Long-Term Stability, Biocompatibility, and Magnetization of Suspensions of Isolated Bacterial Magnetosomes.

Magnetosomes are magnetic nanoparticles biosynthesized by magnetotactic bacteria. Due to a genetically strictly controlled biomineralization process, the ensuing magnetosomes have been envisioned as agents for biomedical and clinical applications. In the present work, different stability parameters of magnetosomes isolated from Magnetospirillum gryphiswaldense upon storage in suspension (HEPES buffer, 4 °C, nitrogen atmosphere) for one year in the absence of antibiotics are examined. The magnetic potency, measured by the saturation magnetization of the particle suspension, drops to one-third of its starting value within this year-about ten times slower than at ambient air and room temperature. The particle size distribution, the integrity of the surrounding magnetosome membrane, the colloidal stability, and the biocompatibility turn out to be not severely affected by long-term storage.

Comparing the Colloidal Stabilities of Commercial and Biogenic Iron Oxide Nanoparticles That Have Potential In Vitro/In Vivo Applications.

For the potential in vitro/in vivo applications of magnetic iron oxide nanoparticles, their stability in different physiological fluids has to be ensured. This important prerequisite includes the preservation of the particles' stability during the envisaged application and, consequently, their invariance with respect to the transfer from storage conditions to cell culture media or even bodily fluids. Here, we investigate the colloidal stabilities of commercial nanoparticles with different coatings as a model system for biogenic iron oxide nanoparticles (magnetosomes) isolated from magnetotactic bacteria. We demonstrate that the stability can be evaluated and quantified by determining the intensity-weighted average of the particle sizes (Z-value) obtained from dynamic light scattering experiments as a simple quality criterion, which can also be used as an indicator for protein corona formation.

Delamination by Repulsive Osmotic Swelling of Synthetic Na-Hectorite with Variable Charge in Binary Dimethyl Sulfoxide-Water Mixtures.

Swelling of clays is hampered by increasing layer charge. With vermiculite-type layer charge densities, crystalline swelling is limited to the two-layer hydrate, while osmotic swelling requires ion exchange with bulky and hydrophilic organic molecules or with Li+ cations to trigger repulsive osmotic swelling. Here, we report on surprising and counterintuitive osmotic swelling behavior of a vermiculite-type synthetic clay [Na0.7]inter[Mg2.3Li0.7]oct[Si4]tetO10F2 in mixtures of water and dimethyl sulfoxide (DMSO). Although swelling in pure water is restricted to crystalline swelling, with the addition of DMSO, osmotic swelling sets in at some threshold composition. Finally, when the DMSO concentration is increased further to 75 vol %, swelling is restricted again to crystalline swelling as expected. Repulsive osmotic swelling thus is observed in a narrow composition range of the binary water-DMSO mixture, where a freezing point suppression is observed. This suppression is related to DMSO and water molecules exhibiting strong interactions leading to stable molecular clusters. Based on this phenomenological observation, we hypothesize that the unexpected swelling behavior might be related to the formation of different complexes of interlayer cations being formed at different compositions. Powder X-ray diffraction and 23Na magic angle spinning-NMR evidence is presented that supports this hypothesis. We propose that the synergistic solvation of the interlayer sodium at favorable compositions exerts a steric pressure by the complexes formed in the interlayer. Concomitantly, the basal spacing is increased to a level, where entropic contributions of interlayer species lead to a spontaneous thermodynamically allowed one-dimensional dissolution of the clay stack.

Tuning Sugar-Based Chiral and Flower-Like Microparticles.

Unique supermolecular structures as chiral and flower-like microparticles and the precise tuning of the morphologies hold immense promise for a variety of applications. Examples of such structures deriving from monosaccharides are still rare, and a general understanding is also lacking. Herein, it is shown that chiral, flower-like, or solid microparticles can be tuned by only using monosaccharide esters without external stimuli. Chiral "left-handed" (counterclockwise) and "right-handed" (clockwise) morphologies can be induced by d- and l-glucose stearoyl esters. In comparison, other monosaccharides, i.e., galactose, mannose, and xylose, cannot formed chiral particles and generated diverse other morphologies of the supermolecular microparticles based on their distinct molecular configurations. Due to the numbers of side chains and the bond orientations, microparticles with solid and porous flower-like morphologies can be obtained. While glucose and xylose esters only lead to solid microparticles, mannose and galactose generate porous flower-like particles. These findings suggest a general method to design and control the superstructures by using monosaccharide backbones with diverse molecular configurations.

Osmotic Delamination: A Forceless Alternative for the Production of Nanosheets Now in Highly Polar and Aprotic Solvents.

