Planar Polymer Membranes Accommodate Functional Self-Assembly of Inserted Resorcinarene Nanocapsules.

Solid-supported polymer membranes (SSPMs) offer great potential in material and life sciences due to their increased mechanical stability and robustness compared to solid-supported lipid membranes. However, there is still a need for expanding the functionality of SSPMs by combining them with synthetic molecular assemblies. In this study, SSPMs served as a flexible matrix for the insertion of resorcinarene monomers and their self-assembly into functional hexameric resorcinarene capsules. Resorcinarene capsules provide a large cavity with affinity specifically for cationic and polyhydroxylated molecules. While the capsules are stable in apolar organic solvents, they disassemble when placed in polar solvents, which limits their application. Here, a solvent-assisted approach was used for copolymer membrane deposition on solid support and simultaneous insertion of the resorcinarene monomers. By investigation of the molecular factors and conditions supporting the codeposition of the copolymer and resorcinarene monomers, a stable hybrid membrane was formed. The hydrophobic domain of the membrane played a crucial role by providing a sufficiently thick and apolar layer, allowing for the self-assembly of the capsules. The capsules were functional inside the membranes by encapsulating cationic guests from the aqueous environment. The amount of resorcinarene capsules in the hybrid membranes was quantified by a combination of quartz-crystal microbalance with dissipation and liquid chromatography-mass spectrometry, while the membrane topography and layer composition were analyzed by atomic force microscopy and neutron reflectometry. Functional resorcinarene capsules inside SSPMs can serve as dynamic sensors and potentially as cross-membrane transporters, thus holding great promise for the development of smart surfaces.

Microfluidic Giant Polymer Vesicles Equipped with Biopores for High-Throughput Screening of Bacteria.

Understanding the mechanisms of antibiotic resistance is critical for the development of new therapeutics. Traditional methods for testing bacteria are often limited in their efficiency and reusability. Single bacterial cells can be studied at high throughput using double emulsions, although the lack of control over the oil shell permeability and limited access to the droplet interior present serious drawbacks. Here, a straightforward strategy for studying bacteria-encapsulating double emulsion-templated giant unilamellar vesicles (GUVs) is introduced. This microfluidic approach serves to simultaneously load bacteria inside synthetic GUVs and to permeabilize their membrane with the pore-forming peptide melittin. This enables antibiotic delivery or the influx of fresh medium into the GUV lumen for highly parallel cultivation and antimicrobial efficacy testing. Polymer-based GUVs proved to be efficient culture and analysis microvessels, as microfluidics allow easy selection and encapsulation of bacteria and rapid modification of culture conditions for antibiotic development. Further, a method for in situ profiling of biofilms within GUVs for high-throughput screening is demonstrated. Conceivably, synthetic GUVs equipped with biopores can serve as a foundation for the high-throughput screening of bacterial colony interactions during biofilm formation and for investigating the effect of antibiotics on biofilms.

Trends in the Synthesis of Polymer Nano- and Microscale Materials for Bio-Related Applications.

Polymeric nano- and microscale materials bear significant potential in manifold applications related to biomedicine. This is owed not only to the large chemical diversity of the constituent polymers, but also to the various morphologies these materials can achieve, ranging from simple particles to intricate self-assembled structures. Modern synthetic polymer chemistry permits the tuning of many physicochemical parameters affecting the behavior of polymeric nano- and microscale materials in the biological context. In this Perspective, an overview of the synthetic principles underlying the modern preparation of these materials is provided, aiming to demonstrate how advances in and ingenious implementations of polymer chemistry fuel a range of applications, both present and prospective.

Structural color from solid-state polymerization-induced phase separation.

Structural colors are produced by wavelength-dependent scattering of light from nanostructures. While living organisms often exploit phase separation to directly assemble structurally colored materials from macromolecules, synthetic structural colors are typically produced in a two-step process involving the sequential synthesis and assembly of building blocks. Phase separation is attractive for its simplicity, but applications are limited due to a lack of robust methods for its control. A central challenge is to arrest phase separation at the desired length scale. Here, we show that solid-state polymerization-induced phase separation can produce stable structures at optical length scales. In this process, a polymeric solid is swollen and softened with a second monomer. During its polymerization, the two polymers become immiscible and phase separate. As free monomer is depleted, the host matrix resolidifies and arrests coarsening. The resulting polymeric composites have a blue or white appearance. We compare these biomimetic nanostructures to those in structurally-colored feather barbs, and demonstrate the flexibility of this approach by producing structural color in filaments and large sheets.

Stable Immobilization of Enzymes in a Macro- and Mesoporous Silica Monolith.

