Production of silver nanoparticles with strong and stable antimicrobial activity against highly pathogenic and multidrug resistant bacteria.
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ABSTRACT:
SUBMITTER: Saeb AT
PROVIDER: S-EPMC4100300 | biostudies-other | 2014
REPOSITORIES: biostudies-other
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Project description:Antibiotic resistance is one of the most important global problems currently confronting the world. Different biomedical applications of silver nanoparticles (AgNPs) have indicated them to be promising antimicrobial agents. In the present study, extracellular extract of an endophytic bacterium, Pantoea ananatis, was used for synthesis of AgNPs. The synthesized AgNPs were characterized by UV⁻Vis spectroscopy, FTIR, transmission electron microscopy (TEM), scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX), and Zeta potential. The antimicrobial potential of the AgNPs against pathogenic Staphylococcus aureus subsp. aureus (ATCC 11632), Bacillus cereus (ATCC 10876), Escherichia coli (ATCC 10536), Pseudomonas aeruginosa (ATCC 10145) and Candida albicans (ATCC 10231), and multidrug resistant (MDR) Streptococcus pneumoniae (ATCC 700677), Enterococcus faecium (ATCC 700221) Staphylococcus aureus (ATCC 33592) Escherichia coli (NCTC 13351) was investigated. The synthesized spherical-shaped AgNPs with a size range of 8.06 nm to 91.32 nm exhibited significant antimicrobial activity at 6 μg/disc concentration against Bacillus cereus (ATCC 10876) and Candida albicans (ATCC 10231) which were found to be resistant to conventional antibiotics. The synthesized AgNPs showed promising antibacterial efficiency at 10 µg/disc concentration against the MDR strains. The present study suggests that AgNPs synthesized by using the endophytic bacterium P. ananatis are promising antimicrobial agent.
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Project description:The present study reported a single step synthesis of silver nanoparticles using ampicillin (Amp-AgNps), a second-generation ? lactam antibiotic to get nanoformulation having dual properties that of antibiotic and silver. The Amp-AgNps was characterized by UV-VIS spectroscopy, TEM, XRD, FTIR and TGA. FTIR and TGA results suggested that amine group of Ampicllin reduce the metalic silver into nano form. These results were further validated by computational molecular dynamics simulation. The antibacterial potential of Amp-AgNps was investigated against sensitive and drug resistant bacteria. MIC of Amp-AgNps against 6 different bacterial strains were in the range of 3-28?µg/ml which is much lower than the MIC of ampicillin (12-720?µg/ml) and chemically synthesized silver nanoparticles (280-640?µg/ml). The repeated exposure to drugs may lead to development of resistance mechanism in bacteria against that drug, so the efficacy of Amp-AgNps after repeated exposure to bacterial strains were also studied. The results indicate that bacterial strains do not show any resistance to these Amp-AgNps even after exposure up to 15 successive cycles. The biocompatibility of these Amp-AgNps was checked against cell lines by using Keratinocytes cell lines (HaCaT).
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Project description:In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA.
