The production of nanomaterials through environmentally friendly methods is a top priority in the sustainable development of nanotechnology. This paper presents data on the synthesis of silver nanoparticles using an aqueous extract of Sphagnum fallax moss at room temperature. The morphology, stability, and size of the nanoparticles were analyzed using various techniques, including transmission electron microscopy, Doppler laser velocimetry, and UV-vis spectroscopy. In addition, Fourier transform infrared spectroscopy was used to analyze the presence of moss metabolites on the surface of nanomaterials. The effects of different concentrations of citrate-stabilized and moss extract-stabilized silver nanoparticles on cell viability, necrosis induction, and cell impedance were compared. The internalization of silver nanoparticles into both monolayers and three-dimensional cells spheroids was evaluated using dark-field microscopy and hyperspectral imaging. An eco-friendly method for the synthesis of silver nanoparticles at room temperature is proposed, which makes it possible to obtain spherical nanoparticles of 20–30 nm in size with high bioavailability and that have potential applications in various areas of human life.
The increase in the resistance of microorganisms to existing antibiotics associated with their ability to survive in biofilms makes the development of new antimicrobial agents a topical question. Of great interest in this field are nanocomposites based on halloysite and silver nanoparticles, since they have a broad antimicrobial activity and can reduce the toxic effects of nanoparticles. In this study, we assessed the effect of the AgHNT nanocomposite at a concentration of 0.5 mg/ml on the pathogenicity factors of Serratia marcescens. The antibiofilm activity of AgHNT was assessed by resazurin assay, and visualized using light and AFM microscopy. AgHNT were found to significantly inhibit biofilm formation in S. marcescens. Although the silver-containing species did not affect protease activity, they greatly reduced the production of prodigiosin, another quorum-sensing factor, by bacteria. The AgHNT nanocomposite was more efficient in suppressing S. marcescens biofilms than the antibiotic streptomycin.
Synthesis of silver nanoparticles using extracts from plants is an advantageous technological alternative to the traditional colloidal synthesis due to its simplicity, low cost, and the inclusion of environmentally friendly processes to obtain a new generation of antimicrobial compounds. The work describes the production of silver and iron nanoparticles using sphagnum extract as well as traditional synthesis. Dynamic light scattering (DLS) and laser doppler velocimetry methods, UV-visible spectroscopy, transmission electron microscopy (TEM) combined with energy dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM), dark-field hyperspectral microscopy, and Fourier-transform infrared spectroscopy (FT-IR) were used to study the structure and properties of synthesized nanoparticles. Our studies demonstrated a high antibacterial activity of the obtained nanoparticles, including the formation of biofilms. Nanoparticles synthesized using sphagnum moss extracts likely have high potential for further research.
The high worldwide consumption of cheap plastic goods has already resulted in a serious environmental plastic pollution, exacerbated by piling of disposed personal protective equipment because of the recent outbreak of COVID-19. The aim of this study was to assess the feasibility of dark-field hyperspectral microscopy in the 400-1000 wavelength range for detection of nanoplastics derived from weathered polypropylene masks. A surgical mask was separated to layers and exposed to UV radiation (254 nm) for 192 h. Oxidative degradation of the polypropylene was evidenced by ATR FT-IR analysis. UV treatment for 192 h resulted in generation of differently shaped micro- and nano-sized particles, visualized by dark-field microscopy. The presence of nanoparticles was confirmed by AFM studies. The hyperspectral profiles (400-1000 nm) were collected after every 48 h of the UV treatment. The distinct hyperspectral features faded after prolonged UV exposure, but the assignment of some particles to either blue or white layers of mask could still be made based on spectral characteristics.
