Small-angle Neutron Scattering Research Articles

Experimental Evidence for the Role of Dynamics in pH-Dependent Enzymatic Activity.

Enzymatic activity is heavily influenced by pH, but the rationale for the dynamical mechanism of pH-dependent enzymatic activity has not been fully understood. In this work, combined neutron scattering techniques, including quasielastic neutron scattering (QENS) and small angle neutron scattering (SANS), are used to study the structural and dynamic changes of a model enzyme, xylanase, under different pH and temperature environments. The QENS results reveal that xylanase at optimal pH exhibits faster relaxational dynamics and a lower energy barrier between conformational substates. The SANS results demonstrate that pH affects both xylanase's stability and monodispersity. Our findings indicate that enzymes have optimized stability and function under their optimal pH conditions, with both structure and dynamics being affected. The current study offers valuable insights into enzymatic functionality mechanisms, allowing for broad industrial applications.

Secondary phases characterization by SANS and XAS of an ODS ferritic steel after thermal aging at 873 K

An ODS steel with nominal composition Fe–14Cr–2W–0.4Ti-0.3Y2O3 (wt.%) was produced by mechanical alloying and compacted by hot isostatic pressing (HIP) followed by hot cross rolling (HCR). To check the effects of thermal aging at relevant temperatures of operation in fusion power plants, the alloy was thermally aged at 873 K for 2000 h. In this work, small-angle neutron scattering (SANS) and X-ray absorption spectroscopy (XAS) techniques are used for the advanced characterization of secondary phases and the oxide nanoparticle dispersion. SANS results show that the oxide nanoparticles remain stable after the thermal aging treatment. Composition of the oxide nanoparticles was identified as Y2TiO5 or Y2Ti2O7 by SANS, while non-stoichiometry was found by XAS analysis. Laves phase precipitation after the thermal aging treatment is further confirmed by SANS, from the magnetic anisotropic contribution to the scattering intensity associated to this metallic phase, and by XANES.

SANS and SAXS: A Love Story to unravel structural evolution of soy proteins and polysaccharide fibres during high moisture extrusion for meat alternatives

Plant-based meat alternatives are seeing considerable interest due to their potential to reduce environmental burden and enhance population health. The food industry, therefore, seeks routes to provide the consumer with whole-cut plant-based products that closely resemble meat products. High-moisture extrusion (HME) of plant proteins enables the industrial manufacturing of meat-like products with highly hierarchical structural organisation of fibres. The major bottleneck in serving the growing market for these products is a lack of insight into how multiscale structures evolve during shear processing. Furthermore, it remains an open question of how two biopolymers, one being a plant protein and the other being a polysaccharide, contribute to the anisotropic structure formation during HME. This study shows how the complementary use of small-angle neutron scattering (SANS) and small-angle X-ray scattering (SAXS) can add clarity to these matters, benefiting from the different contrasts in scattering length density (SLD) encountered with each of these methods. It is demonstrated that two biopolymers have differences in the development of structural anisotropy. The protein fibril alignment starts in the extruder section with its further development along the cooling die. On the other hand, for polysaccharide fibres, the strongest local alignment has been found in the transition zone.

Nanoprecipitation induced giant magnetostriction: A time-resolved small-angle neutron scattering study of the vacancy-assisted kinetics

