Near-complete depolymerization of polyesters with nano-dispersed enzymes.

Successfully interfacing enzymes and biomachinery with polymers affords on-demand modification and/or programmable degradation during the manufacture, utilization and disposal of plastics, but requires controlled biocatalysis in solid matrices with macromolecular substrates1-7. Embedding enzyme microparticles speeds up polyester degradation, but compromises host properties and unintentionally accelerates the formation of microplastics with partial polymer degradation6,8,9. Here we show that by nanoscopically dispersing enzymes with deep active sites, semi-crystalline polyesters can be degraded primarily via chain-end-mediated processive depolymerization with programmable latency and material integrity, akin to polyadenylation-induced messenger RNA decay10. It is also feasible to achieve processivity with enzymes that have surface-exposed active sites by engineering enzyme-protectant-polymer complexes. Poly(caprolactone) and poly(lactic acid) containing less than 2 weight per cent enzymes are depolymerized in days, with up to 98 per cent polymer-to-small-molecule conversion in standard soil composts and household tap water, completely eliminating current needs to separate and landfill their products in compost facilities. Furthermore, oxidases embedded in polyolefins retain their activities. However, hydrocarbon polymers do not closely associate with enzymes, as their polyester counterparts do, and the reactive radicals that are generated cannot chemically modify the macromolecular host. This study provides molecular guidance towards enzyme-polymer pairing and the selection of enzyme protectants to modulate substrate selectivity and optimize biocatalytic pathways. The results also highlight the need for in-depth research in solid-state enzymology, especially in multi-step enzymatic cascades, to tackle chemically dormant substrates without creating secondary environmental contamination and/or biosafety concerns.

Zwitterionic Ammonium Sulfonate Polymers: Synthesis and Properties in Fluids.

Polymer zwitterions continue to emerge as useful materials for numerous applications, ranging from hydrophilic and antifouling coatings to electronic materials interfaces. While several polymer zwitterion compositions are now well established, interest in this field of soft materials science has grown rapidly in recent years due to the introduction of new structures that diversify their chemistry and architecture. Nonetheless, at present, the variation of the chemical composition of the anionic and cationic components of zwitterionic structures remains relatively limited to a few primary examples. In this article, the versatility of 4-vinylbenzyl sultone as a precursor to ammonium sulfonate zwitterionic monomers, which are then used in controlled free radical polymerization chemistry to afford "inverted sulfobetaine" polymer zwitterions, is highlighted. An evaluation of the solubility, interfacial activity, and solution configuration of the resultant polymers reveals the dependence of properties on the selection of tertiary amines used for nucleophilic ring-opening of the sultone precursor, as well as useful properties comparisons across different zwitterionic compositions.

Interfacial Reaction Induced Disruption and Dissolution of Dynamic Polymer Networks.

The reaction of amine-terminated polystyrene (PS-NH2 ) with an epoxy-based dynamic polymer networks (DPNs) above the topology freezing transition temperature of the DPN, results in the disruption of the network by the formation of graft copolymers at the interface between the linear homopolymer and the network. The rate of the disruption decreases with annealing time and is strongly dependent on the molecular weight of the PS-NH2 , with the lower molecular weight PS-NH2 reacting much more rapidly than the higher molecular weight PS-NH2 . A higher catalyst concentration in the DPN also promotes the interfacial reaction, indicating a reaction-rate-controlled process.

Self-assembly of nanomaterials at fluid interfaces.

Recent developments in the field of the self-assembly of nanoscale materials such as nanoparticles, nanorods and nanosheets at liquid/liquid interfaces are reviewed. Self-assembly behavior of both biological and synthetic particles is discussed. For biological nanoparticles, the nanoparticle assembly at fluid interfaces provides a simple route for directing nanoparticles into 2D or 3D constructs with hierarchical ordering. The interfacial assembly of single-walled carbon nanotubes (SWCNTs) at liquid interfaces would play a key role in applications such as nanotube fractionation, flexible electronic thin-film fabrication and synthesis of porous SWCNT/polymer composites foams. Liquids can be structured by the jamming of nanoparticle surfactants at fluid interfaces. By controlling the interfacial packing of nanoparticle surfactants using external triggers, a new class of materials can be generated that combines the desirable characteristics of fluids such as rapid transport of energy carriers with the structural stability of a solid.