Repulsive osmotic delamination is thermodynamically allowed "dissolution" of two-dimensional (2D) materials and therefore represents an attractive alternative to liquid-phase exfoliation to obtain strictly monolayered nanosheets with an appreciable aspect ratio with quantitative yield. However, osmotic delamination was so far restricted to aqueous media, severely limiting the range of accessible 2D materials. Alkali-metal intercalation compounds of MoS2 or graphite are excluded because they cannot tolerate even traces of water. We now succeeded in extending osmotic delamination to polar and aprotic organic solvents. Upon complexation of interlayer cations of synthetic hectorite clay by crown ethers, either 15-crown-5 or 18-crown-6, steric pressure is exerted, which helps in reaching the threshold separation required to trigger osmotic delamination based on translational entropy. This way, complete delamination in water-free solvents like aprotic ethylene and propylene carbonate, N-methylformamide, N-methylacetamide, and glycerol carbonate was achieved.

Self-Assembly of Soft Cellulose Nanospheres into Colloidal Gel Layers with Enhanced Protein Adsorption Capability for Next-Generation Immunoassays.

Soft cationic core/shell cellulose nanospheres can deform and interpenetrate allowing their self-assembly into densely packed colloidal nanogel layers. Taking advantage of their water-swelling capacity and molecular accessibility, the nanogels are proposed as a new and promising type of coating material to immobilize bioactive molecules on thin films and paper. The specific and nonspecific interactions between the cellulosic nanogel and human immunoglobulin G as well as bovine serum albumin (BSA) are investigated. Confocal microscopy, electroacoustic microgravimetry, and surface plasmon resonance are used to access information about the adsorption behavior and viscoelastic properties of self-assembled nanogels. A significant BSA adsorption capacity on nanogel layers (17 mg m-2 ) is measured, 300% higher compared to typical polymer coatings. This high protein affinity further confirms the promise of the introduced colloidal gel layer, in increasing sensitivity and advancing a new generation of substrates for a variety of applications, including immunoassays, as demonstrated in this work.

Osmotic Swelling of Sodium Hectorite in Ternary Solvent Mixtures: Nematic Liquid Crystals in Hydrophobic Media.

The swelling of clay minerals in organic solvents or solvent mixtures is key for the fabrication of polymer nanocomposites with perfectly dispersed filler that contain only individual clay layers. Here, we investigated the swelling behavior of sodium hectorite in different ternary solvent mixtures containing methanol, acetonitrile, ethylene glycol, or glycerol carbonate with minimal amounts of water. We found that in these mixtures, less water is required than in the corresponding binary mixtures to allow for complete delamination by repulsive osmotic swelling. A quantitative study of osmotic swelling in a particular ternary mixture shows that organic solvents resemble swelling behavior in pure water. At hectorite contents larger than 5 vol %, the separation of individual layers scales with ϕ-1. At this concentration, a crossover is observed and swelling continues at a slower pace (ϕ-0.5) below this value.

An automated oxystat fermentation regime for microoxic cultivation of Magnetospirillum gryphiswaldense.

BACKGROUND: Magnetosomes produced by magnetotactic bacteria represent magnetic nanoparticles with unprecedented characteristics. However, their use in many biotechnological applications has so far been hampered by their challenging bioproduction at larger scales. RESULTS: Here, we developed an oxystat batch fermentation regime for microoxic cultivation of Magnetospirillum gryphiswaldense in a 3 L bioreactor. An automated cascade regulation enabled highly reproducible growth over a wide range of precisely controlled oxygen concentrations (1-95% of air saturation). In addition, consumption of lactate as the carbon source and nitrate as alternative electron acceptor were monitored during cultivation. While nitrate became growth limiting during anaerobic growth, lactate was the growth limiting factor during microoxic cultivation. Analysis of microoxic magnetosome biomineralization by cellular iron content, magnetic response, transmission electron microscopy and small-angle X-ray scattering revealed magnetosomal magnetite crystals were highly uniform in size and shape. CONCLUSION: The fermentation regime established in this study facilitates stable oxygen control during culturing of Magnetospirillum gryphiswaldense. Further scale-up seems feasible by combining the stable oxygen control with feeding strategies employed in previous studies. Results of this study will facilitate the highly reproducible laboratory-scale bioproduction of magnetosomes for a diverse range of future applications in the fields of biotechnology and biomedicine.

Direct Measurement of the In-Plane Thermal Diffusivity of Semitransparent Thin Films by Lock-In Thermography: An Extension of the Slopes Method.

