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Green Synthesized Silver Nanoparticles: Antibacterial and Anticancer Activities, Biocompatibility, and Analyses of Surface-Attached Proteins.

ABSTRACT: The increasing number of multi-drug-resistant bacteria and cancer cases, that are a real threat to humankind, forces research world to develop new weapons to deal with it. Biogenic silver nanoparticles (AgNPs) are considered as a solution to this problem. Biosynthesis of AgNPs is regarded as a green, eco-friendly, low-priced process that provides small and biocompatible nanostructures with antimicrobial and anticancer activities and potential application in medicine. The biocompatibility of these nanoparticles is related to the coating with biomolecules of natural origin. The synthesis of AgNPs from actinobacterial strain was confirmed using UV-Vis spectroscopy while their morphology, crystalline structure, stability, and coating were characterized using, transmission electron microscopy (TEM), X-ray diffraction (XRD), Zeta potential and Fourier transform infrared spectroscopy (FTIR). Antibacterial activity of biogenic AgNPs was evaluated by determination of minimum inhibitory and minimum biocidal concentrations (MIC and MBC) against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. The potential mechanism of antibacterial action of AgNPs was determined by measurement of ATP level. Since the use of AgNPs in biomedical applications depend on their safety, the in vitro cytotoxicity of biosynthesized AgNPs on MCF-7 human breast cancer cell line and murine macrophage cell line RAW 264.7 using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, cell lactate dehydrogenase (LDH) release and measurement of reactive oxygen species (ROS) level were assessed. The nanoparticle protein capping agent that can be involved in reduction of silver ions to AgNPs and their stabilization was identified using LC-MS/MS. Nanoparticles were spherical in shape, small in size (mean 13.2 nm), showed crystalline nature, good stability (-18.7 mV) and presence of capping agents. They exhibited antibacterial activity (MIC of 8-128 μg ml^-1, MBC of 64-256 μg ml^-1) and significantly decreased ATP levels in bacterial cells after treatment with different concentrations of AgNPs. The in vitro analysis showed that the AgNPs demonstrated dose-dependent cytotoxicity against RAW 264.7 macrophages and MCF-7 breast cancer cells but higher against the latter than the former. Cell viability decrease was found to be 42.2-14.2 and 38.0-15.5% while LDH leakage 14.6-42.7% and 19.0-45.0%, respectively. IC₅₀ values calculated for MTT assay was found to be 16.3 and 12.0 μg ml^-1 and for LDH assay 102.3 and 76.2 μg ml^-1, respectively. Moreover, MCF-7 cells released a greater amount of ROS than RAW 264.7 macrophages during stimulation with all tested concentrations of AgNPs (1.47-3.13 and 1.02-2.58 fold increase, respectively). The SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) analysis revealed the presence of five protein bands at a molecular weight between 31.7 and 280.9 kDa. These proteins showed the highest homology to hypothetical proteins and porins from E. coli, Delftia sp. and Pseudomonas rhodesiae. Based on obtained results it can be concluded that biogenic AgNPs were capped with proteins and demonstrated potential as antimicrobial and anticancer agent.

