for OXA-23, PhosA3, and VIM and negative for NMD), K. pneumonia (positive for OXA-23 and NMD also negative for VIM). According to the approach previously stated, the MIC was estimated as the maximum dilution of O-AgNPs, demonstrating complete suppression of the pathogens [16,36]. 2.4.2. Minimum bactericidal concentration (MBC) of synthesized AgNPs The minimum bactericidal concentration (MBC) was determined according to the MIC values; the MIC dilutions were taken and grown on Muller Hinton agar medium separately. After 24 hours of incubation, the plates were investigated for bacterial growth. The lowest concentration of NPs that did not show bacteria growth was assigned as the minimum bactericidal concentration (MBC) [16,37]. 2.5. Anticancer activity of synthesized O-AgNPs The MTT assay was performed; Two cancer cell lines comprising AGS and MCF-7 cells were prepared from the pasture Institute, Iran. Briefly, cells in a suspension containing 6-10×103 were seeded to a 96-well plate and incubated for 24 hours at 37 °C in a humidified atmosphere of 95% air and 5% CO2. All tests were performed three times. Cell cultured super- natant was eliminated after 24 hours, and different concent- rations of silver nanoparticles were diluted in culture medium supplemented with 10% fetal bovine serum (FBS) and antibiotics. Treatment was carried out with varying concent- rations of synthesised O-AgNPs (0.1-50 µg/mL) for 24 hours of incubation. One mL of MTT (5 mg/mL, yellow tetrazole) was taken into each well and incubated for 4 hours. The control wells are made of cell culture medium only. In this test, the conversion of MTT to formazan (insoluble) by 'dehydrogenase enzymes in intact mitochondria' of live cells is evaluated. 204 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 Figure 1. Optimization of concentration of silver nitrate in green synthesis of silver nanoparticles. Figure 2. Optimization of temperature in green synthesis of silver nanoparticles. Pipetting two to three times gradually dissolved the created crystals. A Synergy H1 hybrid multimodel microplate reader (Biotek Instruments, Winooski, Vermont, NE, USA), was used to measure the absorbance at 570-620 nm. Equation (1) was used to convert the optical density (OD) value to the percentage of viability [16]: Percentage of cell viability = OD value of samples OD value of control × 100 (1) 3. Results and discussions 3.1. Green synthesis and characterization The green synthesis of metal NPs such as silver, selenium, gold, titanium, etc., makes available advancements over chemical and physical methods because of their cost-effective, eco-friendly responsive, natural origin, and ease of scaled-up synthesis. There is no need for high energy and temperature [38]. The better mechanism for the green synthesis of AgNPs is the plant reduction process related to their phytochemicals [39]. In plant extracts, biomolecules such as alkaloids, flavonoids, terpenoids, amino acids, tannins, saponins, phenols, carbohydrates, etc., play reducing, capping, and stabilising roles. The different parts of a medical plant, such as stem, roots, leaves, flower, fruit, seed, and bark, can be used for the green synthesis of AgNPs [40]. Studies indicated that AgNPs could be prepared using a green reducing agent. The silver salt will be converted to silver nanoparticles. AgNPs have received great attention for their 'unusual chemical, physical, catalytic, electronic, magnetic, and biological activities'. The properties of silver nanoparticles are significantly different from those of bulk silver metal. These are large surface volume ratios that lead to a large fraction of surface atoms, high surface energy, spatial confinement, and fewer imperfections [41]. Silver nanoparticles have strong bactericidal activity against a wide variety of multidrug resistant strains [42,43]. In addition, they exhibited antifungal [40,44], anti-neoplastic, and antiviral activities [45-47]. Various factors affect the synthesis of metal nanoparticles, including temperature, pH, reaction time, extract, and metal salt concentration. UV-vis spectroscopy is a valuable technique for the initial characterisation of NPs. The synthesis of AgNPs by reducing silver ions was evaluated by ultraviolet-visible (UV-vis) spec- troscopy. As a result of surface plasmon vibrations, AgNPs appear brown in an aqueous medium [48]. The shape, tempe- rature, and dielectric constant of the medium could affect the