Nanoparticles In Biological Samples Research Articles

Plastic Analysis with a Plasmonic Nano-Gold Sensor Coated with Plastic-Binding Peptides.

Contamination with plastics of small dimensions (<1 µm) represents a health concern for many terrestrial and aquatic organisms. This study examined the use of plastic-binding peptides as a coating probe to detect various types of plastic using a plasmon nano-gold sensor. Plastic-binding peptides were selected for polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS) based on the reported literature. Using nAu with each of these peptides to test the target plastics revealed high signal, at 525/630 nm, suggesting that the target plastic limited HCl-induced nAu aggregation. Testing with other plastics revealed some lack of specificity but the signal was always lower than that of the target plastic. This suggests that these peptides, although reacting mainly with their target plastic, show partial reactivity with the other target plastics. By using a multiple regression model, the relative levels of a given plastic could be corrected by the presence of other plastics. This approach was tested in freshwater mussels caged for 3 months at sites suspected to release plastic materials: in rainfall overflow discharges, downstream a largely populated city, and in a municipal effluent dispersion plume. The data revealed that the digestive glands of the mussels contained higher levels of PP, PE, and PET plastic particles at the rainfall overflow and downstream city sites compared to the treated municipal effluent site. This corroborated earlier findings that wastewater treatment could remove nanoparticles, at least in part. A quick and inexpensive screening test for plastic nanoparticles in biological samples with plasmonic nAu-peptides is proposed.

Identifying Carbon Nanoparticles in Biological Samples by Means of Transmission Electron Microscopy

Identifying Carbon Nanoparticles in Biological Samples by Means of Transmission Electron Microscopy

Artifacts in magnetic force microscopy of histological sections

Detection of iron-oxide nanoparticles in biological samples has widespread applications. Iron can be naturally present in biological tissues as physiological ferrihydrite particles of ferritin(iron) or as biogenic magnetite nanoparticles in certain neurodegenerative pathologies. In our previous studies, we had reported how magnetic force microscopy (MFM) can be used to map iron deposits in tissue sections in a label-free manner. In this study, to improve the efficiency of MFM for histological analysis, we explored the effect of increasing scan rate. Our results show how MFM images of tissue sections can be contaminated with artifacts due to topographical cross-talk, especially at higher scan rates. These artifacts were observed in rodent spleen as well as in sections of brain tissue obtained from patients with Alzheimer’s Disease. Our results show how topographical cross talk can make it challenging to unambiguously detect histological iron via MFM. Multimodal approaches can help overcome some of these limitations.

Nanoparticles and nanomaterials as inevitable modern toxic agents. Review. Part 2. Main areas of research on toxicity and techniques to measure a content of nanoparticles in tissues

The second part of the review considers the following three main areas of research on the toxicity of nanoparticles (NPs): environmental toxicity, molecular mechanisms of toxicity, and reproductive toxicity. The studies carried out on aquatic and soil model organisms are described, with consideration of the effects of NPs in concentrations close to natural in water with different salinity, as well as in comparison with the effects of ions. The articles devoted to various aspects of NP-induced oxidative stress are listed, the estimations of genotoxicity and mutagenicity of NPs using different standard methods are described, and the currently known data on the formation of protein crowns around NPs are considered. Questions are stated about the dose-dependence of effects and the influence of the applied stabilizing coating. The influence of NPs on the prenatal and postnatal development of various model vertebrate species is considered, which includes morphological disturbances, changes in gene expression and in the behavior of grown animals, as well as the influence on the reproductive system in adult females and males. The main methods for the quantitative measurements of the content of NPs in biological samples are also considered as the necessary stage of research on the toxicity of NPs for humans and animals.

Determination of metallic nanoparticles in biological samples by single particle ICP-MS: a systematic review from sample collection to analysis.