We present an extension of the well-known slopes method for characterization of the in-plane thermal diffusivity of semitransparent polymer films. We introduce a theoretical model which considers heat losses due to convection and radiation mechanisms, as well as semitransparency of the material to the exciting laser heat source (visible range) and multiple reflections at the film surfaces. Most importantly, a potential semitransparency of the material in the IR detection range is also considered. We prove by numerical simulations and by an asymptotic expansion of the surface temperature that the slopes method is also valid for any semitransparent film in the thermally thin regime. Measurements of the in-plane thermal diffusivity performed on semitransparent polymer films covering a wide range of absorption coefficients (to the exciting wavelength and in the IR detection range of our IR camera) validate our theoretical findings.

Probing the Nanostructure and Arrangement of Bacterial Magnetosomes by Small-Angle X-Ray Scattering.

Magnetosomes are membrane-enveloped single-domain ferromagnetic nanoparticles enabling the navigation of magnetotactic bacteria along magnetic field lines. Strict control over each step of biomineralization generates particles of high crystallinity, strong magnetization, and remarkable uniformity in size and shape, which is particularly interesting for many biomedical and biotechnological applications. However, to understand the physicochemical processes involved in magnetite biomineralization, close and precise monitoring of particle production is required. Commonly used techniques, such as transmission electron microscopy (TEM) or Fe measurements, allow only for semiquantitative assessment of the magnetosome formation without routinely revealing quantitative structural information. In this study, lab-based small-angle X-ray scattering (SAXS) is explored as a means to monitor the different stages of magnetosome biogenesis in the model organism Magnetospirillum gryphiswaldense SAXS is evaluated as a quantitative stand-alone technique to analyze the size, shape, and arrangement of magnetosomes in cells cultivated under different growth conditions. By applying a simple and robust fitting procedure based on spheres aligned in linear chains, it is demonstrated that the SAXS data sets contain information on both the diameter of the inorganic crystal and the protein-rich magnetosome membrane. The analyses corroborate a narrow particle size distribution with an overall magnetosome radius of 19 nm in Magnetospirillum gryphiswaldense Furthermore, the averaged distance between individual magnetosomes is determined, revealing a chain-like particle arrangement with a center-to-center distance of 53 nm. Overall, these data demonstrate that SAXS can be used as a novel stand-alone technique allowing for the at-line monitoring of magnetosome biosynthesis, thereby providing accurate information on the particle nanostructure.IMPORTANCE This study explores lab-based small-angle X-ray scattering (SAXS) as a novel quantitative stand-alone technique to monitor the size, shape, and arrangement of magnetosomes during different stages of particle biogenesis in the model organism Magnetospirillum gryphiswaldense The SAXS data sets contain volume-averaged, statistically accurate information on both the diameter of the inorganic nanocrystal and the enveloping protein-rich magnetosome membrane. As a robust and nondestructive in situ technique, SAXS can provide new insights into the physicochemical steps involved in the biosynthesis of magnetosome nanoparticles as well as their assembly into well-ordered chains. The proposed fit model can easily be adapted to account for different particle shapes and arrangements produced by other strains of magnetotactic bacteria, thus rendering SAXS a highly versatile method.

Onset of Osmotic Swelling in Highly Charged Clay Minerals.

Delamination by osmotic swelling of layered materials is generally thought to become increasingly difficult, if not impossible, with increasing layer charge density because of strong Coulomb interactions. Nevertheless, for the class of 2:1 layered silicates, very few examples of delaminating organo-vermiculites were reported in literature. We propose a mechanism for this repulsive osmotic swelling of highly charged vermiculites based on repulsive counterion translational entropy that dominates the interaction of adjacent layers above a certain threshold separation. Based on this mechanistic insight, we were able to identify several organic interlayer cations appropriate to delaminate highly charged, vermiculite-type clay minerals. These findings suggest that the osmotic swelling of highly charged organoclays is a generally applicable phenomenon rather than the odd exemption.

Parallel and Perpendicular Alignment of Anisotropic Particles in Free Liquid Microjets and Emerging Microdroplets.

Liquid microjets play a key role in fiber spinning, inkjet printing, and coating processes. In all of these applications, the liquid jets carry dispersed particles whose spatial and orientational distributions within the jet critically influence the properties of the fabricated structures. Despite its importance, there is currently no knowledge about the orientational distribution of particles within microjets and droplets. Here, we demonstrate a microfluidic device that allows to determine the local particle distribution and orientation by X-ray scattering. Using this methodology, we discovered unexpected changes in the particle orientation upon exiting the nozzle to form a free jet, and upon jet break-up into droplets, causing an unusual biaxial particle orientation. We show how flow and aspect ratio determine the flow orientation of anisotropic particles. Furthermore, we demonstrate that the observed phenomena are a general characteristic of anisotropic particles. Our findings greatly enhance our understanding of particle orientation in free jets and droplets and provide a rationale for controlling particle alignment in liquid jet-based fabrication methodologies.

