A Comparative Study on Biosynthesized Silver Nanoparticles from H. undatus Fruit Peel and its Therapeutic Applications

A Comparative Study on Biosynthesized Silver Nanoparticles from H. undatus Fruit Peel and its Therapeutic Applications | 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 Research Article A Comparative Study on Biosynthesized Silver Nanoparticles from H. undatus Fruit Peel and its Therapeutic Applications Aswini Anguraj, Helan Soundra Rani Michael, Sathish Sugumaran, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3805384/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Mar, 2024 Read the published version in Discover Nano → Version 1 posted 7 You are reading this latest preprint version Abstract The green synthesis of nanoparticles has gained significant impacts in various fields due to their eco-friendly approach. In this study, silver nanoparticles were synthesized from aqueous extract of H. undatus fruit peel. The presence of silver nanoparticles (AgNPs) was analyzed using characterization methods such as UV-Vis, FTIR, GCMS, XRD, EDAX and FESEM. The synthesized AgNPs showed the highest antibacterial activity against E. coli when compared to S. pneumoniae . The highest antifungal activity was observed against C. albicans than C. tropicalis. IC 50 value of antibiofilm activity of AgNPs was recorded as 2.81 µg/ml whereas H. undatus peel extract exhibited the value of 1.34 µg/ml. The invitro antioxidant activity of AgNPs was evaluated using two different methods. A strong DPPH radical scavenging activity of AgNPs and fruit peel extract was observed with the IC 50 values of 3.8 and 2.03 µg/ml respectively. On other hand, nitric oxide radical scavenging activity were recorded and IC 50 values was calculated to be 2.8 and 2.3 µg/ml. The AgNPs demonstrated thrombolytic activity in human blood with 10, 32.36 and 56.25% lysis. The cytotoxicity of AgNPs possessed minimum activity with an IC 50 0.2 µg/ml and peel extract showed the maximum cytotoxicity activity with an IC 50 0.3 µg/ml. The findings of this study demonstrate that the synthesized AgNPs from H. undatus peel extract used as a potential candidate for treating prostate cancer. H. undatus antibacterial antifungal antioxidant thrombolytic anticancer activity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Introduction Nanotechnology is an emerging field that aims to create molecular, atomic and supramolecular properties [ 1 ]. It has gained importance in cosmetics, drug delivery, biomedical, cosmetics and cancer therapy [ 2 ]. Metallic nanoparticles such as silver, copper oxide, zinc oxide and gold nanoparticles are synthesized extensively in nanostructured form [ 3 ]. Among the above-mentioned metals, silver nanoparticles gained more attention due to their unique characteristics. It can be synthesized by physical, chemical and biological methods [ 4 ]. Biological entities such as bacteria, fungi, yeasts and plants have been utilized in the synthesis of silver nanoparticles, resulting in less toxic, cost-efficient, stable and eco-friendly alternatives to chemical methods [ 5 ]. Among the biological methods, microbe-mediated synthesis of nanoparticles is not commercially feasible due to its high aseptic condition requirement. Plant-mediated silver nanoparticles offer several advantages over microorganisms. Because they can be easily scaled up, making them more practical for large-scale production. The nanoparticles can be effectively capped and reduced by extracting active components from plants such as flavonoids, alkaloids, terpenes, tannins and saponins [ 6 ]. Silver nanoparticles from plant extracts have a wide range of applications in the fields of food, health, agriculture, disease control, pharmaceuticals and medicine [ 7 ]. Silver nanoparticles have shown potential antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral and anticancer, anti-plasmodial and anti-platelets effects [ 8 ]. Prostate carcinoma (PC) is the primary reason for cancer in the male reproductive system, affecting a significant number of newly diagnosed cancer patients, accounting for around 25% of cases. When combined with colorectal cancer, PC becomes the second leading cause of male cancer-related deaths in the US, following lung and bronchus cancer. The majority of prostate cancer cases, around 80% are found in the peripheral zone, while 10–20% occur in the transition zone [ 9 ]. Treatment methods like cholesterol-lowering drugs, biopsies, and conventional heating techniques can be costly and may have side effects such as pain or discomfort due to unwanted reflection, scattering or absorption [ 10 ]. It is crucial to explore alternative treatments or medications to address these limitations. Nanoparticles have shown promise in the treatment of prostate cancer patients [ 11 ]. The toxicity associated with this treatment is moderate and only temporarily affects the quality of life. Hence, there is a need to discover new cancer therapies that are both biocompatible and cost-effective [ 12 ]. Dragon fruit ( Hylocereus undatus ) is a cactus species that is commonly consumed in Asian countries. It is a tropical fruit with a red peel and green pins. It is a great source of low-calorie dietary fiber [ 13 ]. It contains antioxidants that can help to prevent various diseases such as cancer, cardiovascular issues, diabetes, urinary and gastrointestinal problems [ 14 ]. The peel of the dragon fruit is composed of pectin, phyllocactin, steroids, betacynanin and betanin. Previous studies reported that it has high antioxidant, antimicrobial, cardioprotective, anti-inflammatory and anticancer properties [ 15 ]. In this study, we focused on the use of Hylocereus undatus for the bioreduction of silver ions in an aqueous medium which leads to the formation of silver nanoparticles. The synthesized nanoparticles were characterized and evaluated for antibacterial, antifungal, antioxidant and thrombolytic activity. Additionally, we examined the potential of these AgNPs to inhibit the growth of prostate cancer cells. Materials and methods Sample collection and extraction In the present study, Hylocereus undatus was collected from the local market, R. S. Puram, Coimbatore. The peel of H. undatus was washed with double distilled water until all the impurities were removed and kept for drying in a shady sterile environment. The dried samples were finely grounded into powder and further processed for extraction. Preparation of aqueous extract The H. undatus aqueous extract was prepared by boiling 20 g of powdered peel in 500 mL of distilled water for 15 minutes. The extracts was agitated and covered until it reached room temperature. The crude extract was filtered through Whatman filter paper No.1. The resulting extract was stored in a container at 2°C for further study. Synthesis of silver nanoparticles A reaction mixture consisting of 220 ml of dragon fruit peel extract and 110 ml of 10 mM silver nitrate was stirred for 24 hours at room temperature using a magnetic stirrer. The reaction mixture was then centrifuged at 12,000 rpm for 15 minutes at 4 o C and the supernatant was discarded. The resulting pellet was dissolved in distilled water and centrifuged again under the same conditions as mentioned before to ensure the purity of the nanosilver. The resultant pellet was dried and stored for further characterization [ 16 ]. Characterization of silver nanoparticles The synthesized AgNPs were characterized using a UV-visible spectrophotometer, Fourier transform infrared spectrophotometer (FTIR), Gas chromatography-mass spectrometry (GCMS), X-ray diffraction (XRD), Energy Dispersive X-ray spectroscopy (EDAX), and Field Emission Scanning Electron Microscopy (FESEM). Antibacterial activity The disc diffusion method was used to evaluate the antibacterial activity of AgNPs against both gram-positive ( Streptococcus pneumoniae ) and gram-negative ( Escherichia coli ) bacteria. The bacterial strains were spread on nutrient agar plates using a sterile cotton swab. The disc was treated with two distinct concentrations (40 and 80 µL) of synthesized AgNPs. The kanamycin was used as a positive control and distilled water was used as a negative control. Then the plates were incubated at 37 o C for 24 hours. The antibacterial screening was evaluated by measuring the zone of inhibition [ 17 ]. The experiments were replicated three times and the diameter of the zone of inhibition was reported as mean ± standard deviation. Antifungal activity The antifungal potential of the biosynthesized AgNPs against two pathogenic strains such as Candida albicans and Candida tropicalis was evaluated using the agar well diffusion method as described by Joseph et al. [ 18 ]. Wells with a diameter of 8 mm were punched in the potato dextrose agar medium and 100 µL of each microorganism was spread using a sterile cotton swab. Distilled water was used as the negative control while Voriconazole as the positive control. Two concentrations such as 40 and 80 µL of AgNPs were added to the wells and incubated at 37 o C for 24 hours. Following, the zones were determined and recorded. Antibiofilm activity A 96-well microtiter plate was used to evaluate the anti-biofilm effectiveness of the AgNPs and H. undatus peel extract. Each well of the plates was filled with 180 µl of Muller Hinton broth and 10 µl of the test pathogens. Then 10 µl of AgNPs and H. undatus peel extract were added and the mixture was thoroughly mixed. The test plates were then incubated for 24 hours at 37 o C. After incubation, the microtiter plates were washed with phosphate buffer saline to remove non-adherent bacteria. The plates were air-dried for 45 minutes and the wells were fixed with sodium acetate. Crystal violet stain was added and incubated in the dark for 30 minutes. Excess dye was removed by washing with water. The plates were air-dried and 200 µl ethanol was added to each well and absorbance was measured at 620 nm [ 19 ] Thrombolytic activity