4D-Printable Photocrosslinkable Polyurethane-Based Inks for Tissue Scaffold and Actuator Applications.

4D printing recently emerges as an exciting evolution of conventional 3D printing, where a printed construct can quickly transform in response to a specific stimulus to switch between a temporary variable state and an original state. In this work, a photocrosslinkable polyethylene-glycol polyurethane ink is synthesized for light-assisted 4D printing of smart materials. The molecular weight distribution of the ink monomers is tunable by adjusting the copolymerization reaction time. Digital light processing (DLP) technique is used to program a differential swelling response in the printed constructs after humidity variation. Bioactive microparticles are embedded into the ink and the improvement of biocompatibility of the printed constructs is demonstrated for tissue engineering applications. Cell studies reveal above 90% viability in 1 week and ≈50% biodegradability after 4 weeks. Self-folding capillary scaffolds, dynamic grippers, and film actuators are made and activated in a humid environment. The approach offers a versatile platform for the fabrication of complex constructs. The ink can be used in tissue engineering and actuator applications, making the ink a promising avenue for future research.

Materials for Smart Soft Actuator Systems.

In contrast to conventional hard actuators, soft actuators offer many vivid advantages, such as improved flexibility, adaptability, and reconfigurability, which are intrinsic to living systems. These properties make them particularly promising for different applications, including soft electronics, surgery, drug delivery, artificial organs, or prosthesis. The additional degree of freedom for soft actuatoric devices can be provided through the use of intelligent materials, which are able to change their structure, macroscopic properties, and shape under the influence of external signals. The use of such intelligent materials allows a substantial reduction of a device's size, which enables a number of applications that cannot be realized by externally powered systems. This review aims to provide an overview of the properties of intelligent synthetic and living/natural materials used for the fabrication of soft robotic devices. We discuss basic physical/chemical properties of the main kinds of materials (elastomers, gels, shape memory polymers and gels, liquid crystalline elastomers, semicrystalline ferroelectric polymers, gels and hydrogels, other swelling polymers, materials with volume change during melting/crystallization, materials with tunable mechanical properties, and living and naturally derived materials), how they are related to actuation and soft robotic application, and effects of micro/macro structures on shape transformation, fabrication methods, and we highlight selected applications.

4D Fabrication of Two-Way Shape Memory Polymeric Composites by Electrospinning and Melt Electrowriting.

This work presents a new method for 4D fabrication of two-way shape memory materials that are capable of reversible shapeshifting right after manufacturing, upon application of proper heating and cooling cycles. The innovative solution presented here consists in the combination of highly stretched electrospun shape memory polymer (SMP) nanofibers with a melt electrowritten elastomer. More specifically, the stretched nanofibers are made of a biocompatible thermoplastic polyurethane (TPU) with crystallizable soft segments, undergoing melt-induced contraction and crystallization-induced elongation upon heating and cooling, respectively. Reversible actuation during crystallization becomes possible due to the elastic recovery of the elastomer component, obtained by melt electrowriting of a commercial TPU filament. Thanks to the design freedom offered by additive manufacturing, the elastomer structure also has the role of guiding the shape transformation. Electrospinning and melt electrowriting process parameters are set up so to obtain smart 4D objects capable of two-way shape memory effect (SME), and the possibility of reversible and repeatable actuation is demonstrated. The two components are then combined in different proportions with the aim of tailoring the two-way SME, taking into account the effect of design parameters such as the SMP content, the elastomer pattern, and the composite thickness.

Self-Healing and Electrical Properties of Viscoelastic Polymer-Carbon Blends.

