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1.  Chrysopogon zizanioides aqueous extract mediated synthesis, characterization of crystalline silver and gold nanoparticles for biomedical applications 
The exploitation of various plant materials for the biosynthesis of nanoparticles is considered a green technology as it does not involve any harmful chemicals. The aim of this study was to develop a simple biological method for the synthesis of silver and gold nanoparticles using Chrysopogon zizanioides. To exploit various plant materials for the biosynthesis of nanoparticles was considered a green technology. An aqueous leaf extract of C. zizanioides was used to synthesize silver and gold nanoparticles by the bioreduction of silver nitrate (AgNO3) and chloroauric acid (HAuCl4) respectively. Water-soluble organics present in the plant materials were mainly responsible for reducing silver or gold ions to nanosized Ag or Au particles. The synthesized silver and gold nanoparticles were characterized by ultraviolet (UV)-visible spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) analysis. The kinetics decline reactions of aqueous silver/gold ion with the C. zizanioides crude extract were determined by UV-visible spectroscopy. SEM analysis showed that aqueous gold ions, when exposed to the extract were reduced and resulted in the biosynthesis of gold nanoparticles in the size range 20–50 nm. This eco-friendly approach for the synthesis of nanoparticles is simple, can be scaled up for large-scale production with powerful bioactivity as demonstrated by the synthesized silver nanoparticles. The synthesized nanoparticles can have clinical use as antibacterial, antioxidant, as well as cytotoxic agents and can be used for biomedical applications.
PMCID: PMC3704452  PMID: 23861583
nanoparticles; bioreduction; SEM; silver; gold
2.  Hyaluronan- and heparin-reduced silver nanoparticles with antimicrobial properties 
Nanomedicine (London, England)  2009;4(4):421-429.
Silver nanoparticles exhibit unique antibacterial properties that make these ideal candidates for biological and medical applications. We utilized a clean method involving a single synthetic step to prepare silver nanoparticles that exhibit antimicrobial activity.
Materials & methods
These nanoparticles were prepared by reducing silver nitrate with diaminopyridinylated heparin (DAPHP) and hyaluronan (HA) polysaccharides and tested for their efficacy in inhibiting microbial growth.
Results & discussion
The resulting silver nanoparticles exhibit potent antimicrobial activity against Staphylococcus aureus and modest activity against Escherichia coli. Silver–HA showed greater antimicrobial activity than silver–DAPHP, while silver–glucose nanoparticles exhibited very weak antimicrobial activity. Neither HA nor DAPHP showed activity against S. aureus or E. coli.
These results suggest that DAPHP and HA silver nanoparticles have potential in antimicrobial therapeutic applications.
PMCID: PMC2717895  PMID: 19505245
antimicrobial; Escherichia coli; silver heparin nanoparticles; silver hyaluronan nanoparticles; silver nanoparticles; Staphylococcus aureus
3.  Gold and silver nanoparticles from Trianthema decandra: synthesis, characterization, and antimicrobial properties 
There is an increasing commercial demand for nanoparticles due to their wide applicability in various markets, including medicine, catalysis, electronics, chemistry, and energy. In this report, a simple and ecofriendly chemical reaction for the synthesis of gold and silver nanoparticles from Trianthema decandra (Aizoaceae) has been developed.
Methods and results
On treatment of aqueous solutions containing chloroauric acid or silver nitrate with root extract of T. decandra, stable gold or silver nanoparticles were rapidly formed. The kinetics of reduction of gold and silver ions during the reaction was analyzed by ultraviolet-visible spectroscopy. Field emission-scanning electron microscopy showed formation of gold nanoparticles in various shapes, including spherical, cubical, triangular, and hexagonal, while silver nanoparticles were spherical. The size of the gold nanoparticles was 33–65 nm and that of the silver nanoparticles was 36–74 nm. Energy dispersive x-ray and Fourier transform infrared spectroscopy confirmed the presence of metallic gold and metallic silver in the respective nanoparticles. The antimicrobial properties of the synthesized nanoparticles were analyzed using the Kirby-Bauer method. The results show varied susceptibility of microorganisms to the gold and silver nanoparticles.
It is believed that phytochemicals present in T. decandra extract reduce the silver and gold ions into metallic nanoparticles. This strategy reduces the cost of production and the environmental impact. The silver and gold nanoparticles formed showed strong activity against all microorganisms tested.
PMCID: PMC3471601  PMID: 23091381
Trianthema decandra; gold; silver; nanoparticles; antimicrobial activity
4.  3, 4-dihydroxy-L-phenylalanine-derived melanin from Yarrowia lipolytica mediates the synthesis of silver and gold nanostructures 
Nanobiotechnology applies the capabilities of biological systems in generating a variety of nano-sized structures. Plants, algae, fungi and bacteria are some systems mediating such reactions. In fungi, the synthesis of melanin is an important strategy for cell-survival under metal-stressed conditions. Yarrowia lipolytica, the biotechnologically significant yeast also produces melanin that sequesters heavy metal ions. The content of this cell-associated melanin is often low and precursors such as L-tyrosine or 3, 4-dihydroxy-L-phenylalanine (L-DOPA) can enhance its production. The induced melanin has not been exploited for the synthesis of nanostructures. In this investigation, we have employed L-DOPA-melanin for the facile synthesis of silver and gold nanostructures. The former have been used for the development of anti-fungal paints.