A systematic review of the use of single particle ICP-MS to analyse engineered nanomaterials (ENMs) in biological samples (plants, animals, body fluids) has highlighted that efforts have focused on a select few types of ENMs (e.g., Ag and TiO2) and there is a lack of information for some important tissues (e.g., reproductive organs, skin and fatty endocrine organs). The importance of sample storage is often overlooked but plays a critical role. Careful consideration of the ENM and matrix composition is required to select an appropriate protocol to liberate ENMs from a tissue whilst not promoting the transformation of them, or genesis of new particulates. A ‘one size fits all’ protocol, applicable to all possible types of ENM and biological matrices, does not seem practical. However, alkaline-based extractions would appear to show greater promise for wide applicability to animal tissues, although enzymatic approaches have a role, especially for plant tissues. There is a lack of consistency in metrics reported and how they are determined (e.g. size limit of detection, and proportions of recovery), making comparison between some studies more difficult. In order to establish standardised protocols for regulatory use, effort is needed to: develop certified reference materials, achieve international agree on nomenclature and the use of control samples, and to create a decision tree to help select the best sample preparation for the type of tissue matrix.

Short-Term Introduction of Fullerene C60 Nanoparticles in Rat Small Intestine Induces the Rapid Development of Hepatocyte Pathology

Fullerenes C60 are used in various fields of industry; therefore, issues of studying their biosafety for living organisms, their biodistribution in organs and tissues organs and tissues of laboratory animals, and their induction of cellular pathologies are very important. In recent years, analytical and transmission electron microscopy (TEM) have proven their efficiency in detecting carbon nanoparticles in biological samples. Combining these methods can reveal pathological changes in cells induced by nanoparticles and detect accumulations of nanoparticles in them. The aim of the study was to reveal cellular pathologies upon direct short-term administration of fullerene C60 nanoparticles into the gastrointestinal tract (GIT) of rats and to detect nanoparticles in samples of the small intestine and liver. The histological and ultrastructural analysis did not reveal pathological changes in the small intestine, but degenerative changes in hepatocytes were indicated in the form of accumulation of lipid inclusions. High-performance liquid chromatography and analytical TEM revealed no accumulations of fullerene C60 nanoparticles in the studied samples. Probably, hepatosteatosis may be a consequence of the indirect influence of C60 nanoparticles on liver cells. Apparently, the surface of fullerene C60 undergoes modifications in the GIT, and its contact with the cells of the small intestine induces the synthesis of proinflammatory cytokines, with their subsequent entry into the liver, leading to the rapid development of fatty degeneration. Revealing and identifying modified C60 nanoparticles and their metabolites, in turn, is a complex problem, which in the future can be solved using radiotracers, mass spectrometry, and modern metabolomic technologies.

Inductively coupled plasma mass spectrometry based platforms for studies involving nanoparticle effects in biological samples.

The widespread application of nanoparticles (NPs) in recent times has caused concern because of their effects in biological systems. Although NPs can be produced naturally, industrially synthesized NPs affect the metabolism of a given organism because of their high reactivity. The biotransformation of NPs involves different processes, including aggregation/agglomeration, and reactions with biomolecules that will be reflected in their toxicity. Several analytical techniques, including inductively coupled plasma mass spectrometry (ICP-MS), have been used for characterizing and quantifying NPs in biological samples. In fact, in addition to providing information regarding the morphology and concentration of NPs, ICP-MS-based platforms, such as liquid chromatography/ICP-MS, single-particle ICP-MS, field-flow fractionation (asymmetrical flow field-flow fractionation)-ICP-MS, and laser ablation-ICP-MS, yield elemental information about molecules. Furthermore, such information together with speciation analysis enlarges our understanding of the interaction between NPs and biological organisms. This study reports the contribution of ICP-MS-based platforms as a tool for evaluating NPs in distinct biological samples by providing an additional understanding of the behavior of NPs and their toxicity in these organisms.

Detection and identification of engineered nanoparticles in exhaled breath condensate, blood serum, and urine of occupationally exposed subjects

The use of nanotechnology and the fields of application of nanomaterials are growing vastly, but the negative health effects on the exposed employees are not well studied. The standardized methods of monitoring of occupational exposure are still absent. The task of occupational physicians is to find the ways of evaluation of potential risks of exposure to engineered nanoparticles and to determine the biomarkers for early diagnostics and prevention of occupational diseases. The aim of our study was to detect and identify engineered nanoparticles in biological samples received from occupationally exposed subjects and to evaluate the association of findings with the results of external aerosol measurements on the workplace. The study cohort consisted of two groups of subjects. The first group was exposed to engineered nanoparticles containing mainly iron, manganese, and carbon compounds; the second group was exposed to the nanoparticles containing copper oxide. The post-shift biological samples (urine, blood serum, and exhaled breath condensate) were collected. The analysis was performed by transmission electron microscopy and energy-dispersive spectroscopy. The nanoparticles were detected in all the biological samples. The most common identified chemical elements were the biogenic ones (carbon, potassium, chlorine, oxygen), but the nanoparticles containing metals were identified in EBC, blood, and urine as well (gold, silver, copper, lanthanum, cerium, and tantalum). Our results demonstrate the possibility of detection of occupational exposure to the engineered nanoparticles in human biological fluids. Further studies are necessary to compare the pre-shift and post-shift burden of samples with engineered nanoparticles and to determine the magnitude of occupational exposure during the shift.