Self-healing polymer-carbon composites are seen as promising materials for future electronic devices, which must be able to restore not only their structural integrity but also electrical performance after cracking and wear. Despite multiple reports about self-healing conductive elements, there is a lack of a broad fundamental understanding of correlation between viscoelasticity of such composites, their electrical properties, and self-healing of their mechanical as well as electrical properties. Here, it is reported thorough investigation of electromechanical properties of blends of carbon black (CB) as conductive filler and viscoelastic polymers (polydimethylsiloxanes (PDMS) and polyborosiloxane (PBS)) with different relaxation times as matrices. It is shown that behavior of composites depends strongly on the viscoelastic properties of polymers. Low molecular polymer composite possesses high conductivity due to strong filler network formation, quick electrical, and mechanical properties restoration, but for this the ability is sacrificed to flow and ductility at large deformation (material is brittle). In contrary, high relaxation time polymer composite behaves elastically on small time and flows at large time scale due to weak filler network and can heal. However, the electrical properties are worse than that of carbon and viscous polymer and degrade with time.

Shape-morphing architectures actuated by Janus fibers.

We describe a combined experimental and theoretical investigation of shape-morphing structures assembled by actuating composite (Janus) fibers, taking into account multiple relevant factors affecting shape transformations, such as strain rate, composition, and geometry of the structures. Starting with simple bending experiments, we demonstrate the ways to attain multiple out-of-plane shapes of closed rings and square frames. Through combining theory and simulation, we examine how the mechanical properties of Janus fibers affect shape transitions. This allows us to control shape changes and to attain target 3D shapes by precise tuning of the material properties and geometry of the fibers. Our results open new perspectives of design of advanced mechanical metamaterials capable to create elaborate structures through sophisticated actuation modes.

Porous Stimuli-Responsive Self-Folding Electrospun Mats for 4D Biofabrication.

We report fabrication and characterization of electrospun, porous multi-layer scaffolds based-on thermo-responsive polymers polycaprolactone (PCL) and poly(N-isopropylacrylamide). We found that the electrospun mats fold into various 3D structures in an aqueous environment at different temperatures. We could determine the mechanism behind different folding behaviors under different conditions by consideration of the properties of the individual polymers. At 37 °C in an aqueous environment, the scaffolds spontaneously rolled into tubular structures with PCL as the inner layer, making them suitable for cell encapsulation. We also demonstrated that the cell adhesion and viability could be improved by coating the polymers with collagen, showing the suitability of this scaffold for several tissue engineering applications.

4D Origami by Smart Embroidery.

There exist many methods for processing of materials: extrusion, injection molding, fibers spinning, 3D printing, to name a few. In most cases, materials with a static, fixed shape are produced. However, numerous advanced applications require customized elements with reconfigurable shape. The few available techniques capable of overcoming this problem are expensive and/or time-consuming. Here, the use of one of the most ancient technologies for structuring, embroidering, is proposed to generate sophisticated patterns of active materials, and, in this way, to achieve complex actuation. By combining experiments and computational modeling, the fundamental rules that can predict the folding behavior of sheets with a variety of stitch-patterns are elucidated. It is demonstrated that theoretical mechanics analysis is only suitable to predict the behavior of the simplest experimental setups, whereas computer modeling gives better predictions for more complex cases. Finally, the applicability of the rules by designing basic origami structures and wrinkling substrates with controlled thermal insulation properties is shown.

Bienzymatic Sequential Reaction on Microgel Particles and Their Cofactor Dependent Applications.

We report, the preparation and characterization of bioconjugates, wherein enzymes pyruvate kinase (Pk) and l-lactic dehydrogenase (Ldh) were covalently bound to poly(N-isopropylacrylamide)-poly(ethylenimine) (PNIPAm-PEI) microgel support using glutaraldehyde (GA) as the cross-linker. The effects of different arrangements of enzymes on the microgels were investigated for the enzymatic behavior and to obtain maximum Pk-Ldh sequential reaction. The dual enzyme bioconjugates prepared by simultaneous addition of both the enzymes immobilized on the same microgel particles (PL), and PiLi, that is, dual enzyme bioconjugate obtained by combining single-enzyme bioconjugates (immobilized pyruvate kinase (Pi) and immobilized lactate dehydrogenase (Li)), were used to study the effect of the assembly of dual enzymes systems on the microgels. The kinetic parameters (Km, kcat), reaction parameters (temperature, pH), stability (thermal and storage), and cofactor dependent applications were studied for the dual enzymes conjugates. The kinetic results indicated an improved turn over number (kcat) for PL, while the kcat and catalytic efficiency was significantly decreased in case of PiLi. For cofactor dependent application, in which the ability of ADP monitoring and ATP synthesis by the conjugates were studied, the activity of PL was found to be nearly 2-fold better than that of PiLi. These results indicated that the influence of spacing between the enzymes is an important factor in optimization of multienzyme immobilization on the support.