Yarrowia lipolytica NCIM 3590 cells were incubated with L-DOPA for 18 h and the resultant dark pigment was subjected to physical and chemical analysis. This biopolymer was used as a reducing and stabilizing agent for the synthesis of silver and gold nanostructures. These nanoparticles were characterized by UV-Visible spectra, X-ray diffraction (XRD) studies, and electron microscopy. Silver nanoparticles were evaluated for anti-fungal activity.
The pigment isolated from Y. lipolytica was identified as melanin. The induced pigment reduced silver nitrate and chloroauric acid to silver and gold nanostructures, respectively. The silver nanoparticles were smaller in size (7 nm) and displayed excellent anti-fungal properties towards an Aspergillus sp. isolated from a wall surface. An application of these nanoparticles as effective paint-additives has been demonstrated.
The yeast mediated enhanced production of the metal-ion-reducing pigment, melanin. A simple and rapid method for the extracellular synthesis of nanoparticles with paint-additive-application was developed.
PMCID: PMC3660187  PMID: 23363424
Yarrowia lipolytica; L-DOPA melanin; Nanoparticles; Anti-fungal activity
5.  PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments 
PVP-capped silver nanoparticles with a diameter of the metallic core of 70 nm, a hydrodynamic diameter of 120 nm and a zeta potential of −20 mV were prepared and investigated with regard to their biological activity. This review summarizes the physicochemical properties (dissolution, protein adsorption, dispersability) of these nanoparticles and the cellular consequences of the exposure of a broad range of biological test systems to this defined type of silver nanoparticles. Silver nanoparticles dissolve in water in the presence of oxygen. In addition, in biological media (i.e., in the presence of proteins) the surface of silver nanoparticles is rapidly coated by a protein corona that influences their physicochemical and biological properties including cellular uptake. Silver nanoparticles are taken up by cell-type specific endocytosis pathways as demonstrated for hMSC, primary T-cells, primary monocytes, and astrocytes. A visualization of particles inside cells is possible by X-ray microscopy, fluorescence microscopy, and combined FIB/SEM analysis. By staining organelles, their localization inside the cell can be additionally determined. While primary brain astrocytes are shown to be fairly tolerant toward silver nanoparticles, silver nanoparticles induce the formation of DNA double-strand-breaks (DSB) and lead to chromosomal aberrations and sister-chromatid exchanges in Chinese hamster fibroblast cell lines (CHO9, K1, V79B). An exposure of rats to silver nanoparticles in vivo induced a moderate pulmonary toxicity, however, only at rather high concentrations. The same was found in precision-cut lung slices of rats in which silver nanoparticles remained mainly at the tissue surface. In a human 3D triple-cell culture model consisting of three cell types (alveolar epithelial cells, macrophages, and dendritic cells), adverse effects were also only found at high silver concentrations. The silver ions that are released from silver nanoparticles may be harmful to skin with disrupted barrier (e.g., wounds) and induce oxidative stress in skin cells (HaCaT). In conclusion, the data obtained on the effects of this well-defined type of silver nanoparticles on various biological systems clearly demonstrate that cell-type specific properties as well as experimental conditions determine the biocompatibility of and the cellular responses to an exposure with silver nanoparticles.
PMCID: PMC4222445  PMID: 25383306
aerosols; biological properties; cell biology; nanoparticles; nanotoxicology; silver
6.  Potential Theranostics Application of Bio-Synthesized Silver Nanoparticles (4-in-1 System) 
Theranostics  2014;4(3):316-335.
In this report, we have designed a simple and efficient green chemistry approach for the synthesis of colloidal silver nanoparticles (b-AgNPs) that is formed by the reduction of silver nitrate (AgNO3) solution using Olax scandens leaf extract. The colloidal b-AgNPs, characterized by various physico-chemical techniques exhibit multifunctional biological activities (4-in-1 system). Firstly, bio-synthesized silver nanoparticles (b-AgNPs) shows enhanced antibacterial activity compared to chemically synthesize silver nanoparticles (c-AgNPs). Secondly, b-AgNPs show anti-cancer activities to different cancer cells (A549: human lung cancer cell lines, B16: mouse melanoma cell line & MCF7: human breast cancer cells) (anti-cancer). Thirdly, these nanoparticles are biocompatible to rat cardiomyoblast normal cell line (H9C2), human umbilical vein endothelial cells (HUVEC) and Chinese hamster ovary cells (CHO) which indicates the future application of b-AgNPs as drug delivery vehicle. Finally, the bio-synthesized AgNPs show bright red fluorescence inside the cells that could be utilized to detect the localization of drug molecules inside the cancer cells (a diagnostic approach). All results together demonstrate the multifunctional biological activities of bio-synthesized AgNPs (4-in-1 system) that could be applied as (i) anti-bacterial & (ii) anti-cancer agent, (iii) drug delivery vehicle, and (iv) imaging facilitator. To the best of our knowledge, there is not a single report of biosynthesized AgNPs that demonstrates the versatile applications (4-in-1 system) towards various biomedical applications. Additionally, a plausible mechanistic approach has been explored for the synthesis of b-AgNPs and its anti-bacterial as well as anti-cancer activity. We strongly believe that bio-synthesized AgNPs will open a new direction towards various biomedical applications in near future.
