Next-Generation Nanopore Sensors Based on Conductive Pulse Sensing for Enhanced Detection of Nanoparticles.

Nanopore sensing has been successfully used to characterize biological molecules with single-molecule resolution based on the resistive pulse sensing approach. However, its use in nanoparticle characterization has been constrained by the need to tailor the nanopore aperture size to the size of the analyte, precluding the analysis of heterogeneous samples. Additionally, nanopore sensors often require the use of high salt concentrations to improve the signal-to-noise ratio, which further limits their ability to study a wide range of nanoparticles that are unstable at high ionic strength. Here, a new paradigm in nanopore research that takes advantage of a polymer electrolyte system to comprise a conductive pulse sensing approach is presented. A finite element model is developed to explain the conductive pulse signals observed and compare these results with experiments. This system enables the analytical characterization of heterogeneous nanoparticle mixtures at low ionic strength . Furthermore, the wide applicability of the method is demonstrated by characterizing metallic nanospheres of varied sizes, plasmonic nanostars with various degrees of branching, and protein-based spherical nucleic acids with different oligonucleotide loadings. This system will complement the toolbox of nanomaterials characterization techniques to enable real-time optimization workflow for engineering a wide range of nanomaterials.

Elucidating the effect of TiO2 nanoparticles on mung bean rhizobia via in vitro assay: Influence on growth, morphology, and plant growth promoting traits.

Titanium dioxide nanoparticles (TiO2 NPs) are among the most commonly used nanomaterials and are most likely to end up in soil. Therefore, it is pertinent to study the interaction of TiO2 NPs with soil microorganisms. The present in vitro broth study evaluates the impacts of low-dose treatments (0, 1.0, 5.0, 10.0, 20.0, and 40.0 mg L-1 ) of TiO2 NPs on cell viability, morphology, and plant growth promoting (PGP) traits of rhizobia isolated from mung bean root nodule. Two types of TiO2 NPs, that is, mixture of anatase and rutile, and anatase alone were used in the study. These TiO2 NPs were supplemented in broth along with a multifunctional isolate (Bradyrhizobium sp.) and two reference cultures. The exposure of TiO2 (anatase+rutile) NPs at low concentrations (less than 20.0 mg L-1 ) enhanced the cell growth, and total soluble protein content, besides improving the phosphate solubilization, Indole-3-acetic acid (IAA) production, siderophore, and gibberellic acid production. The TiO2 (anatase) NPs enhanced exopolysaccharide (EPS) production by the test rhizobial cultures. The radical scavenging assay was performed to reveal the mode of action of the nano-TiO2 particles. The study revealed higher reactive oxygen species (ROS) generation by the TiO2 (anatase) NPs as compared with TiO2 (anatase+rutile) NPs. Exposure to TiO2 NPs also altered the morphology of rhizobial cells. The findings suggest that TiO2 NPs could act as promoters of PGP traits of PGP bacteria when applied at appropriate lower doses.

Basic and advanced spectrometric methods for complete nanoparticles characterization in bio/eco systems: current status and future prospects.

The use of engineered nanoparticles in the environment and human life has increased in the last 20 years. The risk assessment concerning application of nanomaterials in biological systems requires their thorough characterization. Understanding the correlations between physicochemical properties of nanoparticles concerning not only the size, particle size distribution, number concentration, degree of aggregation, or agglomeration but also solubility, stability, binding affinity, surface activity, chemical composition, and nanoparticle synthesis yield allows their reliable characterization. Thus, to find the structure-function/property relationship of nanoparticles, multifaceted characterization approach based on more than one analytical technique is required. On the other hand, the increasing demand for identification and characterization of nanomaterials has contributed to the continuous development of spectrometric techniques which enables for their qualitative and quantitative analysis in complex matrices giving reproducible and reliable results. This review is aimed at providing a discussion concerning four main aspects of nanoparticle characterization: nanoparticle synthesis yield, particle size and number concentration, elemental and isotopic composition of nanoparticles, and their surface properties. The conventional and non-conventional spectrometric techniques such as spectrophotometry UV-Vis, mass spectrometric techniques working in conventional and single-particle mode, or those based on optical emission detection systems are described with special emphasis paid on their advantages and drawbacks. The application and recent advances of these methods are also comprehensively reviewed and critically discussed.