Reversible thermosensitive biodegradable polymeric actuators based on confined crystallization.

We discovered a new and unexpected effect of reversible actuation of ultrathin semicrystalline polymer films. The principle was demonstrated on the example of thin polycaprolactone-gelatin bilayer films. These films are unfolded at room temperature, fold at temperature above polycaprolactone melting point, and unfold again at room temperature. The actuation is based on reversible switching of the structure of the hydrophobic polymer (polycaprolactone) upon melting and crystallization. We hypothesize that the origin of this unexpected behavior is the orientation of polycaprolactone chains parallel to the surface of the film, which is retained even after melting and crystallization of the polymer or the "crystallization memory effect". In this way, the crystallization generates a directed force, which causes bending of the film. We used this effect for the design of new generation of fully biodegradable thermoresponsive polymeric actuators, which are highly desirable for bionano-technological applications such as reversible encapsulation of cells and design of swimmers.

Controlled Retention and Release of Biomolecular Transport Systems Using Shape-Changing Polymer Bilayers.

Biomolecular transport systems based on cytoskeletal filaments and motor proteins have become promising tools for a wide range of nanotechnological applications. In this paper, we report control of such transport systems using substrates with switchable shape. We demonstrate this approach on the example of microtubules gliding on surfaces of self-folding polymer bilayers with adsorbed kinesin motors. The polymer bilayers are able to undergo reversible transitions between flat and tube-like shapes that allow the externally controlled retention and release of gliding microtubules. The demonstrated approach, based on surfaces with reconfigurable topography, opens broad perspectives to control biomolecular transport systems for bioanalytical and sensing applications, as well as for the construction of subcellular compartments in the field of synthetic biology.

Polymeric actuators.

Actuators are materials and devices that are able to change their shape in response to changes in environmental conditions and perform mechanical work on nano-, micro-, and macroscales. Among the huge variety of different actuators, polymer-based ones are highly attractive because of a number of properties such as sensitivity to a broad range of stimuli and good mechanical properties. The goal of this review is to provide a general picture of different mechanisms and working principles of polymeric actuators as well as to show a palette of their applications.

Reversibly Cross-Linkable Thermoresponsive Self-Folding Hydrogel Films.

This paper reports a novel approach for the design of self-folding films using reversibly cross-linkable thermoresponsive polymers with coumarin groups: poly(N-isopropylacrylamide-co-7-(2-methacryloyloxyethoxy)-4-methylcoumarin). We demonstrated that, depending on the structure of the films and the conditions of cross-linking/de-cross-linking, one can fabricate a variety of different forms ranging from simple tubes to complex centipede-like structures. The demonstrated approach opens new perspectives for the design of 3D self-assembling materials.

New insight into icing and de-icing properties of hydrophobic and hydrophilic structured surfaces based on core-shell particles.

