Impact of Protein Corona Formation on the Thermoresponsive Behavior of Acrylamide-Based Nanogels.

The ability to fine-tune the volume phase transition temperature (VPTT) of thermoresponsive nanoparticles is essential to their successful application in drug delivery. The rational design of these materials is limited by our understanding of the impact that nanoparticle-protein interactions have on their thermoresponsive behavior. In this work, we demonstrate how the formation of protein corona impacts the transition temperature values of acrylamide-based nanogels and their reversibility characteristics, in the presence of lysozyme, given its relevance for the ocular and intranasal administration route. Nanogels were synthesized with N-isopropylacrylamide or N-n-propylacrylamide as backbone monomers, methylenebis(acrylamide) (2.5-20 molar %) as a cross-linker, and functionalized with negatively charged monomers 2-acrylamido-2-methylpropanesulfonic acid, N-acryloyl-l-proline, or acrylic acid; characterization showed comparable particle diameter (c.a.10 nm), but formulation-dependent thermoresponsive properties, in the range 28-54 °C. Lysozyme was shown to form a complex with the negatively charged nanogels, lowering their VPTT values; the hydrophilic nature of the charged comonomer controlled the drop in VPTT upon complex formation, while matrix rigidity only had a small, yet significant effect. The cross-linker content was found to play a major role in determining the reversibility of the temperature-dependent transition of the complexes, with only 20 molar % cross-linked-nanogels displaying a fully reversible transition. These results demonstrate the importance of evaluating protein corona formation in the development of drug delivery systems based on thermoresponsive nanoparticles.

Mapping Microplastics in Humans: Analysis of Polymer Types, and Shapes in Food and Drinking Water-A Systematic Review.

Microplastics (MPs) pervade the environment, infiltrating food sources and human bodies, raising concerns about their impact on human health. This review is focused on three key questions: (i) What type of polymers are humans most exposed to? (ii) What are the prevalent shapes of MPs found in food and human samples? (iii) Are the data influenced by the detection limit on the size of particles? Through a systematic literature analysis, we have explored data on polymer types and shapes found in food and human samples. The data provide evidence that polyester is the most commonly detected polymer in humans, followed by polyamide, polyurethane, polypropylene, and polyacrylate. Fibres emerge as the predominant shape across all categories, suggesting potential environmental contamination from the textile industry. Studies in humans and drinking water reported data on small particles, in contrast to larger size MPs detected in environmental research, in particular seafood. Discrepancies in size detection methodologies across different reports were identified, which could impact some of the discussed trends. This study highlights the need for more comprehensive research on the interactions between MPs and biological systems and the effects of MPs on toxicity, together with standardised analytical methodologies to accurately assess contamination levels and human exposure. Understanding these dynamics is essential for formulating effective strategies to mitigate the environmental and health implications of MP pollution.

Protein-Nanoparticle Interactions Govern the Interfacial Behavior of Polymeric Nanogels: Study of Protein Corona Formation at the Air/Water Interface.

Biomedical applications of nanoparticles require a fundamental understanding of their interactions and behavior with biological interfaces. Protein corona formation can alter the morphology and properties of nanomaterials, and knowledge of the interfacial behavior of the complexes, using in situ analytical techniques, will impact the development of nanocarriers to maximize uptake and permeability at cellular interfaces. In this study we evaluate the interactions of acrylamide-based nanogels, with neutral, positive, and negative charges, with serum-abundant proteins albumin, fibrinogen, and immunoglobulin G. The formation of a protein corona complex between positively charged nanoparticles and albumin is characterized by dynamic light scattering, circular dichroism, and surface tensiometry; we use neutron reflectometry to resolve the complex structure at the air/water interface and demonstrate the effect of increased protein concentration on the interface. Surface tensiometry data suggest that the structure of the proteins can impact the interfacial properties of the complex formed. These results contribute to the understanding of the factors that influence the bio-nano interface, which will help to design nanomaterials with improved properties for applications in drug delivery.

Towards point of care systems for the therapeutic drug monitoring of imatinib.