PMCID: PMC3915094  PMID: 24505239
Bio-synthesis; Silver nanoparticle; Green Chemistry; Olax scandens; Multifunctional activities; Antibacterial; anti-cancer.
7.  Oxidative stress mediated cytotoxicity of biologically synthesized silver nanoparticles in human lung epithelial adenocarcinoma cell line 
Nanoscale Research Letters  2014;9(1):459.
The goal of the present study was to investigate the toxicity of biologically prepared small size of silver nanoparticles in human lung epithelial adenocarcinoma cells A549. Herein, we describe a facile method for the synthesis of silver nanoparticles by treating the supernatant from a culture of Escherichia coli with silver nitrate. The formation of silver nanoparticles was characterized using various analytical techniques. The results from UV-visible (UV-vis) spectroscopy and X-ray diffraction analysis show a characteristic strong resonance centered at 420 nm and a single crystalline nature, respectively. Fourier transform infrared spectroscopy confirmed the possible bio-molecules responsible for the reduction of silver from silver nitrate into nanoparticles. The particle size analyzer and transmission electron microscopy results suggest that silver nanoparticles are spherical in shape with an average diameter of 15 nm. The results derived from in vitro studies showed a concentration-dependent decrease in cell viability when A549 cells were exposed to silver nanoparticles. This decrease in cell viability corresponded to increased leakage of lactate dehydrogenase (LDH), increased intracellular reactive oxygen species generation (ROS), and decreased mitochondrial transmembrane potential (MTP). Furthermore, uptake and intracellular localization of silver nanoparticles were observed and were accompanied by accumulation of autophagosomes and autolysosomes in A549 cells. The results indicate that silver nanoparticles play a significant role in apoptosis. Interestingly, biologically synthesized silver nanoparticles showed more potent cytotoxicity at the concentrations tested compared to that shown by chemically synthesized silver nanoparticles. Therefore, our results demonstrated that human lung epithelial A549 cells could provide a valuable model to assess the cytotoxicity of silver nanoparticles.
PMCID: PMC4167841  PMID: 25242904
Adenocarcinoma cells A549; Reactive oxygen species generation (ROS); Lactate dehydrogenase (LDH); Mitochondrial transmembrane potential (MTP); Silver nanoparticles (AgNP)
8.  Antimicrobial efficacy and ocular cell toxicity from silver nanoparticles 
Silver in various forms has long been recognized for antimicrobial properties, both in biomedical devices and in eyes. However, soluble drugs used on the ocular surface are rapidly cleared through tear ducts and eventually ingested, resulting in decreased efficacy of the drug on its target tissue and potential concern for systemic side effects. Silver nanoparticles were studied as a source of anti-microbial silver for possible controlled-release contact lens controlled delivery formulations. Silver ion release over a period of several weeks from nanoparticle sources of various sizes and doses in vitro was evaluated in vitro against Pseudomonas aeruginosa strain PA01. Mammalian cell viability and cytokine expression in response to silver nanoparticle exposure is evaluated using corneal epithelial cells and eye-associated macrophages cultured in vitro in serum-free media. Minimal microcidal and cell toxic effects were observed for several silver nanoparticle suspensions and aqueous extraction times for bulk total silver concentrations commensurate with comparative silver ion (e.g., Ag+(aq)) toxicity. This indicates that (1) silver particles themselves are not microcidal under conditions tested, and (2) insufficient silver ion is generated from these particles at these loadings to produce observable biological effects in these in vitro assays. If dosing allows substantially increased silver particle loading in the lens, the bactericidal efficacy of silver nanoparticles in vitro is one possible approach to limiting bacterial colonization problems associated with extended-wear contact lenses.
PMCID: PMC2768321  PMID: 19865601
silver; nanoparticle; cornea; bacteria; macrophage; antimicrobial; contact lens; infection
9.  Synthesis and antimicrobial effects of silver nanoparticles produced by chemical reduction method 
Background and the purpose of the study
The most prominent nanoparticles for medical uses are nanosilver particles which are famous for their high anti-microbial activity. Silver ion has been known as a metal ion that exhibit anti-mold, anti-microbial and anti-algal properties for a long time. In particular, it is widely used as silver nitrate aqueous solution which has disinfecting and sterilizing actions. The purpose of this study was to evaluate the antimicrobial activity as well as physical properties of the silver nanoparticles prepared by chemical reduction method.
Silver nanoparticles (NPs) were prepared by reduction of silver nitrate in the presence of a reducing agent and also poly [N-vinylpyrolidone] (PVP) as a stabilizer. Two kinds of NPs were synthesized by ethylene glycol (EG) and glucose as reducing agent. The nanostructure and particle size of silver NPs were confirmed by scanning electron microscopy (SEM) and laser particle analyzer (LPA). The formations of the silver NPs were monitored using ultraviolet- visible spectroscopy. The anti-bacterial activity of silver NPs were assessed by determination of their minimum inhibitory concentrations (MIC) against the Gram positive (Staphylococcus aureus and Staphylococcus epidermidis) as well as Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria.