Snapshots of Iron Speciation: Tracking the Fate of Iron Nanoparticle Drugs via a Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometric Approach.

Nanomedicines are nanoparticle-based therapeutic or diagnostic agents designed for targeted delivery or enhanced stability. Nanotechnology has been successfully employed to develop various drug formulations with improved pharmacokinetic characteristics, and current research efforts are focused on the development of new innovator and generic nanomedicines. Nanomedicines, which are often denoted as complex or nonbiological complex drugs, have inherently different physicochemical and pharmacokinetic properties than conventional small molecule drugs. The tools necessary to fully evaluate nanomedicines in clinical settings are limited, which can hamper their development. One of the most successful families of nanomedicines are iron-carbohydrate nanoparticles, which are administered intravenously (IV) to treat iron-deficiency anemia. In the U.S., the FDA has approved six distinct iron-carbohydrate nanoparticles but only one generic version (sodium ferric gluconate for Ferrlecit). There is significant interest in approving additional generic iron-carbohydrate drugs; however, the lack of a direct method to monitor the fate of the iron nanoparticles in clinical samples has impeded this approval. Herein we report a novel liquid chromatography-inductively coupled plasma-mass spectrometry (LC-ICP-MS) method that allows for the direct quantification of the iron-carbohydrate drugs in clinical samples, while simultaneously measuring the speciation of the iron released from the nanoparticles in biological samples. To our knowledge, this is the first time that iron nanoparticles have been observed in clinical samples, opening the door for direct pharmacokinetic studies of this family of drugs. This method has potential applications not only for iron-nanoparticle drugs but also for any nanomedicine with an inorganic component.

Convenient and ultra-sensitive fluorescence detection of bovine serum albumin by using Rhodamine-6G modified gold nanoparticles in biological samples.

This study comprises a convenient, rapid and very sensitive method for the determination of bovine serum albumin (BSA). The technique is based on fluorescence resonance energy transfer (FRET) between Rhodamine-6G (R6G) acting as donor and gold nanoparticles (AuNPs) acting as acceptors. This method takes advantage of AuNPs that have high quenching efficiency, therefore the absorption spectra range shifts from 521to 635nm when aggregation of the AuNPs takes place. Furthermore, when R6G was electrostatically self-adsorbed to the citrate-stabilized AuNPs surface the fluorescence intensity was quenched. After addition of BSA, the fluorescence intensity of the R6G recovered as BSA induced aggregation of the AuNPs and the adsorbed R6G was released to the solution. The recovery of intensity displays a linear relationship with BSA concentration over the range from 0.8×10-11 M to 5.6×10-11 M. The detection limit for BSA was found to be 4.58×10-11 M. The proposed method exhibited rapid analysis with high selectivity for BSA determination in human urine, blood and serum samples.

Biological Monitoring of Inhaled Nanoparticles in Patients: An Appealing Approach To Study Causal Link between Human Respiratory Pathology and Exposure to Nanoparticles

Although necessary, in vitro and in vivo studies are not fully successful at predicting nanomaterials toxicity. We propose to associate such assays to the biological monitoring of nanoparticles in clinical samples to get more relevant data on the chemical and physical nature and dose of nanoparticles found in humans. The concept is to establish the load of nanoparticles in biological samples of patients. Then, by comparing samples from different patient groups, nanoparticles of interest could be identified and a potential link between a given nanoparticle type and toxicity could be suggested. It must be confirmed by investigating the biological effects induced by these nanoparticles using in vitro or in vivo models (mechanistic or dose-response studies). This translational approach from the bedside to the bench and vice versa could allow a better understanding of the nanoparticle effects and mechanisms of toxicity that can contribute, at least in part, to a disease.