Optofluidic force induction as a process analytical technology.

Manufacturers of nanoparticle-based products rely on detailed information about critical process parameters, such as particle size and size distributions, concentration, and material composition, which directly reflect the quality of the final product. These process parameters are often obtained using offline characterization techniques that cannot provide the temporal resolution to detect dynamic changes in particle ensembles during a production process. To overcome this deficiency, we have recently introduced Optofluidic Force Induction (OF2i) for optical real-time counting with single particle sensitivity and high throughput. In this paper, we apply OF2i to highly polydisperse and multi modal particle systems, where we also monitor evolutionary processes over large time scales. For oil-in-water emulsions we detect in real time the transition between high-pressure homogenization states. For silicon carbide nanoparticles, we exploit the dynamic OF2i measurement capabilities to introduce a novel process feedback parameter based on the dissociation of particle agglomerates. Our results demonstrate that OF2i provides a versatile workbench for process feedback in a wide range of applications.

Methanol-based extraction protocol for insoluble and moderately water-soluble nanoparticles in plants to enable characterization by single particle ICP-MS.

The detection and characterization of soluble metal nanoparticles in plant tissues are an analytical challenge, though a scientific necessity for regulating nano-enabled agrichemicals. The efficacy of two extraction methods to prepare plant samples for analysis by single particle ICP-MS, an analytical method enabling both size determination and quantification of nanoparticles (NP), was assessed. A standard enzyme-based extraction was compared to a newly developed methanol-based approach. Au, CuO, and ZnO NPs were extracted from three different plant leaf materials (lettuce, corn, and kale) selected for their agricultural relevance and differing characteristics. The enzyme-based approach was found to be unsuitable because of changes in the recovered NP size distribution of CuO NP. The MeOH-based extraction allowed reproducible extraction of the particle size distribution (PSD) without major alteration caused by the extraction. The type of leaf tissue did not significantly affect the recovered PSD. Total metal losses during the extraction process were largely due to the filtration step prior to analysis by spICP-MS, though this did not significantly affect PSD recovery. The methanol extraction worked with the three different NPs and plants tested and is suitable for studying the fate of labile metal-based nano-enabled agrichemicals.

Hyaluronic Acid-Dexamethasone Nanoparticles for Local Adjunct Therapy of Lung Inflammation.

The delivery of a dexamethasone formulation directly into the lung appears as an appropriate strategy to strengthen the systemic administration, reducing the dosage in the treatment of lung severe inflammations. For this purpose, a hyaluronic acid-dexamethasone formulation was developed, affording an inhalable reconstituted nanosuspension suitable to be aerosolized. The physico-chemical and biopharmaceutical properties of the formulation were tested: size, stability, loading of the spray-dried dry powder, reconstitution capability upon redispersion in aqueous media. Detailed structural insights on nanoparticles after reconstitution were obtained by light and X-ray scattering techniques. (1) The size of the nanoparticles, around 200 nm, is in the proper range for a possible engulfment by macrophages. (2) Their structure is of the core-shell type, hosting dexamethasone nanocrystals inside and carrying hyaluronic acid chains on the surface. This specific structure allows for nanosuspension stability and provides nanoparticles with muco-inert properties. (3) The nanosuspension can be efficiently aerosolized, allowing for a high drug fraction potentially reaching the deep lung. Thus, this formulation represents a promising tool for the lung administration via nebulization directly in the pipe of ventilators, to be used as such or as adjunct therapy for severe lung inflammation.

Facile Biogenic Synthesis and Characterization of Seven Metal-Based Nanoparticles Conjugated with Phytochemical Bioactives Using Fragaria ananassa Leaf Extract.

In this investigation, for the first time, we used Fragaria ananassa (strawberry) leaf extract as a source of natural reducing, capping or stabilizing agents to develop an eco-friendly, cost-effective and safe process for the biosynthesis of metal-based nanoparticles including silver, copper, iron, zinc and magnesium oxide. Calcinated and non-calcinated zinc oxide nanoparticles also synthesized during a method different from our previous study. To confirm the successful formation of nanoparticles, different characterization techniques applied. UV-Vis spectroscopy, X-ray Diffraction (XRD) spectroscopy, Field Emission Scanning Electron Microscopy (FESEM) coupled with Energy Dispersive X-ray Spectroscopy (EDS), Photon Cross-Correlation Spectroscopy (PCCS) and Fourier Transformed Infrared Spectroscopy (FT-IR) were used to study the unique structure and properties of biosynthesized nanoparticles. The results show the successful formation of metal-based particles in the range of nanometer, confirmed by different characterization techniques. Finally, the presented approach has been demonstrated to be effective in the biosynthesis of metal and metal oxide nanoparticles.