The thrombolytic activity of AgNPs and the peel extract of H. undatus was performed according to the method followed by Devi et al. [ 20 ] with slight modification. After determining the weight of the empty Eppendorf tube 20, 40 and 60 µL of blood were drawn in different tubes and incubated at 37◦ C for 30 minutes. After the clot formation, the tubes were re-weighed to determine the weight of the blood clot (X). This was achieved by subtracting the initial weight of the tube. 100 µL of AgNPs at two different concentrations (500 and 1000 µg/mL) was added to the tubes and incubated at 37 o C for 90 minutes. To calculate the weight of the non-lyzed blood clot (Y), the initial weight of the tube was subtracted from the final weight of the tube, which contained the remaining blood clot. 100 µL of distilled water was used as a negative control and streptokinase as a positive control. Thrombolytic activity was calculated using the following equation Thrombolysis (%) = X – Y X 100/X Antioxidant activity DPPH radical scavenging assay The scavenging ability of the AgNPs and H. undatus peel extract was determined using the DPPH method described by Phull et al. [ 21 ]. The DPPH assay was performed in a 96-well plate having ascorbic acid as the reference standard. AgNPs and H. undatus peel extract (20, 40, 60, 80, and 100 µl) was taken from the stock solution and poured into each well of the plate. Then, 100 µl of the DPPH solution was added and incubated at 27°C for 20 minutes. The measurement of the absorbance of the reaction mixture was carried out at 517 nm using a microplate reader. The scavenging activity was calculated as % DPPH radical scavenging activity = control OD − sample OD/control OD x 100 Nitric oxide radical scavenging activity The nitric oxide radical scavenging activity of AgNPs and H. undatus peel extract was carried out using the method outlined by Makhija et al. [ 22 ] with slight modification. Various concentrations of the AgNPs and H. undatus peel extract (20, 40, 60 80 and 100 µL) were combined with 3 mL of sodium nitroprusside (10 mMol/L) in phosphate buffer saline (0.2 mMol/L, pH 7.4). The mixture was then incubated at 25 o C for 150 minutes. Following the addition of 500 µL of Griess reagent (composed of 2% orthophosphoric acid, 1% sulphanilamide, and 0.1% N-1-naphthyl ethylenediamine dihydrochloride) was added. The absorbance was measured at 546 nm. All measurements were conducted in triplicate and the mean values were calculated. Nitric oxide scavenged (%) = A control – A test / A control x 100 Cytotoxicity assay Cell culture The human prostate cancer cell line (PC3) was purchased from NCCS, Pune. The cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic. The cells were kept at a temperature of 37 o C in 5% CO 2 and 95% relative humidity (Devi and Bhimba, 2012). MTT assay The cytotoxic effect of AgNPs and H. undatus peel extract was carried out by MTT assay. PC3 cells were seeded in a 96 well plate and incubated for 24 hours, in 200 µl of DMEM with 10% FBS. Various concentrations (12, 25, and 55 µg/ml) of AgNPs and H. undatus peel extract were added and incubated for 48 hours. After the addition of MTT, the cells were again incubated at 37 o C for 4 hours. The media was then removed, 200 µl of DMSO was added to each well and the absorbance was read at 570 nm [ 23 ]. Results and Discussion Characterization of silver nanoparticles The current study revealed that the H. undatus fruit peel extract to the silver in the form of nitrate. This interaction resulted in the formation of a pale pink to dark brown indicating the presence of silver nanoparticles [ 24 ] as shown in Fig. 1 . Similar color changes have also been observed in the previous study [ 16 ]. The UV-vis spectra were recorded after time intervals of 20 min, 25 min, 30 min, 35 min, 49 min, 45 min, 50 min, 55 min, 60 min, 65 min and 70 min. Figure 2 shows the absorption spectra of AgNPs using H. undatus fruit peel extract. The UV spectrum has exhibited a strong peak at 417 nm which confirms the formation of AgNPs. The purity of the synthesized NPs was checked by FTIR analysis within the range of 400–4000 cm − 1 [ 25 ]. Figure 3 shows the FTIR spectra of biosynthesized AgNPs and the absorption peak bands at 3734.19, 3332.99, 2310.72, 1593.20, 1365.60, 1323.17, 1238.30, 1041.56, 921.97, 821.68, 678.94, 601.79, 559. 36. These absorbance bands are known to be associated with stretching vibrations of O-H (alcohols), O-H (alcohols), C = N (nitriles), C-C (aromatics), C-H (alkanes), C-N (aromatic amines), C-N (aliphatic amines), C-O (alcohols), O-H (carboxylic acids), C-Cl (alkyl halides), C-Br (alkyl halides), C-Br (alkyl halides), C- Br (alkyl halides). H. undatus fruit is primarily composed of betanin, pectin, hylocerenin, betacyanin and phyllocactin [ 26 ]. This study has confirmed the presence of a significant amount of phytocompounds and these substances may potentially be responsible for the reduction, capping and synthesis of AgNPs. The GC-MS analysis of the peel extract of H. undatus was conducted to determine the bioactive chemical compounds present in the extract. Figure 4 displays the GC-MS spectrum with peaks and retention time. The peel extract of H. undatus has shown the presence of the major seven bioactive compounds such as 6-methyl-2, (4-bromophenyl)-7-phenylmethylindolizine, N-[(4,6-Dimethoxynaphthalen-1-yl) methylene], 2,5-dichloro-4-hydroxyphenylamine, Oxacycloheptadec-8-en-2-one, Androstan-17one, 3-ethyl-3-hydroxy, Pyridine-3-carboxamide, 6-chloro-4-trifluoromethyl-N-[2,4-dichloeo-6-methyl]-N-methyl, 1,2-Dipalmitoyl 3-acetyl glycerol Gallic acid, and Triamcinolone acetonide. The peak intensity of N-[(4,6Dimethoxynaphthalen-1-ylmethylene], 2,5-dichloro-4-hydroxyphenylamine was found to be higher with a retention time of 30.12 min followed by Oxacycloheptadec-8-en-2-one with a retention time of 30.74 (Table 1 ). Table 1 Chemical composition of H. undatus peel extract S.No RT Compound name Molecular formula Molecular weight Peak area % 1 27.99 6-methyl-2,(4-bromophenyl)-7- phenylmethylindolizine C 22 H 18 BrN 376.289 7.991989 2 30.12 N-[(4,6Dimethoxynaphthalen-1- ylmethylene], 2,5-dichloro-4- hydroxyphenylamine 𝐶19𝐻15𝐶𝑙2𝑁𝑂3 376.233 8.799596 3 30.74 Oxacycloheptadec-8-en-2-one C16H28O2 252.392 15.430418 4 30.88 Androstan-17one 3-ethyl-3- hydroxy C21H34O2 318.49346 13.414466 5 31.49 Pyridine-3-carboxamide, 6- chloro-4-trifluoromethyl-N-[2,4- dichloeo-6-methyl]-N-methyl C15H10C13F3N2O 397.606 11.854260 6 31.80 1,2-Dipalmitoyl 3-acetyl glycerol Gallic acid C37H70O6 610.948 11.854137 7 32.00 Triamcinolone acetonide. C24H31FO6 434.497 10.116891 The synthesized AgNPs are characterized using XRD to confirm the presence of silver ions and to know the structural information [ 27 ]. Figure 5 shows the XRD pattern of AgNPs which confirmed the crystalline nature of synthesized AgNPs. The peaks at 2𝜃 of 27.57, 32.09, 37.88, 44.65, 46.02, 54.59, 57.19, 64.16, 67.32, 72.62, and 76.42 correspond to the crystalline planes of the face-centered cubic structure of metallic silver. The intense peak at 32.09 possibly suggests the presence of silver ions as the major constituent in the biosynthesized AgNPs. A similar result was observed by Phongtongpasuk et al. [ 16 ] who identified the most intense peak at 32.5 indicated the presence of AgNPs. The elemental analysis of the material is depicted in Fig. 6 , illustrating the synthesized AgNPs through the EDAX spectrum [ 28 ]. This analysis demonstrates a prominent indication of a metallic silver area at 3 KeV and confirms the formation of silver nanoparticles synthesized through the utilization of H. undatus peel (Fig. 6 ). The Ag peak showed a weight percentage of 35.12 and an atomic percentage of 8.02. A low calcium signal was observed and five moderate signals for carbon, oxygen, sodium, chloride and potassium were detected as a result of the chemicals used in the sample preparation. FE-SEM images of AgNPs synthesized using the fruit peel extract of H. undatus are shown in Fig. 7 . The surface morphology of AgNPs showed a spherical shape with agglomeration. In the present study, the histogram of the particle size ranges from 52–79 nm. The increased concentration of bioactive compounds in the colloidal solution could potentially lead to the formation of nanoclusters. The signals produced from the interaction between the electrons and the sample provide valuable information regarding the sample's external morphology, chemical composition, crystalline structure and material orientation. Antibacterial activity The antibacterial activity of synthesized AgNPs were tested at various concentrations by the disc diffusion method. Figure 8 illustrates the zone of inhibition around the individual bacterial culture. The zone of inhibition was measured to be 1.2 ± 0.2 and 1.6 ± 0.1 cm for E. coli while in the case of S. pneumoniae 1.1 ± 0.1 and 1.3 ± 0.1 cm was measured at a concentration of 40 µl and 80 µl of synthesized AgNPs. In the present study, the synthesized AgNPs showed the highest antibacterial activity against E. coli when compared to S . pneumoniae . The potential cause of this occurrence could be attributed to the existence of phytochemicals within the H. undatus. The enhanced antibacterial efficacy of AgNPs can be predominantly attributed to the synergistic interplay between the aforementioned nanoparticles and the naturally occurring chemicals found within the extract as previously substantiated in a prior investigation. It has been determined that AgNPs can liberate silver ions within bacterial cells, thus enhancing their bactericidal properties [ 29 ]. Antifungal activity The antifungal potential of the biosynthesized AgNPs towards two pathogenic strains such as Candida albicans and Candida tropicalis was evaluated using the well diffusion method. The antifungal activity can be identified by the zone of inhibition (Fig. 9 ). The results indicate that the biosynthesized AgNPs possess significant