peak locations of the UV-vis spectra. In contrast, the shape of the UV-Vis peak exhibited a nanoparticle morphology [49]. Surface plasmon resonance (SPR) occurs from the collective oscillations of valence electrons in incident radiation’s electro- magnetic field. This phenomenon plays a key role in determining the absorption spectra of Ag NPs [50]. The λmax for O-AgNPs was observed in the range of 400-500 nm [50]. The UV-Vis spectra of O-AgNPs using a constant concent- ration of the extract (2 mg/mL) with various concentrations of AgNO3 at room temperature are shown in Figure 1. While the concentration of the silver nitrate enhances, the absorption peak gets more sharpness. Furthermore, Figure 2 displays UV- vis spectra of O-AgNPs synthesised at various temperatures. It is clear that as the temperature increases, the absorbance increases with it. So, the best green synthesis was observed at 85 °C. Figure 3 exhibited the effect of pH on the fabrication of AgNPs, raising absorbance with increasing pH at 12. The reaction was monitored for four hours; the UV-vis spectra of O- AgNPs as a function of time are depicted in Figure 4. Further- more, the brown colour of AgNPs appeared shortly after adding the plant extract to the AgNO3 solution. The UV-Vis spectrophotometer records the surface plasmon resonance (SPR) as an absorbance peak around 420 nm. Growth in AgNPs yield led to a higher SPR band by increasing the collision frequency [38]. Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 205 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 Figure 3. Optimization of pH in green synthesis of silver nanoparticles. Figure 4. Optimization of time of incubation in green synthesis of silver nanoparticles. Figure 5. SEM image of AgNPs synthesized by O. corniculata in 500 nm. Furthermore, the creation of bigger nanoparticles generates a redshift in the UV-vis spectrum or a broadening of the peak. On the basis of the SPR bands in our study, the produced O-AgNPs showed small spherical particles. The SEM technique was used to characterise the morphological and structural properties of green synthesised O-AgNPs [50-52]. In Figures 5 and 6, the size and shape of O- AgNPs were seen using the SEM technique. The biosynthesised O-AgNPs had a spherical shape with an average size of 33.57 nm. The EDS results showed the percentage of composition of the mixture. This technique defined Ag0 as the significant element [53]. The elemental silver signal of AgNPs on the vertical axis displays the number of X-ray counts, whereas the horizontal axis exhibits energy in keV [54]. Figure 7 presents the EDS spectrum; A strong peak at 3 keV confirms the formation of AgNPs. X-ray diffraction (XRD) analysis approved the crystallographic structure of green-prepared AgNPs using the characteristic peaks of the X-ray diffraction pattern [42]. An increase or decrease in peak intensity is associated with the amount of constituents [55]. Figure 8 shows the XRD pattern of the prepared O-AgNPs; The diffraction peaks at 2θ = 38.46, 44.5, 64.7, and 77.9° corresponded to Bragg’s from (111), (200), (220) and (311) planes, respectively. The obtained data demonstrated the face-centered cubic (fcc) structure of the synthesized O-AgNPs. Three peaks at 27.94, 32.5, and 46.36° have corresponded to the impurity of AgCl. 3.2. Antibacterial activity of AgNPs AgNPs with multivalent mechanisms display antibacterial activities. Studies have shown that AgNPs possess broad antibacterial activity against infectious microorganisms such as E. coli and S. aureus [56]. The antibacterial effect of AgNPs is related to the oxidation and release of Ag+ ions, so it is an ideal bactericidal agent. 206 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 Figure 6. SEM image of AgNPs synthesized by O. corniculata in 200 nm. Figure 7. EDS spectrum of AgNPs synthesized by O. corniculate. Figure 8. X-ray diffraction analysis of AgNPs synthesised by O. corniculata. The large surface-to-volume ratio and a large fraction of silver nanoparticles’ surface atoms create a tremendous antimicrobial effect compared to silver metal [57]. Further- more, because of their small size, AgNPs quickly penetrate cell membranes and affect intracellular processes. The excellent antibacterial activity of AgNPs is attributed to their well- developed surface, which makes it available to high contacts with the environment. Another considered mechanism is the production of free radicals from the body of the synthesised silver nanoparticles. Free radicals can damage the cell membrane and create a porous layer that eventually kills the cell [58]. Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 207 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 Table 1. ATCC study of green synthesized silver nanoparticles *. MIC (µg/mL) MBC (µg/mL) MIC of extract (µg/mL) Ciprofloxacin (µg/mL) MIC ATCC Bacteria 0.7 12 >4000 0.21 ATCC 29213 S. aureus 10 12 >4000 0.21 ATCC 29212 E. faecalis 1.5 50 >4000 0.23 ATCC27853 P. aeruginosa 1.5 6 >4000 0.25 ATCC 19606 A. baumannii 1.5 1.5 >4000 0.1 ATCC 25922 E. coli 3 12 >4000 0.1 ATCC 700603 K. pneumoniae 3 1.5 >4000 0.1 ATCC 25933 P. mirabilis * MIC: Minimal inhibitory concentration; MBC: Minimal bactericidal concentration. Table 2. Clinical isolated bacteria treatment using green synthesized O-AgNPs *. Susceptibility to antimicrobial agents Treatment with AgNPs Bacteria MC VC OC TC GM PC CIF CEZ ERM CM AK MIC (µg/mL) MBC (µg/mL) R S R R R R R R R R R 6 180 S. aureus R R R R S R R R R R S 6 180 E. faecalis R R R R R R R R R R R 3 45 P. aeruginosa R R R R R R R R R R R 3 90 A. baumannii R R R R R R R R R R R 3 180 E. coli R R R R R R R R R R R 11.5 90 K. pneumoniae R R R R R R R R R R R 6 22.5 P. mirabilis * MIC: Minimal inhibitory concentration; MBC: Minimal bactericidal concentration; MC: Meticillin; VC: Vancomycin; OC: Oxacillin; TC: Tetracycline; GM: Gentamicin; PC: Penicillin; CIF: Ciprofloxacin; CEZ: Ceftazidime; ERM: Erythromycin; CM: Clindamycin; AK: Amikacin. (a) (b) Figure 9. Cell viability of (a) AGS and (b) MCF-7 cancer cell lines (µg/mL). AgNPs cannot simply affect Gram-positive bacteria that have a thick cell wall comprising peptidoglycan. Gram-negative bacteria have a thin lipid layer in their cell wall that causes AgNPs to penetrate them easily [59]. The silver nanoparticles synthesized from O. corniculata showed antibacterial activity against two series of bacteria, including ATCC and clinically isolated strains. The bactericidal mechanism of silver colloid particles against bacteria has not been identified very well until now. The antibacterial activity of AgNPs was organized on P. aeruginosa, K. pneumonia, A. baumannii, P. mirabilis, S. aureus, E. faecalis, and E. coli. In ATCC strains, the lowest MIC was observed for A. baumannii, E. coli, P. mirabilis, which was around 1.5 µg/mL, and MBCs were 6.0, 1.5, and 3.0 µg/mL in treated with O-AgNPs. The results in Table 1 displayed that the AgNPs prepared using O. corniculata affect the growth of the selected bacteria. Subsequently, for further antibacterial evaluation, synthesized nanoparticles were studied against clinically isolated resistant strains (Table 2). Seven clinically isolated bacteria were chosen for this study to test their susceptibility to our manufactured AgNPs. In this regard, A. baumannii and P. aeruginosa were isolated from wounds and phlegm, and the genes encoding carbapene- mase enzymes were found to be resistant to cefepime, piperacillin/tazobactam, ciprofloxacin, aztreonam, ceftazidime, levofloxacin, gentamicin, tobramycin, amikacin, imipenem, and doripenem meropenem. While carbapenems are the last line of treatment, the MIC values for colistin sulphate were reported at 256 and 8 µg/mL, respectively. Our fabricated O-AgNPs considerably inhibited the growth of resistant A. baumannii and P. aeruginosa with a MIC of 3 µg/mL and MBCs of 90 and 45 µg/ml. The isolated K. pneumonia and E. coli from urine were found to be resistant to carbapenems. Colistin sulfate had MICs of 128 and 32 µg/ml, although O-AgNPs had MICs of 11.5 and 3 µg/mL, respectively. Although P. mirabilis isolated urine was resistant to imipenem and meropenem with MICs of 2 and 32 µg/mL, and 208 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 the synthesized AgNPs showed a MIC value of 6 µg/ml with a MBC value of 22.5 µg/mL. E. faecalis isolated from the intestine was resistant to ampicillin, vancomycin, teicoplanin, erythro- mycin, tetracycline, levofloxacin and kanamycin, but was a little sensitive to gentamicin; O-AgNPs could inhibit resistant P. mirabilis with a MIC of 6 µg/mL and a MBC ~180 µg/mL. The S. aureus from the chip was susceptible to vancomycin. Our synthesized O-AgNPs exhibited a MIC of 6 µg/ml and a MBC of 180 µg/mL, respectively. 