SUBMITTER: Wypij M

PROVIDER: S-EPMC8100210 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Green biosynthesized nanoparticles have a bright future because they can be produced using a method that is more energy-efficient, cost-effective, repeatable, and environmentally friendly than physical or chemical synthesis. In this study, silver nanoparticles (AgNPs) were produced using the Fusarium nygamai isolate AJTYC1. Several techniques were used to characterize the synthesized AgNPs, including UV-Vis spectroscopy, transmission electron microscope, zeta potential analysis, X-ray diffraction analysis, energy dispersive X-ray, and Fourier transform-infrared spectroscopy. AgNPs showed a distinctive surface plasmon resonance (SPR) peak in the UV-visible range at 310 nm. The morphology of the biosynthesized AgNPs was spherical, and the TEM image shows that they ranged in size from 27.3 to 53.1 nm. The notable peaks of the FT-IR results show the different groups for the alkane, alkynes, cyclic alkenes, carboxylic, aromatic amine, esters, and phenolics. Additionally, the results showed that AgNPs had superior antioxidant activity when compared to ascorbic acid and butylated hydroxytoluene, which is a powerful antioxidant. Additionally, AgNPs have antibacterial action utilizing agar diffusion against gram-positive bacteria, gram-negative bacteria, and antifungal activity. AgNPs' anticancer activity varied depending on the type of cancer it was used to treat, including hepatocellular cancer (HepG2), colorectal carcinoma (HCT116), and breast cancer of the mammary gland (MCF7). The viability of the cancer cell lines was reduced with increasing AgNP concentration. AgNPs also demonstrated promising photocatalytic activity by reducing methylene blue, safranin, crystal violet, and green malachite by 88.3%, 81.5%, 76.4%, and 78.2%, respectively. In addition, AgNPs significantly affected the Allium cepa plant's mitotic index and resulted in chromosomal abnormalities as compared to the control. Thus, the synthesized AgNPs demonstrated an efficient, eco-friendly, and sustainable method for decolorizing dyes as well as antioxidant, antibacterial, antifungal, and anticancer activities. This could be a huge victory in the fight against numerous dynamic diseases and lessen wastewater dye contamination.

Project description:BackgroundThe overuse of antibiotics leads to the emergence of antibiotic-resistant microbes which causes high mortality worldwide. Therefore, the synthesis of new active compounds has multifunctional activities are the main challenge. Nanotechnology provides a solution for this issue.MethodThe endophytic fungal strain Aspergillus terreus BR.1 was isolated from the healthy root of Allium sativum and identified using internal transcribed spacer (ITS) sequence analysis. The copper oxide nanoparticles (CuO-NPs) were synthesized by harnessing the metabolites of the endophytic fungal strain. The UV-Visble spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), Transmission electron micrscopy (TEM), Energy dispersive X-ray (EDX), X-ray diffraction (XRD), Dynamic light scattering (DLS), and zeta potential (ζ) were used for the characterization of synthesized CuO-NPs. The activity against different pathogenic bacteria and Candida species were investigated by agar well-diffusion method. The biocombatibility and anticancer activity were assessed by MTT assay method. The scavenging of DPPH was used to investigate the antioxidant activity of synthesized CuO-NPs.ResultsData showed the successful formation of crystalline nature and spherical shape CuO-NPs with sizes in the ranges of 15-55 nm. The EDX reveals that the as-formed sample contains ions of C, O, Cl, and Cu with weight percentages of 18.7, 23.82, 11.31, and 46.17%, respectively. The DLS and ζ-potential showed high homogeneity and high stability of synthesized CuO-NPs with a polydispersity index (PDI) of 0.362 and ζ-value of - 26.6 mV. The synthesized CuO-NPs exhibited promising antibacterial and anti-Candida activity (concentration-dependent) with minimum inhibitory concentration (MIC) values in the ranges of 25-50 µg mL-1. Moreover, the fungal mediated-CuO-NPs targeted cancer cells of MCF7 and PC3 at low IC50 concentrations of 159.2 ± 4.5 and 116.2 ± 3.6 µg mL-1, respectively as compared to normal cells (Vero and Wi38 with IC50 value of 220.6 ± 3.7 and 229.5 ± 2.1 µg mL-1, respectively). The biosynthesized CuO-NPs showed antioxidant activity as detected by the DPPH method with scavenging percentages of 80.5 ± 1.2% at a concentration of 1000 µg mL-1 and decreased to 20.4 ± 4.2% at 1.9 µg mL-1 as compared to ascorbic acid (control) with scavenging activity of 97.3 ± 0.2 and 37.5 ± 1.3% at the same concentrations, respectively.ConclusionThe fungal mediated-CuO-NPs exhibited promising activity and can be integrated into various biomedical and theraputic applications.