antifungal activity against two fungal strains that were tested. There is no zone of inhibition was observed in the control. The synthesized AgNPs that could inhibit the fungal growth of C. albicans were found to be 1.1 ± 0.1 and 1.3 ± 0.3, while C. tropicalis was measured at 0.8 ± 0.2 and 1.2 ± 0.2 cm at a concentration of 40 and 80 µL respectively. The highest antifungal activity was observed against C. albicans than C. tropicalis. Numerous studies have shown the antifungal activity of biosynthesized AgNPs against filamentous fungi [ 30 ]. Nanoparticles possess the ability to disrupt the cell walls and cell membranes of fungi, resulting in the release of intracellular components that may the fungal death [ 31 ]. It also ensures fungal death by producing hydroxyl radicals and reactive oxygen species [ 32 ]. Antibiofilm activity According to the findings of the National Institutes of Health and Centre for Disease Control, approximately 65–80% of infections are caused by microbes that form biofilms. Among these, the most prevalent ones are the Gram-negative ( P. aeruginosa and E. coli ) and the Gram-positive bacterium ( Staphylococci and S. aureus ). AgNPs possess a unique capability to disrupt the biofilm formation of various pathogenic bacteria [ 33 ]. In this study, biosynthesized Ag-NPs and H. undatus peel extracts were examined for their potential to inhibit biofilm formation in E. coli . The ELISA reader was used to measure the absorbance at 620 nm and the obtained values were regarded as an indicator of bacterial adhesion to the cell wall surface for the formation of biofilms. The biofilm activity of AgNPs and H. undatus peel extracts were compared based on their resulting IC 50 values. Synthesized AgNPs exhibited maximum inhibitory activity of biofilm (IC 50 2.81 µg/ml) while H. undatus peel extract exhibited the IC 50 value of 1.34 µg/ml (Fig. 10 ). Thrombolytic activity The thrombolytic activity of AgNPs and H. undatus peel extracts was evaluated at three different concentrations. The percentage of thrombolysis was found to be 10, 32.36 and 56.25% for the AgNPs at the concentration of 20, 40 and 80 µg/mL respectively (Fig. 11). Distilled water was taken as a negative control resulting in clot lysis of 6.07% while streptokinase was taken as a positive control exhibiting a clot lysis of 50%. AgNPs may be involved in activating the enzymes that generate plasmin, an enzyme capable of breaking down the cross-links between fibrin molecules and dissolving blood clots [ 34 ]. Limited research has been conducted on the use of AgNPs as a thrombolytic agent. The study revealed that as the concentration of AgNPs increased and the percentage of the clot lysis was also increased and it also highlights the potential of AgNPs suggesting that they could have valuable applications in the clinical field for preventing thrombosis and other related disorders. Figure 11 Thrombolytic activity of AgNPs and H. undatus peel extracts Antioxidant activity The antioxidant activity of silver nanoparticles and H. undatus peel extracts was determined by the DPPH method. The AgNPs and H. undatus peel extracts were compared based on their resulting IC 50 values. Silver nanoparticles exhibited maximum radical scavenging activity (IC 50 3.8 µg/ml), while H. undatus peel extract exhibited the IC 50 value of 2.03 µg/ml (Fig. 12 ). The inhibition % of nitric oxide radical scavenging activity by the AgNPs of H. undatus at different concentrations (20, 40, 60, 80 and 100 µg/mL) was compared based on their resulting IC 50 values. AgNPs exhibited maximum radical scavenging activity (IC50 2.8 µg/mL), while H. undatus peel extract exhibited the IC 50 value of 2.3 µg/ml (Fig. 13 ). There is a growing demand for the development of cost-effective, eco-friendly techniques to synthesize metallic nanoparticles with high yields and low toxicity. Several studies have reported the reduction of silver ions into AgNPs using plant extracts. It contains biologically active phytochemicals like terpenoids, flavonoids, vitamins and phenolics which are known for their antioxidant properties [ 35 ]. These compounds have a diverse range of biological activities and help to protect cells from damage caused by reactive oxygen species. Our study provides clear evidence that the synthesized nanoparticles and fruit peel extract exhibited radical scavenging activity. AgNPs showed promising antioxidant properties when compared to the fruit peel extract. The presence of numerous bioactive compounds in fruit extract may be the reason for its antioxidant activity [ 36 ]. The interaction of plant metabolites with metal ions can lead to the production of enhanced compounds that scavenge free radicals. Negatively charged phytochemicals and positively charged AgNPs work together to enhance the bioactivity of plants through electrostatic attractions [ 35 ]. Previous research has also indicated that antioxidant activity tends to increase as the treatment doses increase [ 36 ]. Anticancer activity The MTT assay was used to determine the cytotoxicity effect of AgNPs and H. undatus peel extract on PC3 cell lines and determine the viability of the cells (Fig. 14 ). This assay was performed using three different concentrations (21, 25 and 55 µl) of AgNPs (Table 2 ) and H. undatus peel extract (Table 2 ). IC 50 results clearly showed the tested AgNPs exhibited the minimum cytotoxic activity (IC 50 0.2 µg/ml), while H. undatus peel extract exhibited the maximum cytotoxic activity (IC 50 0.3 µg/ml) (Fig. 15 ). The presence of bioactive compounds as capping agents in the green synthesis of AgNPs may account for the improved cytotoxic effects. Increasing the concentration of AgNPs and fruit peel extract leads to an increase in cytotoxicity. The anticancer property of AgNPs was due to the activation of reactive oxygen species. This activation leads to oxidative damage to cellular components such as DNA, proteins, and lipids, ultimately resulting in cell death [ 37 ]. The fruit extracts of H. udantus contain polyphenolic compounds that are adsorbed onto the surface of AgNPs. These biomolecules, along with the green synthesis of AgNPs, have been proposed as a potential solution for combating cancer cells in vitro due to their anticancer activity. Table 2 In vitro cytotoxicity activity of the AgNPs against PC3 cell line S.No AgNPs (µg/ml) Absorbance of control at 570 nm Absorbance of sample at 570 nm % of inhibition 1 12 0.568 0.553 2.64 2 25 0.568 0.359 36.79 3 55 0.568 0.235 58.62 Table 3 In vitro cytotoxicity activity of the H. undatus peel extract against PC3 cell line S.No H.undatus peel extract (µg/ml) Absorbance of control at 570 nm Absorbance of sample at 570 nm % of inhibition 1 12 0.568 0.549 3.34 2 25 0.568 0.303 46.65 3 55 0.568 0.223 60.73 Conclusion The nano-sized silver particles gained more attention due to their physiochemical and optical properties. In the current study, AgNps were successfully obtained from the reduction of silver nitrate by H. undatus peel extract. It is observed that the fruit peel extract significantly exhibited the flavonoids and phenolics that are responsible for its antioxidant activity. The potential use of silver nanoparticles obtained from the extract of H. undatus peel extract has been suggested as an effective anticancer agent against PC3 cell lines. The biosynthesized AgNPs from H. undatus peel extract showed significant antibacterial, antioxidant, anticancer, and thrombolytic activity which indicates their potential therapeutic applications. Thereby, further studies are required to discover the medicinal drugs from this source. Declarations The authors declare that they have no known competing financial interests that could have appeared to influence the work reported in this paper. Conflicts of interest/Competing interests The authors declare that there is no conflict of interest. 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Spectrochim Acta A Mol Biomol Spectrosc. 2015; https://doi.org/10.1016/j.saa.2015.07.009 Kumar B, Smita K, Cumbal L, Debut A. Green synthesis of silver nanoparticles using Andean blackberry fruit extract. Saudi J Biol Sci. 2017; https://doi.org/10.1016/j.sjbs.2015.09.006 Dipankar C, Murugan S. The green synthesis, characterization and evaluation of the biological activities of silver nanoparticles synthesized from Iresine herbstii leaf aqueous extracts. Colloids Surf B Biointerfaces. 2012; https://doi.org/10.1016/j.colsurfb.2012.04.006 Ajaykumar AP, Mathew A, Chandni AP, Varma SR, Jayaraj KN, Sabira O, Rasheed VA, Binitha VS, Swaminathan TR, Basheer VS, Giri S, Chatterjee S. Green Synthesis of Silver Nanoparticles Using the Leaf Extract of the Medicinal Plant, Uvaria narum and Its Antibacterial, Antiangiogenic, Anticancer and Catalytic Properties. Antibiotics. 