Lactiplantibacillus plantarum is best known for its significant adaptive potential and ability to colonize different ecological niches. Different strains of L. plantarum are widely used as probiotics. To characterize the probiotic potential of the novel L. plantarum FCa3L strain isolated from fermented cabbage, we sequenced its whole genome using the Illumina MiSeq platform. This bacterial isolate had a circular chromosome of 3,365,929 bp with 44.3% GC content and a cyclic phage phiX174 of 5386 bp with 44.7% GC content. The results of in vitro studies showed that FCa3L was comparable with the reference probiotic strain L. plantarum 8PA3 in terms of acid and bile tolerance, adhesiveness, H2O2 production, and acidification rate. The strain 8PA3 possessed higher antioxidant activity, while FCa3L demonstrated superior antibacterial properties. The antibiotic resistance of FCa3L was more relevant to the probiotic strain than that of 8PA3, although a number of silent antibiotic resistance genes were identified in its genome. Genomic evidence to support adhesive and antibacterial properties, biosynthesis of bioactive metabolites, and safety of FCa3L was also presented. Thus, this study confirmed the safety and probiotic properties of L. plantarum FCa3L via complete genome and phenotype analysis, suggesting its potential as a probiotic, although further in vivo investigations are still necessary.
Here we report a spontaneous electrostatic coating of human hair with aqueous Fe3O4 colloids capable to tailor magnetic properties to hair, orienting and even moving them under the influence of the external magnetic field. Magnetite particles were modified by cationic and anionic polyelectrolytes and then successfully deposited in dense arrays, starting from cuticle gaps and spreading further over a major hair surface. These biocompatible and biodegradable magnetic nanoparticles may serve as carriers for drug loading and delivery for topical pharmaceutical treatments. The deposition process was imaged in real-time using dark-field microscopy. The hair specimens were further studied using a number of characterisation techniques. Under application of an external magnetic field, the nanoparticle magnetic ordering was obtained resulting in the hair alignment and attraction along the field applied. We believe the technology reported here will find a range of applications in topical drug delivery and hair care.
Mesenchymal stem cells (MSCs) have extensive pluripotent potential to differentiate into various cell types, and thus they are an important tool for regenerative medicine and biomedical research. In this work, the differentiation of hTERT-transduced adipose-derived MSCs (hMSCs) into chondrocytes, adipocytes and osteoblasts on substrates with nanotopography generated by magnetic iron oxide nanoparticles (MNPs) and DNA was investigated. Citrate-stabilized MNPs were synthesized by the chemical co-precipitation method and sized around 10 nm according to microscopy studies. It was shown that MNPs@DNA coatings induced chondrogenesis and osteogenesis in hTERT-transduced MSCs. The cells had normal morphology and distribution of actin filaments. An increase in the concentration of magnetic nanoparticles resulted in a higher surface roughness and reduced the adhesion of cells to the substrate. A glass substrate modified with magnetic nanoparticles and DNA induced active chondrogenesis of hTERT-transduced MSC in a twice-diluted differentiation-inducing growth medium, suggesting the possible use of nanostructured MNPs@DNA coatings to obtain differentiated cells at a reduced level of growth factors.
The concerns regarding microplastics and nanoplastics pollution stimulate studies on the uptake and biodistribution of these emerging pollutants in vitro. Atomic force microscopy in nanomechanical PeakForce Tapping mode was used here to visualise the uptake and distribution of polystyrene spherical microplastics in human skin fibroblast. Particles down to 500 nm were imaged in whole fixed cells, the nanomechanical characterization allowed for differentiation between internalized and surface attached plastics. This study opens new avenues in microplastics toxicity research.
Topical administration of drugs is required for the treatment of parasitic diseases and insect infestations; therefore, fabrication of nanoscale drug carriers for effective insecticide topical delivery is needed. Here we report the enhanced immobilization of halloysite tubule nanoclay onto semiaquatic capybaras which have hydrophobic hair surfaces as compared to their close relatives, land-dwelling guinea pigs, and other agricultural livestock. The hair surface of mammals varies in hydrophobicity having a cortex surrounded by cuticles. Spontaneous 1–2 µm thick halloysite hair coverages on the semi-aquatic rodent capybara, non-aquatic rodent guinea pig, and farm goats were compared. The best coating was found for capybara due to the elevated 5 wt% wax content. As a result, we suggest hair pretreatment with diluted wax for enhanced nanoclay adsorption. The formation of a stable goat hair coverage with a 2–3 µm halloysite layer loaded with permethrin insecticide allowed for long-lasting anti-parasitic protection, enduring multiple rain wettings and washings. We expect that our technology will find applications in animal parasitosis protection and may be extended to prolonged human anti-lice treatment.