Solid-state precipitation is an effective strategy for tuning the mechanical and functional properties of advanced alloys. Structure design and modification necessitate good knowledge of the kinetic evolution of precipitates during fabrication, which is strongly correlated with defect concentration. For Fe-Ga alloys, giant magnetostriction can be induced by the precipitation of the nanoscale tetragonal L60 phase. By introducing quenched-in vacancies, we significantly enhance the magnetostriction of the aged Fe81Ga19 polycrystalline alloys to ∼305 ppm, which is close to the level of single crystals. Although vacancies were found to facilitate the generation of the L60 phase, their impact on the precipitation mechanism and kinetics has yet to be revealed. This study combined transmission electron microscopy (TEM) and time-resolved small-angle neutron scattering (SANS) to investigate the precipitation of the L60 phase during the isothermal aging at 350 and 400 °C, respectively. The evolution of L60 nanophase in morphology and number density in as-cast (AC) and liquid nitrogen quenched (LN) Fe81Ga19 alloys with aging time were quantitatively compared. Interestingly, the nucleation of the L60 phase proceeds progressively in AC while suddenly in LN specimens, indicating the homogenous to heterogeneous mechanism switching induced by concentrated vacancies. Moreover, excess vacancies can change the shape of nanoprecipitates and significantly accelerate the growth and coarsening kinetics. The magnetostrictive coefficient is optimized when the size (long-axis) of L60 precipitates lies between 100 and 110 Å with a number density between 3.2–4.3 × 10–7 Å–3. Insight from this study validates the feasibility of achieving high magnetoelastic properties through precise manipulation of the nanostructure.

Impact of thermosonication at neutral pH on the structural characteristics of faba bean protein isolate dispersions and their physicochemical and techno-functional properties

The effect of thermosonication (TS) (90 °C, 10–30 min) on faba bean protein isolate (FPI) at pH 7 was investigated. The microstructural and techno-functional properties of TS-treated FPI were compared with native FPI or FPI treated with conventional prolonged heating (CH, up to 8 h) at 90 °C. TS treatment effectively converted FPI to amorphous aggregates containing predominant β-sheet secondary structures, as determined by Thioflavin T (ThT) fluorescence and circular dichroism (CD). According to sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), these amorphous aggregates could be formed by disulfide bonds. Additionally, TS treatment is efficient in disrupting large protein aggregates of FPI, thus improving their solubility. Both TS and CH treatments induced formation of viscoelastic FPI hydrogels, whose gel strength depends on the type and time of treatment. Hydrogels formation is likely to arise from the entanglement and interaction of protein aggregates as revealed by small angle neutron scattering (SANS) and scanning electron microscopy (SEM). TS-treated FPI was also used to prepare O/W emulsions and whose structural and physical properties were compared with those stabilised by untreated FPI. At all oil volume fractions (φ = 0.2, 0.5, and 0.7) and FPI concentrations (1, 3, and 5 wt %), emulsions stabilised by TS-treated FPI exhibited smaller oil droplet size, greater mechanical strength and superior stability compared to those stabilised by untreated FPI. The study suggests that TS treatment is promising in improving techno-functional properties of FPI; further studies are needed to exploit TS-treated plant proteins as a novel food ingredient in food product development.

Morphology and kinetics of nanoheterogeneities in Fe81Ga19–Tb alloy: A small-angle neutron scattering study

The formation of nanoprecipitates in the Fe81Ga19 alloy doped with Tb (∼0.1 at. %) was studied by small-angle neutron scattering (SANS). Measurements at room temperature for several samples pre-aged at a fixed temperature in the range of (300–700)°C revealed a dilute system of precipitates with a characteristic size at nanoscale (<1000 Å) growing with an increase in ageing temperature. In situ measurements during isothermal ageing at 300 °C for 3 h revealed fast (order of 10 min) formation of precipitates. SANS analysis is consistent with the results of neutron diffraction experiments, which suggest the microstructure of this alloy as a matrix of disordered atomic structure (A2 phase) with embedded precipitates of the structurally and magnetically ordered D03 phase.

Evolution of fractal characteristics in shales with increasing thermal maturity: Evidence from neutron scattering, N2 physisorption, and FE-SEM imaging

Fractal dimension analysis has become an essential tool to assess the pore structure of various porous geomaterials including shale rock. In this regard, changes of fractal dimension during the thermal maturation of organic rich shale could represent the evolution of its pore structure. To this end, complementary tests were conducted on six organic rich shale samples that are in a wide range of thermal maturity (equivalent Ro values of 0.52–3.53 %). Fractal dimensions were obtained from small-angle neutron scattering (SANS), N2 physisorption, and field emission-scanning electron microscopy (FE-SEM) data. Herein, surface fractal dimensions were calculated from the N2 adsorption isotherms using the Frenkel–Halsey–Hill method, which was found between 2.46 and 2.71. Power-law exponents (E) of the SANS profiles varied from 2.75 to 3.48, indicating that both surface fractals and mass fractals would characterize the shale samples. As thermal maturity progressed, the fractal dimension shifted from surface to mass fractal while further analyses suggested this was caused by changes in the number density of pores (NDP) of various pore-size ranges. Ultimately, the pore size of the samples during such transition decreased with the increase of Ro.