3.3. Anticancer activity of O-AgNPs AgNPs have exhibited a wide range of biological activities that make them promising agents for infection, cancer, and multidrug resistant cancer cells [51,60]. AgNPs demonstrated unique anticancer activity against various cancer cells [61]. In this work, the anticancer activity (in vitro) of green synthesized AgNPs by O. corniculata extract against AGS and MCF-7 cell lines was proven by MTT assay. The MTT assay determines cell growth inhibitory activity using mitochondrial dehydrogenase activity in living cells [54]. Active mitochondria have these dehydrogenase enzymes, so the reaction occurs only in living cells [53]. Cell viability (%) treated with various concentrations of silver nanoparticles is shown in Figure 9. This study power- fully revealed the antiproliferative activity of biosynthesized silver nanoparticles with a low effect on normal cells compared to the control. As a result, the inhibitory concentration at 50% (IC50) for MCF-7 and AGS cell lines was found to be 3.3 and 3.2 µg/mL of O-AgNPs, respectively. Cell inhibition of AgNPs was reported in ranging from 3.043 to 25 µg/mL for the MCF-7 cell line [62]. The reported results showed that Ag nanoparticles could have induced reactive oxygen species [63]. The NPs enter the cells, interact with the biological components, and damage DNA and/or the mitochondria-dependent apoptosis pathway leading to cell death. In some cases, the inhibition of the AGS cell line treated with silver nanoparticles was observed, ranging from 10 to 100 µg/mL. Silver nanoparticles in the size of 5-35 nm induce apoptosis through the mitochondrial pathway and target function [61]. Studies reported that silver nanoparticles with spherical shapes and a mean diameter of less than 100 nm indicated considerable toxicity against breast cancer cells. In contrast, it showed less toxicity in normal cells [64,65]. Furthermore, silver nanoparticles can induce the apoptosis pathway lacking p53 and cause reactive oxygen species (ROS) [4,61]. 4. Conclusions In the current research, the one-pot biosynthesis of AgNPs in plant extracts (O. corniculata extract) was explained. The polyphenols contained are reducing and capping agents. Various methods confirmed the synthesis of O-AgNPs; the SEM images exhibited NPs with a spherical shape with an average size of 33.57 nm. In addition, the EDS and XRD results proved the formation of O-AgNPs. The Green prepared NPs demonst- rated antibacterial activity against some pathogenic Gram- positive and negative bacteria. Furthermore, AgNPs indicated anticancer efficacy against MCF-7 and AGS cells, making them a potent chemotherapeutic agent. Therefore, according to the current results, AgNPs synthesized using the plant can be considered effective compounds in cancerous cell treatment. Acknowledgements This research was supported by a grant from the Mazandaran University of Medical Sciences (1397-1294). Disclosure statement Conflict of interest: The authors declare that they have no conflict of interest. Ethical approval: All ethical guidelines have been adhered to. Sample availability: Samples of the compounds are available from the author. CRediT authorship contribution statement Conceptualization: Mohammad Ali Ebrahimzadeh; Methodology: Mohammad Ali Ebrahimzadeh, Seyedeh Roya Alizadeh; Formal analysis: Mohammad Ali Ebrahimzadeh, Seyedeh Roya Alizadeh; Investigation: Mohammad Ali Ebrahimzadeh, Seyedeh Roya Alizadeh; Resources: Zahra Hashemi; Funding: Zahra Hashemi; Supervision: Zahra Hashemi; Writing - Original Draft: Mohammad Ali Ebrahimzadeh, Seyedeh Roya Alizadeh, Zahra Hashemi; Review and Editing: Zahra Hashemi. All authors have read and agreed to the published version of the manuscript. ORCID and Email Mohammad Ali Ebrahimzadeh zadeh20@gmail.com https://orcid.org/0000-0002-8769-9912 Seyedeh Roya Alizadeh r.alizadeh.2019@gmail.com https://orcid.org/0000-0001-7435-4635 Zahra Hashemi sz.hashemi@mazums.ac.ir hngmhashemi@gmail.com https://orcid.org/0000-0002-6889-1976 References [1]. Anu, K.; Devanesan, S.; Prasanth, R.; AlSalhi, M. S.; Ajithkumar, S.; Singaravelu, G. Biogenesis of selenium nanoparticles and their anti- leukemia activity. J. King Saud Univ. Sci. 2020, 32, 2520–2526. [2]. Chang, Y.