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.
Nanobiotechnology is considered to be one of the fastest emerging fields. It is still a relatively new and exciting area of research with considerable potential for development. Among the inorganic nanomaterials, biogenically synthesized silver nanoparticles (bio-AgNPs) have been frequently used due to their unique physicochemical properties that result not only from their shape and size but also from surface coatings of natural origin. These properties determine antibacterial, antifungal, antiprotozoal, anticancer, anti-inflammatory, and many more activities of bio-AgNPs. This review provides the current state of knowledge on the methods and mechanisms of biogenic synthesis of silver nanoparticles as well as their potential applications in different fields such as medicine, food, agriculture, and industries.
Nanotechnology is a new and developing branch that has revolutionized the world by its applications in various fields including medicine and agriculture. In nanotechnology, nanoparticles play an important role in diagnostics, drug delivery, and therapy. The synthesis of nanoparticles by fungi is a novel, cost-effective and eco-friendly approach. Among fungi, Fusarium spp. play an important role in the synthesis of nanoparticles and can be considered as a nanofactory for the fabrication of nanoparticles. The synthesis of silver nanoparticles (AgNPs) from Fusarium, its mechanism and applications are discussed in this review. The synthesis of nanoparticles from Fusarium is the biogenic and green approach. Fusaria are found to be a versatile biological system with the ability to synthesize nanoparticles extracellularly. Different species of Fusaria have the potential to synthesise nanoparticles. Among these, F. oxysporum has demonstrated a high potential for the synthesis of AgNPs. It is hypothesised that NADH-dependent nitrate reductase enzyme secreted by F. oxysporum is responsible for the reduction of aqueous silver ions into AgNPs. The toxicity of nanoparticles depends upon the shape, size, surface charge, and the concentration used. The nanoparticles synthesised by different species of Fusaria can be used in medicine and agriculture.Nanotechnology is a new and developing branch that has revolutionized the world by its applications in various fields including medicine and agriculture. In nanotechnology, nanoparticles play an important role in diagnostics, drug delivery, and therapy. The synthesis of nanoparticles by fungi is a novel, cost-effective and eco-friendly approach. Among fungi, Fusarium spp. play an important role in the synthesis of nanoparticles and can be considered as a nanofactory for the fabrication of nanoparticles. The synthesis of silver nanoparticles (AgNPs) from Fusarium, its mechanism and applications are discussed in this review. The synthesis of nanoparticles from Fusarium is the biogenic and green approach. Fusaria are found to be a versatile biological system with the ability to synthesize nanoparticles extracellularly. Different species of Fusaria have the potential to synthesise nanoparticles. Among these, F. oxysporum has demonstrated a high potential for the synthesis of AgNPs. It is hypothesised that NADH-dependent nitrate reductase enzyme secreted by F. oxysporum is responsible for the reduction of aqueous silver ions into AgNPs. The toxicity of nanoparticles depends upon the shape, size, surface charge, and the concentration used. The nanoparticles synthesised by different species of Fusaria can be used in medicine and agriculture.Fusarium; synthesis; nanoparticles; mechanism; medicine; agriculture; nanofactory; toxicity
Global reports on multidrug resistance (MDR) and life-threatening pathogens such as SARS-CoV-2 and Candida cruris have stimulated researchers to explore new antimicrobials that are eco-friendly and economically viable. In this context, biodegradable polymers such as nisin, chitin, and pullulan play an important role in solving the problem. Pullulan is an important edible, biocompatible, water-soluble polymer secreted by Aureobasidium pullulans that occurs ubiquitously. It consists of maltotriose units linked with α-1,6 glycosidic bonds and is classed as Generally Regarded as Safe (GRAS) by the Food and Drug Administration (FDA) in the USA. Pullulan is known for its antibacterial, antifungal, antiviral, and antitumor activities when incorporated with other additives such as antibiotics, drugs, nanoparticles, and so on. Considering the importance of its antimicrobial activities, this polymer can be used as a potential antimicrobial agent against various pathogenic microorganisms including the multidrug-resistant (MDR) pathogens. Moreover, pullulan has ability to synthesize biogenic silver nanoparticles (AgNPs), which are remarkably efficacious against pathogenic microbes. The pullulan-based nanocomposites can be applied for wound healing, food packaging, and also enhancing the shelf-life of fruits and vegetables. In this review, we have discussed biosynthesis of pullulan and its role as antibacterial, antiviral, and antifungal agent. Pullulan-based films impregnated with different antimicrobials such as AgNPs, chitosan, essential oils, and so on, forming nanocomposites have also been discussed as natural alternatives to combat the problems posed by pathogens.
Natural polymer-based nanocomposites have received significant attention in both scientific and industrial research in recent years. They can help to eliminate the consequences of application of petroleum-derived polymeric materials and related environmental concerns. Such nanocomposites consist of natural biopolymers (e.g., chitosan, starch, cellulose, alginate and many more) derived from plants, microbes and animals that are abundantly available in nature, biodegradable and thus eco-friendly, and can be used for developing nanocomposites for agriculture and food industry applications. Biopolymer-based nanocomposites can act as slow-release nanocarriers for delivering agrochemicals (fertilizers/nutrients) or pesticides to crop plants to increase yields. Similarly, biopolymer-based nanofilms or hydrogels may be used as direct product coating to extend product shelf life or improve seed germination or protection from pathogens and pests. Biopolymers have huge potential in food-packaging. However, their packaging properties, such as mechanical strength or gas, water or microbial barriers can be remarkably improved when combined with nanofillers such as nanoparticles. This article provides an overview of the strategic applications of natural polymer nanocomposites in food and agriculture as nanocarriers of active compounds, polymer-based hydrogels, nanocoatings and nanofilms. However, the risk, challenges, chances, and consumers’ perceptions of nanotechnology applications in agriculture and food production and packaging have been also discussed.