Acute toxicity of nanoparticles of aluminum oxide (Al2O3) with a size of 13–16 nm was investigated by the biotesting method using Paramecium caudatum ciliates in the concentration range of 10–100 μg/mL. Aluminum oxide has an acute toxic effect on paramecium at concentrations of 20–100 μg/mL. The mean lethal dose (LD50) is equal to the concentration of nanoparticles at which the mortality of ciliates in relation to the control reached 50%. The LD50 for Al2O3 nanoparticles is 23 μg/mL at a 24-h exposure. According to published data, the toxic effect of Al2O3 nanoparticles is specific and depends on the size and surface charge of the particles and on the interfacial interaction of nanoparticles with the cell surface, as well as on the concentration and exposure time.
The use, in veterinary practice, of a three-dimensional biopolymer matrix (based on chitosan, agarose, and gelatin and doped with halloysite nanotubes) as a vehicle for mesenchymal stem cells (MSCs) to repair bone defects is reported here. The nanocomposite, combined with allogenic adipose-derived stem cells, was suitable for the repair of bone defects in dogs when paired with standard surgery involving metal Kirshner wires. The absence of inflammatory reactions to biopolymer matrices with allogenic stem cells was revealed in the case of an animal prone to inflammatory and allergic reactions. In addition, positive dynamics in the fusion of chronic bone defects without rejection reactions was observed after using a biopolymer matrix with MSCs.
Enhanced dark-field hyperspectral imaging (EDF-HSI) integrates advanced spectroscopy and imaging techniques, enabling fast and quantitative insights into the interactions between nanomaterials and organisms at the cellular or invertebrate level. However, the usage of biomolecule-based nanomaterials as contrast agents for EDF-HSI is highly challenging because these nanomaterials usually lack the characteristic optical scattering properties that can be readily distinguished from the scattering signals of organisms. Herein, we report peptide nanoparticles fabricated through covalent self-assembly of dipeptides and genipin for EDF-HSI in vitro and in vivo. Two dipeptides with either C-terminal amidation or not were used as the building blocks for covalent self-assembly. Significantly, the optical scattering intensity of the resulting nanoparticles is enhanced by the amidation. The amidation also enhances the stability of the peptide nanoparticles both in solutions and inside cells or organisms due to inhibition of electrostatic repulsion among the building blocks. Therefore, the nanoparticles fabricated from the dipeptide with uncharged C-terminal amide can be facilely visualized and tracked inside cells and nematodes through EDF-HSI. These results demonstrate that peptide nanomaterials can be endowed with optical scattering properties suitable for EDF-HSI, providing a promising approach for visualization and analysis of the activities of biomaterials in vitro and in vivo.
Fly ash produced during coal combustion is one of the major sources of air and water pollution, but the data on the impact of micrometer-size fly ash particles on human cells is still incomplete. Fly ash samples were collected from several electric power stations in the United States (Rockdale, TX; Dolet Hill, Mansfield, LA; Rockport, IN; Muskogee, OK) and from a metallurgic plant located in the Russian Federation (Chelyabinsk Electro-Metallurgical Works OJSC). The particles were characterized using dynamic light scattering, atomic force, and hyperspectral microscopy. According to chemical composition, the fly ash studied was ferro-alumino-silicate mineral containing substantial quantities of Ca, Mg, and a negligible concentration of K, Na, Mn, and Sr. The toxicity of the fly ash microparticles was assessed in vitro using HeLa cells (human cervical cancer cells) and Jurkat cells (immortalized human T lymphocytes). Incubation of cells with different concentrations of fly ash resulted in a dose-dependent decrease in cell viability for all fly ash variants. The most prominent cytotoxic effect in HeLa cells was produced by the ash particles from Rockdale, while the least was produced by the fly ash from Chelyabinsk. In Jurkat cells, the lowest toxicity was observed for fly ash collected from Rockport, Dolet Hill and Muscogee plants. The fly ash from Rockdale and Chelyabinsk induced DNA damage in HeLa cells, as revealed by the single cell electrophoresis, and disrupted the normal nuclear morphology. The interaction of fly ash microparticles of different origins with cells was visualized using dark-field microscopy and hyperspectral imaging. The size of ash particles appeared to be an important determinant of their toxicity, and the smallest fly ash particles from Chelyabinsk turned out to be the most cytotoxic to Jukart cells and the most genotoxic to HeLa cells.