Results and Conclusion
The silver nanoparticles were spherical with particle size between 10 to 250 nm. Analysis of the theoretical (Mie light scattering theory) and experimental results showed that the silver NPs in colloidal solution had a diameter of approximately 50 nm.
Both colloidal silver NPs showed high anti-bacterial activity against Gram positive and Gram negative bacteria. Glucose nanosilver colloids showed a shorter killing time against most of the tested bacteria which could be due to their nanostructures and uniform size distribution patterns.
PMCID: PMC3304363  PMID: 22615613
Nanoparticles; Silver; Ethylene glycol; Glucose; Colloids; Reduction method
10.  Potential of Microalgae and Lactobacilli in Biosynthesis of Silver Nanoparticles  
BioImpacts : BI  2011;1(3):149-152.
Application of nanoparticles has been extensively increased in last decades. Nanoparticles of noble metals such as gold, platinum and especially silver are widely applied in medical and pharmaceutical applications. Although, variety of physical and chemical methods has been developed for production of metal nanoparticles, because of destructive effects of them on environment, biosynthetic methods have been suggested as a novel alternative. Some bacteria and microalgae have different ranges of potentiality to uptake metal ions and produce nanoparticles during detoxification process. In the present work, we study the potential of three Lactobacilli and three algal species in production of AgNPs in different concentrations of silver nitrate.
Utilizing AAS, XRD and TEM methods, Nannochloropsis oculata, Dunaliella salina and Chlorella vulgaris as three algal species in addition to three Lactobacilli including L. acidophilus, L. casei, L. reuteri were monitored for production of silver nanoparticles. Three concentrations of AgNO3 (0.001, 0.002, 0.005 M) and two incubation times (24h and 48h) were included in this study.
Our findings demonstrated that C. vulgaris, N. oculata and L. acidophilus have the potential of nanosilver production in a culture medium containing 0.001 M of AgNO3 within 24 hours. Also L. casei and L. reuteri species exhibited their potential for production of silver nanoparticles in 0.002 M concentration of AgNO3 in 24 hours. The size range of particles was approximately less than 15 nm. The uptake rate of silver in the five species was between 1.0 to 2.7 mg/g of dry weight. Nanoparticle production was not detected in other treatments and the algae Dunaliella.
The biosynthesis of silver nanoparticles in all of three Lactobacilli and two algal species including N. oculata and C. vulgaris was confirmed.
PMCID: PMC3648959  PMID: 23678420
Silver nanoparticles; Biosynthesis; Lactobacilli; Microalgae
11.  Memecylon edule leaf extract mediated green synthesis of silver and gold nanoparticles 
We used an aqueous leaf extract of Memecylon edule (Melastomataceae) to synthesize silver and gold nanoparticles. To our knowledge, this is the first report where M. edule leaf broth was found to be a suitable plant source for the green synthesis of silver and gold nanoparticles. On treatment of aqueous solutions of silver nitrate and chloroauric acid with M. edule leaf extract, stable silver and gold nanoparticles were rapidly formed. The gold nanoparticles were characterized by UV-visible spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray analysis (EDAX) and Fourier transform infra-red spectroscopy (FTIR). The kinetics of reduction of aqueous silver and gold ions during reaction with the M. edule leaf broth were easily analyzed by UV-visible spectroscopy. SEM analysis showed that aqueous gold ions, when exposed to M. edule leaf broth, were reduced and resulted in the biosynthesis of gold nanoparticles in the size range 20–50 nm. TEM analysis of gold nanoparticles showed formation of triangular, circular, and hexagonal shapes in the size range 10–45 nm. The resulting silver nanoparticles were predominantly square with uniform size range 50–90 nm. EDAX results confirmed the presence of triangular nanoparticles in the adsorption peak of 2.30 keV. Further FTIR analysis was also done to identify the functional groups in silver and gold nanoparticles. The characterized nanoparticles of M. edule have potential for various medical and industrial applications. Saponin presence in aqueous extract of M. edule is responsible for the mass production of silver and gold nanoparticles.
PMCID: PMC3131193  PMID: 21753878
Memecylon edule; nanoparticles; bioreduction; electron microscopy; FTIR
12.  Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria 
Nanoscale Research Letters  2014;9(1):365.
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.
PMCID: PMC4114801  PMID: 25114655
Green synthesis; Macrophomina phaseolina; Silver nanoparticles; Antimicrobial; Capping; DNA fragmentation
13.  A Preliminary Assessment of Silver Nanoparticle Inhibition of Monkeypox Virus Plaque Formation 
Nanoscale Research Letters  2008;3(4):129-133.
