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Characterization, Antimicrobial and Antioxidant Evaluation of Biofabricated Silver Nanoparticles from Endophytic Pantoea anthophila.

ABSTRACT: Endophyte mediated nanoparticles fabrication were emerging as a new frontier in nanomedicines that produce high biocompatible and functionalized silver nanoparticles. In this study, silver nanoparticles were successfully biosynthesized from the extracellular extract of endophytic bacterium Pantoea anthophila isolated from the stem of Waltheria indica for the first time. The synthesized nanoparticles showed a strong absorption band at 410 nm in the UV-Visible range. The dynamic light scattering and zeta potential analysis indicated that the average particle size was 16 nm at 5.30 mV. FTIR spectrum displayed the presence of various functional groups at 3423.65, 1633.71, 1022.27, 607.58 cm^-1 that stabilised the nanoparticle. X-ray diffraction peaks were conferred to 100, 200, 220 and 311 planes of a face centred cubic structure. TEM and SEM micrograph revealed the spherical-shaped, polycrystalline nature with the presence of elemental silver analysed by EDAX. Selected area electron diffraction also confirms the orientation of silver nanoparticles with X-ray diffraction analysis. Antimicrobial activity against 10 different human pathogenic bacteria and fungi showed a broad spectrum inhibition against both Gram-positive and Gram-negative bacteria. Among the bacterial pathogens, B. Subtilis exhibited low activity compared to other pathogens. C. albicans was greatly controlled than other fungal species. A strong free radical scavenging activity of silver nanoparticles with IC₅₀ values 31.29 ± 0.73, 19.83 ± 1.57, 35.64 ± 0.94, 42.07 ± 1.30, 29.70 ± 2.26, 29.10 ± 0.82, 36.80 ± 0.63 μg/ml was obtained in different antioxidant assays that were comparable to the reference. The study suggests that the silver nanoparticles can be biosynthesized from endophytic P. anthophila metabolites with significant therapeutic potential. With proper validation, the biosynthesized silver nanoparticles can be developed as a promising antiviral and anticancer drug candidate.

SUBMITTER: Nirmala C

PROVIDER: S-EPMC8006880 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:An endophytic fungal strain isolated from the leaves of Gymnema sylvestre was identified as Pestalotiopsis microspora VJ1/VS1 based on nucleotide sequencing of internal transcribed spacer region (ITS 1-5.8S-ITS 2) of 18S rRNA gene (NCBI accession number KX213894). In this study, an efficient and ecofriendly approach has been reported for the synthesis of silver nanoparticles (AgNPs) using aqueous culture filtrate of P. microspora. Ultraviolet-visible analysis confirmed the synthesis of AgNPs by showing characteristic absorption peak at 435 nm. Fourier transform infrared spectroscopy analysis revealed the presence of phenolic compounds and proteins in the fungal filtrate, which are plausibly involved in the biosynthesis and capping of AgNPs. Transmission electron microscopy (TEM) showed that the AgNPs were spherical in shape of 2-10 nm in size. Selected area electron diffraction and X-ray diffraction studies determined the crystalline nature of AgNPs with face-centered cubic (FCC) lattice phase. Dynamic light scattering analysis showed that the biosynthesized AgNPs possess high negative zeta potential value of -35.7 mV. Biosynthesized AgNPs were proved to be potential antioxidants by showing effective radical scavenging activity against 2,2'-diphenyl-1-picrylhydrazyl and H2O2 radicals with IC50 values of 76.95±2.96 and 94.95±2.18 µg/mL, respectively. The biosynthesized AgNPs exhibited significant cytotoxic effects against B16F10 (mouse melanoma, IC50 =26.43±3.41 µg/mL), SKOV3 (human ovarian carcinoma, IC50 =16.24±2.48 µg/mL), A549 (human lung adenocarcinoma, IC50 =39.83±3.74 µg/mL), and PC3 (human prostate carcinoma, IC50 =27.71±2.89 µg/mL) cells. The biosynthesized AgNPs were found to be biocompatible toward normal cells (Chinese hamster ovary cell line, IC50 =438.53±4.2 µg/mL). Cytological observations on most susceptible SKOV3 cells revealed concentration-dependent apoptotic changes that include cell membrane blebbing, cell shrinkage, pyknotic nuclei, karyorrhexis followed by destructive fragmentation of nuclei. The results together in this study strongly provided a base for the development of potential and versatile biomedical applications of biosynthesized AgNPs in the near future.

Project description:An accumulating body of evidence reports the synthesis and biomedical applications of silver nanoparticles. However, the studies regarding the use of maleic acid and citric acid in the synthesis of nano-sized silver particles (AgNPs) and micro-sized silver particles (AgMPs) as well as their antibacterial, antifungal, and anticancer activities have not been reported. In the current study, we synthesized AgNPs and AgMPs using maleic acid and citric acid as capping agents and have characterized them by UV-Vis, energy-dispersive X-Ray spectroscopy (EDS), X-Ray diffraction (XRD), and scanning electron microscope (SEM) analysis. The capped silver particles were examined for their antimicrobial activity and cytotoxicity against bacteria, fungi, and brine shrimp. Additionally, the anticancer activity of these particles was tested against human breast and liver cancer cell lines. The free radical scavenging activity of capped silver particles was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. SEM analysis revealed a round plate-like morphology of maleic acid capped particles with an average size of 39 ± 4 nm, whereas citric acid capped particles display flower-shaped morphology with rough surfaces and an average size of 250 ± 5 nm. The uncapped AgMPs were hexagonal with 500 ± 4 nm size. EDS and XRD analysis confirmed the presence of Ag and face-centered cubic crystalline nature, respectively. Functionally, capped silver particles exhibited antibacterial activity against Gram-positive (Staphylococcus aureus, Bacillus subtilis, and Micrococcus luteus) and Gram-negative bacteria (Salmonellasetubal, Enterobacter aerogenes, and Agrobacterium tumefaciens). The bactericidal activity was more active against Gram-negative bacteria with minimum inhibitory concentration (MIC) as low as 5 ppm as compared to 25 ppm for Gram-positive. Similarly, the silver particles demonstrated antifungal activity by inhibiting the growth of five fungal strains (Mucor species, Aspergillusniger, Aspergillus flavus, Aspergillus fumigatus, and Fusariumsolani) up to 50% at the concentration of 500 ppm. Additionally, these particles showed substantial toxicity against brine shrimp and also significantly inhibited the proliferation of breast cancer (MCF7) and liver cancer (HePG2) cell lines (IC50 8.9-18.56 µM). Uncapped AgMPs were less effective, inhibiting only the proliferation of MCF7 cells with IC50 46.54 µM. Besides cytotoxicity, these particles acted as potential antioxidants, showing free radical scavenging up to 74.4% in a concentration-dependent manner. Taken together, our results showed that the modifiers affect the shape and size of silver particles and may, in part, contribute to the antimicrobial and antioxidant activity of silver particles. However, the contribution of maleic acid and citric acid in enhancing the antimicrobial, anticancer, and antioxidant potential independent of silver nano and microparticles needs to be studied further. In vivo experiments may determine the therapeutic effectiveness of silver particles capped with these modifiers.

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Characterization, Antimicrobial and Antioxidant Evaluation of Biofabricated Silver Nanoparticles from Endophytic Pantoea anthophila.

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