Assembly of acid-functionalized single-walled carbon nanotubes at oil/water interfaces.

The efficient segregation of water-soluble, acid-functionalized, single-walled carbon nanotubes (SWCNTs) at the oil/water interface was induced by dissolving low-molecular-weight amine-terminated polystyrene (PS-NH2) in the oil phase. Salt-bridge interactions between carboxylic acid groups of SWCNTs and amine groups of PS drove the assembly of SWCNTs at the interface, monitored by pendant drop tensiometry and laser scanning confocal microscopy. The impact of PS end-group functionality, PS and SWCNT concentrations, and the degree of SWCNT acid modification on the interfacial activity was assessed, and a sharp drop in interfacial tension was observed above a critical SWCNT concentration. Interfacial tensions were low enough to support stable oil/water emulsions. Further experiments, including potentiometric titrations and the replacement of SWCNTs by other carboxyl-containing species, demonstrated that the interfacial tension drop reflects the loss of SWCNT charge as the pH falls near/below the intrinsic carboxyl dissociation constant; species lacking multivalent carboxylic acid groups are inactive. The trapped SWCNTs appear to be neither ordered nor oriented.

Reconfigurable Magnetic Liquid Building Blocks for Constructing Artificial Spinal Column Tissues.

All-liquid molding can be used to transform a liquid into free-form solid constructs, while maintaining internal fluidity. Traditional biological scaffolds, such as cured pre-gels, are normally processed in solid state, sacrificing flowability and permeability. However, it is essential to maintain the fluidity of the scaffold to truly mimic the complexity and heterogeneity of natural human tissues. Here, this work molds an aqueous biomaterial ink into liquid building blocks with rigid shapes while preserving internal fluidity. The molded ink blocks for bone-like vertebrae and cartilaginous-intervertebral-disc shapes, are magnetically manipulated to assemble into hierarchical structures as a scaffold for subsequent spinal column tissue growth. It is also possible to join separate ink blocks by interfacial coalescence, different from bridging solid blocks by interfacial fixation. Generally, aqueous biomaterial inks are molded into shapes with high fidelity by the interfacial jamming of alginate surfactants. The molded liquid blocks can be reconfigured using induced magnetic dipoles, that dictated the magnetic assembly behavior of liquid blocks. The implanted spinal column tissue exhibits a biocompatibility based on in vitro seeding and in vivo cultivating results, showing potential physiological function such as bending of the spinal column.

Kinetically trapped co-continuous polymer morphologies through intraphase gelation of nanoparticles.

We describe an approach to prepare co-continuous microstructured blends of polymers and nanoparticles by formation of a percolating network of particles within one phase of a polymer mixture undergoing spinodal decomposition. Nanorods or nanospheres of CdSe were added to near-critical blends of polystyrene and poly(vinyl methyl ether) quenched to above their lower critical solution temperature. Beyond a critical loading of nanoparticles, phase separation is arrested due to the aggregation of particles into a network (or colloidal gel) within the poly(vinyl methyl ether) phase, yielding a co-continuous spinodal-like structure with a characteristic length scale of several micrometers. The critical concentration of nanorods to achieve kinetic arrest is found to be smaller than for nanospheres, which is in qualitative agreement with the expected dependence of the nanoparticle percolation threshold on aspect ratio. Compared to structural arrest by interfacial jamming, our approach avoids the necessity for neutral wetting of particles by the two phases, providing a general pathway to co-continuous micro- and nanoscopic structures.

Aquabots.