Therapeutic drug monitoring is used in the clinical setting in the optimisation of dosages to overcome inter-patient pharmacokinetic variability, increasing efficacy whilst reducing toxicity. Imatinib is a tyrosine kinase inhibitor, displaying large variations in plasma concentrations that impact therapeutic success. As a result, imatinib has been the focus in the development of innovative techniques, aimed at its quantification in plasma. Liquid chromatography coupled with tandem mass spectrometry is currently the gold standard; however, cost and availability of the equipment limit its wider application in clinical settings. Recent advances in the field have shown Raman spectroscopy and electrochemistry to be key techniques for the development of promising analytical tools. This article reviews the latest advances towards less costly, more portable solutions that can be used at the point of care. Graphical abstract.

Microbial Degradation of Plastic in Aqueous Solutions Demonstrated by CO2 Evolution and Quantification.

The environmental accumulation of plastics worldwide is a consequence of the durability of the material. Alternative polymers, marketed as biodegradable, present a potential solution to mitigate their ecological damage. However, understanding of biodegradability has been hindered by a lack of reproducible testing methods. We developed a novel method to evaluate the biodegradability of plastic samples based on the monitoring of bacterial respiration in aqueous media via the quantification of CO2 produced, where the only carbon source available is from the polymer. Rhodococcus rhodochrous and Alcanivorax borkumensis were used as model organisms for soil and marine systems, respectively. Our results demonstrate that this approach is reproducible and can be used with a variety of plastics, allowing comparison of the relative biodegradability of the different materials. In the case of low-density polyethylene, the study demonstrated a clear correlation between the molecular weight of the sample and CO2 released, taken as a measure of biodegradability.

Prediction of self-assembly of adenosine analogues in solution: a computational approach validated by isothermal titration calorimetry.

The recent discovery of the role of adenosine-analogues as neuroprotectants and cognitive enhancers has sparked interest in these molecules as new therapeutic drugs. Understanding the behavior of these molecules in solution and predicting their ability to self-assemble will accelerate new discoveries. We propose a computational approach based on density functional theory, a polarizable continuum solvation description of the aqueous environment, and an efficient search procedure to probe the potential energy surface, to determine the structure and thermodynamic stability of molecular clusters of adenosine analogues in solution, using caffeine as a model. The method was validated as a tool for the prediction of the impact of small structural variations on self-assembly using paraxanthine. The computational results were supported by isothermal titration calorimetry experiments. The thermodynamic parameters enabled the quantification of the actual percentage of dimer present in solution as a function of concentration. The data suggest that both caffeine and paraxanthine are present at concentrations comparable to the ones found in biological samples.

Adsorption versus aggregation of NIPAM nanogels: new insight into their behaviour at the air/water interface as a function of concentration.

We have used neutron reflectivity (NR) measurements in combination with dynamic light scattering (DLS), surface tension and ellipsometry, to study the adsorption behaviour at the air/water interface of N-isopropylacrylamide-based nanogels as a function of concentration. The data provide clear evidence that the nanogels are adsorbed at the interface in a strongly deformed shape and forming a multi-layer where the thickness increases with nanogel concentration in the bulk. The combination of surface characterisation techniques and bulk studies indicate that interfacial film formation is preferred over bulk aggregation. This observation at the air/water interface supports the Derjaguin prediction, that a sphere's interaction with a plane (the thick adsorbed nanogel layer at interface) is much larger than nanogel-nanogel (sphere-sphere) association in the bulk. These findings, in particular the changes in conformations and the thick layer adsorption at the interface as a function of concentration, impact significantly on a number of applications for which nanogels are currently being investigated. These results contribute to the understanding of the behaviour of soft colloids at the interfaces.

The 'Leaky Pipeline'.

Gender bias is widespread and needs to be acknowledged and addressed by the scientific community. In this Guest Editorial, M. Resmini, Professor of Materials Chemistry at the Queen Mary University of London, describes how supervisors can play an important role in addressing the 'leaky pipeline', namely, the progressive loss of capable women from STEM disciplines.

Smart Polymeric Nanoparticles as Emerging Tools for Imaging--The Parallel Evolution of Materials.

The field of imaging has developed considerably over the past decade and recent advances in the area of nanotechnology, in particular nanomaterials, have opened new opportunities. Polymeric nanoparticles are particularly interesting and a number of novel materials, characterized by stimuli-responsive characteristics and fluorescent tagging, have allowed visualization, intracellular labeling and real-time tracking. In some of the latest applications the nanoparticles have been used for imagining of tumor cells, both in vivo and ex vivo.