Green Silver and Gold Nanoparticles: Biological Synthesis Approaches and Potentials for Biomedical Applications.

The nanomaterial industry generates gigantic quantities of metal-based nanomaterials for various technological and biomedical applications; however, concomitantly, it places a massive burden on the environment by utilizing toxic chemicals for the production process and leaving hazardous waste materials behind. Moreover, the employed, often unpleasant chemicals can affect the biocompatibility of the generated particles and severely restrict their application possibilities. On these grounds, green synthetic approaches have emerged, offering eco-friendly, sustainable, nature-derived alternative production methods, thus attenuating the ecological footprint of the nanomaterial industry. In the last decade, a plethora of biological materials has been tested to probe their suitability for nanomaterial synthesis. Although most of these approaches were successful, a large body of evidence indicates that the green material or entity used for the production would substantially define the physical and chemical properties and as a consequence, the biological activities of the obtained nanomaterials. The present review provides a comprehensive collection of the most recent green methodologies, surveys the major nanoparticle characterization techniques and screens the effects triggered by the obtained nanomaterials in various living systems to give an impression on the biomedical potential of green synthesized silver and gold nanoparticles.

Measuring the Hydrodynamic Size of Nanoparticles Using Fluctuation Correlation Spectroscopy.

Fluctuation correlation spectroscopy (FCS) is a well-established analytical technique traditionally used to monitor molecular diffusion in dilute solutions, the dynamics of chemical reactions, and molecular processes inside living cells. In this review, we present the recent use of FCS for measuring the size of colloidal nanoparticles in solution. We review the theoretical basis and experimental implementation of this technique and its advantages and limitations. In particular, we show examples of the use of FCS to measure the size of gold nanoparticles, monitor the rotational dynamics of gold nanorods, and investigate the formation of protein coronas on nanoparticles.

Weighing nanoparticles in solution at the attogram scale.

Physical characterization of nanoparticles is required for a wide range of applications. Nanomechanical resonators can quantify the mass of individual particles with detection limits down to a single atom in vacuum. However, applications are limited because performance is severely degraded in solution. Suspended micro- and nanochannel resonators have opened up the possibility of achieving vacuum-level precision for samples in the aqueous environment and a noise equivalent mass resolution of 27 attograms in 1-kHz bandwidth was previously achieved by Lee et al. [(2010) Nano Lett 10(7):2537-2542]. Here, we report on a series of advancements that have improved the resolution by more than 30-fold, to 0.85 attograms in the same bandwidth, approaching the thermomechanical noise limit and enabling precise quantification of particles down to 10 nm with a throughput of more than 18,000 particles per hour. We demonstrate the potential of this capability by comparing the mass distributions of exosomes produced by different cell types and by characterizing the yield of self-assembled DNA nanoparticle structures.

Detection of zinc oxide and cerium dioxide nanoparticles during drinking water treatment by rapid single particle ICP-MS methods.

Nanoparticles (NPs) entering water systems are an emerging concern as NPs are more frequently manufactured and used. Single particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) methods were validated to detect Zn- and Ce-containing NPs in surface and drinking water using a short dwell time of 0.1 ms or lower, ensuring precision in single particle detection while eliminating the need for sample preparation. Using this technique, information regarding NP size, size distribution, particle concentration, and dissolved ion concentrations was obtained simultaneously. The fates of Zn- and Ce-NPs, including those found in river water and added engineered NPs, were evaluated by simulating a typical drinking water treatment process. Lime softening, alum coagulation, powdered activated carbon sorption, and disinfection by free chlorine were simulated sequentially using river water. Lime softening removed 38-53 % of Zn-containing and ZnO NPs and >99 % of Ce-containing and CeO2 NPs. Zn-containing and ZnO NP removal increased to 61-74 % and 77-79 % after alum coagulation and disinfection, respectively. Source and drinking water samples were collected from three large drinking water treatment facilities and analyzed for Zn- and Ce-containing NPs. Each facility had these types of NPs present. In all cases, particle concentrations were reduced by a minimum of 60 % and most were reduced by >95 % from source water to finished drinking water. This study concludes that uncoated ZnO and CeO2 NPs may be effectively removed by conventional drinking water treatments including lime softening and alum coagulation.