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Project description:BackgroundCyanobacteria are considered as green nano-factories. Manipulation of the size of biogenic silver nanoparticles is needed to produce particles that suit the different applications such as the use as antibacterial agents. The present study attempts to manipulate the size of biosynthesized silver nanoparticles produced by cyanobacteria and to test the different-sized nanoparticles against pathogenic clinical bacteria.MethodsCyanothece-like. coccoid unicellular cyanobacterium was tested for its ability to biosynthesize nanosilver particles of different sizes. A stock solution of silver nitrate was prepared from which three different concentrations were added to cyanobacterial culture. UV-visible spectroscopy and FTIR were conducted to characterize the silver nanoparticles produced in the cell free filtrate. Dynamic Light Scattering (DLS) was performed to determine the size of the nanoparticles produced at each concentration. The antimicrobial bioassays were conducted on broad host methicillin-resistant Staphylococcus aureus (MRSA), and Streptococcus sp., was conducted to detect the nanoparticle size that was most efficient as an antimicrobial agent.ResultsThe UV-Visible spectra showed excellent congruence of the plasmon peak characteristic of nanosilver at 450 nm for all three different concentrations, varying peak heights were recorded according to the concentration used. The FTIR of the three solutions revealed the absence of characteristic functional groups in the solution. All three concentrations showed spectra at 1636 and 2050-2290 nm indicating uniformity of composition. Moreover, DLS analysis revealed that the silver nanoparticles produced with lowest concentration of precursor AgNO3 had smallest size followed by those resulting from the higher precursor concentration. The nanoparticles resulting from highest concentration of precursor AgNO3 were the biggest in size and tending to agglomerate when their size was above 100 nm. The three types of differently-sized silver nanoparticles were used against two bacterial pathogenic strains with broad host range; MRSA-(Methicillin-resistant Staphylococcus aureus) and Streptococcus sp. The three types of nanoparticles showed antimicrobial effects with the smallest nanoparticles being the most efficient in inhibiting bacterial growth.DiscussionNanosilver particles biosynthesized by Cyanothece-like cyanobacterium can serve as antibacterial agent against pathogens including multi-drug resistant strains. The most appropriate nanoparticle size for efficient antimicrobial activity had to be identified. Hence, size-manipulation experiment was conducted to find the most effective size of nanosilver particles. This size manipulation was achieved by controlling the amount of starting precursor. Excessive precursor material resulted in the agglomeration of the silver nanoparticles to a size greater than 100 nm. Thereby decreasing their ability to penetrate into the inner vicinity of microbial cells and consequently decreasing their antibacterial potency.ConclusionAntibacterial nanosilver particles can be biosynthesized and their size manipulated by green synthesis. The use of biogenic nanosilver particles as small as possible is recommended to obtain effective antibacterial agents.
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Project description:Conventional antibiotics are facing strong microbial resistance that has recently reached critical levels. This situation is leading to significantly reduced therapeutic potential of a huge proportion of antimicrobial agents currently used in clinical settings. Antimicrobial peptides (AMPs) could provide the medical community with an alternative strategy to traditional antibiotics for combating microbial resistance. However, the development of AMPs into clinically useful antibiotics is hampered by their relatively low stability, toxicity, and high manufacturing costs. In this study, a novel in-house-designed potent ultrashort AMP named RBRBR was encapsulated into chitosan-based nanoparticles (CS-NPs) based on the ionotropic gelation method. The encapsulation efficacy reported for RBRBR into CS-NPs was 51.33%, with a loading capacity of 10.17%. The release kinetics of RBRBR from the nanocarrier exhibited slow release followed by progressive linear release for 14 days. The antibacterial kinetics of RBRBR-CS-NPs was tested against four strains of Staphylococcus aureus for 4 days, and the developed RBRBR-CS-NPs exhibited a 3-log decrease in the number of colonies when compared to CS-NP and a 5-log decrease when compared to control bacteria. The encapsulated peptide NP formulation managed to limit the toxicity of the free peptide against both mammalian cells and human erythrocytes. Additionally, the peptide NPs demonstrated up to 98% inhibition of biofilm formation when tested against biofilm-forming bacteria. Loading RBRBR into CS-NPs could represent an innovative approach to develop delivery systems based on NP technology for achieving potent antimicrobial effects against multidrug-resistant and biofilm-forming bacteria, with negligible systemic toxicity and reduced synthetic costs, thereby overcoming the obstructions to clinical development of AMPs.
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Project description:The anti-microbial properties of acetone extracts from Hibiscus sabdariffa calyces, fractions isolated by silica gel chromatography and hibiscus acid purified from some of these fractions and additionally identified by nuclear magnetic resonance spectroscopy, mid-infrared spectroscopy and X-ray diffraction, were studied against both multidrug-resistant Salmonella strains and pathogenic Escherichia coli bacteria. Gel diffusion was used to determine the anti-microbial effects. The mode of action of hibiscus acid was determined by crystal violet assay. Hibiscus acid and 17 of the 25 chromatographic fractions obtained, displayed an anti-microbial effect against all bacterial strains tested. Hibiscus acid showed a greater anti-microbial effect than the acetone extract against most of the bacteria strains, while chromatographic fractions IX-XIV exerted the greatest anti-microbial effect against all bacteria. The minimum inhibitory concentration of the acetone extract was 7 mg/mL, and the minimum bactericidal concentration was 10 mg/mL, while the corresponding values for hibiscus acid were 4-7 and 7 mg/mL, respectively. The results of the crystal violet assay indicate that hibiscus acid alters membrane permeability. Hibiscus acid is a potential alternative to control multidrug-resistant bacteria. Due to its ready availability and easy extraction from H. sabdariffa, hibiscus acid is potentially useful in the food industries.