-N.; Zhang, M.; Xia, L.; Zhang, J.; Xing, G. The toxic effects and mechanisms of CuO and ZnO nanoparticles. Materials (Basel) 2012, 5, 2850–2871. [3]. Majumdar, M.; Shivalkar, S.; Pal, A.; Verma, M. L.; Sahoo, A. K.; Roy, D. N. Nanotechnology for enhanced bioactivity of bioactive compounds. In Biotechnological Production of Bioactive Compounds; Elsevier, 2020; pp. 433–466. [4]. Ebrahimzadeh, M. A.; Tafazoli, A.; Akhtari, J.; Biparva, P.; Eslami, S. Engineered silver nanoparticles, A new nanoweapon against cancer. Anticancer Agents Med. Chem. 2019, 18, 1962–1969. [5]. Agarwal, M.; Murugan, M. S.; Sharma, A.; Rai, R.; Kamboj, A.; Sharma, H.; Roy, S. K. Nanoparticles and its toxic effects: A review. Int. J. Curr. Microbiol. App. Sci. 2013, 2, 76–82. [6]. Siegel, R.; DeSantis, C.; Virgo, K.; Stein, K.; Mariotto, A.; Smith, T.; Cooper, D.; Gansler, T.; Lerro, C.; Fedewa, S.; Lin, C.; Leach, C.; Cannady, R. S.; Cho, H.; Scoppa, S.; Hachey, M.; Kirch, R.; Jemal, A.; Ward, E. Cancer treatment and survivorship statistics, 2012. CA Cancer J. Clin. 2012, 62, 220–241. [7]. Misra, R.; Acharya, S.; Sahoo, S. K. Cancer nanotechnology: application of nanotechnology in cancer therapy. Drug Discov. Today 2010, 15, 842–850. [8]. Alizadeh, S. R.; Ebrahimzadeh, M. A. Characterization and anticancer activities of green synthesized CuO nanoparticles, A review. Anticancer Agents Med. Chem. 2021, 21, 1529–1543. [9]. ICGC/TCGA Pan-Cancer Analysis of Whole Genomes Consortium Pan- cancer analysis of whole genomes. Nature 2020, 578, 82–93. [10]. Caputo, F.; De Nicola, M.; Ghibelli, L. Pharmacological potential of bioactive engineered nanomaterials. Biochem. Pharmacol. 2014, 92, 112–130. [11]. Vinardell, M. P.; Mitjans, M. Antitumor activities of metal oxide nanoparticles. Nanomaterials (Basel) 2015, 5, 1004–1021. [12]. Hashemi, Z.; Ebrahimzadeh, M. A.; Biparva, P.; Mortazavi-Derazkola, S.; Goli, H. R.; Sadeghian, F.; Kardan, M.; Rafiei, A. Biogenic silver and zero-Valent iron nanoparticles by Feijoa: Biosynthesis, characterization, cytotoxic, antibacterial and antioxidant activities. Anticancer Agents Med. Chem. 2020, 20, 1673–1687. [13]. Ebrahimzadeh, M. A.; Naghizadeh, A.; Amiri, O.; Shirzadi-Ahodashti, M.; Mortazavi-Derazkola, S. Green and facile synthesis of Ag nanoparticles using Crataegus pentagyna fruit extract (CP-AgNPs) for organic pollution dyes degradation and antibacterial application. Bioorg. Chem. 2020, 94, 103425. [14]. Mortazavi-Derazkola, S.; Ebrahimzadeh, M. A.; Amiri, O.; Goli, H. R.; Rafiei, A.; Kardan, M.; Salavati-Niasari, M. Facile green synthesis and characterization of Crataegus microphylla extract-capped silver nanoparticles (CME@Ag-NPs) and its potential antibacterial and mailto:zadeh20@gmail.com https://orcid.org/0000-0002-8769-9912 mailto:r.alizadeh.2019@gmail.com https://orcid.org/0000-0001-7435-4635 mailto:sz.hashemi@mazums.ac.ir mailto:hngmhashemi@gmail.com https://orcid.org/0000-0002-6889-1976 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 209 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 anticancer activities against AGS and MCF-7 human cancer cells. J. Alloys Compd. 2020, 820, 153186. [15]. Shirzadi-Ahodashti, M.; Mizwari, Z. M.; Hashemi, Z.; Rajabalipour, S.; Ghoreishi, S. M.; Mortazavi-Derazkola, S.; Ebrahimzadeh, M. A. Discovery of high antibacterial and catalytic activities of biosynthesized silver nanoparticles using C. fruticosus (CF-AgNPs) against multi-drug resistant clinical strains and hazardous pollutants. Environ. Technol. Innov. 2021, 23, 101607. [16]. Hashemi, Z.; Mohammadyan, M.; Naderi, S.; Fakhar, M.; Biparva, P.; Akhtari, J.; Ebrahimzadeh, M. A. Green synthesis of silver nanoparticles using Ferula persica extract (Fp-NPs): Characterization, antibacterial, antileishmanial, and in vitro anticancer activities. Mater. Today Commun. 2021, 27, 102264. [17]. Shirzadi-Ahodashti, M.; Hashemi, Z.; Mortazavi, Y.; Khormali, K.; Mortazavi-Derazkola, S.; Ebrahimzadeh, M. A. Discovery of high antibacterial and catalytic activities against multi-drug resistant clinical bacteria and hazardous pollutants by biosynthesized of silver nanoparticles using Stachys inflata extract (AgNPs@SI). Colloids Surf. A Physicochem. Eng. Asp. 2021, 617, 126383. [18]. Ebrahimzadeh, M. A.; Hashemi, Z.; Mohammadyan, M.; Fakhar, M.; Mortazavi-Derazkola, S. In vitro cytotoxicity against human cancer cell lines (MCF-7 and AGS), antileishmanial and antibacterial activities of green synthesized silver nanoparticles using Scrophularia striata extract. Surf. Interfaces 2021, 23, 100963. [19]. Ranjbar, T.; Hashemi, Z.; Sadeghian, F.; Goli, H. R.; Ahanjan, M.; Ebrahimzadeh, M. A. Green Synthesis of Silver Nanoparticles with Allium paradoxum Extract and Evaluation of their Antibacterial Activities. Journal of Mazandaran University