Fibrillisation of faba bean protein isolate by thermosonication for process efficacy: Microstructural characteristics, assembly behaviour, and physicochemical properties

The effect of thermosonication (TS) (90 °C, 10–30 min) on the fibrillisation of faba bean protein isolate (FPI) was studied. The self-assembly behaviour, microstructural characteristics and techno-functional (gelation and emulsification) properties of FPI fibrils obtained from TS treatment were compared with those obtained from conventional prolonged heating (CH) at 90 °C up to 8 h. Compared to CH treatment, TS treatment was shown to significantly accelerate the formation of FPI fibrils with prominent β-sheet structures as revealed by Thioflavin T (ThT) fluorescence, Fourier-transform infrared spectroscopy (FTIR) and circular dichroism (CD). The characteristics of fibril building blocks were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography linked to tandem mass spectrometry (LC-MS/MS) to obtain the differences between TS and CH induced fibrillisation of FPI. Transmission electron microscopy (TEM) and small-angle neutron scattering (SANS) showed that 4 h CH and 10 min TS treatments resulted in the fibrils with similar radius (from 5 to 10 nm). Furthermore, SANS indicated that TS treatment induced the formation of an entangled FPI fibrillar network, which could lead to the observed viscoelastic properties of FPI at a high concentration (10 wt%). Finally, high internal phase O/W emulsions (HIPE, φ = 0.75) stabilised by 30 min TS induced FPI fibrils (3 wt%) demonstrated a stronger gel strength and smaller oil droplet size compared to those prepared with untreated FPI, suggesting a superior emulsification capability of FPI fibrils. This finding demonstrates that TS treatment is a promising and efficient method for fibrillisation of plant proteins with the resultant fibrils generating excellent gelation and emulsification properties.

Alcohol-Induced Denaturation of Hen Egg White Lysozyme Studied by Infrared, Circular Dichroism, and Small-Angle Neutron Scattering.

In aqueous binary solvents with fluorinated alcohols, 2,2,2-trifluoroethanol (TFE) and 1,1,1,3,3,3-hexafluoroisopropanol (HFIP), and aliphatic alcohols, ethanol (EtOH) and 2-propanol (2-PrOH), the denaturation of hen egg white lysozyme (HEWL) with increasing alcohol mole fraction xA has been investigated in a wide view from the molecular vibration to the secondary and ternary structures. Circular dichroism (CD) measurement showed that the secondary structure of α-helix content of HEWL increases on adding a small amount of the fluorinated alcohol to the aqueous solution, while the β-sheet content decreases. On the contrary, the secondary structure does not significantly change by the addition of the aliphatic alcohols. Correspondingly, the infrared (IR) spectroscopic measurements revealed that the amide I band red-shifts on the addition of the fluorinated alcohol. However, the band remains unchanged in the aliphatic alcohol systems with increasing alcohol content. To observe the ternary structure of HEWL, small-angle neutron scattering (SANS) experiments with H/D substitution technique have been applied to the HEWL solutions. The SANS experiments were successful in revealing the details of how the geometry of the HEWL changes as a function of xA. The SANS profiles indicated the spherical structure of HEWL in all of the alcohol systems in the xA range examined. The mean radius of HEWL in the two fluorinated alcohol systems increases from ∼16 to ∼18 Å during the change in the secondary structure against the increase in the fluorinated alcohol content. On contrast, the radius does not significantly change in both aliphatic alcohol systems below xA = 0.3 but expands to ∼19 Å as the alcohol content is close to the limitation of the HEWL solubility. According to the present results, together with our knowledge of the alcohol cluster formation and the interaction of the trifluoromethyl (CF3) groups with the hydrophobic moieties of biomolecules, the effects of alcohols on the denaturation of the protein have been discussed on a molecular scale.