Soft robots, made from elastomers, easily bend and flex, but deformability constraints severely limit navigation through and within narrow, confined spaces. Using aqueous two-phase systems we print water-in-water constructs that, by aqueous phase-separation-induced self-assembly, produce ultrasoft liquid robots, termed aquabots, comprised of hierarchical structures that span in length scale from the nanoscopic to microsciopic, that are beyond the resolution limits of printing and overcome the deformability barrier. The exterior of the compartmentalized membranes is easily functionalized, for example, by binding enzymes, catalytic nanoparticles, and magnetic nanoparticles that impart sensitive magnetic responsiveness. These ultrasoft aquabots can adapt their shape for gripping and transporting objects and can be used for targeted photocatalysis, delivery, and release in confined and tortuous spaces. These biocompatible, multicompartmental, and multifunctional aquabots can be readily applied to medical micromanipulation, targeted cargo delivery, tissue engineering, and biomimetics.

Photocontrol over the disorder-to-order transition in thin films of polystyrene-block-poly(methyl methacrylate) block copolymers containing photodimerizable anthracene functionality.

Reversible photocontrol over the ordering transition of block copolymers (BCPs) from a disordered state to an ordered state, namely the disorder-to-order transition (DOT), can be used to create long-range ordered nanostructures in self-assembled BCPs over macroscopic distances by photocombing, similar to the classic zone refining used to produce highly pure, large single crystals. Here, we have designed and synthesized an anthracene-functionalized tri-BCP containing deuterated polystyrene (d(8)-PS) and poly(methyl methacrylate) (PMMA) blocks, as well as a short middle block of poly(2-hydroxyethyl methacrylates) (PHEMA) that is randomly functionalized by anthracene. This tri-BCP maintains the order-to-disorder transition-type phase behavior of its parent d(8)-PS-b-PMMA di-BCPs. Under 365 nm UV irradiation, the junction between d(8)-PS and PMMA blocks is photocoupled through the anthracene photodimers, leading to a significant increase in the total molecular weight of the tri-BCP. As a consequence, when the tri-BCP is phase-mixed but close to the boundary of the ordering transition, it undergoes the DOT, as evidenced by small-angle neutron scattering and transmission electron microscopy. The tri-BCP could be reversibly brought through the DOT in thin films by taking advantage of photodimerization and thermal dissociation of anthracene. Currently, anthracene-functionalized d(8)-PS-b-PMMA BCP is one of the most promising candidates for the photocombing process to promote long-range laterally ordered nanostructures over macroscopic distances in a noninvasive manner.

Temperature-triggered micellization of block copolymers on an ionic liquid surface.

In situ neutron reflectivity was used to study thermally induced structural changes of the lamellae-forming polystyrene-block-poly(2-vinylpyridine) (PS-b-P2VP) block copolymer thin films floating on the surface of an ionic liquid (IL). The IL, 1-butyl-3-methylimidazolium trifluoromethanesulfonate, is a nonsolvent for PS and a temperature-tunable solvent for P2VP, and, as such, micellization can be induced at the air-IL interface by changing the temperature. Transmission electron microscopy and scanning force microscopy were used to investigate the resultant morphologies of the micellar films. It was found that highly ordered nanostructures consisting of spherical micelles with a PS core surrounded by a P2VP corona were produced. In addition, bilayer films of PS homopolymer on top of a PS-b-P2VP layer also underwent micellization with increasing temperature but the micellization was strongly dependent on the thickness of the PS and PS-b-P2VP layers.

Density fluctuations and phase transitions of ferroelectric polymer nanowires.

Phase transitions of polymeric materials are accompanied by changes in density as a function of temperature. Being able to measure these changes in polymeric systems in one, two or three dimensions on the nanoscopic length-scale is a challenge, but it would provide a simple route to assess phase transitions in nanoscopically confined systems. It is shown that the measurement of the dielectric permittivity in the high frequency limit (in spectral regions not affected by dielectric dispersions) offers an effective and very sensitive means to assess density fluctuations, and hence phase transitions, in nanoscopic systems. The sensitivity of this approach is demonstrated by assessing the phase transition behavior of ferroelectric polymer nanowires confined within alumina membranes. No significant shifts in the Curie transition are observed down to pore diameters as small as 15 nm.