of Medical Sciences 2020, 29, 1–11. [20]. Rastogi, L.; Arunachalam, J. Sunlight based irradiation strategy for rapid green synthesis of highly stable silver nanoparticles using aqueous garlic (Allium sativum) extract and their antibacterial potential. Mater. Chem. Phys. 2011, 129, 558–563. [21]. Kanipandian, N.; Kannan, S.; Ramesh, R.; Subramanian, P.; Thirumurugan, R. Characterization, antioxidant and cytotoxicity evaluation of green synthesized silver nanoparticles using Cleistanthus collinus extract as surface modifier. Mater. Res. Bull. 2014, 49, 494–502. [22]. Reddy, N. J.; Nagoor Vali, D.; Rani, M.; Rani, S. S. Evaluation of antioxidant, antibacterial and cytotoxic effects of green synthesized silver nanoparticles by Piper longum fruit. Mater. Sci. Eng. C Mater. Biol. Appl. 2014, 34, 115–122. [23]. Dinesh, D.; Murugan, K.; Madhiyazhagan, P.; Panneerselvam, C.; Kumar, P. M.; Nicoletti, M.; Jiang, W.; Benelli, G.; Chandramohan, B.; Suresh, U. Mosquitocidal and antibacterial activity of green- synthesized silver nanoparticles from Aloe vera extracts: towards an effective tool against the malaria vector Anopheles stephensi? Parasitol. Res. 2015, 114, 1519–1529. [24]. Sukirtha, R.; Priyanka, K. M.; Antony, J. J.; Kamalakkannan, S.; Thangam, R.; Gunasekaran, P.; Krishnan, M.; Achiraman, S. Cytotoxic effect of Green synthesized silver nanoparticles using Melia azedarach against in vitro HeLa cell lines and lymphoma mice model. Process Biochem. 2012, 47, 273–279. [25]. Kaur, G.; Kalia, A.; Sodhi, H. S. Size controlled, time-efficient biosynthesis of silver nanoparticles from Pleurotus florida using ultra- violet, visible range, and microwave radiations. Inorg. Nano-met. Chem. 2020, 50, 35–41. [26]. Das, B.; De, A.; Podder, S.; Das, S.; Ghosh, C. K.; Samanta, A. Green biosynthesis of silver nanoparticles using Dregea volubilis flowers: Characterization and evaluation of antioxidant, antidiabetic and antibacterial activity. Inorg. Nano-met. Chem. 2021, 51, 1066–1079. [27]. Khorrami, S.; Zarrabi, A.; Khaleghi, M.; Danaei, M.; Mozafari, M. R. Selective cytotoxicity of green synthesized silver nanoparticles against the MCF-7 tumor cell line and their enhanced antioxidant and antimicrobial properties. Int. J. Nanomedicine 2018, 13, 8013–8024. [28]. Kirtikar, K.; Basu, B.; Blatter, E. Indian medicinal plants. International Book Distributors, Delhi, India, Lalit Mohan Basu, Allahabad, India, 1935. https://www.indianculture.gov.in/ebooks/indian-medicinal- plants (accessed January 10, 2022). [29]. Sarkar, T.; Ghosh, P.; Poddar, S.; Choudhury, S.; Sarkar, A.; Chatterjee, S. Oxalis corniculata Linn. (Oxalidaceae): A brief review. J. Pharmacogn. Phytochem. 2020, 9, 651–655. [30]. Khan, M. R.; Marium, A.; Shabbir, M.; Saeed, N.; Bokhari, J. Antioxidant and hepatoprotective effects of Oxalis corniculata against carbon tetrachloride (CCl4) induced injuries in rat. Afr. J. Pharm. Pharmacol. 2012, 6, 2255–2267. [31]. Salahuddin, H.; Mansoor, Q.; Batool, R.; Farooqi, A. A.; Mahmood, T.; Ismail, M. Anticancer activity of Cynodon dactylon and Oxalis corniculata on Hep2 cell line. Cell. Mol. Biol. (Noisy-le-grand) 2016, 62, 60–63. [32]. Rehman, A.; Rehman, A.; Ahmad, I. Antibacterial, antifungal, and insecticidal potentials ofOxalis corniculataand its isolated compounds. Int. J. Anal. Chem. 2015, 2015, 1–5. [33]. Rahman, M. S.; Khan, M. M. H.; Jamal, M. A. H. M. Anti-bacterial Evaluation and Minimum Inhibitory Concentration Analysis of Oxalis corniculata and Ocimum santum against Bacterial Pathogens. Biotechnology (Faisalabad) 2010, 9, 533–536. [34]. Durgawale, P. P.; Hendre, A. S.; Phatak, R. S. Gc/ms characterization, antioxidant and free radical scavenging capacities of methanolic extract of Oxalis corniculata Linn: An ayurvedic herb. Rasayan J. Chem. 2015, 8, 271–278. [35]. Kritikar, K. R.; Basu, B. D. Indian Medicinal Plants. In: Compositae, Vol. 2; nternational Book Distributors: Dehradun, 1987. [36]. Hashemi, Z.; Mizwari, Z. M.; Mohammadi-Aghdam, S.; Mortazavi- Derazkola, S.; Ali Ebrahimzadeh, M. Sustainable green synthesis of silver nanoparticles using Sambucus ebulus phenolic extract (AgNPs@SEE): Optimization and assessment of photocatalytic degradation of methyl orange and their in vitro antibacterial and anticancer activity. Arab. J. Chem. 2022, 15, 103525. [37]. Alizadeh, S. R.; Seyedabadi, M.; Montazeri, M.; Khan, B. A.; Ebrahimzadeh, M. A. Allium paradoxum extract mediated green synthesis of