Icing is an important problem, which often leads to emergency situations in northern countries. The reduction of icing requires a detailed understanding of this process. In this work, we report on a systematic investigation of the effects of geometry and chemical properties of surfaces on the formation of an ice layer, its properties, and thawing. We compare in detail icing and ice thawing on flat and rough hydrophilic and hydrophobic surfaces. We also show advantages and disadvantages of the surfaces of each kind. We demonstrate that water condenses in a liquid form, leading to the formation of a thin continuous water layer on a hydrophilic surface. Meanwhile, separated rounded water droplets are formed on hydrophobic surfaces. As a result of slower heat exchange, the freezing of rounded water droplets on a hydrophobic surface occurs later than the freezing of the continuous water layer on a hydrophilic one. Moreover, growth of ice on hydrophobic surfaces is slower than on the hydrophilic ones, because ice grows due to the condensation of water vapor on already formed ice crystals, and not due to the condensation on the polymer surface. Rough hydrophobic surfaces also demonstrate a very low ice adhesion value, which is because of the reduced contact area with ice. The main disadvantage of hydrophobic and superhydrophobic surfaces is the pinning of water droplets on them after thawing. Flat hydrophilic poly(ethylene glycol)-modified surfaces also exhibit very low ice adhesion, which is due to the very low freezing point of the water-poly(ethylene glycol) mixtures. Water easily leaves from flat hydrophilic poly(ethylene glycol)-modified surfaces, and they quickly become dry. However, the ice growth rate on poly(ethylene glycol)-modified hydrophilic surfaces is the highest. These results indicate that neither purely (super)hydrophobic polymeric surfaces, nor "antifreeze" hydrophilic ones provide an ideal solution to the problem of icing.

Programmable patterning of protein bioactivity by visible light.

The simple and quick patterning of functional proteins on engineered surfaces affords an opportunity to fabricate protein microarrays in lab-on-chip systems. We report on the programmable patterning of proteins as well as the local activation of enzymes by visible light. We successfully generated functional protein patterns with different geometries in situ and demonstrated the specific patterning of multiple kinds of proteins side-by-side without the need for specific linker molecules or elaborate surface preparation.

Smart core-shell microgel support for acetyl coenzyme A synthetase: a step toward efficient synthesis of polyketide-based drugs.

The flexibility in tuning the structure and charge properties of PNIPAm microgels during their synthesis makes them a suitable choice for various biological applications. Two-step free radical polymerization, a common method employed for synthesis of core-shell microgel has been well adopted to obtain cationic poly(N-isopropylacrylamide-aminoethyl methacrylate) (PNIPAm-AEMA) shell and PNIPAm core. Scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, and ninhydrin assay suggests nearly monodispersed particles of cationic nature. Amino groups on the microgel provides suitable attachment point for covalent immobilization of acetyl coenzyme A synthetase (Acs) via 1-ethyl-3-(3-N,N- dimethylaminopropyl) carbodiimide (EDC) chemistry. On immobilization, 61.55% of initial activity of Acs has been retained, while Michaelis-Menten kinetics of the immobilized Acs indicates identical K(m) (Michaelis constant) but decrease in the V(max) (maximum substrate conversion rate) compared to free enzyme. Immobilized Acs shows an improvement in activity at wide temperature and pH range and also demonstrates good thermal, storage, and operational stability. The Acs-microgel bioconjugate has been successfully reused for four consecutive operation cycles with more than 50% initial activity.

Stimuli-responsive microjets with reconfigurable shape.

Flexible thermoresponsive polymeric microjets are formed by the self-folding of polymeric layers containing a thin Pt film used as catalyst for self-propulsion in solutions containing hydrogen peroxide. The flexible microjets can reversibly fold and unfold in an accurate manner by applying changes in temperature to the solution in which they are immersed. This effect allows microjets to rapidly start and stop multiple times by controlling the radius of curvature of the microjet. This work opens many possibilities in the field of artificial nanodevices, for fundamental studies on self-propulsion at the microscale, and also for biorelated applications.

Highly efficient phase boundary biocatalysis with enzymogel nanoparticles.

The enzymogel nanoparticle made of a magnetic core and polymer brush shell demonstrates a novel type of remote controlled phase-boundary biocatalysis that involves remotely directed binding to and engulfing insoluble substrates, high mobility, and stability of the catalytic centers. The mobile enzymes reside in the polymer brush scaffold and shuttle between the enzymogel interior and surface of the engulfed substrate in the bioconversion process. Biocatalytic activity of the mobile enzymes is preserved in the enzymogel while the brush-like architecture favors the efficient interfacial interaction when the enzymogel spreads over the substrate and extends substantially the reaction area as compared with rigid particles.

Biofabrication of Composite Bioink-Nanofiber Constructs: Effect of Rheological Properties of Bioinks on 3D (Bio)Printing and Cells Interaction with Aligned Touch Spun Nanofibers.

This paper reports on a novel approach for the fabrication of composite multilayered bioink-nanofibers construct. This work achieves this by using a hands-free 3D (bio)printing integrated touch-spinning approach. Additionally, this work investigates the interaction of fibroblasts in different bioinks with the highly aligned touch-spun nanofibers. This work conducts a comprehensive characterization of the rheological properties of the inks, starting with low-strain oscillatory rheology to analyze the viscoelastic behavior, when the material structure remains intact. Moreover, this work performs amplitude sweeps to investigate the stability of the inks under large deformations, rotational rheology to examine the shear thinning profile, and a three-step creep experiment to study time-dependent rheological behavior. The obtained rheological results are correlated to visual observation of the flow behavior of inks. These behaviors span from an ink with zero-shear viscosity, very weak shear thinning, and no thixotropic behavior to inks exhibiting flow stress, pronounced shear thinning, and thixotropy. It is demonstrated that inks have an essential effect on cell behavior. While all bioinks allow a preferred directionality of the fibroblasts along the fiber direction, cells tend to form aggregates in bioinks with higher viscosity, and a considerable number of agglomerates are observed in the presence of laponite-RD.

Modular photoorigami-based 4D manufacturing of vascular junction elements.

Four-dimensional (4D) printing, combining three-dimensional (3D) printing with time-dependent stimuli-responsive shape transformation, eliminates the limitations of the conventional 3D printing technique for the fabrication of complex hollow constructs. However, existing 4D printing techniques have limitations in terms of the shapes that can be created using a single shape-changing object. In this paper, we report an advanced 4D fabrication approach for vascular junctions, particularly T-junctions, using the 4D printing technique based on coordinated sequential folding of two or more specially designed shape-changing elements. In our approach, the T-junction is split into two components, and each component is 4D printed using different synthesized shape memory polyurethanes and their nanohybrids, which have been synthesized with varying hard segment contents and by incorporating different weight percentages of photo-responsive copper sulfide-polyvinyl pyrrolidone nanoparticles. The formation of a T-junction is demonstrated by assigning different shape memory behaviors to each component of the T-junction. A cell culture study with human umbilical vein endothelial cells reveals that the cells proliferate over time, and almost 90% of cells remain viable on day 7. Finally, the formation of the T-junction in the presence of near-infrared light has been demonstrated after seeding the endothelial cells on the programmed flat surface of the two components and fluorescence microscopy at day 3 and 7 reveals that the cells adhered well and continue to proliferate over time. Hence, the proposed alternative approach has huge potential and can be used to fabricate vascular junctions in the future.

Electrospun cellulose acetate nanofibrous composites for multi-responsive shape memory actuators and self-powered pressure sensors.

Soft actuators and sensors have attracted extensive scientific interest attributed to their great potential applications in various fields, but the integration of actuating and sensing functions in one material is still a big challenge. Here, we developed an electrospun cellulose acetate (CA)/carbon nanotube nanofiborous composite with both functional applications as multi-responsive shape memory actuators and triboelectric nanogenerator (TENG) based sensors. Attributed to excellent thermo- and light-induced shape memory performance, the CA nanofiborous composites showed high heavy-lift capability as light driven actuators, able to lift burdens 1050 times heavier than their own weight. The CA nanofiborous membranes based TENG exhibited high output performance with open-circuit voltage, short-circuit density, and instantaneous power density about 103.2 V, 7.93 mA m-2 and 0.74 W m-2, respectively. The fabricated TENG based pressure sensor exhibited a high sensitivity of 3.03 V kPa-1 below 6.8 kPa and 0.11 V kPa-1 in the pressure range from 6.8 to 65 kPa, which can be effectively used to monitor human motion state and measure wind velocity. It is expected that the electrospun composites with actuating and sensing functions will show prosperous applications prospects in soft robotics.