Enhanced Laser Desorption/Ionization Mass Spectrometric Detection of Gold Nanoparticles in Biological Samples Using the Synergy between Added Matrix and the Gold Core.

Laser desorption/ionization mass spectrometry (LDI-MS) has been used to detect gold nanoparticles (AuNPs) in biological samples, such as cells and tissues, by ionizing their attached monolayer ligands. Many NP-attached ligands, however, are difficult to ionize by LDI, making it impossible to track these NPs in biological samples. In this work, we demonstrate that concentrations of matrix-assisted LDI (MALDI) matrices an order of magnitude below the values typically used in MALDI can facilitate the selective detection of AuNPs with these ligands, even in samples as complex as cell lysate. This enhanced sensitivity arises from a synergistic relationship between the gold core and the matrix that helps to selectively ionize ligands attached to the AuNPs.

Synthesis and characterization of paclitaxel-imprinted nanoparticles for recognition and controlled release of an anticancer drug

Imprinted nanoparticles as drug delivery carriers have been considered because owing to their cross-linked network, they act as the drug reservoir for controlled release. In this study, selective MIPs nanoparticles of paclitaxel (PTX) were successfully developed for application in the biological molecular recognition and in the design of new anticancer drug delivery systems. The MIPs nanoparticles prepared by miniemulsion polymerization technique using methacrylic acid (MAA) and methyl methacrylate as non-covalent functional monomer, ethylene glycol dimethacrylate and trimethylolpropane trimethacrylate (TRIM) as cross-linker agent, azobisisobutyronitrile as initiator, and hexadecane as hydrophobic agent. In order to prepare of MIP nanoparticles, the synthesis conditions and effective parameters, such as: cross-linker agent, different molar ratios of template–functional monomer–cross-linker agent, were investigated. In addition, the effect of different molar ratios of template and monomers on polymers binding and morphology were characterized. Structure and thermal properties of MIPs were confirmed by FT-IR spectroscopy and thermogravimetric analysis. Imprinted nanoparticles showed significant drug loading and encapsulation efficiency, 17.8 and 100 %, respectively. The particle size of MIP nanoparticles varies between 187 and 726 nm, according the SEM images and laser light scattering data. The imprinted nanoparticles showed satisfactory affinity (84 %) to PTX with a binding of 12 times higher than non-imprinted nanoparticles in biological samples when MAA and TRIM were used as functional and cross-linker monomer, respectively. Results from release experiments of MIPs showed a very slow and controlled release of PTX which would be helpful for sustained drug delivery.

Optical imaging of individual plasmonic nanoparticles in biological samples.

Imaging of plasmonic nanoparticles (PNP) with optical microscopy has aroused considerable attention in recent years. The unique localized surface plasmon resonance (LSPR) from metal nanoparticles facilitates the transduction of a chemical or physical stimulus into optical signals in a highly efficient way. It is therefore possible to perform chemical or biological assays at the single object level with the help of standard optical microscopes. Because the source of background noise from different samples is different, distinct imaging modalities have been developed to discern the signals of interest in complex surroundings. With these convenient yet powerful techniques, great improvements in chemical and biological assays have been demonstrated, and many interesting phenomena and dynamic processes have also been elucidated. Further development and application of optical imaging methods for plasmonic probes should lead to many exciting results in chemistry and biology in the future.

Mapping nanoparticles injected into a biological tissue using laser-induced breakdown spectroscopy

We describe a setup for LIBS mapping of nanoparticles and trace metallic elements in biological organ as well as the followed experimental procedure. Mapping was performed for metallic elements such as Gd, Si, Ca and Fe, with a resolution of 100μm on kidney slices sampled from a mouse 24h after intravenous injection of a solution of gadolinium-based nanoparticles. An approach for quantifying Gd in the tissue is also presented with a good agreement with measurement performed by ICP-OES. We demonstrate that LIBS offers a simple and robust method to study the distribution of gadolinium-based nanoparticles in biological samples, without any labeling of the nanoparticles. The used bench-top instrumentation is fully compatible with the standard optical microscopy, which shows its large potential use in Biology and Medicine as a tool for complementary observation of trace metallic elements with respect to the classical optical observations which are generally based on the responses of biomolecules or cells.