The use of nanotechnology and nanomaterials in medical research is growing. Silver-containing nanoparticles have previously demonstrated antimicrobial efficacy against bacteria and viral particles. This preliminary study utilized an in vitro approach to evaluate the ability of silver-based nanoparticles to inhibit infectivity of the biological select agent, monkeypox virus (MPV). Nanoparticles (10–80 nm, with or without polysaccharide coating), or silver nitrate (AgNO3) at concentrations of 100, 50, 25, and 12.5 μg/mL were evaluated for efficacy using a plaque reduction assay. Both Ag-PS-25 (polysaccharide-coated, 25 nm) and Ag-NP-55 (non-coated, 55 nm) exhibited a significant (P ≤ 0.05) dose-dependent effect of test compound concentration on the mean number of plaque-forming units (PFU). All concentrations of silver nitrate (except 100 μg/mL) and Ag-PS-10 promoted significant (P ≤ 0.05) decreases in the number of observed PFU compared to untreated controls. Some nanoparticle treatments led to increased MPV PFU ranging from 1.04- to 1.8-fold above controls. No cytotoxicity (Vero cell monolayer sloughing) was caused by any test compound, except 100 μg/mL AgNO3. These results demonstrate that silver-based nanoparticles of approximately 10 nm inhibit MPV infection in vitro, supporting their potential use as an anti-viral therapeutic.
PMCID: PMC3244796
Nanoparticle; Monkeypox virus; Silver; Anti-viral therapeutic; Plaque reduction assay
14.  Facile Synthesis of Gold Nanoparticles with Narrow Size Distribution by Using AuCl or AuBr as the Precursor 
Gold(I) halides, including AuCl and AuBr, were employed for the first time as precursors in the synthesis of Au nanoparticles. The synthesis was accomplished by dissolving AuI halides in chloroform in the presence of alkylamines, followed by decomposition at 60 °C. The relative low stability of the AuI halides and there derivatives eliminated the need for a reducing agent, which is usually required for AuIII-based precursors to generate Au nanoparticles. Controlled growth of Au nanoparticles with a narrow size distribution was achieved when AuCl and oleylamine were used for the synthesis. FTIR and mass spectra revealed that a complex, [AuCl(oleylamine)], was formed through coordination between oleylamine and AuCl. Thermolysis of the complex in chloroform led to the formation of dioleylamine and Au nanoparticles. When oleylamine was replaced with octadecylamine, much larger nanoparticles were obtained due to the lower stability of [AuCl(octadecylamine)] complex relative to [AuCl(oleylamine)]. Au nanoparticles can also be prepared from AuBr through thermolysis of the [AuBr(oleylamine)] complex. Due to the oxidative etching effect caused by Br−, the nanoparticles obtained from AuBr exhibited an aspect ratio of 1.28, in contrast to 1.0 for the particles made from AuCl. Compared to the existing methods for preparing Au nanoparticles through the reduction of AuIII compounds, this new approach based on AuI halides offers great flexibility in terms of size control.
PMCID: PMC2492888  PMID: 18058964
alkylamines; complexes; gold; nanoparticles
15.  Biosynthesis of silver nanoparticles from Schizophyllum radiatum HE 863742.1: their characterization and antimicrobial activity 
3 Biotech  2013;4(3):227-234.
Development of reliable and eco-friendly process for synthesis of silver nanoparticles is an important step in the field of application in nanotechnology. One of the options to achieve this objective is to use natural biological processes. They have an advantage over conventional methods involving chemical agents associated with environmental toxicity. This study demonstrates the extra-cellular synthesis of stable silver nanoparticles using the white rot fungus, Schizophyllum radiatum with GenBank Accession no HE 863742.1. The supernatant of the seed media obtained after separating the cells has been used for the synthesis of silver nanoparticles. The morphology and structure of synthesized silver nanoparticles were characterized using FT-IR, XRD, UV–visible spectrum of the aqueous medium containing silver ion showed a peak in the range of 420–430 nm corresponding to the Plasmon absorbance of silver nanoparticles. Scanning electron microscopy micrograph showed formation of well-dispersed silver nanoparticles in the range of 10–40 nm. The effect of different carbon sources and the time taken for formation particles and the anti-microbial activity of synthesized nanoparticles were carried and compared with silver nitrate solution and with standard streptomycin. The process of reduction being extra-cellular and fast may lead to the development of an easy bioprocess for synthesis of silver nanoparticles.
PMCID: PMC4026449
White rot fungi; Silver nanoparticles; Antimicrobial activity; Scanning electron microscopy
16.  The morphology of silver nanoparticles prepared by enzyme-induced reduction 
Silver nanoparticles were synthesized by an enzyme-induced growth process on solid substrates. In order to customize the enzymatically grown nanoparticles (EGNP) for analytical applications in biomolecular research, a detailed study was carried out concerning the time evolution of the formation of the silver nanoparticles, their morphology, and their chemical composition. Therefore, silver-nanoparticle films of different densities were investigated by using scanning as well as transmission electron microscopy to examine their structure. Cross sections of silver nanoparticles, prepared for analysis by transmission electron microscopy were additionally studied by energy-dispersive X-ray spectroscopy in order to probe their chemical composition. The surface coverage of substrates with silver nanoparticles and the maximum particle height were determined by Rutherford backscattering spectroscopy. Variations in the silver-nanoparticle films depending on the conditions during synthesis were observed. After an initial growth state the silver nanoparticles exhibit the so-called desert-rose or nanoflower-like structure. This complex nanoparticle structure is in clear contrast to the auto-catalytically grown spherical particles, which maintain their overall geometrical appearance while increasing their diameter. It is shown, that the desert-rose-like silver nanoparticles consist of single-crystalline plates of pure silver. The surface-enhanced Raman spectroscopic (SERS) activity of the EGNP structures is promising due to the exceptionally rough surface structure of the silver nanoparticles. SERS measurements of the vitamin riboflavin incubated on the silver nanoparticles are shown as an exemplary application for quantitative analysis.