2023; https://doi.org/10.3390/antibiotics12030564 Additional Declarations No competing interests reported. Supplementary Files GA.jpg Cite Share Download PDF Status: Published Journal Publication published 18 Mar, 2024 Read the published version in Discover Nano → Version 1 posted Editorial decision: Revision requested 25 Jan, 2024 Reviews received at journal 17 Jan, 2024 Reviewers agreed at journal 10 Jan, 2024 Reviewers invited by journal 10 Jan, 2024 Editor assigned by journal 01 Jan, 2024 Submission checks completed at journal 29 Dec, 2023 First submitted to journal 25 Dec, 2023 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-3805384","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":264289620,"identity":"358e3a7e-dd46-4d7d-b9d9-357e757ddd4f","order_by":0,"name":"Aswini Anguraj","email":"","orcid":"","institution":"Sri Ramakrishna College of Arts \u0026 Science","correspondingAuthor":false,"prefix":"","firstName":"Aswini","middleName":"","lastName":"Anguraj","suffix":""},{"id":264289621,"identity":"1d7228b5-dde7-4fbb-8f70-6bb6bd4aba62","order_by":1,"name":"Helan Soundra Rani Michael","email":"","orcid":"","institution":"Sri Ramakrishna College of Arts \u0026 Science","correspondingAuthor":false,"prefix":"","firstName":"Helan","middleName":"Soundra Rani","lastName":"Michael","suffix":""},{"id":264289622,"identity":"c36ebc85-89d2-4359-8df3-0d5448b14b26","order_by":2,"name":"Sathish Sugumaran","email":"","orcid":"","institution":"MVJ College of Engineering, Bangalore","correspondingAuthor":false,"prefix":"","firstName":"Sathish","middleName":"","lastName":"Sugumaran","suffix":""},{"id":264289623,"identity":"dbcbc77f-6403-474f-a0f3-1531d20a2e0e","order_by":3,"name":"Gogul Ramnath Madhusudhanan","email":"","orcid":"","institution":"Sri Ramakrishna College of Arts \u0026 Science","correspondingAuthor":false,"prefix":"","firstName":"Gogul","middleName":"Ramnath","lastName":"Madhusudhanan","suffix":""},{"id":264289624,"identity":"744735f3-33b7-45b0-a1cc-cfa417ba7b96","order_by":4,"name":"Rathish Kumar Sivaraman","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/UlEQVRIiWNgGAWjYBACA2YwdQCIeRgOMDbYABmMjQcIaGFsAGthA2tJA2lpwK+FAUkLkHkYbilOYM7OfPzBh5o7+fzzew8e/LnjvN3a9sNAW2psonFpsWxmS2ycceyZ5YxjfAmHec/cTt52JhGo5VhabgMuhx3mMWzmbThswHCMx+AwY9vtZLMDQC1AF+LX8heoRR6o5eDPtnPJZucfEqEFqMDAAKjlAG/bATuzGwRsAfllZs+xwwaGx3IMDvO2JSeY3QDakoDHL+b8hw98+FFz2EDu8Bnjjz/b7OzNzqc/BIahDU4tGCARrDKBWOUgYE+K4lEwCkbBKBgZAACoTmu6nGuQAgAAAABJRU5ErkJggg==","orcid":"","institution":"Sri Ramakrishna College of Arts \u0026 Science","correspondingAuthor":true,"prefix":"","firstName":"Rathish","middleName":"Kumar","lastName":"Sivaraman","suffix":""}],"badges":[],"createdAt":"2023-12-25 18:29:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3805384/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3805384/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s11671-024-03995-w","type":"published","date":"2024-03-18T15:01:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":49041395,"identity":"6e7bb36d-93e2-407d-9a81-c9e8ab3d94b7","added_by":"auto","created_at":"2024-01-02 05:22:53","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":260427,"visible":true,"origin":"","legend":"\u003cp\u003eGreen synthesis of silver nanoparticles from \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/a7fcea31a0bcf48e022ea3ac.jpeg"},{"id":49041762,"identity":"8f38d0b7-b3c6-4f1c-a2a4-509adf4d752c","added_by":"auto","created_at":"2024-01-02 05:30:53","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":76540,"visible":true,"origin":"","legend":"\u003cp\u003eUV-Visible spectra of biosynthesized AgNPs using \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/a33265cc8d7e49a460a12715.jpeg"},{"id":49041391,"identity":"30e99630-9743-44fe-b41f-70c57e4a1a26","added_by":"auto","created_at":"2024-01-02 05:22:53","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":103629,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR analysis of biosynthesized AgNPs using \u003cem\u003eH. undatus\u003c/em\u003e fruit peel 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biosynthesized AgNPs using \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/0bbe52387e3dce15fd453b8b.png"},{"id":49041393,"identity":"09871d6c-0130-41ed-9a3b-8a99a2c1ec01","added_by":"auto","created_at":"2024-01-02 05:22:53","extension":"jpeg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":62135,"visible":true,"origin":"","legend":"\u003cp\u003eEDAX analysis of biosynthesized AgNPs using \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract\u003c/p\u003e","description":"","filename":"floatimage7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/67e7a10353f43b298c4ca990.jpeg"},{"id":49041405,"identity":"bc1a59d0-cc3a-4559-aa04-197b2ee6f5c8","added_by":"auto","created_at":"2024-01-02 05:22:53","extension":"jpeg","order_by":7,"title":"Figure 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pneumoniae\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage9.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/339767055ca5ef4b25d6fc10.jpeg"},{"id":49041855,"identity":"4fdd9149-4bed-46dd-b43c-d50b97f19f70","added_by":"auto","created_at":"2024-01-02 05:38:53","extension":"jpeg","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":288046,"visible":true,"origin":"","legend":"\u003cp\u003eAntifungal activity of AgNPs against \u003cem\u003eC. albicans\u003c/em\u003e and \u003cem\u003eC. tropicalis\u003c/em\u003e\u003c/p\u003e","description":"","filename":"floatimage10.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/4d0279769790ca1915371c0f.jpeg"},{"id":49041853,"identity":"bf1118ff-3231-4c9d-835f-88b721524b36","added_by":"auto","created_at":"2024-01-02 05:38:53","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":24823,"visible":true,"origin":"","legend":"\u003cp\u003eDetermination of % antibiofilm activity of AgNPs and \u003cem\u003eH. undatus \u003c/em\u003epeel extracts on \u003cem\u003eE. coli\u003c/em\u003e\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/9730e1d9ff3e8c1bbf65d90c.png"},{"id":49041854,"identity":"93007825-8578-434b-abc4-d3d4c18f74df","added_by":"auto","created_at":"2024-01-02 05:38:53","extension":"jpg","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":10030,"visible":true,"origin":"","legend":"\u003cp\u003eThrombolytic activity of AgNPs and \u003cem\u003eH. undatus \u003c/em\u003epeel extracts\u003c/p\u003e","description":"","filename":"11.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/6533d6a56d52a11d604332c4.jpg"},{"id":49041764,"identity":"92adfcb3-38e7-4ccd-9ae9-a632ff7b5063","added_by":"auto","created_at":"2024-01-02 05:30:53","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":30392,"visible":true,"origin":"","legend":"\u003cp\u003eDPPH radical scavenging activity of \u003cem\u003eH. undatus \u003c/em\u003eand AgNPs\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/ed353fb85c4f93df30931415.png"},{"id":49041766,"identity":"f021355d-bc0e-439c-b7f9-27fc2140eb7e","added_by":"auto","created_at":"2024-01-02 05:30:53","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":35872,"visible":true,"origin":"","legend":"\u003cp\u003eNitric oxide radical scavenging activity of \u003cem\u003eH. undatus \u003c/em\u003eand AgNPs\u003c/p\u003e","description":"","filename":"13.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/bf837ff9b10fbc2fb2de0f34.png"},{"id":49041406,"identity":"7e20b888-e677-4565-95b2-2d50f9050182","added_by":"auto","created_at":"2024-01-02 05:22:54","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":664573,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eA, B \u0026amp; C \u003c/strong\u003eDifferent concentrations (21, 25 and 55 µl) of \u003cem\u003eH. undatus\u003c/em\u003e peel extract treated on PC3 cell line\u003c/p\u003e","description":"","filename":"14.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/05c4bbc9f7a105383aa9c8ee.png"},{"id":49041769,"identity":"da148265-55b5-4370-92b9-822656826bec","added_by":"auto","created_at":"2024-01-02 05:30:53","extension":"png","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":23797,"visible":true,"origin":"","legend":"\u003cp\u003eIn vitro cytotoxic activity of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts on PC3 cell line\u003c/p\u003e","description":"","filename":"15.png","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/f90b9006354317933db38f7f.png"},{"id":53403534,"identity":"56514ce7-8e63-40d6-b799-487e89b706f7","added_by":"auto","created_at":"2024-03-25 15:12:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1719423,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/93055717-2f0a-45c3-931f-c83de1d13ca0.pdf"},{"id":49041763,"identity":"ea40ffd8-ffb3-49c5-a57b-5fe45ac80778","added_by":"auto","created_at":"2024-01-02 05:30:53","extension":"jpg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":122908,"visible":true,"origin":"","legend":"","description":"","filename":"GA.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3805384/v1/84b1404a14f5e6ebc6557d4a.jpg"}],"financialInterests":"No competing interests reported.","formattedTitle":"A Comparative Study on Biosynthesized Silver Nanoparticles from H. undatus Fruit Peel and its Therapeutic Applications","fulltext":[{"header":"Introduction","content":"\u003cp\u003eNanotechnology is an emerging field that aims to create molecular, atomic and supramolecular properties [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. It has gained importance in cosmetics, drug delivery, biomedical, cosmetics and cancer therapy [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Metallic nanoparticles such as silver, copper oxide, zinc oxide and gold nanoparticles are synthesized extensively in nanostructured form [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Among the above-mentioned metals, silver nanoparticles gained more attention due to their unique characteristics. It can be synthesized by physical, chemical and biological methods [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Biological entities such as bacteria, fungi, yeasts and plants have been utilized in the synthesis of silver nanoparticles, resulting in less toxic, cost-efficient, stable and eco-friendly alternatives to chemical methods [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Among the biological methods, microbe-mediated synthesis of nanoparticles is not commercially feasible due to its high aseptic condition requirement. Plant-mediated silver nanoparticles offer several advantages over microorganisms. Because they can be easily scaled up, making them more practical for large-scale production. The nanoparticles can be effectively capped and reduced by extracting active components from plants such as flavonoids, alkaloids, terpenes, tannins and saponins [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Silver nanoparticles from plant extracts have a wide range of applications in the fields of food, health, agriculture, disease control, pharmaceuticals and medicine [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Silver nanoparticles have shown potential antibacterial, antifungal, anti-inflammatory, antioxidant, antiviral and