Screening test of silver nanoparticles in biological samples by graphite furnace-atomic absorption spectrometry

A simple, rapid and inexpensive screening test is presented to determine the presence of silver (Ag) nanoparticles in biological samples. The method is based on graphite furnace-atomic absorption spectrometry (Zeeman background correction) where an increase in the atomization temperature is observed with an increase in the particle size of Ag. The method is able to determine the presence of Ag ions from the presence of nano-Ag of 20, 60 and 80 nm, but the methodology was less apt to resolve nanoparticles between 20 and 60 nm. The proposed methodology was capable of determining the presence of dissolved Ag(+) from 20 nm in prepared mixtures, and in the liver of rainbow trout exposed to either dissolved or 20-nm nano-Ag.

Nanobarcoding: detecting nanoparticles in biological samples using in situ polymerase chain reaction.

BackgroundDetermination of the fate of nanoparticles (NPs) in a biological system, or NP biodistribution, is critical in evaluating an NP formulation for nanomedicine. Current methods to determine NP biodistribution are greatly inadequate, due to their limited detection thresholds. Herein, proof of concept of a novel method for improved NP detection based on in situ polymerase chain reaction (ISPCR), coined “nanobarcoding,” is demonstrated.MethodsNanobarcoded superparamagnetic iron oxide nanoparticles (NB-SPIONs) were characterized by dynamic light scattering, zeta potential, and hyperspectral imaging measurements. Cellular uptake of Cy5-labeled NB-SPIONs (Cy5-NB-SPIONs) was imaged by confocal microscopy. The feasibility of the nanobarcoding method was first validated by solution-phase PCR and “pseudo”-ISPCR before implementation in the model in vitro system of HeLa human cervical adenocarcinoma cells, a cell line commonly used for ISPCR-mediated detection of human papilloma virus (HPV).ResultsDynamic light-scattering measurements showed that NB conjugation stabilized SPION size in different dispersion media compared to that of its precursor, carboxylated SPIONs (COOH-SPIONs), while the zeta potential became more positive after NB conjugation. Hyperspectral imaging confirmed NB conjugation and showed that the NB completely covered the SPION surface. Solution-phase PCR and pseudo-ISPCR showed that the expected amplicons were exclusively generated from the NB-SPIONs in a dose-dependent manner. Although confocal microscopy revealed minimal cellular uptake of Cy5-NB-SPIONs at 50 nM over 24 hours in individual cells, ISPCR detected definitive NB-SPION signals inside HeLa cells over large sample areas.ConclusionProof of concept of the nanobarcoding method has been demonstrated in in vitro systems, but the technique needs further development before its widespread use as a standardized assay.

Detection, separation, and quantification of unlabeled silica nanoparticles in biological media using sedimentation field-flow fractionation

A rapid, high-resolution methodology for characterization, separation, and quantification of unlabeled inorganic nanoparticles extracted from biological media, based on sedimentation field-flow fractionation and light scattering detection is presented. Silica nanoparticles were added to either human endothelial cell lysate or rat lung tissue homogenate and incubated. The nanoparticles were extracted by acid digestion and then separated and characterized by sedimentation field-flow fractionation. Fractions collected at the peak maxima were analyzed by transmission electron microscopy (TEM) to verify the size and shape of the isolated nanoparticles. Using the linear relationship between the particle number and the area under the fractogram, the recoveries of particles from the tissue homogenate and cell lysate were calculated as 25% and 79%, respectively. The presented methodology facilitates detection, separation, size characterization, and quantification of inorganic nanoparticles in biological samples, within one experimental run.

Magnetic Force Microscopy of an Oxygen‐Sensing Spin‐Probe

AbstractIn the present work we provide magnetic force microscopy (MFM) analysis on particles of a paramagnetic spin‐probe lithium octa‐n‐butoxy‐naphthalo‐cyanine (LiNc‐BuO) used for EPR oximetry. We demonstrate how MFM can generate a magnetic contrast in ambient air for LiNc‐BuO particles when they assemble into structures &gt;50 nm in size. The same sample when assembled into particles &lt;10 nm in size failed to show contrast in MFM images. The potential for utilizing MFM to understand the magnetic signature of paramagnetic nanoparticles in biological samples is discussed.