Horseradish peroxidase isoenzyme C (HRP) and Engyodontium album proteinase K (proK) were immobilized inside macro- and mesoporous silica monoliths. Stable immobilization was achieved through simple noncovalent adsorption of conjugates, which were prepared from a polycationic, water-soluble second generation dendronized polymer (denpol) and the enzymes. Conjugates prepared from three denpols with the same type of repeating unit (r.u.), but different average lengths were compared. It was shown that there is no obvious advantage of using denpols with very long chains. Excellent results were achieved with denpols having on average 750 or 1000 r.u. The enzyme-loaded monoliths were tested as flow reactors. Comparison was made with microscopy glass coverslips onto which the conjugates were immobilized and with glass micropipettes containing adsorbed conjugates. High enzyme loading was achieved using the monoliths. Monoliths containing immobilized denpol-HRP conjugates exhibited good operational stability at 25 °C (for at least several hours), and good storage stability at 4 °C (at least for weeks) was demonstrated. Such HRP-containing monoliths were applied as continuous flow reactors for the quantitative determination of hydrogen peroxide in aqueous solution between 1 µM (34 ng/mL) and 50 µM (1.7 µg/mL). Although many methods for immobilizing enzymes on silica surfaces exist, there are only a few approaches with porous silica materials for the development of flow reactors. The work presented is a promising contribution to this field of research toward bioanalytical and biosynthetic applications.

Can one determine the density of an individual synthetic macromolecule?

Dendronized polymers (DPs) are large and compact main-chain linear polymers with a cylindrical shape and cross-sectional diameters of up to ∼15 nm. They are therefore considered molecular objects, and it was of interest whether given their experimentally accessible, well-defined dimensions, the density of individual DPs could be determined. We present measurements on individual, deposited DP chains, providing molecular dimensions from scanning and transmission electron microscopy and mass-per-length values from quantitative scanning transmission electron microscopy. These results are compared with density values obtained from small-angle X-ray scattering on annealed bulk specimen and with classical envelope density measurements, obtained using hydrostatic weighing or a density gradient column. The samples investigated comprise a series of DPs with side groups of dendritic generations g = 1-8. The key findings are a very large spread of the density values over all samples and methods, and a consistent increase of densities with g over all methods. While this work highlights the advantages and limitations of the applied methods, it does not provide a conclusive answer to the question of which method(s) to use for the determination of densities of individual molecular objects. We are nevertheless confident that these first attempts to answer this challenging question will stimulate more research into this important aspect of polymer and soft matter science.

Main-chain scission of individual macromolecules induced by solvent swelling.

We present a comprehensive investigation of main-chain scission processes affecting peripherally charged and neutral members of a class of dendronized polymers (DPs) studied in our laboratory. In these thick, sterically highly congested macromolecules, scission occurs by exposure to solvents, in some cases at room temperature, in others requiring modest heating. Our investigations rely on gel permeation chromatography and atomic force microscopy and are supported by molecular dynamics simulations as well as by electron paramagnetic resonance spectroscopy. Strikingly, DP main-chain scission depends strongly on two factors: first the solvent, which must be highly polar to induce scission of the DPs, and second the dendritic generation g. In DPs of generations 1 ≤ g ≤ 8, scission occurs readily only for g = 5, no matter whether the polymer is charged or neutral. Much more forcing conditions are required to induce degradation in DPs of g ≠ 5. We propose solvent swelling as the cause for the main-chain scission in these individual polymer molecules, explaining in particular the strong dependence on g: g < 5 DPs resemble classical polymers and are accessible to the strongly interacting, polar solvents, whereas g > 5 DPs are essentially closed off to solvent due to their more closely colloidal character. g = 5 DPs mark the transition between these two regimes, bearing strongly sterically congested side chains which are still solvent accessible to some degree. Our results suggest that, even in the absence of structural elements which favour scission such as cross-links, solvent swelling may be a generally applicable mechanochemical trigger. This may be relevant not only for DPs, but also for other types of sterically strongly congested macromolecules.

Excitonic channels from bio-inspired templated supramolecular assembly of J-aggregate nanowires.

Supramolecular assemblies with controlled morphology are of paramount importance for energy transport in organic semiconductors. Despite considerable freedom in molecular design, the preparation of dyes that form one dimensional J-aggregates is challenging. Here, we demonstrate a simple and effective route to functionalize dendronized polymers (DPs) with J-aggregates to construct tubular DP/J-aggregate nanowires. When J-aggregates are adsorbed onto DPs anchored to glass substrates, they assemble into microcrystalline domains typical for J-aggregates adsorbed on functionalized surfaces. Differently, the complexation between the dendronized polymer and J-aggregates in solution leads to dense packing of J-aggregate strands on the periphery of the DPs. Using a layer-by-layer (LBL) technique, DPs loaded with J-aggregates can also be adsorbed onto a DP monolayer. In this case, the thin film absorption spectra are narrower and indicate higher ratios of J-aggregate to monomer and dimer absorption than bare J-aggregates deposited similarly. The demonstration of J-aggregate adsorption on filamentous polymeric templates is a promising step toward artificial 1D light harvesting antennas, with potential applications in opto-electronic devices.

3D Conformations of Thick Synthetic Polymer Chains Observed by Cryogenic Electron Microscopy.