Single particle ICP-MS method development for the determination of plant uptake and accumulation of CeO2 nanoparticles.

Cerium dioxide nanoparticles (CeO2NPs) are among the most broadly used engineered nanoparticles that will be increasingly released into the environment. Thus, understanding their uptake, transportation, and transformation in plants, especially food crops, is critical because it represents a potential pathway for human consumption. One of the primary challenges for the endeavor is the inadequacy of current analytical methodologies to characterize and quantify the nanomaterial in complex biological samples at environmentally relevant concentrations. Herein, a method was developed using single particle-inductively coupled plasma-mass spectrometry (SP-ICP-MS) technology to simultaneously detect the size and size distribution of particulate Ce, particle concentration, and dissolved cerium in the shoots of four plant species including cucumber, tomato, soybean, and pumpkin. An enzymatic digestion method with Macerozyme R-10 enzyme previously used for gold nanoparticle extraction from the tomato plant was adapted successfully for CeO2NP extraction from all four plant species. This study is the first to report and demonstrate the presence of dissolved cerium in plant seedling shoots exposed to CeO2NPs hydroponically. The extent of plant uptake and accumulation appears to be dependent on the plant species, requiring further systematic investigation of the mechanisms.

Quantification of the cellular dose and characterization of nanoparticle transport during in vitro testing.

BACKGROUND: The constant increase of the use of nanomaterials in consumer products is making increasingly urgent that standardized and reliable in vitro test methods for toxicity screening be made available to the scientific community. For this purpose, the determination of the cellular dose, i.e. the amount of nanomaterials effectively in contact with the cells is fundamental for a trustworthy determination of nanomaterial dose responses. This has often been overlooked in the literature making it difficult to undertake a comparison of datasets from different studies. Characterization of the mechanisms involved in nanomaterial transport and the determination of the cellular dose is essential for the development of predictive numerical models and reliable in vitro screening methods. RESULTS: This work aims to relate key physico-chemical properties of gold nanoparticles (NPs) to the kinetics of their deposition on the cellular monolayer. Firstly, an extensive characterization of NPs in complete culture cell medium was performed to determine the diameter and the apparent mass density of the formed NP-serum protein complexes. Subsequently, the kinetics of deposition were studied by UV-vis absorbance measurements in the presence or absence of cells. The fraction of NPs deposited on the cellular layer was found to be highly dependent on NP size and apparent density because these two parameters influence the NP transport. The NP deposition occurred in two phases: phase 1, which consists of cellular uptake driven by the NP-cell affinity, and phase 2 consisting mainly of NP deposition onto the cellular membrane. CONCLUSION: The fraction of deposited NPs is very different from the initial concentration applied in the in vitro assay, and is highly dependent of the size and density of the NPs, on the associated transport rate and on the exposure duration. This study shows that an accurate characterization is needed and suitable experimental conditions such as initial concentration of NPs and liquid height in the wells has to be considered since they strongly influence the cellular dose and the nature of interactions of NPs with the cells.

Nanophotonic force microscopy: characterizing particle-surface interactions using near-field photonics.

Direct measurements of particle-surface interactions are important for characterizing the stability and behavior of colloidal and nanoparticle suspensions. Current techniques are limited in their ability to measure pico-Newton scale interaction forces on submicrometer particles due to signal detection limits and thermal noise. Here we present a new technique for making measurements in this regime, which we refer to as nanophotonic force microscopy. Using a photonic crystal resonator, we generate a strongly localized region of exponentially decaying, near-field light that allows us to confine small particles close to a surface. From the statistical distribution of the light intensity scattered by the particle we are able to map out the potential well of the trap and directly quantify the repulsive force between the nanoparticle and the surface. As shown in this Letter, our technique is not limited by thermal noise, and therefore, we are able to resolve interaction forces smaller than 1 pN on dielectric particles as small as 100 nm in diameter.