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Project description:In this study, the cell free modified tryptone soya broth (pH 7.4 ± 0.2) of Bacillus subtilis URID 12.1 showed significant antimicrobial activity against multidrug-resistant strains of Staphylococcus aureus, S. epidermidis, Streptococcus pyogenes and Enterococcus faecalis. The partially purified antimicrobial molecule was found to be resistant to extremes of pH and temperatures and also to higher concentrations of trypsin and proteinase K. The antimicrobial molecule was purified by a three-step method that included reversed-phase high performance liquid chromatography (RP-HPLC). The minimum inhibitory concentration (MIC) values were determined for 14 species of bacteria using a microbroth dilution technique. The HPLC-purified fraction showed the MICs ranging from 0.5 to 16 ?g/ml for methicillin and vancomycin-resistant Staphylococcus aureus (MVRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE) strains. The molecular mass of the antimicrobial compound was determined to be 842.37 Da. The same antimicrobial fraction showed negligible haemolytic activity against human red blood cells even at a concentration as high as 100 ?g/ml. Because of its significant antimicrobial activity at low MIC values coupled with its non-haemolytic property, it may prove to be a novel antimicrobial lead molecule.
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Project description:Objectives:Due to the expanded bacterial genetic tolerance to antibiotics through different mechanisms, infectious diseases of MDR bacteria are difficult for treatment. Consequently, we synthesized drug conjugated nanoparticles to dissolve this problem. Moreover, the present study aims to display the cell death status treated with cefotaxime-CS-AgNPs and also, apoptosis pathways of human RPE-1 normal cells and human MCF-7 breast cancer cells. Methods:Here, we demonstrate the possibility to synthesize AgNPs and conjugate them with cefotaxime to survey the probability of cefotaxime-CS-AgNPs as an antimicrobial agent against cefotaxime-resistant strains E. coli and MRSA. Results:TEM showed the size of AgNPs, CS-AgNPs and cefotaxime-CS-AgNPs ranged from 7.42 to 18.3 nm, 8.05-23.89 nm and 8.48-25.3 nm, respectively, with a spherical shape. The cefotaxime-CS-AgNPs enhanced the high antimicrobial properties compared to AgNPs or pure antibiotic. The MIC of Cefotaxime-CS-AgNPs ranged from 3 µg/mL to 8 µg/mL against tested E. coli and MRSA bacteria. Consequently, the highest reduction in the MIC of cefotaxime-CS-AgNPs was noted against tested strains ranging from 22% to 96%. Comparing cefotaime-CS-AgNPs to AgNPs we showed that cefotaime-CS-AgNPs have no cytotoxic effect on normal cells at even 12 µg/mL for 24 hrs. The IC50 for the AgNPs and cefotaxime-CS-AgNPs was 12 µg/mL for human RPE-1 normal cells and human MCF-7 breast cancer cell lines. The pro-apoptotic genes p53, p21, and Bax of cancer cell lines significantly upregulated followed by downregulated by anti-apoptotic gene Bcl-2 after 48 hrs at 24 µg/mL, and this concentration represents the most effective dose. Conclusion:Results enhanced the conjugating utility in old unresponsive cefotaxime to AgNPs to restore its efficiency against previous strains and demonstrated potential therapeutic applications of cefotaxime-CS-AgNPs. Moreover, this research gives remarkable insights for designing nanoscale delivery and curative systems that have a pronounced cytotoxic activity on cancer cells and are safe to normal cells.
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