Anion Architecture Controls Structure and Electroresponsivity of Anhalogenous Ionic Liquids in a Sustainable Fluid.

Three nonhalogenated ionic liquids (ILs) dissolved in 2-ethylhexyl laurate (2-EHL), a biodegradable oil, are investigated in terms of their bulk and electro-interfacial nanoscale structures using small-angle neutron scattering (SANS) and neutron reflectivity (NR). The ILs share the same trihexyl(tetradecyl)phosphonium ([P6,6,6,14]+) cation paired with different anions, bis(mandelato)borate ([BMB]-), bis(oxalato)borate ([BOB]-), and bis(salicylato)borate ([BScB]-). SANS shows a high aspect ratio tubular self-assembly structure characterized by an IL core of alternating cations and anions with a 2-EHL-rich shell or corona in the bulk, the geometry of which depends upon the anion structure and concentration. NR also reveals a solvent-rich interfacial corona layer. Their electro-responsive behavior, pertaining to the structuring and composition of the interfacial layers, is also influenced by the anion identity. [P6,6,6,14][BOB] exhibits distinct electroresponsiveness to applied potentials, suggesting an ion exchange behavior from cation-dominated to anion-rich. Conversely, [P6,6,6,14][BMB] and [P6,6,6,14][BScB] demonstrate minimal electroresponses across all studied potentials, related to their different dissociative and diffusive behavior. A mixed system is dominated by the least soluble IL but exhibits an increase in disorder. This work reveals the subtlety of anion architecture in tuning bulk and electro-interfacial properties, offering valuable molecular insights for deploying nonhalogenated ILs as additives in biodegradable lubricants and supercapacitors.

ABC block copolymer micelles driving the thermogelation: Scattering, imaging and spectroscopy

Thermoresponsive polymers have attracted much scientific attention due to their capacity for temperature-driven hydrogel formation. For biomedical applications, such as drug delivery, this transition should be tuned below body temperature to facilitate controlled and targeted drug release. We have recently developed a thermoresponsive polymer that forms gel at low concentrations (2 w/w%) in aqueous media and offers a cost-effective alternative to thermoresponsive systems currently being applied in clinics. This polymer is an ABC triblock terpolymer, where A, B, and C correspond to oligo(ethylene glycol) methyl ether methacrylate with average Mn 300 g mol−1 (OEGMA300), n-butyl methacrylate (BuMA), and di(ethylene glycol) methyl ether methacrylate (DEGMA). To investigate the self-assembly and the gelation mechanism in diluted solutions, we used small-angle neutron scattering (SANS) on 1 w/w% (below the gelation concentration) and 5 w/w% solutions (above the gelation concentration). As a comparison, we also investigated the solutions of the most studied thermoresponsive polymer, namely, Pluronic F127, an ABA triblock bipolymer made of ethylene glycol (A) and propylene glycol (B) blocks. SANS revealed that the in-house synthesised polymer forms elliptical cylinders, whose length increases significantly with temperature. In contrast, Pluronic F127 solutions form core-shell spherical micelles, which slightly elongate with temperature. Transmission electron microscopy images support the SANS findings, with tubular/worm structures being present. Variable-temperature circular dichroism (CD) and proton nuclear magnetic resonance (1H NMR) spectroscopy experiments reveal insights on the tacticity, structural changes, and molecular origin of the self-assembly.