Self-assembly of smallest magnetic particles.

The assembly of tiny magnetic particles in external magnetic fields is important for many applications ranging from data storage to medical technologies. The development of ever smaller magnetic structures is restricted by a size limit, where the particles are just barely magnetic. For such particles we report the discovery of a kind of solution assembly hitherto unobserved, to our knowledge. The fact that the assembly occurs in solution is very relevant for applications, where magnetic nanoparticles are either solution-processed or are used in liquid biological environments. Induced by an external magnetic field, nanocubes spontaneously assemble into 1D chains, 2D monolayer sheets, and large 3D cuboids with almost perfect internal ordering. The self-assembly of the nanocubes can be elucidated considering the dipole-dipole interaction of small superparamagnetic particles. Complex 3D geometrical arrangements of the nanodipoles are obtained under the assumption that the orientation of magnetization is freely adjustable within the superlattice and tends to minimize the binding energy. On that basis the magnetic moment of the cuboids can be explained.

Two-Step Delamination of Highly Charged, Vermiculite-like Layered Silicates via Ordered Heterostructures.

Because of strong Coulomb interactions, the delamination of charged layered materials becomes progressively more difficult with increasing charge density. For instance, highly charged sodium fluorohectorite (Na0.6Mg2.4Li0.6Si4O10F2, Na-Hec) cannot be delaminated directly by osmotic swelling in water because its layer charge exceeds the established limit for osmotic swelling of 0.55 per formula unit Si4O10F2. Quite surprisingly, we found that this hectorite at the border of the smectite and vermiculite group can, however, be utterly delaminated into 1-nm-thick platelets with a high aspect ratio (24 000) in a two-step process. The hectorite is first converted by partial ion exchange into a one-dimensionally ordered, interstratified heterostructure with strictly alternating Na+ and n-butylammonium (C4) interlayers. This heterostructure then spontaneously delaminates into uniform single layers upon immersion in water whereas neither of the homoionic phases (Na-Hec and C4-Hec) swells osmotically. The delamination of more highly charged synthetic layered silicates is a key step to push the aspect ratio beyond the current limits.

Thermodynamic Study of the Interaction of Bovine Serum Albumin and Amino Acids with Cellulose Nanocrystals.

The interaction of bovine serum albumin (BSA) with sulfated, carboxylated, and pyridinium-grafted cellulose nanocrystals (CNCs) was studied as a function of the degree of substitution by determining the adsorption isotherm and by directly measuring the thermodynamics of interaction. The adsorption of BSA onto positively charged pyridinium-grafted cellulose nanocrystals followed Langmuirian adsorption with the maximum amount of adsorbed protein increasing linearly with increasing degree of substitution. The binding mechanism between the positively charged pyridinum-grafted cellulose nanocrystals and BSA was found to be endothermic and based on charge neutralization. A positive entropy of adsorption associated with an increase of the degree of disorder upon addition of BSA compensated for the unfavorable endothermic enthalpy and enabled formation of pyridinium-g-CNC-BSA complexes. The endothermic enthalpy of adsorption was further found to decrease as a function of increasing degree of substitution. Negatively charged cellulose nanocrystals bearing sulfate and/or carboxylic functionalities were found to not interact significantly with the BSA protein. To investigate in more detail the role of single amino acids in the adsorption of proteins onto cellulose nanocrystals, we also studied the interaction of different types of amino acids with CNCs, i.e., charged (lysine, aspartic acid), aromatic (tryptophan, tyrosine), and polar (serine) amino acids. We found that none of the single amino acids bound with CNCs irrespective of surface charge and that therefore the binding of proteins with CNCs appears to require larger amino acid sequences that induce a greater entropic contribution to stabilize binding. Single amino acids are thus not adsorbed onto cellulose nanocrystals.

In-Depth Insights into the Key Steps of Delamination of Charged 2D Nanomaterials.

Delamination is a key step to obtain individual layers from inorganic layered materials needed for fundamental studies and applications. For layered van der Waals materials such as graphene, the adhesion forces are small, allowing for mechanical exfoliation, whereas for ionic layered materials such as layered silicates, the energy to separate adjacent layers is considerably higher. Quite counterintuitively, we show for a synthetic layered silicate (Na0.5-hectorite) that a scalable and quantitative delamination by simple hydration is possible for high and homogeneous charge density, even for aspect ratios as large as 20000. A general requirement is the separation of adjacent layers by solvation to a distance where layer interactions become repulsive (Gouy-Chapman length). Further hydration up to 34 nm leads to the formation of a highly ordered lamellar liquid crystalline phase (Wigner crystal). Up to eight higher-order reflections indicate excellent positional order of individual layers. The Wigner crystal melts when the interlayer separation reaches the Debye length, where electrostatic interactions between adjacent layers are screened. The layers become weakly charge-correlated. This is indicated by fulfilling the classical Hansen-Verlet and Lindeman criteria for melting. We provide insight into the requirements for layer separation and controlling the layer distances for a broad range of materials and outline an important pathway for the integration of layers into devices for advanced applications.

Controlled Exfoliation of Layered Silicate Heterostructures into Bilayers and Their Conversion into Giant Janus Platelets.

Ordered heterostructures of layered materials where interlayers with different reactivities strictly alternate in stacks offer predetermined slippage planes that provide a precise route for the preparation of bilayer materials. We use this route for the synthesis of a novel type of reinforced layered silicate bilayer that is 15 % stiffer than the corresponding monolayer. Furthermore, we will demonstrate that triggering cleavage of bilayers by osmotic swelling gives access to a generic toolbox for an asymmetrical modification of the two vis-à-vis standing basal planes of monolayers. Only two simple steps applying arbitrary commercial polycations are needed to obtain such Janus-type monolayers. The generic synthesis route will be applicable to many other layered compounds capable of osmotic swelling, rendering this approach interesting for a variety of materials and applications.

In situ formation of a MoS2 -based inorganic-organic nanocomposite by directed thermal decomposition.

Nanocomposites based on molybdenum disulfide (MoS2 ) and different carbon modifications are intensively investigated in several areas of applications due to their intriguing optical and electrical properties. Addition of a third element may enhance the functionality and application areas of such nanocomposites. Herein, we present a facile synthetic approach based on directed thermal decomposition of (Ph4 P)2 MoS4 generating MoS2 nanocomposites containing carbon and phosphorous. Decomposition at 250 °C yields a composite material with significantly enlarged MoS2 interlayer distances caused by in situ formation of Ph3 PS bonded to the MoS2 slabs through MoS bonds and (Ph4 P)2 S molecules in the van der Waals gap, as was evidenced by (31) P solid-state NMR spectroscopy. Visible-light-driven hydrogen generation demonstrates a high catalytic performance of the materials.

Simultaneous SAXS/WAXS/UV-Vis Study of the Nucleation and Growth of Nanoparticles: A Test of Classical Nucleation Theory.

Despite the increasing interest in the applications of functional nanoparticles, a comprehensive understanding of the formation mechanism starting from the precursor reaction with subsequent nucleation and growth is still a challenge. We for the first time investigated the kinetics of gold nanoparticle formation systematically by means of a lab-based in situ small-angle X-ray scattering (SAXS)/wide-angle X-ray scattering (WAXS)/UV-vis absorption spectroscopy experiment using a stopped-flow apparatus. We thus could systematically investigate the influence of all major factors such as precursor concentration, temperature, the presence of stabilizing ligands and cosolvents on the temporal evolution of particle size, size distribution, and optical properties from the early prenucleation state to the late growth phase. We for first time formulated and numerically solved a closed nucleation and growth model including the precursor reaction. We observe that the results can be well described within the framework of classical nucleation and growth theory, including also results of previous studies by other research groups. From the analysis, we can quantitatively derive values for the rate constants of precursor reaction and growth together with their activation free enthalpies. We find the growth process to be surface-reaction limited with negligible influence of Ostwald ripening yielding narrow disperse gold nanoparticles.

Plasmonic gold-poly(N-isopropylacrylamide) core-shell colloids with homogeneous density profiles: a small angle scattering study.

Coating metal nanocrystals with responsive polymers provides a model case of smart, functional materials, where the optical properties can be modulated by external stimuli. However the optical response is highly sensitive to the polymer shell morphology, thickness and dielectric contrast. In this paper we study the nature of cross-linked, thermoresponsive polymer shells for the first time using four different scattering approaches to elucidate the density profile of the shells. Each scattering method provides unique information about the temperature-induced changes of shell thickness in terms of hydrodynamic radius and radius of gyration, the pair-distance distribution functions of the shells as well as the dynamic network fluctuations. Only a combination of these different scattering techniques allows to develop a morphological model of the core-shell particles. We further demonstrate control of the cross-linker distribution in core-shell synthesis by semi-batch precipitation copolymerization. Conducting the polymerization in three steps, we show for the first time that the polymer shell thickness can be successively increased without affecting the shell morphology and response behavior.