anticancer, anti-plasmodial and anti-platelets effects [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eProstate carcinoma (PC) is the primary reason for cancer in the male reproductive system, affecting a significant number of newly diagnosed cancer patients, accounting for around 25% of cases. When combined with colorectal cancer, PC becomes the second leading cause of male cancer-related deaths in the US, following lung and bronchus cancer. The majority of prostate cancer cases, around 80% are found in the peripheral zone, while 10\u0026ndash;20% occur in the transition zone [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Treatment methods like cholesterol-lowering drugs, biopsies, and conventional heating techniques can be costly and may have side effects such as pain or discomfort due to unwanted reflection, scattering or absorption [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. It is crucial to explore alternative treatments or medications to address these limitations. Nanoparticles have shown promise in the treatment of prostate cancer patients [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The toxicity associated with this treatment is moderate and only temporarily affects the quality of life. Hence, there is a need to discover new cancer therapies that are both biocompatible and cost-effective [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eDragon fruit (\u003cem\u003eHylocereus undatus\u003c/em\u003e) is a cactus species that is commonly consumed in Asian countries. It is a tropical fruit with a red peel and green pins. It is a great source of low-calorie dietary fiber [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. It contains antioxidants that can help to prevent various diseases such as cancer, cardiovascular issues, diabetes, urinary and gastrointestinal problems [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The peel of the dragon fruit is composed of pectin, phyllocactin, steroids, betacynanin and betanin. Previous studies reported that it has high antioxidant, antimicrobial, cardioprotective, anti-inflammatory and anticancer properties [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. In this study, we focused on the use of \u003cem\u003eHylocereus undatus\u003c/em\u003e for the bioreduction of silver ions in an aqueous medium which leads to the formation of silver nanoparticles. The synthesized nanoparticles were characterized and evaluated for antibacterial, antifungal, antioxidant and thrombolytic activity. Additionally, we examined the potential of these AgNPs to inhibit the growth of prostate cancer cells.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSample collection and extraction\u003c/h2\u003e \u003cp\u003eIn the present study, \u003cem\u003eHylocereus undatus\u003c/em\u003e was collected from the local market, R. S. Puram, Coimbatore. The peel of \u003cem\u003eH. undatus\u003c/em\u003e was washed with double distilled water until all the impurities were removed and kept for drying in a shady sterile environment. The dried samples were finely grounded into powder and further processed for extraction.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of aqueous extract\u003c/h2\u003e \u003cp\u003eThe \u003cem\u003eH. undatus\u003c/em\u003e aqueous extract was prepared by boiling 20 g of powdered peel in 500 mL of distilled water for 15 minutes. The extracts was agitated and covered until it reached room temperature. The crude extract was filtered through Whatman filter paper No.1. The resulting extract was stored in a container at 2\u0026deg;C for further study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eSynthesis of silver nanoparticles\u003c/h2\u003e \u003cp\u003eA reaction mixture consisting of 220 ml of dragon fruit peel extract and 110 ml of 10 mM silver nitrate was stirred for 24 hours at room temperature using a magnetic stirrer. The reaction mixture was then centrifuged at 12,000 rpm for 15 minutes at 4\u003csup\u003eo\u003c/sup\u003eC and the supernatant was discarded. The resulting pellet was dissolved in distilled water and centrifuged again under the same conditions as mentioned before to ensure the purity of the nanosilver. The resultant pellet was dried and stored for further characterization [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of silver nanoparticles\u003c/h2\u003e \u003cp\u003eThe synthesized AgNPs were characterized using a UV-visible spectrophotometer, Fourier transform infrared spectrophotometer (FTIR), Gas chromatography-mass spectrometry (GCMS), X-ray diffraction (XRD), Energy Dispersive X-ray spectroscopy (EDAX), and Field Emission Scanning Electron Microscopy (FESEM).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eAntibacterial activity\u003c/h2\u003e \u003cp\u003eThe disc diffusion method was used to evaluate the antibacterial activity of AgNPs against both gram-positive (\u003cem\u003eStreptococcus pneumoniae\u003c/em\u003e) and gram-negative (\u003cem\u003eEscherichia coli\u003c/em\u003e) bacteria. The bacterial strains were spread on nutrient agar plates using a sterile cotton swab. The disc was treated with two distinct concentrations (40 and 80 \u0026micro;L) of synthesized AgNPs. The kanamycin was used as a positive control and distilled water was used as a negative control. Then the plates were incubated at 37\u003csup\u003eo\u003c/sup\u003e C for 24 hours. The antibacterial screening was evaluated by measuring the zone of inhibition [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The experiments were replicated three times and the diameter of the zone of inhibition was reported as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eAntifungal activity\u003c/h2\u003e \u003cp\u003eThe antifungal potential of the biosynthesized AgNPs against two pathogenic strains such as \u003cem\u003eCandida albicans\u003c/em\u003e and \u003cem\u003eCandida tropicalis\u003c/em\u003e was evaluated using the agar well diffusion method as described by Joseph et al. [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Wells with a diameter of 8 mm were punched in the potato dextrose agar medium and 100 \u0026micro;L of each microorganism was spread using a sterile cotton swab. Distilled water was used as the negative control while Voriconazole as the positive control. Two concentrations such as 40 and 80 \u0026micro;L of AgNPs were added to the wells and incubated at 37\u003csup\u003eo\u003c/sup\u003e C for 24 hours. Following, the zones were determined and recorded.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eAntibiofilm activity\u003c/h2\u003e \u003cp\u003eA 96-well microtiter plate was used to evaluate the anti-biofilm effectiveness of the AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract. Each well of the plates was filled with 180 \u0026micro;l of Muller Hinton broth and 10 \u0026micro;l of the test pathogens. Then 10 \u0026micro;l of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract were added and the mixture was thoroughly mixed. The test plates were then incubated for 24 hours at 37\u003csup\u003eo\u003c/sup\u003e C. After incubation, the microtiter plates were washed with phosphate buffer saline to remove non-adherent bacteria. The plates were air-dried for 45 minutes and the wells were fixed with sodium acetate. Crystal violet stain was added and incubated in the dark for 30 minutes. Excess dye was removed by washing with water. The plates were air-dried and 200 \u0026micro;l ethanol was added to each well and absorbance was measured at 620 nm [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003eThrombolytic activity\u003c/h2\u003e \u003cp\u003eThe thrombolytic activity of AgNPs and the peel extract of \u003cem\u003eH. undatus\u003c/em\u003e was performed according to the method followed by Devi et al. [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e] with slight modification. After determining the weight of the empty Eppendorf tube 20, 40 and 60 \u0026micro;L of blood were drawn in different tubes and incubated at 37◦ C for 30 minutes. After the clot formation, the tubes were re-weighed to determine the weight of the blood clot (X). This was achieved by subtracting the initial weight of the tube. 100 \u0026micro;L of AgNPs at two different concentrations (500 and 1000 \u0026micro;g/mL) was added to the tubes and incubated at 37\u003csup\u003eo\u003c/sup\u003eC for 90 minutes. To calculate the weight of the non-lyzed blood clot (Y), the initial weight of the tube was subtracted from the final weight of the tube, which contained the remaining blood clot. 100 \u0026micro;L of distilled water was used as a negative control and streptokinase as a positive control. Thrombolytic activity was calculated using the following equation\u003c/p\u003e \u003cp\u003eThrombolysis (%)\u0026thinsp;=\u0026thinsp;X \u0026ndash; Y X 100/X\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eAntioxidant activity\u003c/h2\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003eDPPH radical scavenging assay\u003c/h2\u003e \u003cp\u003eThe scavenging ability of the AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract was determined using the DPPH method described by Phull et al. [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. The DPPH assay was performed in a 96-well plate having ascorbic acid as the reference standard. AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract (20, 40, 60, 80, and 100 \u0026micro;l) was taken from the stock solution and poured into each well of the plate. Then, 100 \u0026micro;l of the DPPH solution was added and incubated at 27\u0026deg;C for 20 minutes. The measurement of the absorbance of the reaction mixture was carried out at 517 nm using a microplate reader. The scavenging activity was calculated as\u003c/p\u003e \u003cp\u003e% DPPH radical scavenging activity\u0026thinsp;=\u0026thinsp;control OD\u0026thinsp;\u0026minus;\u0026thinsp;sample OD/control OD x 100\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eNitric oxide radical scavenging activity\u003c/h2\u003e \u003cp\u003eThe nitric oxide radical scavenging activity of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract was carried out using the method outlined by Makhija et al. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] with slight modification. Various concentrations of the AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract (20, 40, 60 80 and 100 \u0026micro;L) were combined with 3 mL of sodium nitroprusside (10 mMol/L) in phosphate buffer saline (0.2 mMol/L, pH 7.4). The mixture was then incubated at 25\u003csup\u003eo\u003c/sup\u003eC for 150 minutes. Following the addition of 500 \u0026micro;L of Griess reagent (composed of 2% orthophosphoric acid, 1% sulphanilamide, and 0.1% N-1-naphthyl ethylenediamine dihydrochloride) was added. The absorbance was measured at 546 nm. All measurements were conducted in triplicate and the mean values were calculated.\u003c/p\u003e \u003cp\u003eNitric oxide scavenged (%)\u0026thinsp;=\u0026thinsp;A\u003csub\u003econtrol\u003c/sub\u003e \u0026ndash; A\u003csub\u003etest\u003c/sub\u003e / A\u003csub\u003econtrol\u003c/sub\u003e x 100\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eCytotoxicity assay\u003c/h2\u003e \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e \u003ch2\u003eCell culture\u003c/h2\u003e \u003cp\u003eThe human prostate cancer cell line (PC3) was purchased from NCCS, Pune. The cells were cultured in Dulbecco\u0026rsquo;s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic. The cells were kept at a temperature of 37\u003csup\u003eo\u003c/sup\u003e C in 5% CO\u003csub\u003e2\u003c/sub\u003e and 95% relative humidity (Devi and Bhimba, 2012).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eMTT assay\u003c/h2\u003e \u003cp\u003eThe cytotoxic effect of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract was carried out by MTT assay. PC3 cells were seeded in a 96 well plate and incubated for 24 hours, in 200 \u0026micro;l of DMEM with 10% FBS. Various concentrations (12, 25, and 55 \u0026micro;g/ml) of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract were added and incubated for 48 hours. After the addition of MTT, the cells were again incubated at 37\u003csup\u003eo\u003c/sup\u003eC for 4 hours. The media was then removed, 200 \u0026micro;l of DMSO was added to each well and the absorbance was read at 570 nm [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of silver nanoparticles\u003c/h2\u003e \u003cp\u003eThe current study revealed that the \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract to the silver in the form of nitrate. This interaction resulted in the formation of a pale pink to dark brown indicating the presence of silver nanoparticles [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Similar color changes have also been observed in the previous study [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The UV-vis spectra were recorded after time intervals of 20 min, 25 min, 30 min, 35 min, 49 min, 45 min, 50 min, 55 min, 60 min, 65 min and 70 min. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e shows the absorption spectra of AgNPs using \u003cem\u003eH. undatus\u003c/em\u003e fruit peel extract. The UV spectrum has exhibited a strong peak at 417 nm which confirms the formation of AgNPs.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe purity of the synthesized NPs was checked by FTIR analysis within the range of 400\u0026ndash;4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e shows the FTIR spectra of biosynthesized AgNPs and the absorption peak bands at 3734.19, 3332.99, 2310.72, 1593.20, 1365.60, 1323.17, 1238.30, 1041.56, 921.97, 821.68, 678.94, 601.79, 559. 36. These absorbance bands are known to be associated with stretching vibrations of O-H (alcohols), O-H (alcohols), C\u0026thinsp;=\u0026thinsp;N (nitriles), C-C (aromatics), C-H (alkanes), C-N (aromatic amines), C-N (aliphatic amines), C-O (alcohols), O-H (carboxylic acids), C-Cl (alkyl halides), C-Br (alkyl halides), C-Br (alkyl halides), C- Br (alkyl halides). \u003cem\u003eH. undatus\u003c/em\u003e fruit is primarily composed of betanin, pectin, hylocerenin, betacyanin and phyllocactin [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. This study has confirmed the presence of a significant amount of phytocompounds and these substances may potentially be responsible for the reduction, capping and synthesis of AgNPs.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe GC-MS analysis of the peel extract of \u003cem\u003eH. undatus\u003c/em\u003e was conducted to determine the bioactive chemical compounds present in the extract. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e displays the GC-MS spectrum with peaks and retention time. The peel extract of \u003cem\u003eH. undatus\u003c/em\u003e has shown the presence of the major seven bioactive compounds such as 6-methyl-2, (4-bromophenyl)-7-phenylmethylindolizine, N-[(4,6-Dimethoxynaphthalen-1-yl) methylene], 2,5-dichloro-4-hydroxyphenylamine, Oxacycloheptadec-8-en-2-one, Androstan-17one, 3-ethyl-3-hydroxy, Pyridine-3-carboxamide, 6-chloro-4-trifluoromethyl-N-[2,4-dichloeo-6-methyl]-N-methyl, 1,2-Dipalmitoyl 3-acetyl glycerol Gallic acid, and Triamcinolone acetonide. The peak intensity of N-[(4,6Dimethoxynaphthalen-1-ylmethylene], 2,5-dichloro-4-hydroxyphenylamine was found to be higher with a retention time of 30.12 min followed by Oxacycloheptadec-8-en-2-one with a retention time of 30.74 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChemical composition of \u003cem\u003eH. undatus\u003c/em\u003e peel extract\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eRT\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCompound name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMolecular\u003c/p\u003e \u003cp\u003eformula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMolecular\u003c/p\u003e \u003cp\u003eweight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePeak area\u003c/p\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6-methyl-2,(4-bromophenyl)-7-\u003c/p\u003e \u003cp\u003ephenylmethylindolizine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC\u003csub\u003e22\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003eBrN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e376.289\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7.991989\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eN-[(4,6Dimethoxynaphthalen-1-\u003c/p\u003e \u003cp\u003eylmethylene], 2,5-dichloro-4- hydroxyphenylamine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026#119862;19\u0026#119867;15\u0026#119862;\u0026#119897;2\u0026#119873;\u0026#119874;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e376.233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8.799596\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxacycloheptadec-8-en-2-one\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC16H28O2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e252.392\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e15.430418\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAndrostan-17one 3-ethyl-3-\u003c/p\u003e \u003cp\u003ehydroxy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC21H34O2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e318.49346\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13.414466\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e31.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePyridine-3-carboxamide, 6- chloro-4-trifluoromethyl-N-[2,4-\u003c/p\u003e \u003cp\u003edichloeo-6-methyl]-N-methyl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC15H10C13F3N2O\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e397.606\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e11.854260\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e31.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1,2-Dipalmitoyl 3-acetyl\u003c/p\u003e \u003cp\u003eglycerol Gallic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC37H70O6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e610.948\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e11.854137\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e32.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTriamcinolone acetonide.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eC24H31FO6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e434.497\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10.116891\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe synthesized AgNPs are characterized using XRD to confirm the presence of silver ions and to know the structural information [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Figure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e shows the XRD pattern of AgNPs which confirmed the crystalline nature of synthesized AgNPs. The peaks at 2\u0026#120579; of 27.57, 32.09, 37.88, 44.65, 46.02, 54.59, 57.19, 64.16, 67.32, 72.62, and 76.42 correspond to the crystalline planes of the face-centered cubic structure of metallic silver. The intense peak at 32.09 possibly suggests the presence of silver ions as the major constituent in the biosynthesized AgNPs. A similar result was observed by Phongtongpasuk et al. [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] who identified the most intense peak at 32.5 indicated the presence of AgNPs.