Incorporation of Cobalt-Cyclen Complexes into Templated Nanogels Results in Enhanced Activity.

Recent advances in nanomaterials have identified nanogels as an excellent matrix for novel biomimetic catalysts using the molecular imprinting approach. Polymerisable Co-cyclen complexes with phosphonate and carbonate templates have been prepared, fully characterised and used to obtain nanogels that show high activity and turnover with low catalytic load, compared to the free complex, in the hydrolysis of 4-nitrophenyl phosphate, a nerve agent simulant. This work demonstrates that the chemical structure of the template has an impact on the coordination geometry and oxidation state of the metal centre in the polymerisable complex resulting in very significant changes in the catalytic properties of the polymeric matrix. Both pseudo-octahedral cobalt(III) and trigonal-bipyramidal cobalt(II) structures have been used for the synthesis of imprinted nanogels, and the catalytic data demonstrate that: i) the imprinted nanogels can be used in 15 % load and show turnover; ii) the structural differences in the polymeric matrices resulting from the imprinting approach with different templates are responsible for the molecular recognition capabilities and the catalytic activity. Nanogel P1, imprinted with the carbonate template, shows >50 % higher catalytic activity than P2 imprinted with the phosphonate.

Smart coumarin-tagged imprinted polymers for the rapid detection of tamoxifen.

A signalling molecularly imprinted polymer was synthesised for easy detection of tamoxifen and its metabolites. 6-Vinylcoumarin-4-carboxylic acid (VCC) was synthesised from 4-bromophenol to give a fluorescent monomer, designed to switch off upon binding of tamoxifen. Clomiphene, a chlorinated analogue, was used as the template for the imprinting, and its ability to quench the coumarin fluorescence when used in a 1:1 ratio was demonstrated. Tamoxifen and 4-hydroxytamoxifen were also shown to quench coumarin fluorescence. Imprinted and non-imprinted polymers were synthesised using VCC, methacrylic acid as a backbone monomer and ethylene glycol dimethacrylate as cross-linker, and were ground and sieved to particle sizes ranging between 45 and 25 µm. Rebinding experiments demonstrate that the imprinted polymer shows very strong affinity for both clomiphene and tamoxifen, while the non-imprinted polymer shows negligible rebinding. The fluorescence of the imprinted polymer is quenched by clomiphene, tamoxifen and 4-hydroxytamoxifen. The switch off in fluorescence of the imprinted polymer under these conditions could also be detected under a UV lamp with the naked eye, making this matrix suitable for applications when coupled with a sample preparation system.

Molecularly Imprinted Polymer Coated Quantum Dots for Multiplexed Cell Targeting and Imaging.

Advanced tools for cell imaging are of great interest for the detection, localization, and quantification of molecular biomarkers of cancer or infection. We describe a novel photopolymerization method to coat quantum dots (QDs) with polymer shells, in particular, molecularly imprinted polymers (MIPs), by using the visible light emitted from QDs excited by UV light. Fluorescent core-shell particles specifically recognizing glucuronic acid (GlcA) or N-acetylneuraminic acid (NANA) were prepared. Simultaneous multiplexed labeling of human keratinocytes with green QDs conjugated with MIP-GlcA and red QDs conjugated with MIP-NANA was demonstrated by fluorescence imaging. The specificity of binding was verified with a non-imprinted control polymer and by enzymatic cleavage of the terminal GlcA and NANA moieties. The coating strategy is potentially a generic method for the functionalization of QDs to address a much wider range of biocompatibility and biorecognition issues.

The Role of Crosslinker Content of Positively Charged NIPAM Nanogels on the In Vivo Toxicity in Zebrafish.