Project description:The prospective application of probiotics is an adjuvant for the advancement of novel antimicrobial and wound-healing agents. Currently, probiotic bacteria are utilized for the biosynthesis of nanoparticles in the development of innovative therapeutics. The present study aimed at using nanoparticle-conjugated probiotic bacteria for enhanced antibacterial and wound-healing activity. In the present investigation, the probiotic bacteria were isolated from a dairy source (milk from domestic herbivores). They screened for antibacterial activity against infection-causing Gram-negative (Pseudomonas aeruginosa and Escherichia coli) and Gram-positive (Bacillus subtilis and Staphylococcus aureus) pathogens. Further, the probiotic strain with higher bactericidal activity was used to synthesize silver, selenium, and copper nanoparticles. The isolated strain was found to be Lactiplantibacillus plantarum and it only has the ability to synthesize silver nanoparticles. This was verified using Ultra violet-Visible (UV-Vis) spectroscopy, where the test solution turned brown and the greatest UV-Vis absorptions peaked at 425 nm. Optimization studies on the synthesis of AgNPs (silver nanoparticles) are presented and the results show that stable synthesis was obtained by using a concentration of 1mM silver nitrate (AgNO3) at a temperature of 37 °C with pH 8. The FTIR (Fourier transform infrared spectroscopy) study confirmed the involvement of functional groups from the cell biomass that were involved in the reduction process. Additionally, biosynthesized AgNPs showed increased antioxidant and antibacterial activities. The nano silver had a size distribution of 14 nm and was recorded with HR-TEM (high-resolution transmission electron microscopy) examination. The EDX (energy dispersive X-ray) analysis revealed 57% of silver groups found in the nanoparticle production. The biosynthesized AgNPs show significant wound-healing capabilities with 96% of wound closure (fibroblast cells) being observed through an in vitro scratch-wound assay. The cytotoxic experiments demonstrated that the biosynthesized AgNPs are not extremely hazardous to the fibroblast cells. The present study provides a new platform for the green synthesis of AgNPs using probiotic bacteria, showing significant antibacterial and wound-healing potentials against infectious pathogens.

Project description:Silver nanoparticles (Ag-NPs) have a unique mode of action as antibacterial agents in addition to their anticancer and antioxidant properties. In this study, microbial nanotechnology is employed to synthesize Ag-NPs using the cell filtrate of Streptomyces enissocaesilis BS1. The synthesized Ag-NPs are confirmed by ultraviolet-visible (UV-Vis), Fourier transform infrared (FT-IR), X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Also, the effects of different factors on Ag-NPs synthesis were evaluated to set the optimum synthesis conditions. Also, the antibacterial, antibiofilm, and anticancer activity of Ag-NPs was assessed. The X-ray diffraction (XRD) analysis confirmed the crystalline nature of the sample and validated that the crystal structure under consideration is a face-centered cubic (FCC) pattern. The TEM examination displayed the spherical particles of the Ag-NPs and their average size, which is 32.2 nm. Fourier transform infrared spectroscopy (FTIR) revealed significant changes in functionality after silver nanoparticle dispersion, which could be attributed to the potency of the cell filtrate of Streptomyces enissocaesilis BS1 to act as both a reducing agent and a capping agent. The bioactivity tests showed that our synthesized Ag-NPs exhibited remarkable antibacterial activity against different pathogenic strains. Also, when the preformed biofilms of Pseudomonas aeruginosa ATCC 9027, Salmonella typhi ATCC 12023, Escherichia coli ATCC 8739, and Staphylococcus aureus ATCC 6598 were exposed to Ag NPs 50 mg/ml for 24 hours, the biofilm biomass was reduced by 10.7, 34.6, 34.75, and 39.08%, respectively. Furthermore, the Ag-NPs showed in vitro cancer-specific sensitivity against human breast cancer MCF-7 cell lines and colon cancer cell line Caco-2, and the IC50 was 0.160 mg/mL and 0.156 mg/mL, respectively. The results of this study prove the ease and efficiency of the synthesis of Ag-NPs using actinomycetes and demonstrate the significant potential of these Ag-NPs as anticancer and antibacterial agents.

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Green Synthesized Silver Nanoparticles: Antibacterial and Anticancer Activities, Biocompatibility, and Analyses of Surface-Attached Proteins.

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