SeNPs: Assessment of their anticancer, antioxidant, iron chelating activities, and antimicrobial activities against fungi, ATCC bacterial strains, Leishmania parasite, and catalytic reduction of methylene blue. Mater. Chem. Phys. 2023, 296, 127240. [38]. Sandeep, S.; S. Santhosh, A.; Kumara Swamy, N.; S. Suresh, G.; S. Melo, J.; Mallu, P. Biosynthesis of silver nanoparticles using Convolvulus pluricaulis leaf extract and assessment of their catalytic, electrocatalytic and phenol remediation properties. Adv. Mater. Lett. 2016, 7, 383–389. [39]. Pandit, R. Green synthesis of silver nanoparticles from seed extract of Brassica nigra and its antibacterial activity. Nusant. Biosci. 1970, 7, 15–19. [40]. Elumalai, K.; Velmurugan, S. Green synthesis, characterization and antimicrobial activities of zinc oxide nanoparticles from the leaf extract of Azadirachta indica (L.). Appl. Surf. Sci. 2015, 345, 329–336. [41]. Kumar, B.; Kumari, S.; Cumbal, L.; Debut, A. Lantana camara berry for the synthesis of silver nanoparticles. Asian Pac. J. Trop. Biomed. 2015, 5, 192–195. [42]. Sankar, R.; Karthik, A.; Prabu, A.; Karthik, S.; Shivashangari, K. S.; Ravikumar, V. Origanum vulgare mediated biosynthesis of silver nanoparticles for its antibacterial and anticancer activity. Colloids Surf. B Biointerfaces 2013, 108, 80–84. [43]. Bose, D.; Chatterjee, S. Antibacterial activity of green synthesized silver nanoparticles using Vasaka (Justicia adhatoda L.) leaf extract. Indian J. Microbiol. 2015, 55, 163–167. [44]. Nasrollahi, A.; Pourshamsian, K.; Mansourkiaee, P. Antifungal activity of silver nanoparticles on some of fungi. International journal of nano dimension 2011, 1, 233–239. [45]. Mori, Y.; Ono, T.; Miyahira, Y.; Nguyen, V. Q.; Matsui, T.; Ishihara, M. Antiviral activity of silver nanoparticle/chitosan composites against H1N1 influenza A virus. Nanoscale Res. Lett. 2013, 8, 93. [46]. Haggag, E. G.; Elshamy, A. M.; Rabeh, M. A.; Gabr, N. M.; Salem, M.; Youssif, K. A.; Samir, A.; Bin Muhsinah, A.; Alsayari, A.; Abdelmohsen, U. R. Antiviral potential of green synthesized silver nanoparticles of Lampranthus coccineus and Malephora lutea. Int. J. Nanomedicine 2019, 14, 6217–6229. [47]. Jacob, S. J. P.; Prasad, V. L. S.; Sivasankar, S.; Muralidharan, P. Biosynthesis of silver nanoparticles using dried fruit extract of Ficus carica - Screening for its anticancer activity and toxicity in animal models. Food Chem. Toxicol. 2017, 109, 951–956. [48]. Lakshmanan; Sathiyaseelan; Kalaichelvan; Murugesan Plant- mediated synthesis of silver nanoparticles using fruit extract of Cleome viscosa L.: Assessment of their antibacterial and anticancer activity. Karbala Int. J. Mod. Sci. 2018, 4, 61–68. [49]. Dhand, V.; Soumya, L.; Bharadwaj, S.; Chakra, S.; Bhatt, D.; Sreedhar, B. Green synthesis of silver nanoparticles using Coffea arabica seed extract and its antibacterial activity. Mater. Sci. Eng. C Mater. Biol. Appl. 2016, 58, 36–43. [50]. Al-Sheddi, E. S.; Farshori, N. N.; Al-Oqail, M. M.; Al-Massarani, S. M.; Saquib, Q.; Wahab, R.; Musarrat, J.; Al-Khedhairy, A. A.; Siddiqui, M. A. Anticancer potential of green synthesized silver nanoparticles using extract of Nepeta deflersiana against human cervical cancer cells (HeLA). Bioinorg. Chem. Appl. 2018, 2018, 9390784. [51]. Jadhav, K.; Deore, S.; Dhamecha, D.; H R, R.; Jagwani, S.; Jalalpure, S.; Bohara, R. Phytosynthesis of silver nanoparticles: Characterization, biocompatibility studies, and anticancer activity. ACS Biomater. Sci. Eng. 2018, 4, 892–899. [52]. Mollick, M. M. R.; Rana, D.; Dash, S. K.; Chattopadhyay, S.; Bhowmick, B.; Maity, D.; Mondal, D.; Pattanayak, S.; Roy, S.; Chakraborty, M.; Chattopadhyay, D. Studies on green synthesized silver nanoparticles using Abelmoschus esculentus (L.) pulp extract having anticancer (in vitro) and antimicrobial applications. Arab. J. Chem. 2019, 12, 2572– 2584. [53]. Naraginti, S.; Li, Y. Preliminary investigation of catalytic, antioxidant, anticancer and bactericidal activity of green synthesized silver and gold nanoparticles using Actinidia deliciosa. J. Photochem. Photobiol. B 2017, 170, 225–234. [54]. Kummara, S.; Patil, M. B.; Uriah, T. Synthesis, characterization, biocompatible and anticancer activity of green and chemically https://www.indianculture.gov.in/ebooks/indian-medicinal-plants https://www.indianculture.gov.in/ebooks/indian-medicinal-plants 210 Ebrahimzadeh et al. / European Journal of Chemistry 14 (2) (2023) 202-210 2023 – European Journal of Chemistry – CC BY NC – DOI: 10.5155/eurjchem.14.2.202-210.2406 synthesized silver nanoparticles - A comparative study. Biomed. Pharmacother. 