Highly ordered gold nanotubes using thiols at a cleavable block copolymer interface.

We have prepared functionalized nanoporous thin films from a polystyrene-block-polyethylene oxide block copolymer, which was made cleavable due to the intervening disulfide bond. The cleavage reaction of the disulfide bond leaves behind free thiol groups inside the nanopores of polystyrene thin film. This nanoporous thin film can be used as a template for generating gold nanoring structures. This strategy can provide a facile method to form a highly ordered array of biopolymer or metal-polymer composite structures.

Preparation of metallic line patterns from functional block copolymers.

A simple route for the preparation of nanoscopic metallic line patterns from functional block copolymers (BCPs) containing poly(2-vinylpyridine) or poly(methyl methacrylate) blocks is demonstrated. The time evolution of the surface morphologies of BCP thin films exposed to solvent vapors is studied to optimize the conditions for generating BCP microdomains oriented parallel or normal to the substrate. BCP templates are prepared by film reconstruction or by removal of one of the copolymer microdomains, depending on the properties of the functional BCPs. Finally, metallic line patterns are prepared by either electrochemical etching or direct metal deposition and lift-off processes using the BCP templates.

A simple top-down/bottom-up approach to sectored, ordered arrays of nanoscopic elements using block copolymers.

A top-down/bottom-up approach is demonstrated by combining electron-beam (e-beam) lithography and a solvent annealing process. Micellar arrays of polystyrene-block-poly(4-vinylpyridine) (PS-b-P4VP) with a high degree of lateral order can be produced on a surface where sectoring is defined by e-beam patterning. The e-beam is used to crosslink the block copolymer (BCP) film immediately after spin-coating when the BCP is disordered or in a highly ordered solvent-annealed film. Any patterns can be written into the BCP by crosslinking. Upon exposure to a preferential solvent for the minor component block followed by drying, cylindrical nanopores are generated within the nonexposed areas by a surface reconstruction process, while, in the exposed areas, the films remain unchanged. Nickel nanodot arrays can be placed over selected areas on a surface by thermal evaporation and lift-off process.

Contrasting Chemistry of Block Copolymer Films Controls the Dynamics of Protein Self-Assembly at the Nanoscale.

Biological systems are able to control the assembly and positioning of proteins with nanoscale precision, as exemplified by the intricate molecular structures within cell membranes, virus capsids, and collagen matrices. Controlling the assembly of biomolecules is critical for the use of biomaterials in artificial systems such as antibacterial coatings, engineered tissue samples, and implanted medical devices. Furthermore, understanding the dynamics of protein assembly on heterogeneous templates will ultimately enable the control of protein crystallization in general. Here, we show a biomimetic, hierarchical bottom-up approach to direct the self-assembly of crystalline S-layers through nonspecific interactions with nanostructured block copolymer (BCP) thin-film templates. A comparison between physically and chemically patterned BCP substrates shows that chemical heterogeneity is required to confine the adhesion and self-assembly of S-layers to specific BCP domains. Furthermore, we show that this mechanism can be extended to direct the formation of collagen fibers along the principal direction of the underlying BCP substrate. The dynamics of protein self-assembly at the solid-liquid interface are followed using in situ high-resolution atomic force microscopy under continuous flow conditions, allowing the determination of the rate constants of the self-assembly. A pattern of alternating, chemically distinct nanoscale domains drastically increases the rate of self-assembly compared to non-patterned chemically homogeneous substrates.

Novel transparent nano- to micro-heterogeneous substrates for in-situ cell migration study.