The backbone conformations of individual, unperturbed synthetic macromolecules have so far not been observed directly in spite of their fundamental importance to polymer physics. Here we report the dilute solution conformations of two types of linear dendronized polymers, obtained by cryogenic transmission electron stereography and tomography. The three-dimensional trajectories show that the wormlike chain model fails to adequately describe the scaling of these thick macromolecules already beyond a few nanometers in chain length, in spite of large apparent persistence lengths and long before a signature of self-avoidance appears. This insight is essential for understanding the limitations of polymer physical models, and it motivated us to discuss the advantages and disadvantages of this approach in comparison to the commonly applied scattering techniques.

Tailoring polymer dispersity and shape of molecular weight distributions: methods and applications.

The width and shape of molecular weight distributions can significantly affect the properties of polymeric materials and thus are key parameters to control. This mini-review aims to critically summarise recent approaches developed to tailor molecular weight distributions and highlights the strengths and limitations of each technique. Special emphasis will also be given to applications where tuning the molecular weight distribution has been used as a strategy to not only enhance polymer properties but also to increase the fundamental understanding behind complex mechanisms and phenomena.

Effect of Polymerization Components on Oxygen-Tolerant Photo-ATRP.

Photo-ATRP has recently emerged as a powerful technique that allows for oxygen-tolerant polymerizations and the preparation of polymers with low dispersity and high end-group fidelity. However, the effect of various photo-ATRP components on oxygen consumption and polymerization remains elusive. Herein, we employ an in situ oxygen probe and UV-vis spectroscopy to elucidate the effects of ligand, initiator, monomer, and solvent on oxygen consumption. We found that the choice of photo-ATRP components significantly impacts the rate at which the oxygen is consumed and can subsequently affect both the polymerization time and the dispersity of the resulting polymer. Importantly, we discovered that using the inexpensive ligand TREN results in the fastest oxygen consumption and shortest polymerization time, even though no appreciable reduction of CuBr2 is observed. This work provides insight into oxygen consumption in photo-ATRP and serves as a guideline to the judicious selection of photo-ATRP components for the preparation of well-defined polymers.

Dual, Site-Specific Modification of Antibodies by Using Solid-Phase Immobilized Microbial Transglutaminase.

Microbial transglutaminase (MTG) was stably solid-phase immobilized on glass microbeads by using a second-generation dendronized polymer. Immobilized MTG enabled the efficient generation of site-specifically conjugated proteins, including antibody fragments, as well as whole antibodies through distinct glutamines and, unprecedentedly, also through lysines with various bifunctional substrates with defined stoichiometries. With this method, we generated dual, site-specifically modified antibodies comprising a fluorescent probe and a metal chelator for radiolabeling-a strategy anticipated to design antibodies for imaging and simultaneous therapy. Furthermore, we provide evidence that immobilized MTG features higher siteselectivity than soluble MTG.

Preparation and Applications of Dendronized Polymer-Enzyme Conjugates.

Dendronized polymer-enzyme conjugates are large, water-soluble macromolecular structures built from a linear, fully synthetic, dendronized polymer (denpol), and several copies of enzyme molecules covalently bound to the peripheral functional groups of the denpol. Since denpol chains comprise repeating units with regularly branched side chains (dendrons), denpols have a cylindrical shape and are much thicker than conventional linear polymers. Depending on the dendron generation and chemical structure, denpols may have a large number of functional groups on their surface, exposed to the aqueous medium in which they are dissolved. Enzymes (and also other molecules) can be attached to these functional groups, for example, via a stable bis-aryl hydrazone (BAH) bond. The dendronized polymer scaffold might also serve as a nanoarmor and stabilize the delicate enzymes. One of the denpols which can be used for the preparation of denpol-enzyme conjugates is de-PG2. It has a poly(methacrylate) backbone and consists of second-generation dendrons with four peripheral amino groups in each repeating unit. The synthesis of de-PG2 and the preparation of a de-PG2 conjugate carrying BAH-linked proteinase K (proK), as an example, are described here for applications in the field of enzyme immobilization on solid surfaces. The nanoarmored enzyme-polymer conjugate indicated high stability and retention of enzymatic activity.

Exploring the Loading Capacity of Generation Six to Eight Dendronized Polymers in Aqueous Solution.

Aspects of size, structural (im)perfection, inner density, and guest molecule loading capacity of dendronized polymers (DPs) of high generation (6≤g≤8) in aqueous solution are studied using electron paramagnetic resonance spectroscopy on amphiphilic, spin-labeled guest molecules. The results show that the interior of the charged DPs is strongly polar, especially in comparison to their lower generation (1-4) analogues. This is a direct sign that large amounts of water penetrate the DP surface, reflecting the structural (im)perfections of these high-generation DPs and much lower segmental densities than theoretically achievable. Images obtained with atomic force microscopy reveal that the high-generation DPs do not aggregate and give further insights into the structural imperfections. Electron paramagnetic resonance spectroscopic data further show that despite their structural imperfections, these DPs can bind and release large numbers of amphiphilic molecules. It is concluded that attention should be paid to their synthesis, for which a protocol needs to be developed that avoids the relatively large amount of defects generated in the direct conversion of a generation g=4 DP to a generation g=6 DP, which had to be used here.