Transport of Nanoparticles into Plants and Their Detection Methods.

Nanoparticle transport into plants is an evolving field of research with diverse applications in agriculture and biotechnology. This article provides an overview of the challenges and prospects associated with the transport of nanoparticles in plants, focusing on delivery methods and the detection of nanoparticles within plant tissues. Passive and assisted delivery methods, including the use of roots and leaves as introduction sites, are discussed, along with their respective advantages and limitations. The barriers encountered in nanoparticle delivery to plants are highlighted, emphasizing the need for innovative approaches (e.g., the stem as a new recognition site) to optimize transport efficiency. In recent years, research efforts have intensified, leading to an evendeeper understanding of the intricate mechanisms governing the interaction of nanomaterials with plant tissues and cells. Investigations into the uptake pathways and translocation mechanisms within plants have revealed nuanced responses to different types of nanoparticles. Additionally, this article delves into the importance of detection methods for studying nanoparticle localization and quantification within plant tissues. Various techniques are presented as valuable tools for comprehensively understanding nanoparticle-plant interactions. The reliance on multiple detection methods for data validation is emphasized to enhance the reliability of the research findings. The future outlooks of this field are explored, including the potential use of alternative introduction sites, such as stems, and the continued development of nanoparticle formulations that improve adhesion and penetration. By addressing these challenges and fostering multidisciplinary research, the field of nanoparticle transport in plants is poised to make significant contributions to sustainable agriculture and environmental management.

Quantifying Bound Proteins on Pegylated Gold Nanoparticles Using Infrared Spectroscopy.

Protein-nanoparticle (NP) complexes are nanomaterials that have numerous potential uses ranging from biosensing to biomedical applications such as drug delivery and nanomedicine. Despite their extensive use quantifying the number of bound proteins per NP remains a challenging characterization step that is crucial for further developments of the conjugate, particularly for metal NPs that often interfere with standard protein quantification techniques. In this work, we present a method for quantifying the number of proteins bound to pegylated thiol-capped gold nanoparticles (AuNPs) using an infrared (IR) spectrometer, a readily available instrument. This method takes advantage of the strong IR bands present in proteins and the capping ligands to quantify protein-NP ratios and circumvents the need to degrade the NPs prior to analysis. We show that this method is generalizable where calibration curves made using inexpensive and commercially available proteins such as bovine serum albumin (BSA) can be used to quantify protein-NP ratios for proteins of different sizes and structures.

The vast majority of nucleic acid-loaded lipid nanoparticles contain cargo.

Nucleic acid-based therapies are transforming medicine, but rely on an efficient delivery vehicle such as lipid nanoparticles (LNPs). Concerns exists in the nanomedicine field, that a large fraction of the LNPs in the ensemble does not contain any nucleic acid cargo and thus exert no functional effect. Nevertheless, how LNP lipid formulation, the LNP preparation method employed and nucleic acid cargo size correlates with the proportion of empty LNPs remains largely unexplored. Here we employ a well-established single particle based method to study nucleic acid loading heterogeneity in LNPs. We find that only a minor fraction of LNPs are "empty", both for LNPs loaded with siRNA, mRNA and plasmids. For clinically relevant LNPs for mRNA delivery, we never detected more than 16% empty nanoparticles in the ensemble. Thus employing standard LNP lipid-cargo combinations and preparation schemes results in LNPs with the potential to serve their biomedical function.

Green-synthesized CuO and ZnO nanoparticles derived from Calotropis gigantea (Apple of Sodom): enhancing plant growth, efficient dye removal, and potent antibacterial applications.