IntroductionDue to the increasing resistance of bacteria and fungi to antimicrobials, it is necessary to search for effective alternatives to prevent and treat pathogens causing diseases in humans, animals, and plants. In this context, the mycosynthesized silver nanoparticles (AgNPs) are considered as a potential tool to combat such pathogenic microorganisms.MethodsAgNPs were synthesized from Fusarium culmorum strain JTW1 and characterized by Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), Fourier Transform Infrared (FTIR) spectroscopy, Nanoparticle Tracking Analysis (NTA), Dynamic Light Scattering (DLS) and Zeta potential measurement. The minimum inhibitory (MIC) and biocidal concentrations (MBC) were determined against 13 bacterial strains. Moreover, the combined effect of AgNPs with antibiotics (streptomycin, kanamycin, ampicillin, tetracycline) was also studied by determining the Fractional Inhibitory Concentration (FIC) index. The anti-biofilm activity was examined by crystal violet and fluorescein diacetate (FDA) assays. Furthermore, antifungal activity of AgNPs was evaluated against a panel of phytopathogenic fungi viz., Botrytis, Colletotrichum, Fusarium, Phoma, Sclerotinia, and an oomycete pathogen Phytophthora by agar well-diffusion and micro-broth dilution method to evaluate the minimal AgNPs concentrations that inhibit fungal spore germination.ResultsFungi-mediated synthesis resulted in the formation of small (15.56 ± 9.22 nm), spherical and stable (zeta potential of – 38.43 mV) AgNPs with good crystallinity. The results of FTIR spectroscopy indicated the presence of various functional groups, namely hydroxyl, amino, and carboxyl ones, from the biomolecules on the surface of AgNPs. The AgNPs showed antimicrobial and antibiofilm formation activities against Gram-positive and Gram-negative bacteria. The values of MIC and MBC ranged between 16–64 and 32–512 μg mL−1, respectively. The enhanced effect of AgNPs in combination with antibiotics was confirmed against human pathogens. The highest synergistic effect (FIC = 0.0625) was demonstrated by the combination of AgNPs with streptomycin against two strains of Escherichia coli (ATCC 25922 and ATCC 8739), followed by Klebsiella pneumoniae and Pseudomonas aeruginosa (FIC = 0.125). Enhanced effects of AgNPs with ampicillin were also shown against Staphylococcus aureus ATCC 25923 (FIC = 0.125) and P. aeruginosa (FIC = 0.25), as well as kanamycin against S. aureus ATCC 6538 (FIC = 0.25). The crystal violet assay revealed that the lowest concentration of AgNPs (0.125 μg mL−1) reduced the development of biofilms of Listeria monocytogenes and Salmonella enterica, while the maximum resistance was shown by Salmonella infantis, its biofilm was reduced after exposure to a concentration of 512 μg mL−1. A high inhibitory effect on the activity of bacterial hydrolases was observed by the FDA assay. AgNPs at a concentration of 0.125 μg mL−1 reduced the hydrolytic activity of all biofilms formed by the tested pathogens, except E. coli ATCC 25922, P. aeruginosa, and Pectobacterium carotovorum (efficient concentration was 2-fold higher, at 0.25 μg mL−1), while the hydrolytic activity of E. coli ATCC 8739, Salmonella infantis and S. aureus ATCC 6538 was suppressed after treatment with AgNPs at concentrations of 0.5, 2 and 8 μg mL−1, respectively. Moreover, AgNPs inhibited fungal growth and spore germination of Botrytis cinerea, Phoma lingam, and Sclerotinia sclerotiorum. MIC and MFC values of AgNPs against spores of these fungal strains were determined at 64, 256, and 32 μg mL−1, and zones of growth inhibition were 4.93, 9.54, and 3.41 mm, respectively.DiscussionFusarium culmorum strain JTW1 was found to be an eco-friendly biological system for an easy, efficient and inexpensive synthesis of AgNPs. In our study, the mycosynthesised AgNPs demonstrated remarkable antimicrobial (antibacterial and antifungal) and antibiofilm activities against a wide range of human and plant pathogenic bacteria and fungi singly and in combination with antibiotics. These AgNPs could be applied in medicine, agriculture, and food industry to control such pathogens that cause numerous human diseases and crop losses. However, before using them extensive animal studies are required to evaluate the toxicity, if any.
A novel composite based on biocompatible hydroxyapatite (HA) nanoparticles and Cu-HKUST-1 (Cu-HKUST-1@HA) has been prepared following a layer-by-layer strategy. Cu-HKUST-1 was carefully selected from a group of four Cu-based metal−organic frameworks as the material with the most promising antimicrobial activity. The formation of a colloidal Cu-HKUST-1 layer on HA nanoparticles was confirmed by various techniques, e.g., infrared spectroscopy, powder X-ray diffraction, N 2 sorption, transmission electron microscopy imaging, electron paramagnetic resonance, and X-ray absorption spectroscopy. Importantly, such a Cu-HKUST-1 layer significantly improved the nanomechanical properties of the composite, with Young's modulus equal to that of human cortical bone (13.76 GPa). At the same time, Cu-HKUST-1@HA has maintained the negative zeta potential (−16.3 mV in pH 7.4) and revealed biocompatibility toward human dermal fibroblasts up to a concentration of 1000 μg/ mL, without inducing ex vivo hemolysis. Chemical stability studies of the composite over 21 days in a buffer-simulated physiological fluid allowed a detailed understanding of the transformations that the Cu-HKUST-1@HA undergoes over time. Finally, it has been confirmed that the Cu-HKUST-1 layer provides antibacterial properties to HA, and the synergism reached in this way makes it promising for bone tissue regeneration.
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