PMCID: PMC3388365  PMID: 23016145
EGNP; enzymatically grown silver nanoparticles; enzyme-induced deposition; nanoflower; SERS
17.  New Paradigm Shift for the Green Synthesis of Antibacterial Silver Nanoparticles Utilizing Plant Extracts 
Toxicological Research  2014;30(3):169-178.
This review covers general information regarding the green synthesis of antibacterial silver nanoparticles. Owing to their antibacterial properties, silver nanoparticles are widely used in many areas, especially biomedical applications. In green synthesis practices, the chemical reducing agents are eliminated, and biological entities are utilized to convert silver ions to silver nanoparticles. Among the various biological entities, natural plant extracts have emerged as green reducing agents, providing eco-friendly routes for the preparation of silver nanomaterials. The most obvious merits of green synthesis are the increased biocompatibility of the resulting silver nanoparticles and the ease with which the reaction can be carried out. This review summarizes some of the plant extracts that are used to produce antibacterial silver nanoparticles. Additionally, background information regarding the green synthesis and antibacterial activity of silver nanoparticles is provided. Finally, the toxicological aspects of silver nanoparticles are briefly mentioned.
PMCID: PMC4206743  PMID: 25343010
Plant extracts; Green synthesis; Silver nanoparticles; Antibacterial activity
18.  SERS Properties of Different Sized and Shaped Gold Nanoparticles Biosynthesized under Different Environmental Conditions by Neurospora crassa Extract 
PLoS ONE  2013;8(10):e77486.
Surface-enhanced Raman scattering (SERS) is a surface-sensitive technique that enhances Raman scattering by molecules adsorbed on rough metal surfaces. It is known that metal nanoparticles, especially gold and silver nanoparticles, exhibit great SERS properties, which make them very attractive for the development of biosensors and biocatalysts. On the other hand, the development of ecofriendly methods for the synthesis of metallic nanostructures has become the focus of research in several countries, and many microorganisms and plants have already been used to biosynthesize metallic nanostructures. However, the majority of these are pathogenic to plants or humans. Here, we report gold nanoparticles with good SERS properties, biosynthesized by Neurospora crassa extract under different environmental conditions, increasing Raman signals up to 40 times using methylene blue as a target molecule. Incubation of tetrachloroauric acid solution with the fungal extract at 60°C and a pH value of a) 3, b) 5.5, and c) 10 resulted in the formation of gold nanoparticles of a) different shapes like triangles, hexagons, pentagons etc. in a broad size range of about 10-200 nm, b) mostly quasi-spheres with some different shapes in a main size range of 6-23 nm, and c) only quasi-spheres of 3-12 nm. Analyses included TEM, HRTEM, and EDS in order to corroborate the shape and the elemental character of the gold nanoparticles, respectively. The results presented here show that these ‘green’ synthesized gold nanoparticles might have potential applicability in the field of biological sensing.
PMCID: PMC3793955  PMID: 24130891
19.  Sunlight-induced rapid and efficient biogenic synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum Linn. with enhanced antibacterial activity 
Nanotechnology is now regarded as a distinct field of research in modern science and technology with multifaceted areas including biomedical applications. Among the various approaches currently available for the generation of metallic nanoparticles, biogenic synthesis is of increasing demand for the purpose of green nanotechnology. Among various natural sources, plant materials are the most readily available template-directing matrix offering cost-effectiveness, eco-friendliness, and easy handling. Moreover, the inherent pharmacological potentials of these medicinal plant extracts offer added biomedical implementations of the synthesized metal nanoparticles.
A robust practical method for eco-friendly synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum (Tulsi) as both reducing and capping agent, under the influence of direct sunlight has been developed without applying any other chemical additives. The nanoparticles were characterized with the help of UV-visible spectrophotometer and transmission electron microscopy (TEM). The prepared silver nanoparticles exhibited considerable antibacterial activity. The effects were more pronounced on non-endospore-forming Gram-positive bacteria viz., Staphylococcus aureus, Staphylococcus epidermidis, and Listeria monocytogenes than endospore-forming species Bacillus subtilis. The nanoparticles also showed prominent activity on Gram-negative human pathogenic Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, and plant pathogenic Pantoea ananatis. A bactericidal mode of action was observed for both Gram-positive and Gram-negative bacteria by the nanoparticles.
We have developed a very simple, efficient, and practical method for the synthesis of silver nanoparticles using aqueous leaf extract of O. sanctum under the influence of direct sunlight. The biosynthesis of silver nanoparticles making use of such a traditionally important medicinal plant without applying any other chemical additives, thus offers a cost-effective and environmentally benign route for their large-scale commercial production. The nanoparticles dispersed in the mother solution showed promising antibacterial efficacy.