Simulation of spin-echo SANS (SESANS) using McStas on monochromatic and time of flight instruments

We conduct simulations of Spin Echo Small Angle Neutron Scattering (SESANS) by employing Monte Carlo methods to a setup using four magnetic Wollaston prisms. Our primary focus involves the validation of these models, encompassing monochromatic scenarios across various neutron wavelengths to ascertain the reliability of the simulations. Subsequently, we extend this validation to encompass simulations in time-of-flight mode. Our model consistently and precisely predicts the scattering patterns emanating from dilute spheres in both monochromatic and time-of-flight modes. Notably, it also accurately reproduces the intricate encoding associated with scattering occurring between the third and fourth magnetic Wollaston prism, which provides us with another approach to increase the solid angle coverage of a SESANS instrument. This validation process conclusively demonstrates the efficacy of our simulation methods. Importantly, it paves the way for simulating more intricate and realistic instrumental configurations, broadening the horizons for future research endeavours.

The Nanoscale Structure and Stability of Organic Photovoltaic Blends Processed with Solvent Additives.

Controlling the nanomorphology in bulk heterojunction photoactive blends is crucial for optimizing the performance and stability of organic photovoltaic (OPV) technologies. A promising approach is to alter the drying dynamics and consequently, the nanostructure of the blend film using solvent additives such as 1,8-diiodooctane (DIO). Although this approach is demonstrated extensively for OPV systems incorporating fullerene-based acceptors, it is unclear how solvent additive processing influences the morphology and stability of nonfullerene acceptor (NFA) systems. Here, small angle neutron scattering (SANS) is used to probe the nanomorphology of two model OPV systems processed with DIO: a fullerene-based system (PBDB-T:PC71BM) and an NFA-based system (PBDB-T:ITIC). To overcome the low intrinsic neutron scattering length density contrast in polymer:NFA blend films, the synthesis of a deuterated NFA analog (ITIC-d52) is reported. Using SANS, new insights into the nanoscale evolution of fullerene and NFA-based systems are provided by characterizing films immediately after fabrication, after thermal annealing, and after aging for 1year. It is found that DIO processing influences fullerene and NFA-based systems differently with NFA-based systems characterized by more phase-separated domains. After long-term aging, SANS reveals both systems demonstrate some level of thermodynamic induced domain coarsening.

Simulation of neutron dark-field data for grating-based interferometers

Hierarchical structures and heterogeneous materials are found in many natural and engineered systems including additive manufacturing, alternative energy, biology and polymer science. Though the structure–function relationship is important for developing more advanced materials, structural characterization over broad length scales often requires multiple complementary measurements. Neutron far-field interferometry aims to enable multi-scale characterization by combining the best of neutron imaging with small-angle neutron scattering (SANS) via dark-field imaging. The microstructure, nominally from 1 nm to 10 µm, is averaged over each volume element ∼(50 µm)3 in the sample, resulting in a `tomographic SANS' measurement. Unlike in small-angle scattering, there are few analytical models to fit dark-field imaging data to extract properties of the microstructure. Fortunately, the dark field and SANS are related through a single Hankel transform. In this work, we discuss the development of a Python-based library, correlogram-tools, that makes use of existing small-angle scattering models and a numerical implementation of the Hankel transform to simulate dark-field interferometry data. We demonstrate how this software can be used to inform researchers of viable sample sets for interferometry experiments, analyze interferometry data, and simulate raw and reconstructed interferometry images for the training of more advanced segmentation models and analysis protocols.

3,4-Ethylenedioxythiophene Hydrogels: Relating Structure and Charge Transport in Supramolecular Gels.

Ionic charge transport is a ubiquitous language of communication in biological systems. As such, bioengineering is in constant need of innovative, soft, and biocompatible materials that facilitate ionic conduction. Low molecular weight gelators (LMWGs) are complex self-assembled materials that have received increasing attention in recent years. Beyond their biocompatible, self-healing, and stimuli responsive facets, LMWGs can be viewed as a "solid" electrolyte solution. In this work, we investigate 3,4-ethylenedioxythiophene (EDOT) as a capping group for a small peptide library, which we use as a system to understand the relationship between modes of assembly and charge transport in supramolecular gels. Through a combination of techniques including small-angle neutron scattering (SANS), NMR-based Van't Hoff analysis, atomic force microscopy (AFM), rheology, four-point probe, and electrochemical impedance spectroscopy (EIS), we found that modifications to the peptide sequence result in distinct assembly pathways, thermodynamic parameters, mechanical properties, and ionic conductivities. Four-point probe conductivity measurements and electrochemical impedance spectroscopy suggest that ionic conductivity is approximately doubled by programmable gel assemblies with hollow cylinder morphologies relative to gels containing solid fibers or a control electrolyte. More broadly, it is hoped this work will serve as a platform for those working on charge transport of aqueous soft materials in general.

