Exploring the Antibacterial and Antibiofilm Efficacy of Silver Nanoparticles Biosynthesized Using Punica granatum Leaves.
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ABSTRACT:
SUBMITTER: Singhal M
PROVIDER: S-EPMC8510064 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:This study aimed to determine phytochemical contents, antibacterial properties, and antibiotic modulating potential of Punica granatum leaf extracts: hexane, chloroform, ethyl acetate, ethanol, and aqueous extracts as well as an extract enriched with total oligomer flavonoids (TOFs). The TOF extract contained the highest value of phenols and flavonoids. Rutin, luteolin, gallic acid, and ellagic acid were determined by HPLC analysis of this extract. The antibacterial activity was assayed by the disc diffusion method and microdilution method against Staphylococcus aureus and Escherichia coli standard ATCC strains and clinical isolates resistant strains. The TOF extract was the most active against all tested strains. The checkerboard method was used for the determination of synergy between two antibiotics (amoxicillin and cefotaxime) and P. granatum leaf extracts. The best synergistic interaction was found with TOF extract combined with amoxicillin for penicillin-resistant E. coli and penicillin-resistant S. aureus. These results can be assigned to tannins, flavonoids, and phenolic acids found in P. granatum leaf extracts. Pomegranate leaf extracts or active compounds isolated from these extracts could be used to fight the emergence and spread of resistant bacterial strains.
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Project description:Microbial resistance and biofilm formation have been considered as the main problems associated with microbial resistance. Several antimicrobial agents cannot penetrate biofilm layers and cannot eradicate microbial infection. Therefore, the aim of this study is the biological synthesis of silver and copper nanoparticles to assess their activities on bacterial attachment and on the viability of dormant cells within the biofilm matrix. Ag-NPs and Cu-NPs were biosynthesized using Streptomyces isolate S29. The biologically synthesized Ag-NPs and Cu-NPs exhibited brown and blue colors and were detected by UV/Vis spectrophotometry at 476 and 594 nm, respectively. The Ag-NPs showed an average size of 10-20 nm as indicated by TEM, and 25-35 nm for Cu-NPs. Both Ag-NPs and Cu-NPs were monodispersed with a polydispersity index of 0.1-0.546 and zeta potential were - 29.7, and - 33.7 mv, respectively. The biologically synthesized Ag-NPs and Cu-NPs significantly eliminated bacterial attachment and decreased the viable cells in the biofilm matrix as detected by using crystal violet and tri-phenyl tetrazolium chloride assays. Furthermore, Ag-NPs and Cu-NPs significantly eradicated mature biofilms developed by various Gram-negative pathogens, including A. baumannii, K. pneumoniae and P. aeruginosa standard strains and clinical isolates. Data were also confirmed at the molecular level with prominent elimination of biofilm gene expression carO, bssS and pelA in A. baumannii, K. pneumoniae and P. aeruginosa, respectively compared to untreated cells under the same conditions. As indicated, Ag-NPs and Cu-NPs could be used as adjuvant therapy in eradication of antibiotic resistance and biofilm matrix associated with Gram-negative bacterial infection.
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Project description:Silver nanoparticles were produced with AgF as the starting Ag(I) salt, with pectin as the reductant and protecting agent. While the obtained nanoparticles (pAgNP-F) have the same dimensional and physicochemical properties as those already described by us and obtained from AgNO3 and pectin (pAgNP-N), the silver nanoparticles from AgF display an increased antibacterial activity against E. coli PHL628 and Staphylococcus epidermidis RP62A (S. epidermidis RP62A), both as planktonic strains and as their biofilms with respect to pAgNP-N. In particular, a comparison of the antimicrobial and antibiofilm action of pAgNP-F has been carried out with pAgNP-N, pAgNP-N and added NaF, pure AgNO3, pure AgF, AgNO3 and added NaF and pure NaNO3 and NaF salts. By also measuring the concentration of the Ag+ cation released by pAgNP-F and pAgNP-N, we were able to unravel the separate contributions of each potential antibacterial agent, observing an evident synergy between p-AgNP and the F- anion: the F- anion increases the antibacterial power of the p-AgNP solutions even when F- is just 10 µM, a concentration at which F- alone (i.e., as its Na+ salt) is completely ineffective.