Transparent substrates having heterogeneities ranging from nanometer to micrometer lateral length scale were fabricated to study cell migration. The surfaces were generated using thin films of block copolymers and homopolymer blends on ultra smooth transparent polyethylene terephthalate films. Results show that the lateral size scale of the surface heterogeneities affects fibroblast (NIH-3T3) adhesion, spreading and motility. More specifically, fibroblasts migrate faster on micron-sized than on nanometer-sized heterogeneities. Cell movements and morphology on the micron patterned surfaces resemble cells cultured in a 3D environment. These surfaces, therefore, can potentially be utilized as models to study cell behavior in physiologically relevant conditions which can add to our fundamental understanding of cell-substrate interactions and facilitate development of surfaces for medical devices.

Printing Fabrication of Bulk Heterojunction Solar Cells and In Situ Morphology Characterization.

Polymer-based materials hold promise as low-cost, flexible efficient photovoltaic devices. Most laboratory efforts to achieve high performance devices have used devices prepared by spin coating, a process that is not amenable to large-scale fabrication. This mismatch in device fabrication makes it difficult to translate quantitative results obtained in the laboratory to the commercial level, making optimization difficult. Using a mini-slot die coater, this mismatch can be resolved by translating the commercial process to the laboratory and characterizing the structure formation in the active layer of the device in real time and in situ as films are coated onto a substrate. The evolution of the morphology was characterized under different conditions, allowing us to propose a mechanism by which the structures form and grow. This mini-slot die coater offers a simple, convenient, material efficient route by which the morphology in the active layer can be optimized under industrially relevant conditions. The goal of this protocol is to show experimental details of how a solar cell device is fabricated using a mini-slot die coater and technical details of running in situ structure characterization using the mini-slot die coater.

An optical waveguide study on the nanopore formation in block copolymer/homopolymer thin films by selective solvent swelling.

Thin films of mixtures of asymmetric poly(styrene-block-methyl methacrylate) (PS-b-PMMA) diblock copolymers and PMMA homopolymers with cylindrical PMMA microdomains oriented normal to the substrate surface were used to couple optical modes in the Kretschmann configuration, and their optical properties were investigated by optical waveguide spectroscopy (OWS). The nanopore formation in the block copolymer (BCP) waveguide layer via selective solvent swelling and subsequent reannealing was monitored in terms of shifts in the coupling mode angles. The sequential swelling/reannealing of the initial mixture film resulted in a number of discrete or partially interconnected pores instead of cylindrical pores with a high aspect ratio. The simultaneous processes occurring inside and on top of the BCP waveguide layer were discerned independently with high selectivity for p- and s-polarization.

Fibroblast adhesion to micro- and nano-heterogeneous topography using diblock copolymers and homopolymers.

Polymeric substrates of different surface chemistry and length scales were found to have profound influence on cell adhesion. The adhesion of fibroblasts on surfaces of oxidized polystyrene (PS), on surfaces modified with random copolymers of PS and poly(methyl methacrylate) [P(S-r-MMA)] with topographic features, and chemically patterned surfaces that varied in lateral length scales from nanometers to microns were studied. Surfaces with heterogeneous topographies were generated from thin film mixtures of a block copolymer, PS-b-MMA, with homopolymers of PS and PMMA. The two homopolymers macroscopically phase separated and, with the addition of diblock copolymer, the size scales of the phases decreased to nanometer dimensions. Cell spreading area analysis showed that a thin film of oxidized PS surface promoted adhesion whereas a thin film of P(S-r-MMA) surface did not. Fibroblast adhesion was examined on surfaces in which the lateral length scale varied from 60 nm to 6 microm. It was found that, as the lateral length scale between the oxidized PS surfaces decreased, cell spreading area and degree of actin stress fiber formation increased. In addition, scanning electron microscopy was used to evaluate the location of filopodia and lamellipodia. It was found that most of the filopodia and lamellipodia interacted with the oxidized PS surfaces. This can be attributed to both chemical and topographic surface interactions that prevent cells from interacting with the P(S-r-MMA) at the base of the topographic features.