Graphical AbstractSunlight-induced rapid and efficient biogenic synthesis of silver nanoparticles using aqueous leaf extract of Ocimum sanctum Linn. with enhanced antibacterial activity.
Electronic supplementary material
The online version of this article (doi:10.1186/s13588-014-0018-6) contains supplementary material, which is available to authorized users.
PMCID: PMC4297304  PMID: 25621198
Silver nanoparticles; Ocimum sanctum; Sunlight; Antibacterial activity; Mode of action; Nanomedicine
20.  Nanosilver Particle Production Using Juglans Regia L. (Walnut) Leaf Extract 
The production of nanoparticles using a biosystem is considered green chemistry. Application of plant extracts as a biological process has been proven to be suitable for synthesis of nanoparticles.
This study designed in order to evaluate the production of silver nanoparticles using Juglans regia leaf extract and to compare the outcome of different preparation methods of plant extracts (ethanolic extract, boiling water extract and plant powder) for the generation of nanoparticles.
Materials and Methods
The reaction mixture contained the following ingredients: AgNO3 (10 mM) as the biotransformation substrate, plant extract or powder as the biocatalyst, glucose (560 mM) as the electron donor, phosphate buffer (pH = 7, 100 mM) and ethanol 70% as the solvent in the reaction mixture. The samples were taken from the reaction mixtures at different times, and the absorbance (450 nm) of the colloidal suspensions of silver nanoparticle hydrosols was recorded immediately following dilution (1:80) so as to preserve its freshness.
UV-visible spectrophotometer analysis revealed that the direct application of powder of the walnut leaf was the most efficient technique. TEM (Transmission electron microscopy) micrograph obtained by using this method revealed the generation of aggregated polydisperse, quasi-spherical nanoparticles in sizes of 10-50 nm. Ethonolic extract resulted in single silver nanoparticles which were nearly monodisperse, spherical, and individual nanoparticles ranged in size from 1-5 nm. Therefore, using direct powder of Walnut created more particles but applying ethanolic extract synthesized particles with smaller dimensions and no aggregation.
Different preparation methods of Juglans regia influence silver nanoparticles formation.
PMCID: PMC3941878  PMID: 24624182
Silver; Nanoparticles; Biosynthesis; Plant Extract; 
21.  Analysis of SiO2 nanoparticles binding proteins in rat blood and brain homogenate 
International Journal of Nanomedicine  2014;9(Suppl 2):207-215.
A multitude of nanoparticles, such as titanium oxide (TiO2), zinc oxide, aluminum oxide, gold oxide, silver oxide, iron oxide, and silica oxide, are found in many chemical, cosmetic, pharmaceutical, and electronic products. Recently, SiO2 nanoparticles were shown to have an inert toxicity profile and no association with an irreversible toxicological change in animal models. Hence, exposure to SiO2 nanoparticles is on the increase. SiO2 nanoparticles are routinely used in numerous materials, from strengthening filler for concrete and other construction composites, to nontoxic platforms for biomedical application, such as drug delivery and theragnostics. On the other hand, recent in vitro experiments indicated that SiO2 nanoparticles were cytotoxic. Therefore, we investigated these nanoparticles to identify potentially toxic pathways by analyzing the adsorbed protein corona on the surface of SiO2 nanoparticles in the blood and brain of the rat. Four types of SiO2 nanoparticles were chosen for investigation, and the protein corona of each type was analyzed using liquid chromatography-tandem mass spectrometry technology. In total, 115 and 48 plasma proteins from the rat were identified as being bound to negatively charged 20 nm and 100 nm SiO2 nanoparticles, respectively, and 50 and 36 proteins were found for 20 nm and 100 nm arginine-coated SiO2 nanoparticles, respectively. Higher numbers of proteins were adsorbed onto the 20 nm sized SiO2 nanoparticles than onto the 100 nm sized nanoparticles regardless of charge. When proteins were compared between the two charges, higher numbers of proteins were found for arginine-coated positively charged SiO2 nanoparticles than for the negatively charged nanoparticles. The proteins identified as bound in the corona from SiO2 nanoparticles were further analyzed with ClueGO, a Cytoscape plugin used in protein ontology and for identifying biological interaction pathways. Proteins bound on the surface of nanoparticles may affect functional and conformational properties and distributions in complicated biological processes.
PMCID: PMC4279766  PMID: 25565838
silica; nanoparticles; protein corona; plasma; brain homogenate; nanotoxicity
22.  A novel bacterial isolate Stenotrophomonas maltophilia as living factory for synthesis of gold nanoparticles 
The synthesis of gold nanoparticles (GNPs) has received considerable attention with their potential applications in various life sciences related applications. Recently, there has been tremendous excitement in the study of nanoparticles synthesis by using some natural biological system, which has led to the development of various biomimetic approaches for the growth of advanced nanomaterials. In the present study, we have demonstrated the synthesis of gold nanoparticles by a novel bacterial strain isolated from a site near the famous gold mines in India. A promising mechanism for the biosynthesis of GNPs by this strain and their stabilization via charge capping was investigated.