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Project description:The present study demonstrates an economical and environmental affable approach for the synthesis of "protein-capped" silver nanoparticles in aqueous solvent system. A variety of standard techniques viz. UV-visible spectroscopy, transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD) measurements were employed to characterize the shape, size and composition of nanoparticles. The synthesized nanoparticles were found to be homogenous, spherical, mono-dispersed and covered with multi-layered protein shell. In order to prepare bare silver nanoparticles, the protein shell was removed from biogenic nanoparticles as confirmed by UV-visible spectroscopy, FTIR and photoluminescence analysis. Subsequently, the antibacterial efficacy of protein-capped and bare silver nanoparticles was compared by bacterial growth rate and minimum inhibitory concentration assay. The results revealed that bare nanoparticles were more effective as compared to the protein-capped silver nanoparticles with varying antibacterial potential against the tested Gram positive and negative bacterial species. Mechanistic studies based on ROS generation and membrane damage suggested that protein-capped and bare silver nanoparticles demonstrate distinct mode of action. These findings were strengthened by the TEM imaging along with silver ion release measurements using inductively coupled plasma atomic emission spectroscopy (ICP-AES). In conclusion, our results illustrate that presence of protein shell on silver nanoparticles can decrease their bactericidal effects. These findings open new avenues for surface modifications of nanoparticles to modulate and enhance their functional properties.
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Project description:The provision of nanoparticles using biogenic material as a part of green chemistry is an attractive nanotechnology. The current research aimed to test the antimicrobial and cytotoxic efficacy of silver nanoparticles synthesized by extracts of Phoenix dactylifera, Ferula asafetida, and Acacia nilotica as reductant and stabilizing agents in silver nanoparticle formation. Synthesized nanoparticles were evaluated for their antimicrobial activity against Staphylococcus aureus (Gram-positive) and Pseudomonas aeruginosa and Escherichia coli (Gram-negative) using an agar well diffusion assay. Furthermore, cytotoxic ability was investigated against LoVo cells. The potential phyto-constituents of plant extracts were identified by Fourier-transform infrared spectroscopy (FT-IR) techniques. Field emission scanning electron microscopy (FE-SEM), transmission electron microscope (TEM), and zeta potential analyzed the size and morphology of the biogenic nanoparticles. The current study revealed the ability of the tested plant extract to convert silver ions to silver nanoparticles with an average size that ranged between 67.8 ± 0.3 and 155.7 ± 1.5 nm in diameter. Biogenic AgNPs showed significant antibacterial ability (10 to 32 mm diameter) and anticancer ability against a LoVo cell with IC50 ranged between 35.15⁻56.73 μg/mL. The innovation of the present study is that the green synthesis of NPs, which is simple and cost effective, provides stable nano-materials and can be an alternative for the large-scale synthesis of silver nanoparticles.