Nanoparticles, owing to their unique physicochemical properties, have garnered significant attention in various scientific disciplines, including materials science, chemistry, biology, and environmental engineering. In recent years, the synthesis of metal oxide nanoparticles, such as NiO, Fe2O3, ZnO, SnO2, and CuO via green routes, has gained attraction due to their diverse applications in fields ranging from catalysis and electronics to medicine and environmental remediation. This study focuses on the green synthesis of copper oxide (CuO) and zinc oxide (ZnO) nanoparticles using Calotropis gigantea (Apple of Sodom) leaf extract as a reducing agent and stabilizer, with zinc nitrate (ZnNO3.6H2O) and copper nitrate (CuNO3.3H2O) as precursors. The hexagonal phase of ZnO and monoclinic plan structure of CuO with high crystallinity was confirmed by XRD and elemental composition by EDX analysis. With the help of an SEM image, particle size measured for CuO and ZnO using ImageJ software was found to be 56.08 nm and 46.49 nm, respectively. This study investigates the efficacy of nanoparticles in wastewater treatment, particularly focusing on methylene blue dye decolorization using the statistical processing of response surface methodology (RSM) using the Box-Behnken method. Additionally, it explores the impact of synthesized nanoparticles on seed growth enhancement, using Vigna radiata (green gram) seeds immersed in various doses of nanoparticles (0, 0.5, 1, 1.5, 2 mg/30 mL). Furthermore, the antibacterial activity of the nanoparticles against both gram-positive and gram-negative bacteria is evaluated. The results confirm the effectiveness of the materials for methylene blue dye removal, achieving 80.53% with CuO and 78.25% with ZnO. Significant seed growth was observed with a low nanoparticle dosage of 1.5 mg/30 mL, resulting in the highest seedling vigour index and germination percentage. This reduces the need for fertilizers and lessens environmental impact.

Combining orthogonal measurements to unveil diclofenac encapsulation into polymeric and lipid nanocarriers.

The quantification of the drug associated to nanoparticle carriers, often expressed in terms of encapsulation efficiency, is a regulatory requirement. The establishment of independent methods to evaluate this parameter provides a means for measurement validation, which is critical in providing confidence in the methods and enabling the robust characterization of nanomedicines. Chromatography is traditionally used to measure drug encapsulation into nanoparticles. Here, we describe an additional independent strategy based on analytical centrifugation. The encapsulation of diclofenac into nanocarriers was quantified based on the mass difference between placebo (i.e. unloaded) and loaded nanoparticles. This difference was estimated using particle densities measured by differential centrifugal sedimentation (DCS) and size and concentration values measured by particle tracking analysis (PTA). The proposed strategy was applied to two types of formulations, namely poly(lactic-co-glycolic acid) (PLGA) nanoparticles and nanostructured lipid carriers, which were analysed by DCS operated in sedimentation and flotation modes, respectively. The results were compared to those from high performance liquid chromatography (HPLC) measurements. Additionally, X-ray photoelectron spectroscopy analysis was used to elucidate the surface chemical composition of the placebo and loaded nanoparticles. The proposed approach enables the monitoring of batch-to-batch consistency and the quantification of diclofenac association to PLGA nanoparticles from 0.7 ng to 5 ng of drug per 1 µg of PLGA, with good linear correlation between DCS and HPLC results (R2 = 0.975). Using the same approach, similar quantification in lipid nanocarriers was possible for a loading of diclofenac ≥1.1 ng per 1 µg of lipids, with results in agreement with the HPLC method (R2 = 0.971). Hence, the strategy proposed here expands the analytical tools available for evaluating nanoparticles encapsulation efficiency, being thus significant for increasing the robustness of drug-delivery nanocarriers characterization.

Solid Lipid Nanoparticles vs. Nanostructured Lipid Carriers: A Comparative Review.

Solid-lipid nanoparticles and nanostructured lipid carriers are delivery systems for the delivery of drugs and other bioactives used in diagnosis, therapy, and treatment procedures. These nanocarriers may enhance the solubility and permeability of drugs, increase their bioavailability, and extend the residence time in the body, combining low toxicity with a targeted delivery. Nanostructured lipid carriers are the second generation of lipid nanoparticles differing from solid lipid nanoparticles in their composition matrix. The use of a liquid lipid together with a solid lipid in nanostructured lipid carrier allows it to load a higher amount of drug, enhance drug release properties, and increase its stability. Therefore, a direct comparison between solid lipid nanoparticles and nanostructured lipid carriers is needed. This review aims to describe solid lipid nanoparticles and nanostructured lipid carriers as drug delivery systems, comparing both, while systematically elucidating their production methodologies, physicochemical characterization, and in vitro and in vivo performance. In addition, the toxicity concerns of these systems are focused on.