Simple Scattering: Lipid nanoparticle structural data repository.

Lipid nanoparticles (LNPs) are being intensively researched and developed to leverage their ability to safely and effectively deliver therapeutics. To achieve optimal therapeutic delivery, a comprehensive understanding of the relationship between formulation, structure, and efficacy is critical. However, the vast chemical space involved in the production of LNPs and the resulting structural complexity make the structure to function relationship challenging to assess and predict. New components and formulation procedures, which provide new opportunities for the use of LNPs, would be best identified and optimized using high-throughput characterization methods. Recently, a high-throughput workflow, consisting of automated mixing, small-angle X-ray scattering (SAXS), and cellular assays, demonstrated a link between formulation, internal structure, and efficacy for a library of LNPs. As SAXS data can be rapidly collected, the stage is set for the collection of thousands of SAXS profiles from a myriad of LNP formulations. In addition, correlated LNP small-angle neutron scattering (SANS) datasets, where components are systematically deuterated for additional contrast inside, provide complementary structural information. The centralization of SAXS and SANS datasets from LNPs, with appropriate, standardized metadata describing formulation parameters, into a data repository will provide valuable guidance for the formulation of LNPs with desired properties. To this end, we introduce Simple Scattering, an easy-to-use, open data repository for storing and sharing groups of correlated scattering profiles obtained from LNP screening experiments. Here, we discuss the current state of the repository, including limitations and upcoming changes, and our vision towards future usage in developing our collective knowledge base of LNPs.

Contrast dependence of scattering profiles for poly(ethylene glycol) in water: Investigation by small-angle neutron scattering with 3He spin filter and small-angle x-ray scattering.

The small-angle neutron scattering (SANS) and small-angle x-ray scattering (SAXS) measurements were performed for deuterated and non-deuterated poly(ethylene glycol) (d-PEG and h-PEG, respectively) in D2O and a D2O/H2O mixed solvent (Mix) to compare the scattering profiles. To determine the coherent scattering intensity of SANS, a 3He spin filter was utilized. The scattering profiles determined by the SANS measurements were analyzed in terms of the wormlike chain model with touched beads along the contour of the chain. However, the SAXS profiles were not explained by the same model with uniform beads but with beads each consisting of a core and a shell having different electron densities. To explore the chain thickness determined from the SANS profile, the scattering intensities for different combinations of d-PEG/D2O, d-PEG/Mix, h-PEG/D2O, and h-PEG/Mix were also examined.

Wetting of alginate aerogels, from mesoporous solids to hydrogels: a small-angle scattering analysis

Mesoporous polysaccharide aerogels are versatile functional materials for drug delivery and wound dressing devices. The hydration and wetting of these aerogels control their application-related performance, e.g. the release of encapsulated drugs. Reported here is a detailed small-angle neutron scattering (SANS) analysis of the hydration mechanism of a calcium alginate aerogel, based on mathematical modelling of the scattering. The model accounts for the hierarchical structure of the material comprising a mesoporous structure, the solid skeleton of which is made up of water-swollen polymers. At large scale, the mesoporous structure is modelled as a random collection of elongated cylinders, which grow in size as they absorb water and aggregate. The small-scale inner structure of the skeleton is described as a Boolean model of polymer coils, which captures the progressive transition from a dense dry polymer to a fully hydrated gel. Using known physico-chemical characteristics of the alginate, the SANS data are fitted using the size of the cylinders as the only adjustable parameter. The alginate aerogel maintains a nanometre-scale, albeit altered, structure for low water contents but it collapses into micrometre-sized structures when the water content approaches one gram of water per gram of alginate. In addition to the wetting of aerogels, the model might be useful for the small-angle scattering analysis of the supercritical drying of gels.