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Project description:Nanotechnology is being investigated for its potential to improve nanomedicine for human health. The purpose of this study was to isolate carbapenemase-producing Gram-negative bacilli (CPGB), investigate the presence of carbapenemase resistance genes, determine their antibiogram and ability to biosynthesise silver nanoparticles (Ag NPs), and estimate the antibacterial activity of Acinetobacter baumannii-biosynthesised Ag NPs on CPGB alone and in combination with antibiotics. A total of 51 CPGBs were isolated from various specimens in the study. The automated Vitek-2 system was used to identify and test these strains' antimicrobial susceptibilities. The carbapenemase resistance genes were identified using a polymerase chain reaction (PCR). Under the CPGB, A. baumannii could biosynthesise Ag NPs. X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), and field emission scanning electron were used to characterise Ag NPs. The antibacterial activity of Ag NP alone and in combination with antibiotics against CPGB was determined using the broth microdilution method, and their synergistic effect was determined using the checkerboard assay. bla NDM and bla OXA-48 were the most commonly reported, and 90% of the isolates produced multiple carbapenemase genes. Tigecycline proved to be the most effective anti-CPGB antibiotic. Isolates with more resistance genes were more resistant to antibiotics, and isolates with three genes (42%) had the most extensively drug-resistant patterns (38%). A significant relationship was discovered between genetic and antibiotic resistance patterns. Only A. baumannii produced Ag NPs out of all the isolates tested. Ag NPs with a size of 10 nm were confirmed by UV-visible spectroscopy, FT-IR, XRD, and TEM analysis. The Ag NPs were effective against CPGB, with minimum inhibitory concentrations ranging from 64 to 8 μg/ml on average. Surprisingly, the combination of Ag NPs and antibiotics demonstrated synergistic and partial synergistic activity (fractional inhibitory concentration between 0.13 and 0.56) against CPGB, as well as a significant reduction in antibiotic concentrations, particularly in the case of A. baumanii versus ceftriaxone (1024 to 4 μg/ml). The notable synergistic activity of Ag NPs with antibiotics represents a valuable nanomedicine that may find clinical application in the future as a combined remedy.
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Project description:The biogenic synthesis of silver nanoparticles (AgNPs) has a wide range of applications in the pharmaceutical industry. Here, we synthesized AgNPs using the aqueous flower extract of Bauhinia tomentosa Linn. Formation of AgNPs was observed using ultraviolet-visible light spectrophotometry at different time intervals. Maximum absorption was observed after 4 h at 420 nm due to the reduction of Ag+ to Ag0. The stabilizing activity of functional groups was identified by Fourier-transform infrared spectroscopy. Size and surface morphology were also analyzed using scanning electron microscopy. The present study revealed the AgNPs were spherical in form with a diameter of 32 nm. The face-centered cubic structure of AgNPs was indexed using X-ray powder diffraction with peaks at 2θ = 37°, 49°, 63°, and 76° (corresponding to the planes of silver 111, 200, 220, 311), respectively. Energy-dispersive X-ray spectroscopy revealed that pure reduced silver (Ag0) was the major constituent (59.08%). Antimicrobial analyses showed that the biosynthesized AgNPs possess increased antibacterial activity (against Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative), with larger zone formation against S. aureus (9.25 mm) compared with that of E. coli (6.75 mm)) and antifungal activity (against Aspergillus flavus and Candida albican (with superior inhibition against A. flavus (zone of inhibition: 7 mm) compared with C. albicans (zone of inhibition: 5.75 mm)). Inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was found to be dose-dependent with half-maximal inhibitory concentration (IC50) values of 56.77 μg/mL and 43.03 μg/mL for AgNPs and ascorbic acid (control), respectively, thus confirming that silver nanoparticles have greater antioxidant activity than ascorbic acid. Molecular docking was used to determine the mode of antimicrobial interaction of our biosynthesized B. tomentosa Linn flower-powder extract-derived AgNPs. The biogenic AgNPs preferred hydrophobic contacts to inhibit bacterial and fungal sustainability with reducing antioxidant properties, suggesting that biogenic AgNPs can serve as effective medicinal agents.