The problem of purifying domestic and hospital wastewater from pharmaceutical compounds is becoming more and more urgent every year, because of the continuous accumulation of chemical pollutants in the environment and the limited availability of freshwater resources. Clay adsorbents have been repeatedly proposed as adsorbents for treatment purposes, but natural clays are hydrophilic and can be inefficient for catching hydrophobic pharmaceuticals. In this paper, a comparison of adsorption properties of pristine montmorillonite (MMT) and montmorillonite modified with stearyl trimethyl ammonium (hydrophobic MMT-STA) towards carbamazepine, ibuprofen, and paracetamol pharmaceuticals was performed. The efficiency of adsorption was investigated under varying solution pH, temperature, contact time, initial concentration of pharmaceuticals, and adsorbate/adsorbent mass ratio. MMT-STA was better than pristine MMT at removing all the pharmaceuticals studied. The adsorption capacity of hydrophobic montmorillonite to pharmaceuticals decreased in the following order: carbamazepine (97%) > ibuprofen (95%) > paracetamol (63–67%). Adsorption isotherms were best described by Freundlich model. Within the pharmaceutical concentration range of 10–50 µg/mL, the most optimal mass ratio of adsorbates to adsorbents was 1:300, pH 6, and a temperature of 25 °C. Thus, MMT-STA could be used as an efficient adsorbent for deconta×ating water of carbamazepine, ibuprofen, and paracetamol.
The environmental hazards of oil spills cannot be underestimated. Bioremediation holds promise among various approaches to tackle oil spills in soils and sediments. In particular, using oil-degrading bacteria is an efficient and self-regulating way to remove oil spills. Using animals for oil spills remediation is in its infancy, mostly due to the lack of efficient oil-degrading capabilities in eukaryotes. Here we show that Caenorhabditis elegans nematodes survive for extended periods (up to 22 days) on pure crude oil diet. Moreover, we report for the first time the use of Alcanivorax borkumensis hydrocarbonoclastic bacteria for colonisation of C. elegans intestines, which allows for effective digestion of crude oil by the nematodes. The worms fed and colonised by A. borkumensis demonstrated the similar or even better longevity, resistance against oxidative and thermal stress and reproductivity as those animals fed with Escherichia coli bacteria (normal food). Importantly, A. borkumensis-carrying nematodes were able to accumulate oil droplet from oil-contaminated soils. Artificial colonisation of soil invertebrates with oil-degrading bacteria will be an efficient way to distribute microorganisms in polluted soil, thus opening new avenues for oil spills zooremediation.
Polycations are an essential part of layer-by-layer (LbL)-assembled drug delivery systems, especially for gene delivery. In addition, they are used for other related applications, such as cell surface engineering. As a result, an assessment of the cytotoxicity of polycations and elucidation of the mechanisms of polycation toxicity is of paramount importance. In this study, we examined in detail the effects of a variety of water-soluble, positively charged synthetic polyelectrolytes on in vitro cytotoxicity, cell and nucleus morphology, and monolayer expansion changes. We have ranked the most popular cationic polyelectrolytes from the safest to the most toxic in relation to cell cultures. 3D cellular cluster formation was disturbed by addition of polyelectrolytes in most cases in a dose-dependent manner. Atomic force microscopy allowed us to visualize in detail the structures of the polyelectrolyte–DNA complexes formed due to electrostatic interactions. Our results indicate a relationship between the structure of the polyelectrolytes and their toxicity, which is necessary for optimization of drug and gene delivery systems.