A bacterial isolate capable of gold nanoparticle synthesis was isolated and identified as a novel strain of Stenotrophomonas malophilia (AuRed02) based on its morphology and an analysis of its 16S rDNA gene sequence. After 8 hrs of incubation, monodisperse preparation of gold nanoparticles was obtained. Gold nanoparticles were characterized and found to be of ~40 nm size. Electrophoresis, Zeta potential and FTIR measurements confirmed that the particles are capped with negatively charged phosphate groups from NADP rendering them stable in aqueous medium.
The process of synthesis of well-dispersed nanoparticles using a novel microorganism isolated from the gold enriched soil sample has been reported in this study, leading to the development of an easy bioprocess for synthesis of GNPs. This is the first study in which an extensive characterization of the indigenous bacterium isolated from the actual gold enriched soil was conducted. Promising mechanism for the biosynthesis of GNPs by the strain and their stabilization via charge capping is suggested, which involves an NADPH-dependent reductase enzyme that reduces Au3+ to Au0 through electron shuttle enzymatic metal reduction process.
PMCID: PMC2719591  PMID: 19619318
23.  Green Synthesis of Robust, Biocompatible Silver Nanoparticles Using Garlic Extract 
Journal of nanomaterials  2012;2012:730746.
This paper details a facile approach for the synthesis of stable and monodisperse silver nanoparticles performed at ambient/low temperature where Allium sativum (garlic) extract functions as the silver salt reducing agent during nanoparticle synthesis as well as the post-synthesis stabilizing ligands. Varying the synthesis conditions provides control of particle size, size-distribution, and kinetics of particle formation. Infrared spectroscopy, energy dispersive x-ray chemical analysis, and high performance liquid chromatography indicated that the carbohydrates present in the garlic extract are the most likely nanoparticle stabilizing chemistry. The synthesized silver nanoparticles also demonstrate potential for biomeical applications, owing to the 1) enhanced stability in biological media, 2) resistance to oxidation by the addition of H2O2, 3) ease and scalability of synthesis, and 4) lack of harsh chemicals required for synthesis. Cytotoxicity assays indicated no decrease in cellular proliferation for vascular smooth muscle cells and 3T3 fibroblasts at a concentration of 25 μg/ml, confirming that garlic extract prepared silver nanoparticles are ideal candidates for future experimentation and implementation into biomedical applications.
PMCID: PMC3966315  PMID: 24683414
green nanotechnology; garlic; silver nanoparticles; cytotoxicity; 3T3 fibroblasts
24.  Enhanced oligonucleotide–nanoparticle conjugate stability using thioctic acid modified oligonucleotides 
Nucleic Acids Research  2007;35(11):3668-3675.
Metallic nanoparticles of gold functionalized with oligonucleotides conventionally use a terminal thiol modification and have been used in a wide range of applications. Although readily available, the oligonucleotide–nanoparticle conjugates prepared in this way suffer from a lack of stability when exposed to a variety of small molecules or elevated temperatures. If silver is used in place of gold then this lack of stability is even more pronounced. In this study we report the synthesis of highly stabilized oligonucleotide–nanoparticle conjugates using a simple oligonucleotide modification. A modified solid support was used to generate 3′-thioctic acid modified oligonucleotides by treatment with an N-hydroxysuccimidyl ester of thioctic acid. Unusually, both gold and silver nanoparticles have been investigated in this study and show that these disulphide-modified oligonucleotide probes offer significant improvements in nanoparticle stability when treated with dithiothreitol (DTT) compared with monothiol analogues. This is a significant advance in oligonucleotide–nanoparticle conjugate stability and for the first time allows silver nanoparticles to be prepared that are more stable than standard gold-thiol functionalized nanoparticles. This opens up the possibility of using silver nanoparticles functionalized with oligonucleotides as an alternative to gold.
PMCID: PMC1920241  PMID: 17488844
25.  Synthesis of Silver Nanostructures by Multistep Methods 
Sensors (Basel, Switzerland)  2014;14(4):5860-5889.
The shape of plasmonic nanostructures such as silver and gold is vital to their physical and chemical properties and potential applications. Recently, preparation of complex nanostructures with rich function by chemical multistep methods is the hotspot of research. In this review we introduce three typical multistep methods to prepare silver nanostructures with well-controlled shapes, including the double reductant method, etching technique and construction of core-shell nanostructures. The growth mechanism of double the reductant method is that different favorable facets of silver nanocrystals are produced in different reductants, which can be used to prepare complex nanostructures such as nanoflags with ultranarrow resonant band bandwidth or some silver nanostructures which are difficult to prepare using other methods. The etching technique can selectively remove nanoparticles to achieve the aim of shape control and is widely used for the synthesis of nanoflowers and hollow nanostructures. Construction of core-shell nanostructures is another tool to control shape and size. The three methods can not only prepare various silver nanostructures with well-controlled shapes, which exhibit unique optical properties, such as strong surface-enhanced Raman scattering (SERS) signal and localized surface plasmon resonance (LSPR) effect, but also have potential application in many areas.
PMCID: PMC4029645  PMID: 24670722
reductant; nanoparticle etching; composited nanostructures; shape control; plasmonic sensors

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