Polymeric nanogels as drug delivery systems offer great advantages, such as high encapsulation capacity and easily tailored formulations; however, data on biocompatibility are still limited. We synthesized N-isopropylacrylamide nanogels, with crosslinker content between 5 and 20 mol%, functionalized with different positively charged co-monomers, and investigated the in vivo toxicity in zebrafish. Our results show that the chemical structure of the basic unit impacts the toxicity profile depending on the degree of ionization and hydrogen bonding capability. When the degree of crosslinking of the polymer was altered, from 5 mol% to 20 mol%, the distribution of the positively charged monomer 2-tert-butylaminoethyl methacrylate was significantly altered, leading to higher surface charges for the more rigid nanogels (20 mol% crosslinker), which resulted in >80% survival rate (48 h, up to 0.5 mg/mL), while the more flexible polymers (5 mol% crosslinker) led to 0% survival rate (48 h, up to 0.5 mg/mL). These data show the importance of tailoring both chemical composition and rigidity of the formulation to minimize toxicity and demonstrate that using surface charge data to guide the design of nanogels for drug delivery may be insufficient.

Modulation of imprinting efficiency in nanogels with catalytic activity in the Kemp elimination.

The interactions between the template and the functional monomer are a key to the formation of cavities in the imprinted nanogels with high molecular recognition properties. Nanogels with enzyme-like activity for the Kemp elimination have been synthesized using 4-vinylpyridine as the functional monomer and indole as the template. The weak hydrogen bond interaction in the complex is shown to be able to induce very distinctive features in the cavities of the imprinted nanogels. The percentage of initiator used in the polymerisation, ranging from 1% to 3%, although it does not have a substantial effect on the catalytic rate, reduces considerably the imprinting efficiency. The alteration of the template/monomer ratio is also investigated, and the data show that there is considerable loss of imprinting efficiency. In terms of substrate selectivity, a number of experiments have been performed using 5-Cl-benzisoxazole as substrate analogue, as well as 5-nitro-indole as template analogue for the preparation of a different set of nanogels. All the kinetic data demonstrate that the chemical structure of the template is key to the molecular recognition properties of the imprinted nanogels that are closely tailored and able to differentiate among small structural changes.

Molecularly imprinted polymers as biomimetic catalysts.

The quest for synthetic biomimetic catalysts able to complement the activity of enzymes has attracted substantial research efforts, and the molecular imprinting approach is one of the attractive techniques that are currently being investigated. In the last 3 years, there has been considerable interest in studying in greater detail the parameters that control and influence the catalytic activity of imprinted polymers and applying molecular imprinting to a wider range of polymeric matrices. This article reports on some of the interesting examples available in the literature regarding the use of metal-containing polymers, microgels and nanogels and thermoresponsive polymers.

Adsorption of soft NIPAM nanogels at hydrophobic and hydrophilic interfaces: Conformation of the interfacial layers determined by neutron reflectivity.

The application of stimuli-responsive microgels and nanogels in drug delivery, catalysis, sensing, and coatings is restricted currently by the limited understanding of the factors influencing their adsorption dynamics and structural changes at interfaces. We have used neutron reflectivity to resolve, on the Ångström scale, the structure of 5% crosslinked N-isopropylacrylamide nanogels at both hydrophobic and hydrophilic interfaces in situ, as a function of temperature and bulk nanogel concentration. Our results show that the higher flexibility given by the low crosslinker content allows for a more ordered structure and packing. The adsorption of the thermoresponsive nanogels is primarily driven by temperature, more specifically its proximity to its volume phase transition temperature, while concentration plays a secondary role. Hydrophobic interactions drive the conformation of the first layer at the interface, which plays a key role in influencing the overall nanogel structure. The mobility of the first layer at the air-water interface as opposed to the interfacial confinement at the solid (SiC8)-liquid interface, results in a different conformation, a more compact and less deformed packing structure, which ultimately drives the structure of the subsequent layers. The evidence for the different structural conformations determined by the degree of hydrophobicity of the interface provides new knowledge, which is essential for the development of further applications. The key role of hydrophobic interactions in driving adsorption and interfacial behavior was also confirmed by fluid AFM experiments which visualized adherence of the nanogels to SiC8 modified surfaces.

Influence of surface chemistry and morphology of nanoparticles on protein corona formation.