Insights into primary carbides and nanoparticles in an additively manufactured high-alloy steel

This paper reports the use of combined electron microscopy and small-angle neutron scattering (SANS) to probe into the compositional and size evolution of the primary carbides and nanoparticles (defined by a size threshold of 100 nm) in an additively manufactured high-alloy steel before and after heat treatment. The primary carbides exhibit marginal changes in their total volume fraction and size after the austenitising and tempering. However, those carbides located at the prior-austenite boundaries provide a pinning effect to impede grain growth during the austenitising. The grain size increases from 1.7 µm in the as-manufactured to 2.8 µm in the as-tempered conditions, resulting in a limited strength loss of 33–126 MPa as estimated by using the Hall-Petch relationship. Atom probe tomography, which offers atomic spatial resolution examining a sample volume of 6.6 × 105 nm3, reveals the presence of numerous V and Cr-enriched nanoparticles with sizes of 1 to 10 nm within the steel matrix after the tempering. In contrast, the complementary SANS which examines a significantly larger sample volume of 0.9 mm3 but lacking spatial resolution, provides nanoparticle size information. It reveals a radius reduction from 7.6 to 1.0 nm and a volume fraction increase from 1.6 % to 2.3 %, in the as-manufactured and as-tempered conditions, respectively. The nanoparticles induced during tempering can contribute to a strength enhancement of 691 MPa, primarily through the Orowan bypass mechanism. This suggests that the combination of limited prior-austenite grain growth and the presence of nanoparticles is the key factor responsible for the unprecedented material strength.

Iron-carbohydrate complexes treating iron anaemia: Understanding the nano-structure and interactions with proteins through orthogonal characterisation

Intravenous (IV) iron-carbohydrate complexes are widely used nanoparticles (NPs) to treat iron deficiency anaemia, often associated with medical conditions such as chronic kidney disease, heart failure and various inflammatory conditions. Even though a plethora of physicochemical characterisation data and clinical studies are available for these products, evidence-based correlation between physicochemical properties of iron-carbohydrate complexes and clinical outcome has not fully been elucidated yet. Studies on other metal oxide NPs suggest that early interactions between NPs and blood upon IV injection are key to understanding how differences in physicochemical characteristics of iron-carbohydrate complexes cause variance in clinical outcomes. We therefore investigated the core-ligand structure of two clinically relevant iron-carbohydrate complexes, iron sucrose (IS) and ferric carboxymaltose (FCM), and their interactions with two structurally different human plasma proteins, human serum albumin (HSA) and fibrinogen, using a combination of cryo-scanning transmission electron microscopy (cryo-STEM), x-ray diffraction (XRD), small-angle x-ray scattering (SAXS) and small-angle neutron scattering (SANS). Using this orthogonal approach, we defined the nano-structure, individual building blocks and surface morphology for IS and FCM. Importantly, we revealed significant differences in the surface morphology of the iron-carbohydrate complexes. FCM shows a localised carbohydrate shell around its core, in contrast to IS, which is characterised by a diffuse and dynamic layer of carbohydrate ligand surrounding its core. We hypothesised that such differences in carbohydrate morphology determine the interaction between iron-carbohydrate complexes and proteins and therefore investigated the NPs in the presence of HSA and fibrinogen. Intriguingly, IS showed significant interaction with HSA and fibrinogen, forming NP-protein clusters, while FCM only showed significant interaction with fibrinogen. We postulate that these differences could influence bio-response of the two formulations and their clinical outcome. In conclusion, our study provides orthogonal characterisation of two clinically relevant iron-carbohydrate complexes and first hints at their interaction behaviour with proteins in the human bloodstream, setting a prerequisite towards complete understanding of the correlation between physicochemical properties and clinical outcome.