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe elemental analysis of the material is depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e, illustrating the synthesized AgNPs through the EDAX spectrum [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. This analysis demonstrates a prominent indication of a metallic silver area at 3 KeV and confirms the formation of silver nanoparticles synthesized through the utilization of \u003cem\u003eH. undatus\u003c/em\u003e peel (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The Ag peak showed a weight percentage of 35.12 and an atomic percentage of 8.02. A low calcium signal was observed and five moderate signals for carbon, oxygen, sodium, chloride and potassium were detected as a result of the chemicals used in the sample preparation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFE-SEM images of AgNPs synthesized using the fruit peel extract of \u003cem\u003eH. undatus\u003c/em\u003e are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e. The surface morphology of AgNPs showed a spherical shape with agglomeration. In the present study, the histogram of the particle size ranges from 52\u0026ndash;79 nm. The increased concentration of bioactive compounds in the colloidal solution could potentially lead to the formation of nanoclusters. The signals produced from the interaction between the electrons and the sample provide valuable information regarding the sample's external morphology, chemical composition, crystalline structure and material orientation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eAntibacterial activity\u003c/h2\u003e \u003cp\u003eThe antibacterial activity of synthesized AgNPs were tested at various concentrations by the disc diffusion method. Figure\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003e illustrates the zone of inhibition around the individual bacterial culture. The zone of inhibition was measured to be 1.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 and 1.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 cm for \u003cem\u003eE. coli\u003c/em\u003e while in the case of \u003cem\u003eS. pneumoniae\u003c/em\u003e 1.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 and 1.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 cm was measured at a concentration of 40 \u0026micro;l and 80 \u0026micro;l of synthesized AgNPs. In the present study, the synthesized AgNPs showed the highest antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e when compared to \u003cem\u003eS\u003c/em\u003e. \u003cem\u003epneumoniae\u003c/em\u003e. The potential cause of this occurrence could be attributed to the existence of phytochemicals within the \u003cem\u003eH. undatus.\u003c/em\u003e The enhanced antibacterial efficacy of AgNPs can be predominantly attributed to the synergistic interplay between the aforementioned nanoparticles and the naturally occurring chemicals found within the extract as previously substantiated in a prior investigation. It has been determined that AgNPs can liberate silver ions within bacterial cells, thus enhancing their bactericidal properties [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eAntifungal activity\u003c/h2\u003e \u003cp\u003eThe antifungal potential of the biosynthesized AgNPs towards two pathogenic strains such as \u003cem\u003eCandida albicans\u003c/em\u003e and \u003cem\u003eCandida tropicalis\u003c/em\u003e was evaluated using the well diffusion method. The antifungal activity can be identified by the zone of inhibition (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e9\u003c/span\u003e). The results indicate that the biosynthesized AgNPs possess significant antifungal activity against two fungal strains that were tested. There is no zone of inhibition was observed in the control. The synthesized AgNPs that could inhibit the fungal growth of \u003cem\u003eC. albicans\u003c/em\u003e were found to be 1.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 and 1.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3, while \u003cem\u003eC. tropicalis\u003c/em\u003e was measured at 0.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 and 1.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 cm at a concentration of 40 and 80 \u0026micro;L respectively. The highest antifungal activity was observed against \u003cem\u003eC. albicans\u003c/em\u003e than \u003cem\u003eC. tropicalis.\u003c/em\u003e Numerous studies have shown the antifungal activity of biosynthesized AgNPs against filamentous fungi [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Nanoparticles possess the ability to disrupt the cell walls and cell membranes of fungi, resulting in the release of intracellular components that may the fungal death [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. It also ensures fungal death by producing hydroxyl radicals and reactive oxygen species [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eAntibiofilm activity\u003c/h2\u003e \u003cp\u003eAccording to the findings of the National Institutes of Health and Centre for Disease Control, approximately 65\u0026ndash;80% of infections are caused by microbes that form biofilms. Among these, the most prevalent ones are the Gram-negative (\u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e) and the Gram-positive bacterium (\u003cem\u003eStaphylococci\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e). AgNPs possess a unique capability to disrupt the biofilm formation of various pathogenic bacteria [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. In this study, biosynthesized Ag-NPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts were examined for their potential to inhibit biofilm formation in \u003cem\u003eE. coli\u003c/em\u003e. The ELISA reader was used to measure the absorbance at 620 nm and the obtained values were regarded as an indicator of bacterial adhesion to the cell wall surface for the formation of biofilms. The biofilm activity of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts were compared based on their resulting IC\u003csub\u003e50\u003c/sub\u003e values. Synthesized AgNPs exhibited maximum inhibitory activity of biofilm (IC\u003csub\u003e50\u003c/sub\u003e2.81 \u0026micro;g/ml) while \u003cem\u003eH. undatus\u003c/em\u003e peel extract exhibited the IC\u003csub\u003e50\u003c/sub\u003e value of 1.34 \u0026micro;g/ml (Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eThrombolytic activity\u003c/h2\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe thrombolytic activity of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts was evaluated at three different concentrations. The percentage of thrombolysis was found to be 10, 32.36 and 56.25% for the AgNPs at the concentration of 20, 40 and 80 \u0026micro;g/mL respectively (Fig.\u0026nbsp;11). Distilled water was taken as a negative control resulting in clot lysis of 6.07% while streptokinase was taken as a positive control exhibiting a clot lysis of 50%. AgNPs may be involved in activating the enzymes that generate plasmin, an enzyme capable of breaking down the cross-links between fibrin molecules and dissolving blood clots [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Limited research has been conducted on the use of AgNPs as a thrombolytic agent. The study revealed that as the concentration of AgNPs increased and the percentage of the clot lysis was also increased and it also highlights the potential of AgNPs suggesting that they could have valuable applications in the clinical field for preventing thrombosis and other related disorders.\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure\u0026nbsp;11\u003c/b\u003e Thrombolytic activity of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts\u003c/p\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eAntioxidant activity\u003c/h2\u003e \u003cp\u003eThe antioxidant activity of silver nanoparticles and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts was determined by the DPPH method. The AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extracts were compared based on their resulting IC\u003csub\u003e50\u003c/sub\u003e values. Silver nanoparticles exhibited maximum radical scavenging activity (IC\u003csub\u003e50\u003c/sub\u003e 3.8 \u0026micro;g/ml), while \u003cem\u003eH. undatus\u003c/em\u003e peel extract exhibited the IC\u003csub\u003e50\u003c/sub\u003e value of 2.03 \u0026micro;g/ml (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e12\u003c/span\u003e). The inhibition % of nitric oxide radical scavenging activity by the AgNPs of \u003cem\u003eH. undatus\u003c/em\u003e at different concentrations (20, 40, 60, 80 and 100 \u0026micro;g/mL) was compared based on their resulting IC\u003csub\u003e50\u003c/sub\u003e values. AgNPs exhibited maximum radical scavenging activity (IC50 2.8 \u0026micro;g/mL), while \u003cem\u003eH. undatus\u003c/em\u003e peel extract exhibited the IC\u003csub\u003e50\u003c/sub\u003e value of 2.3 \u0026micro;g/ml (Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e13\u003c/span\u003e). There is a growing demand for the development of cost-effective, eco-friendly techniques to synthesize metallic nanoparticles with high yields and low toxicity. Several studies have reported the reduction of silver ions into AgNPs using plant extracts. It contains biologically active phytochemicals like terpenoids, flavonoids, vitamins and phenolics which are known for their antioxidant properties [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. These compounds have a diverse range of biological activities and help to protect cells from damage caused by reactive oxygen species. Our study provides clear evidence that the synthesized nanoparticles and fruit peel extract exhibited radical scavenging activity. AgNPs showed promising antioxidant properties when compared to the fruit peel extract. The presence of numerous bioactive compounds in fruit extract may be the reason for its antioxidant activity [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The interaction of plant metabolites with metal ions can lead to the production of enhanced compounds that scavenge free radicals. Negatively charged phytochemicals and positively charged AgNPs work together to enhance the bioactivity of plants through electrostatic attractions [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Previous research has also indicated that antioxidant activity tends to increase as the treatment doses increase [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eAnticancer activity\u003c/h2\u003e \u003cp\u003eThe MTT assay was used to determine the cytotoxicity effect of AgNPs and \u003cem\u003eH. undatus\u003c/em\u003e peel extract on PC3 cell lines and determine the viability of the cells (Fig.