2016, 84, 10–21. [55]. Abdelghany, T. M.; Al-Rajhi, A. M. H.; Al Abboud, M. A.; Alawlaqi, M. M.; Ganash Magdah, A.; Helmy, E. A. M.; Mabrouk, A. S. Recent advances in green synthesis of silver nanoparticles and their applications: About future directions. A review. Bionanoscience 2018, 8, 5–16. [56]. Tang, S.; Zheng, J. Antibacterial activity of silver nanoparticles: Structural effects. Adv. Healthc. Mater. 2018, 7, e1701503. [57]. Jayashree, S.; Vani, G. S. In vitro study on antibacterial activity of aqueous extract and silver nanoparticles of Andrographis paniculata. Int. J. Curr. Microbiol. Appl. Sci. 2016, 5, 400–406. [58]. Elgorban, A. M.; Al-Rahmah, A. N.; Sayed, S. R.; Hirad, A.; Mostafa, A. A.- F.; Bahkali, A. H. Antimicrobial activity and green synthesis of silver nanoparticles usingTrichoderma viride. Biotechnol. Biotechnol. Equip. 2016, 30, 299–304. [59]. Prasannaraj, G.; Venkatachalam, P. Green engineering of biomolecule- coated metallic silver nanoparticles and their potential cytotoxic activity against cancer cell lines. Adv. Nat. Sci. Nanosci. Nanotechnol. 2017, 8, 025001. [60]. Abdel-Fattah, W. I.; W Ali, G. On the anti-cancer activities of silver nanoparticles. J. Appl. Biotechnol. Bioeng. 2018, 5, 43–46. [61]. Firdhouse, J.; Lalitha, P. Apoptotic efficacy of biogenic silver nanoparticles on human breast cancer MCF-7 cell lines. Prog. Biomater. 2015, 4, 113–121. [62]. Salehi, S.; Shandiz, S. A. S.; Ghanbar, F.; Darvish, M. R.; Ardestani, M. S.; Mirzaie, A.; Jafari, M. Phytosynthesis of silver nanoparticles using Artemisia marschalliana Sprengel aerial part extract and assessment of their antioxidant, anticancer, and antibacterial properties. Int. J. Nanomedicine 2016, 11, 1835–1846. [63]. Saravanan, M.; Barabadi, H.; Ramachandran, B.; Venkatraman, G.; Ponmurugan, K. Emerging plant-based anti-cancer green nanomaterials in present scenario. In Engineered Nanomaterials and Phytonanotechnology: Challenges for Plant Sustainability; Elsevier, 2019; pp. 291–318. [64]. Barabadi, H.; Hosseini, O.; Damavandi Kamali, K.; Jazayeri Shoushtari, F.; Rashedi, M.; Haghi-Aminjan, H.; Saravanan, M. Emerging theranostic silver nanomaterials to combat lung cancer: A systematic review. J. Cluster Sci. 2020, 31, 1–10. Copyright © 2023 by Authors. This work is published and licensed by Atlanta Publishing House LLC, Atlanta, GA, USA. The full terms of this license are available at http://www.eurjchem.com/index.php/eurjchem/pages/view/terms and incorporate the Creative Commons Attribution-Non Commercial (CC BY NC) (International, v4.0) License (http://creativecommons.org/licenses/by-nc/4.0). By accessing the work, you hereby accept the Terms. This is an open access article distributed under the terms and conditions of the CC BY NC License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited without any further permission from Atlanta Publishing House LLC (European Journal of Chemistry). No use, distribution, or reproduction is permitted which does not comply with these terms. Permissions for commercial use of this work beyond the scope of the License (http://www.eurjchem.com/index.php/eurjchem/pages/view/terms) are administered by Atlanta Publishing House LLC (European Journal of Chemistry). http://www.eurjchem.com/index.php/eurjchem/pages/view/terms http://creativecommons.org/licenses/by-nc/4.0 http://www.eurjchem.com/index.php/eurjchem/pages/view/terms 1. Introduction 2. Experimental 2.1. Preparation of O. corniculata extract 2.2. Biosynthesis of O-AgNPs 2.3. Characterization of O-AgNPs and instrumentation 2.4. Antibacterial activity 2.4.1. Minimal inhibitory concentration (MIC) of synthesized O-AgNPs 2.4.2. Minimum bactericidal concentration (MBC) of synthesized AgNPs 2.5. Anticancer activity of synthesized O-AgNPs 3. Results and discussions 3.1. Green synthesis and characterization 3.2. Antibacterial activity of AgNPs 3.3. Anticancer activity of O-AgNPs 4. Conclusions Acknowledgements Disclosure statement CRediT authorship contribution statement ORCID and Email References PrintField10: PrintField11: PrintField12: PrintField13: PrintField14: PrintField15: PrintField16: PrintField17: PrintField18: PrintField20: PrintField21: PrintField22: PrintField23: PrintField24: PrintField25: PrintField26: PrintField27: PrintField28: