Photocleavable Polymer Cubosomes: Synthesis, Self-Assembly, and Photorelease.

Polymer cubosomes (PCs) are a recent class of self-assembled block copolymer (BCP) microparticles with an accessible periodic channel system. Most reported PCs consist of a polystyrene scaffold, which provides mechanical stability for templating but has a limited intrinsic functionality. Here, we report the synthesis of photocleavable BCPs with compositions suitable for PC formation. We analyze the self-assembly mechanism and study the model release of dyes during irradiation, where the transition of the BCPs from amphiphilic to bishydrophilic causes the rapid disassembly of the PCs. A combination of modeling and experiment shows that the evolution of PCs proceeds first via liquid-liquid phase separation into polymer-rich droplets, followed by microphase separation within this droplet confinement, and finally, membrane reorganization into high internal order. This insight may encourage exploration of alternative preparation strategies to better control the size and homogeneity of PCs.

Volume Phase Transition of Thermoresponsive Microgels Scrutinized by Dynamic Light Scattering and Turbidity: Correlations Depend on Microgel Homogeneity.

Thermoresponsive microgels experience a volume phase transition triggered by temperature changes, a phenomenon often analyzed using dynamic light scattering to observe overall size alterations via the diffusion coefficient. However, local structural changes are typically assessed using more intricate and expensive techniques like small-angle neutron or X-ray scattering. In our research, we investigate the volume phase transition of poly-N-isopropylacrylamide (PNIPAM)-based microgels by employing a combination of temperature-dependent dynamic light scattering and simpler, faster, and more efficient attenuation measurements. We utilize attenuation at a fixed wavelength as a direct measure of dispersion turbidity, linking the absolute changes in hydrodynamic radius to the absolute changes in turbidity. This approach allows us to compare "classical" PNIPAM microgels from precipitation polymerization, charged copolymer microgels from precipitation copolymerization, and core-shell microgels from seeded precipitation polymerization. Our study includes a systematic analysis and comparison of 30 different microgels. By directly comparing data from dynamic light scattering and attenuation spectroscopy, we gain insights into structural heterogeneity and deviations from the established fuzzy sphere morphology. Furthermore, we demonstrate how turbidity data can be converted to swelling curves.

A review of stimuli-responsive polymer-based gating membranes.

The formation and properties of smart (stimuli-responsive) membranes are reviewed, with a special focus on temperature and pH triggering of gating to water, ions, polymers, nanoparticles, or other molecules of interest. The review is organized in two parts, starting with all-smart membranes based on intrinsically smart materials, in particular of the poly(N-isopropylacrylamide) family and similar polymers. The key steps of membrane fabrication are discussed, namely the deposition into thin films, functionalization of pores, and the secondary crosslinking of pre-existing microgel particles into membranes. The latter may be free-standing and do not necessitate the presence of a porous support layer. The temperature-dependent swelling properties of polymers provide a means of controlling the size of pores, and thus size-sensitive gating. Throughout the review, we highlight "positive" (gates open) or "negative" (closed) gating effects with respect to increasing temperature. In the second part, the functionalization of porous organic or inorganic membranes of various origins by either microgel particles or linear polymer brushes is discussed. In this case, the key steps are the adsorption or grafting mechanisms. Finally, whenever provided by the authors, the suitability of smart gating membranes for specific applications is highlighted.

Growth of Smart Microgels in a Flow Reactor Scrutinized by In-Line SAXS.

In this work, a continuous flow setup for in situ investigation of microgel growth with small-angle X-ray scattering (SAXS) is established. Poly(N-n-propylacrylamide) (PNNPAM) and poly(N-isopropylacrylamide) (PNIPAM) microgels are synthesized in H2O at different residence times inside a continuous flow reactor. The microgels are investigated by in situ SAXS and ex situ photon correlation spectroscopy. The size of the microgels was found to be reproducible in independent experiments with run times of up to 7 h. Already the scattering curves of the microgels with a time of residence of 15 min show a well-defined form factor. Further analysis of the scattering profiles confirms the spherical shape of the microgels. At a residence time of 2 min, the scattering intensity is significantly lower corresponding to a smaller particle size. The experimental conditions remain constant over time, which is crucial for long-time experiments. The PNNPAM system is found to be more suitable for the flow reactor experiment with in-line SAXS as it shows less polymer deposition in the tubing and forms particles with lower polydispersity. The presented reactor is characterized by a compact design and offers a plug-and-play setup close to the sample environment. This work paves the way for investigations of microgel growth at e.g. synchrotron X-ray beamlines.

Adhesion, proliferation, and detachment of various cell types on thermoresponsive microgel coatings.

Cutting-edge biomedical applications require increasingly complex and fastidious cell systems, for example, various classes of primary or stem cells. Their cultivation, however, still differs little from 30 years ago. This especially applies to the use of indiscriminative proteases for nonspecific cell detachment. A far more gentle alternative changes the adhesive properties of the cell culture substrates through coatings based on thermoresponsive polymers. Such polymers mediate cell adhesion at 37°C, but become repulsive upon a cell-compatible temperature drop to, for example, 32°C. While the high functionality of this method has already been well proven, it must also be easy and reproducible to apply. Here, we emphasize the potential of standard cell culture materials coated by spraying with thermoresponsive microgels for routine cultivation and beyond. On these surfaces, we successfully cultivated and detached various cell types, including induced pluripotent stem cells and cells in serum-free culture. In addition, we evaluated the compatibility of the microgel-sprayed surfaces with adhesion-promoting proteins, which are essential for, for example, stem cells or neuronal cells. Finally, we demonstrate that the microgel surfaces do not impair proliferation and show their long-term stability. We conclude that for cell detachment, thermoresponsive cell culture substrates can fully substitute proteases, like trypsin, by employing a comparably straightforward protocol that is compatible with many industrial processing lines.

Thermoresponsive Microgel-Based Free-Standing Membranes: Influence of Different Microgel Cross-Linkers on Membrane Function.

In this study we show a possibility to produce thermoresponsive, free-standing microgel membranes based on N-isopropylacrylamide (NIPAM) and the UV-sensitive comonomer 2-hydroxy-4-(methacryloyloxy)benzophenone (HMABP). To influence the final network structure and functionality of the membranes, we use different cross-linkers in the microgel syntheses and characterize the resulting structural microgel properties and the swelling behavior by means of AFM, FTIR, and PCS measurements. Varying the cross-linker results in significant changes in the structure and swelling behavior of the individual microgels and has an influence on the incorporation of the comonomer, which is essential for subsequent photochemical membrane formation. We investigate the ion transport through the different membranes by temperature-dependent resistance measurements revealing a sharp increase in resistance when the copolymer microgels reach their collapsed state. The resistance of the membranes can be adjusted by different cross-linkers and the associated incorporation of the comonomer. Furthermore, we show that transferring a reversible cross-linker from a cross-linked state to an un-cross-linked state strongly influences the membrane properties and even reverses the switching behavior, while the mechanical stability of the membrane is maintained.

Tuning Xanthan Viscosity by Directed Random Coil-to-Helix Transition.

Xanthan gum is a polysaccharide that is widely used as a thickening agent in numerous food, cosmetic, and technical applications. Therefore, the knowledge of the molecular interplay that builds up and stabilizes water-binding networks is crucial for the optimization of xanthan thickening performance. Using atomic force microscopy, rheometry, and inductively coupled plasma optical emission spectroscopy, we show a clear correlation between xanthan thickening properties and the ability to form characteristic secondary structures as well as the valence and amount of cations coordinated at the polysaccharide side chain. Based on these findings and the Debye-Hückel theory, we derive an ion-interaction model in which divalent cations mediate bridging of adjacent single polymer strands due to chelate-like coordination building stable secondary structures. We furthermore demonstrate in a cation exchange assay that xanthan secondary structures can be modified in a directed and reversible manner, which, in turn, alters its thickening properties.

Core-shell microgels synthesized in continuous flow: deep insight into shell growth using temperature-dependent FTIR.

While core-shell microgels have been intensively studied in their fully synthesized state, the formation mechanism of the shell has not been completely understood. Such insight is decisive for a customization of microgel properties for applications. In this work, microgels based on a N-isopropylmethacrylamide (NiPMAM) core and a N-n-propylacrylamide (NnPAM) shell are synthesized in a continuous flow reactor. The shell growth is studied depending on the solution's time of residence inside the reactor. PCS experiments reveal a significant decrease of the volume phase transition temperatures of the core and the shell, with increasing residence time. At early stages, a decreased swelling capacity is found before a discrete NnPAM shell is formed. Temperature-dependent FTIR spectroscopy shows that the decreased swelling capacity originates from a pronounced interpenetrated network (IPN) between NnPAM and NiPMAM. AFM images resolve heterogeneously distributed shell material after 3 min, pointing to an aggregation of NnPAM domains before the distinct shell forms. The combination of diffusional properties, AFM images and vibrational information confirms a deeply interpenetrated network already at early stages of the precipitation polymerization, in which the shell material heavily influences the swelling properties.

Protein Nanopore Membranes Prepared by a Simple Langmuir-Schaefer Approach.

Filtration through membranes with nanopores is typically associated with high transmembrane pressures and high energy consumption. This problem can be addressed by reducing the respective membrane thickness. Here, a simple procedure is described to prepare ultrathin membranes based on protein nanopores, which exhibit excellent water permeance, two orders of magnitude superior to comparable, industrially applied membranes. Furthermore, incorporation of either closed or open protein nanopores allows tailoring the membrane's ion permeability. To form such membranes, the transmembrane protein ferric hydroxamate uptake protein component A (FhuA) or its open-pore variant are assembled at the air-water interface of a Langmuir trough, compressed to a dense film, crosslinked by glutaraldehyde, and transferred to various support materials. This approach allows to prepare monolayer or multilayer membranes with a very high density of protein nanopores. Freestanding membranes covering holes up to 5 µm in diameter are visualized by atomic force microscopy (AFM), helium ion microscopy, and transmission electron microscopy. AFM PeakForce quantitative nanomechanical property mapping (PeakForce QNM)  demonstrates remarkable mechanical stability and elastic properties of freestanding monolayer membranes with a thickness of only 5 nm. The new protein membrane can pave the way to energy-efficient nanofiltration.

Smart membranes by electron beam cross-linking of copolymer microgels.

Poly(N-isopropylacrylamide) (pNIPAM) based copolymer microgels were used to create free-standing, transferable, thermoresponsive membranes. The microgels were synthesized by copolymerization of NIPAM with N-benzylhydrylacrylamide (NBHAM). Monolayers of these colloidal gels were subsequently cross-linked using an electron gun leading to the formation of a connected monolayer. Furthermore, the cross-linked microgel layer is detached from the supporting material by dissolving the substrate. These unique systems can be used as transferable, thermoresponsive coatings and as thermoresponsive membranes. As a proof of principle for the use of such membranes we studied the ion transport through them at different temperatures revealing drastic changes when the lower critical solution temperature of the copolymer microgels is reached.

Aescin - a natural soap for the formation of lipid nanodiscs with tunable size.

The saponin ß-aescin from the seed extract of the horse chestnut tree Aesculus hippocastanum has demonstrated a beneficial role in clinical therapy which is in part related to its strong interaction with biological membranes. In this context the present work investigates the self-assembly of nm-sized discoidal lipid nanoparticles composed of ß-aescin and the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). The discoidal lipid nanoparticles reassemble from small discs into larger discs, ribbons and finally stacks of sheets upon heating from gel-phase to fluid phase DMPC. The morphological transition of the lipid nano-particles is mainly triggered by the phospholipid phase state change. The final morphology depends on the phospholipid-to-saponin ratio and the actual temperature. The study is conducted by small-angle X-ray scattering (SAXS) and transmission (TEM) and freeze fracture electron microscopy (FFEM) are used to cover larger length scales. Two different models, representing a disc and ribbon-like shape are applied to the SAXS data, evaluating possible geometries and molecular mixing of the nano-particles. The stacked sheets are analysed by the Caillé theory.

Temperature Controlled Loading and Release of the Anti-Inflammatory Drug Cannabidiol by Smart Microgels.

CBD is a promising candidate for treatment of many diseases and plays a major role in the growing trend to produce high-end drugs from natural, renewable resources. In the present work, we demonstrate a way to incorporate the anti-inflammatory drug CBD into smart microgel particles. The copolymer microgels that we chose as carrier systems exhibit a volume phase transition temperature of 39 ∘C, which is just above normal body temperature and makes them ideal candidates for hyperthermia treatment. While a simple loading route of CBD was not successful due to the enormous hydrophobicity of CBD, an alternative route was developed by immersing the microgels in ethanol. Despite the expected loss of thermoresponsive behaviour of the microgel matrix due to the solvent exchange, a temperature-dependent release of CBD was detected by the material, creating an interesting question of interactions between CBD and the microgel particles in ethanol. Furthermore, the method developed for loading of the microgel particles with CBD in ethanol was further improved by a subsequent transfer of the loaded particles into water, which proves to be an even more promising approach due to the successful temperature-dependent release of the drug above the collapse temperature of the microgels.

Stable DOPG/Glycyrrhizin Vesicles with a Wide Range of Mixing Ratios: Structure and Stability as Seen by Scattering Experiments and Cryo-TEM.

Phosphatidylglycerols represent a large share of the lipids in the plasmamembrane of procaryotes. Therefore, this study investigates the role of charged lipids in the plasma membrane with respect to the interaction of the antiviral saponin glycyrrhizin with such membranes. Glycyrrhizin is a natural triterpenic-based surfactant found in licorice. Vesicles made of 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1'-glycerol) (DOPG)/glycyrrhizin are characterized by small-angle scattering with neutrons and X-rays (SANS and SAXS). Small-angle scattering data are first evaluated by the model-independent modified Kratky-Porod method and afterwards fitted by a model describing the shape of small unilamellar vesicles (SUV) with an internal head-tail contrast. Complete miscibility of DOPG and glycyrrhizin was revealed even at a ratio of lipid:saponin of 1:1. Additional information about the chain-chain correlation distance of the lipid/saponin mixtures in the SUV structures is obtained from wide-angle X-ray scattering (WAXS).

Heating-Induced DMPC/Glycyrrhizin Bicelle-to-Vesicle Transition: A X-Ray Contrast Variation Study.

In this study, we investigated the conversion of lipid bicelles into vesicles in the case of a system composed of the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and the saponin glycyrrhizin in the presence of sucrose. Glycyrrhizin is a biosurfactant present in the licorice root and possesses a triterpenic hydrophobic backbone and a hydrophilic headgroup built from two sugar molecules. The aim of this study is to determine the initial bicelle size at temperatures below the lipid's main phase transition temperature Tm and, based on these results, characteristics of the temperature-induced bicelle-to-vesicle transition. Moreover, the influence of the heating rate on this transition is followed. The general picture concluded from photon correlation spectroscopy and small angle X-ray scattering was confirmed by additional imaging with cryogenic transmission electron microscopy. Small angle X-ray scattering was especially used to determine size parameters of the existing structures. To enhance the contrast for X-rays, a buffer containing 25 wt% sucrose was used. It was found that larger vesicles were formed from smaller precursor particles and that monodisperse precursors are required for formation of very monodisperse vesicles upon temperature increase. At high glycyrrhizin contents and above a critical heating rate of ∼5°C min-1, the polydispersity of these vesicles is decoupled from both parameters, glycyrrhizin content and heating rate. However, the vesicle size stays tunable by the glycyrrhizin content and increases upon increasing the glycyrrhizin concentration. Therefore, vesicles of defined size and with a rather low polydispersity of ∼12-14% can be formed.

Suzuki-Miyaura Cross-Coupling of Bromotryptophan Derivatives at Ambient Temperature.

Mild reaction conditions are highly desirable for bio-orthogonal side chain derivatizations of amino acids, peptides or proteins due to the sensitivity of these substrates. Transition metal catalysed cross-couplings such as Suzuki-Miyaura reactions are highly versatile, but usually require unfavourable reaction conditions, in particular, when applied with aryl bromides. Ligand-free solvent-stabilised Pd-nanoparticles represent an efficient and sustainable alternative to conventional phosphine-based catalysts, because the cross-coupling can be performed at considerably lower temperature. We report on the application of such a highly reactive heterogeneous catalyst for the Suzuki-Miyaura cross-coupling of brominated tryptophan derivatives. The solvent-stabilised Pd-nanoparticles are even more efficient than the literature-known ADHP-Pd precatalyst. Interestingly, the latter also leads to the formation of quasi-homogeneous Pd-nanoparticles as the catalytic species. One advantage of our approach is the compatibility with aqueous and aerobic conditions at near-ambient temperatures and short reaction times of only 2 h. The influence of different Nα -protecting groups, boronic acids as well as the impact of different amino acid side chains in bromotryptophan-containing peptides has been studied. Notably, a surprising acceleration of the catalysis was observed when palladium-coordinating side chains were present in proximal positions.

Smart microgels investigated by super-resolution fluorescence microscopy: influence of the monomer structure on the particle morphology.

In a recent publication [Bergmann et al. Phys. Chem. Chem. Phys., 2018, 20, 5074-5083] we presented a method which enables to investigate the morphology of microgels by superresolution fluorescence microscopy. Here, this method is applied to three microgel species, based on N-isopropylmethacrylamide (NIPMAM), N-n-propylacrylamide (NNPAM) and N-n-propylmethacrylamide (NNPMAM)) with 5, 7.5 and 10 mol% cross-linker, respectively. Super-resolution microscopy reveals differences of the network morphology of the synthesized particles showing the importance of the monomer structure.

Core-shell microgels as thermoresponsive carriers for catalytic palladium nanoparticles.

Responsive core-shell microgels are promising systems for a stabilization of Pd nanoparticles and control of their catalytic activity. Here, poly-N-n-propylacrylamide (PNNPAM) was copolymerized with methacrylic acid to yield microgel core particles, which were subsequently coated with an additional, acid-free poly-N-isopropylmethacrylamide (PNIPMAM) shell. Both core and core-shell systems were used as pH- and temperature-responsive carrier systems for the incorporation of palladium nanoparticles. The embedded nanoparticles were found to have a uniform size distribution with diameters at around 20 nm. Their catalytic activity was investigated by following the kinetics of the reduction of p-nitrophenol to p-aminophenol using UV-vis spectroscopy. For the PNNPAM microgel core, the temperature dependence of the rate constant followed the Arrhenius equation, which is an unusual behaviour for thermoresponsive carrier systems but common for passive systems such as polyelectrolyte brushes. In contrast, the catalytic activity of nanoparticles embedded in microgel core-shell systems decreased drastically at the volume phase transition temperature (44 °C) of the PNIPMAM shell. Accordingly, a promising architecture of passive nanoparticle-carrying core and thermoresponsive shell was realized successfully.

Contrast variation SANS measurement of shell monomer density profiles of smart core-shell microgels.

The radial density profile of deuterated poly(N,n-propyl acrylamide) shell monomers within core-shell microgels has been studied by small-angle neutron scattering in order to shed light on the origin of their linear thermally-induced swelling. The poly(N-isopropyl methacrylamide) core monomers have been contrast-matched by the H2O/D2O solvent mixture, and the intensity thus provides a direct measurement of the spatial distribution of the shell monomers. Straightforward modelling shows that their structure does not correspond to the expected picture of a well-defined external shell. A multi-shell model solved by a reverse Monte Carlo approach is then applied to extract the monomer density as a function of temperature and of the core crosslinking. It is found that most shell monomers fill the core at high temperatures approaching synthesis conditions of collapsed particles, forming only a dilute corona. As the core monomers tend to swell at lower temperatures, a skeleton of insoluble shell monomers hinders swelling, inducing the progressive linear thermoresponse.

Shear-Induced Transformation of Polymer-Rich Lamellar Phases to Micron-Sized Vesicles.

In the present work, we study the shear-induced transformation of polymer-rich lamellar phases into vesicles. The evolution of vesicle size is studied by different scattering techniques, rheology, and microscopy methods. The lamellar phase found in the system D2O/ o-xylene/ Pluronic PE9400/C8TAB can be fully transformed to multilamellar vesicles (MLVs) by applying shear. The size of the MLVs is proportional to the inverse square root of the shear rate. Hence, the polymer-based quaternary system behaves similar to lamellar phases based on small surfactant molecules. Additionally, we found a growth effect leading to a size increase of the vesicles after shearing was stopped.

Aescin-Induced Conversion of Gel-Phase Lipid Membranes into Bicelle-like Lipid Nanoparticles.

Mixtures of the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and the saponin ß-aescin spontaneously form monodisperse, bilayered discoidal micelles (also known as "bicelles" or "nanodisks") in aqueous solution. Such bicelles form below the melting temperature of DMPC when the phospholipids are in the rigid Lß' state and are precursors of spontaneously formed vesicles. The aescin concentration must be far above the cmcaescin (≈0.3-0.4 mM). It was found that the shape and size of the bicelles are tunable by composition. High amounts of aescin decrease the size of the bicelles from diameters of ∼300 Å at 7 mol % to ∼120 Å at 30 mol % ß-aescin. The structures are scrutinized by complementary small-angle X-ray and neutron scattering experiments. The scattering curves are subsequently analyzed by a model-independent (indirect Fourier transform analysis) and a model-based approach where bicelles are described as polydisperse bilayer disks encircled by a ß-aescin rim. Moreover, the monomodal distribution and low polydispersity of the samples were confirmed by photon correlation spectroscopy. The discoidal structures were visualized by transmission electron microscopy.