\u0026nbsp;\u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e14\u003c/span\u003e). This assay was performed using three different concentrations (21, 25 and 55 \u0026micro;l) of AgNPs (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) and \u003cem\u003eH. undatus\u003c/em\u003e peel extract (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). IC\u003csub\u003e50\u003c/sub\u003e results clearly showed the tested AgNPs exhibited the minimum cytotoxic activity (IC\u003csub\u003e50\u003c/sub\u003e 0.2 \u0026micro;g/ml), while \u003cem\u003eH. undatus\u003c/em\u003e peel extract exhibited the maximum cytotoxic activity (IC\u003csub\u003e50\u003c/sub\u003e 0.3 \u0026micro;g/ml) (Fig.\u0026nbsp;\u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e15\u003c/span\u003e). The presence of bioactive compounds as capping agents in the green synthesis of AgNPs may account for the improved cytotoxic effects. Increasing the concentration of AgNPs and fruit peel extract leads to an increase in cytotoxicity. The anticancer property of AgNPs was due to the activation of reactive oxygen species. This activation leads to oxidative damage to cellular components such as DNA, proteins, and lipids, ultimately resulting in cell death [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. The fruit extracts of \u003cem\u003eH. udantus\u003c/em\u003e contain polyphenolic compounds that are adsorbed onto the surface of AgNPs. These biomolecules, along with the green synthesis of AgNPs, have been proposed as a potential solution for combating cancer cells in vitro due to their anticancer activity.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eIn vitro cytotoxicity activity of the AgNPs against PC3 cell line\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAgNPs\u003c/p\u003e \u003cp\u003e(\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAbsorbance of\u003c/p\u003e \u003cp\u003econtrol at 570 nm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAbsorbance of\u003c/p\u003e \u003cp\u003esample at 570 nm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% of inhibition\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.553\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.64\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.359\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e36.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.235\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e58.62\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eIn vitro cytotoxicity activity of the \u003cem\u003eH. undatus\u003c/em\u003e peel extract against PC3 cell line\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eH.undatus\u003c/em\u003e peel extract (\u0026micro;g/ml)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAbsorbance of control at 570\u003c/p\u003e \u003cp\u003enm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAbsorbance of sample at 570\u003c/p\u003e \u003cp\u003enm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e% of inhibition\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.549\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.303\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e46.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.223\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e60.73\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe nano-sized silver particles gained more attention due to their physiochemical and optical properties. In the current study, AgNps were successfully obtained from the reduction of silver nitrate by \u003cem\u003eH. undatus\u003c/em\u003e peel extract. It is observed that the fruit peel extract significantly exhibited the flavonoids and phenolics that are responsible for its antioxidant activity. The potential use of silver nanoparticles obtained from the extract of \u003cem\u003eH. undatus\u003c/em\u003e peel extract has been suggested as an effective anticancer agent against PC3 cell lines. The biosynthesized AgNPs from \u003cem\u003eH. undatus\u003c/em\u003e peel extract showed significant antibacterial, antioxidant, anticancer, and thrombolytic activity which indicates their potential therapeutic applications. Thereby, further studies are required to discover the medicinal drugs from this source.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe authors declare that they have no known competing financial interests that could have appeared to influence the work reported in this paper.\u003c/p\u003e\u003cp\u003e \u003ch2\u003eConflicts of interest/Competing interests\u003c/h2\u003e \u003cp\u003eThe authors declare that there is no conflict of interest.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding \u0026ndash;\u003c/h2\u003e \u003cp\u003eNot applicable\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eSRK conceived the idea. The literature was searched and the manuscript was written by AA and SRK, Corrected by MHS, edited by SS, GRM and SRK, and supervised by SRK and SS. All authors reviewed the manuscript.\u003c/p\u003e\u003ch2\u003eData availability\u003c/h2\u003e \u003cp\u003e(data transparency) - Not applicable\u003c/p\u003e\u003ch2\u003eCode availability \u0026ndash;\u003c/h2\u003e \u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eVennila K, Chitra L, Balagurunathan R, Palvannan T. Comparison of biological activities of selenium and silver nanoparticles attached with bioactive phytoconstituents: green synthesized using Spermacocehispida extract. Adv Nat Sci Nano Sci Nanotechnol. 2018; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1088/2043-6254/aa9f4d\u003c/span\u003e\u003cspan address=\"10.1088/2043-6254/aa9f4d\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNirmala C, Sridevi M. 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Antibiotics. 2023; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/antibiotics12030564\u003c/span\u003e\u003cspan address=\"10.3390/antibiotics12030564\" 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":"H. undatus, antibacterial, antifungal, antioxidant, thrombolytic, anticancer activity","lastPublishedDoi":"10.21203/rs.3.rs-3805384/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3805384/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe green synthesis of nanoparticles has gained significant impacts in various fields due to their eco-friendly approach. In this study, silver nanoparticles were synthesized from aqueous extract of \u003cem\u003eH. undatus\u003c/em\u003e fruit peel. The presence of silver nanoparticles (AgNPs) was analyzed using characterization methods such as UV-Vis, FTIR, GCMS, XRD, EDAX and FESEM. The synthesized AgNPs showed the highest antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e when compared to \u003cem\u003eS. pneumoniae\u003c/em\u003e. The highest antifungal activity was observed against \u003cem\u003eC. albicans\u003c/em\u003e than \u003cem\u003eC. tropicalis.\u003c/em\u003e IC\u003csub\u003e50\u003c/sub\u003e value of antibiofilm activity of AgNPs was recorded as 2.81 \u0026micro;g/ml whereas \u003cem\u003eH. undatus\u003c/em\u003e peel extract exhibited the value of 1.34 \u0026micro;g/ml. The invitro antioxidant activity of AgNPs was evaluated using two different methods. A strong DPPH radical scavenging activity of AgNPs and fruit peel extract was observed with the IC\u003csub\u003e50\u003c/sub\u003e values of 3.8 and 2.03 \u0026micro;g/ml respectively. On other hand, nitric oxide radical scavenging activity were recorded and IC\u003csub\u003e50\u003c/sub\u003e values was calculated to be 2.8 and 2.3 \u0026micro;g/ml. The AgNPs demonstrated thrombolytic activity in human blood with 10, 32.36 and 56.25% lysis. The cytotoxicity of AgNPs possessed minimum activity with an IC\u003csub\u003e50\u003c/sub\u003e 0.2 \u0026micro;g/ml and peel extract showed the maximum cytotoxicity activity with an IC\u003csub\u003e50\u003c/sub\u003e 0.3 \u0026micro;g/ml. The findings of this study demonstrate that the synthesized AgNPs from \u003cem\u003eH. undatus\u003c/em\u003e peel extract used as a potential candidate for treating prostate cancer.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e","manuscriptTitle":"A Comparative Study on Biosynthesized Silver Nanoparticles from H. undatus Fruit Peel and its Therapeutic Applications","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-02 05:22:48","doi":"10.21203/rs.3.rs-3805384/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-01-25T09:36:19+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-01-18T04:08:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"3260f51b-b549-4cdd-b099-3f51d95072bf","date":"2024-01-10T08:49:56+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-01-10T07:23:25+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-01-02T00:56:55+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-12-29T09:51:40+00:00","index":"","fulltext":""},{"type":"submitted","content":"Discover Nano","date":"2023-12-25T18:13:01+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":"47f361af-60f5-46ef-956f-a6fbfe40c441","owner":[],"postedDate":"January 2nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-03-25T15:04:45+00:00","versionOfRecord":{"articleIdentity":"rs-3805384","link":"https://doi.org/10.1186/s11671-024-03995-w","journal":{"identity":"discover-nano","isVorOnly":false,"title":"Discover Nano"},"publishedOn":"2024-03-18 15:01:00","publishedOnDateReadable":"March 18th, 2024"},"versionCreatedAt":"2024-01-02 05:22:48","video":"","vorDoi":"10.1186/s11671-024-03995-w","vorDoiUrl":"https://doi.org/10.1186/s11671-024-03995-w","workflowStages":[]},"version":"v1","identity":"rs-3805384","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3805384","identity":"rs-3805384","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}