Nanomaterials offer promising solutions as drug delivery systems and imaging agents in response to the demand for better therapeutics and diagnostics. However, the limited understanding of the interaction between nanoparticles and biological entities is currently hampering the development of new systems and their applications in clinical settings. Proteins and lipids in biological fluids are known to complex with nanoparticles to form a "biomolecular corona". This has been shown to affect particles' morphology and behavior in biological systems and their interactions with cells. Hence, understanding how nanomaterials' physicochemical properties affect the formation and composition of this biocorona is a crucial step. This work evaluates existing literature on how morphology (size and shape), and surface chemistry (charge and hydrophobicity) of nanoparticles influence the formation of protein corona. The latest evidence suggest that although surface charge promotes the interaction with proteins and lipids, surface chemistry plays a leading role in determining the affinity of the nanoparticle for biomolecules and, ultimately, the composition of the corona. More recently the study of additional nanoparticles' properties like shape and surface chirality have demonstrated a significant effect on protein corona architecture, providing new tools to tailor biomolecular corona formation. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials.

Identification of interactions involved in the generation of nucleophilic reactivity and of catalytic competence in the catalytic site Cys/His ion pair of papain.

Understanding the roles of noncovalent interactions within the enzyme molecule and between enzyme and substrate or inhibitor is an essential goal of the investigation of active center chemistry and catalytic mechanism. Studies on members of the papain family of cysteine proteinases, particularly papain (EC 3.4.22.2) itself, continue to contribute to this goal. The historic role of the catalytic site Cys/His ion pair now needs to be understood within the context of multiple dynamic phenomena. Movement of Trp177 may be necessary to expose His159 to solvent with consequent decrease in its degree of electrostatic solvation of (Cys25)-S(-). Here we report an investigation of this possibility using computer modeling of quasi-transition states and pH-dependent kinetics using 3,3'-dipyridazinyl disulfide, its n-propyl and phenyl derivatives, and 4,4'-dipyrimidyl disulfide as reactivity probes that differ in the location of potential hydrogen-bonding acceptor atoms. Those interactions that influence ion pair geometry and thereby catalytic competence, including by transmission of the modulatory effect of a remote ionization with pK(a) 4, were identified. A key result is the correlation between the kinetic influence of the modulatory trigger of pK(a) 4 and disruption of the hydrogen bond donated by the indole N-H of Trp177, the hydrophobic shield of the initial "intimate" ion pair. This hydrogen bond is accepted by the amide O of Gln19-a component of the oxyanion hole that binds the tetrahedral species formed from the substrate during the catalytic act. The disruption would be expected to contribute to the mobility of Trp177 and possibly to the effectiveness of the binding of the developing oxyanion.

Tuning molecular recognition in water-soluble nanogels with enzyme-like activity for the kemp elimination.

The synthesis and characterization of water-soluble imprinted nanogels with enzyme-like activity in the Kemp elimination is reported together with studies that demonstrate how the recognition properties, morphology, and catalytic activity of the nanoparticles can be tuned by the use of surfactants, such as Tween 20. A detailed kinetic investigation is carried out, which shows clear evidence of saturation kinetics and rule out the effects of mass transfer. This is supported by characterization of the polymeric materials that confirms the morphological changes resulting from the use of surfactants. These results provide an important tool for the development of nanoparticle-based, new catalyst-mimicking enzymes.

A Triple Culture Cell System Modeling the Human Blood-Brain Barrier.

The delivery of drugs to the brain remains a challenge due to the blood-brain barrier's (BBB) highly specific and restrictive properties, which controls and restrict access to the brain parenchyma. However, with the development of nanotechnologies, large panels of new nanomaterials were developed to improve drug delivery, highlighting the need for reliable in vitro microsystems to predict brain penetration in the frame of preclinical assays. Here is a straightforward method to set up a microphysiological system to model the BBB using solely human cells. In its configuration, the model consists of a triple culture including brain-like endothelial cells (BLECs), pericytes, and astrocytes, the three main BBB cellular actors necessary to induce and regulate the BBB properties in a more physiological manner without the requirement of tightening compounds. The model developed in a 12-well plate format, ready after 6 days of triple culture, is characterized in physical properties, gene, and protein expressions and used for polymeric nanogel transport measurement. The model can be used for an extensive range of experiments in healthy and pathological conditions and represents a valuable tool for preclinical assessments of molecule and particle transport, as well as inter-and intracellular trafficking.