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Project description:Metallic nanoparticles (NPs) especially silver (Ag) NPs have shown immense potential in medical applications due to their distinctive physio-chemical and biological properties. This article reports the conjugation of Ag NPs with Rubus fairholmianus extract. The modification of Ag NPs was confirmed using various physico-chemical characterization techniques. The cytotoxic effect of Rubus-conjugated Ag NPs (RAgNPs) was studied by LDH assay and proliferation by ATP assay. The apoptotic inducing ability of the NPs were investigated by Annexin V/PI staining, caspase 3/7 analysis, cytochrome c release, intracellular ROS analysis, Hoechst staining and mitochondrial membrane potential analysis using flow cytometry. The expression of apoptotic proteins caspase 3, Bax and P53 were analyzed using ELISA and caspase 3, Bax using western blotting. Cells treated with 10 µg/mL RAgNPs showed an increased number of cell death by microscopic analysis compared to untreated control cells. The RAgNPs induced a statistically significant dose-dependent decrease in proliferation (p < 0.001 for 5 and 10 µg/mL) and increased cytotoxicity in MCF-7 cells. A 1.83 fold increase in cytotoxicity was observed in cells treated with 10 µg/mL (p < 0.05) compared to the untreated cells. Nuclear damage and intracellular ROS production were observed upon treatment with all tested concentrations of RAgNPs and the highest concentrations (5 and 10 µg/mL) showed significant (p < 0.05, p < 0.01) expression of caspase 3, Bax and P53 proteins. The data strongly suggest that RAgNPs induces cell death in MCF-7 cells through the mitochondrial-mediated intrinsic apoptosis pathway. The present investigation supports the potential of RAgNPs in anticancer drug development.
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Project description:The current study demonstrates the synthesis of fatty acids (FAs) capped silver nanoparticles (AgNPs) using aqueous poly-herbal drug Liv52 extract (PLE) as a reducing, dispersing and stabilizing agent. The NPs were characterized by various techniques and used to investigate their potent antibacterial, antibiofilm, antifungal and anticancer activities. GC-MS analysis of PLE shows a total of 37 peaks for a variety of bio-actives compounds. Amongst them, n-hexadecanoic acid (21.95%), linoleic acid (20.45%), oleic acid (18.01%) and stearic acid (13.99%) were found predominately and most likely acted as reducing, stabilizing and encapsulation FAs in LIV-AgNPs formation. FTIR analysis of LIV-AgNPs shows some other functional bio-actives like proteins, sugars and alkenes in the soft PLE corona. The zone of inhibition was 10.0 ± 2.2-18.5 ± 1.0 mm, 10.5 ± 2.5-22.5 ± 1.5 mm and 13.7 ± 1.0-16.5 ± 1.2 against P. aeruginosa, S. aureus and C. albicans, respectively. LIV-AgNPs inhibit biofilm formation in a dose-dependent manner i.e., 54.4% ± 3.1%-10.12% ± 2.3% (S. aureus), 72.7% ± 2.2%-23.3% ± 5.2% (P. aeruginosa) and 85.4% ± 3.3%-25.6% ± 2.2% (C. albicans), and SEM analysis of treated planktonic cells and their biofilm biomass validated the fitness of LIV-AgNPs in future nanoantibiotics. In addition, as prepared FAs rich PLE capped AgNPs have also exhibited significant (p < 0.05 *) antiproliferative activity against cultured HCT-116 cells. Overall, this is a very first demonstration on employment of FAs rich PLE for the synthesis of highly dispersible, stable and uniform sized AgNPs and their antibacterial, antifungal, antibiofilm and anticancer efficacy.
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Project description:The production of cheap and effective compound for medicinal application is a major challenge for scientific community. So, several biological materials have been explored for the possible application in material synthesis which are useful in biomedical uses. Here, biomolecules from green tea leaves were functionalized on the surface of silicon dioxide nanoparticles (GSiO2 NPs). Next, the decoration silver (Ag) NPs on the surface of the GSiO2 NPs was observed in very short time of incubation in aqueous AgNO3. Ultraviolet-visible spectroscopy confirmed the formation of Ag NPs and the high-resolution transmission and scanning electron microscopies confirmed the decoration of spherical Ag NPs of 10 to 15 nm size on the surface of GSiO2 NPs. The antimicrobial activity of the Ag-GSiO2 NPs was determined against Staphylococcus aureus and Escherichia coli. The Ag-GSiO2 NPs displayed sustainable antimicrobial activity compared to Ag ions. The results indicate the potential value of Ag-GSiO2 NPs in surgical material and food processing.
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