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.

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.

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.

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.

Ink Material Selection and Optical Design Considerations in DLP 3D Printing.

Digital light processing (DLP) 3D printing has become a powerful manufacturing tool for the fast fabrication of complex functional structures. The rapid progress in DLP printing has been linked to research on optical design factors and ink selection. This critical review highlights the main challenges in the DLP printing of photopolymerizable inks. The kinetics equations of photopolymerization reaction in a DLP printer are solved, and the dependence of curing depth on the process optical parameters and ink chemical properties are explained. Developments in DLP platform design and ink selection are summarized, and the roles of monomer structure and molecular weight on DLP printing resolution are shown by experimental data. A detailed guideline is presented to help engineers and scientists to select inks and optical parameters for fabricating functional structures for multi-material and 4D printing applications.

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.

4D Biofabrication of Mechanically Stable Tubular Constructs Using Shape Morphing Porous Bilayers for Vascularization Application.

This study reports the fabrication of highly porous electrospun self-folding bilayers, which fold into tubular structures with excellent mechanical stability, allowing them to be easily manipulated and handled. Two kinds of bilayers based on biocompatible and biodegradable soft (PCL, polycaprolactone) and hard (PHB, poly-hydroxybutyrate) thermoplastic polymers have been fabricated and compared. Multi-scroll structures with tunable diameter are obtained after the shape transformation of the bilayer in aqueous media, where PCL-based bilayer rolled longitudinally and PHB-based one rolled transversely with respect to the fiber direction. A combination of higher elastic modulus and transverse orientation of fibers with respect to rolling direction allowed precise temporal control of shape transformation of PHB-bilayer - stress produced by swollen methacrylated hyaluronic acid (HA-MA) do not relax with time and folding is not affected by the fact that bilayer is fixed in unfolded state in cell culture medium for more than 1 h. This property of PHB-bilayer allowed cell culturing without a negative effect on its shape transformation ability. Moreover, PHB-based tubular structure demonstrated superior mechanical stability compared to PCL-based ones and do not collapse during manipulations that happened to PCL-based one. Additionally, PHB/HA-MA bilayers showed superior biocompatibility, degradability, and long-term stability compared to PCL/HA-MA.

Self-healable printed magnetic field sensors using alternating magnetic fields.

We employ alternating magnetic fields (AMF) to drive magnetic fillers actively and guide the formation and self-healing of percolation networks. Relying on AMF, we fabricate printable magnetoresistive sensors revealing an enhancement in sensitivity and figure of merit of more than one and two orders of magnitude relative to previous reports. These sensors display low noise, high resolution, and are readily processable using various printing techniques that can be applied to different substrates. The AMF-mediated self-healing has six characteristics: 100% performance recovery; repeatable healing over multiple cycles; room-temperature operation; healing in seconds; no need for manual reassembly; humidity insensitivity. It is found that the above advantages arise from the AMF-induced attraction of magnetic microparticles and the determinative oscillation that work synergistically to improve the quantity and quality of filler contacts. By virtue of these advantages, the AMF-mediated sensors are used in safety application, medical therapy, and human-machine interfaces for augmented reality.

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.

Smart Mechanically Tunable Surfaces with Shape Memory Behavior and Wetting-Programmable Topography.

This paper reports for the first time the fabrication and investigation of wetting properties of structured surfaces formed by lamellae with an exceptionally high aspect ratio of up to 57:1 and more. The lamellar surfaces were fabricated using a polymer with tunable mechanical properties and shape-memory behavior. It was found that wetting properties of such structured surfaces depend on temperature, and thermal treatment history-structured surfaces are wetted easier at elevated temperature or after cooling to room temperature when the polymer is soft because of the easier deformability of lamellae. The shape of lamellae deformed by droplets can be temporarily fixed at low temperature and remains fixed upon heating to room temperature. Heating above the transition temperature of the shape-memory polymer restores the original shape. The high aspect ratio allows tuning of geometry not only manually, as it is done in most works reported previously but can also be made by a liquid droplet and is controlled by temperature. This behavior opens new opportunities for the design of novel smart elements for microfluidic devices such as smart valves, whose state and behavior can be switched by thermal stimuli: valves that can or cannot be opened that are able to close or can be fixed in an open or closed states.

Digital Light Processing Bioprinting Advances for Microtissue Models.

Digital light processing (DLP) bioprinting has been widely introduced as a fast and robust biofabrication method in tissue engineering. The technique holds a great promise for creating tissue models because it can replicate the resolution and complexity of natural tissues and constructs. A DLP system projects 2D images onto layers of bioink using a digital photomask. The resolution of DLP bioprinting strongly depends on the characteristics of the projected light and the photo-cross-linking response of the bioink microenvironment. In this review, we present a summary of DLP fundamentals with a focus on bioink properties, photoinitiator selection, and light characteristics in resolution of bioprinted constructs. A simple guideline is provided for bioengineers interested in using DLP platforms and customizing technical specifications for its design. The literature review reveals the promising future of DLP bioprinting for disease modeling and biofabrication.

Fibrous Scaffolds for Muscle Tissue Engineering Based on Touch-Spun Poly(Ester-Urethane) Elastomer.

Development of fiber-spinning technologies and materials with proper mechanical properties is highly important for the manufacturing of aligned fibrous scaffolds mimicking structure of the muscle tissues. Here, the authors report touch spinning of a thermoplastic poly(1,4-butylene adipate)-based polyurethane elastomer, obtained via solvent-free polymerization. This polymer possesses a combination of important advantages such as 1) low elastic modulus in the range of a few MPa, 2) good recovery ratio and 3) resilience, 4) processability, 5) nontoxicity, 6) biocompatibility, and 7) biodegradability that makes it suitable for fabrication of structures mimicking extracellular matrix of muscle tissue. Touch spinning allows fast and precise deposition of highly aligned micro- and nano-fibers without use of high voltage. C2C12 myoblasts readily align along soft polymer fibers and demonstrate high viability as well as proliferation that make proposed combination of polymer and fabrication method highly suitable for engineering skeletal muscles.

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 Biofabrication Using a Combination of 3D Printing and Melt-Electrowriting of Shape-Morphing Polymers.

We report the fabrication of scroll-like scaffolds with anisotropic topography using 4D printing based on a combination of 3D extrusion printing of methacrylated alginate, melt-electrowriting of polycaprolactone fibers, and shape-morphing of the fabricated object. A combination of 3D extrusion printing and melt-electrowriting allows programmed deposition of different materials and fabrication of structures with high resolution. Shape-morphing allows the transformation of a patterned surface of a printed structure in a pattern on inner surface of a folded object that is used to align cells. We demonstrate that the concentration of calcium ions, the environment media, and the geometrical shape of the scaffold influences shape-morphing that allows it to be efficiently programmed. Myoblasts cultured inside a scrolled bilayer scaffold demonstrate excellent viability and proliferation. Moreover, the patterned surface generated by PCL fibers allow a very high degree of orientation of cells, which cannot be achieved on the alginate layer without fibers.

Soft Elastic Fibrous Scaffolds for Muscle Tissue Engineering by Touch Spinning.

This paper reports an approach for the fabrication of highly aligned soft elastic fibrous scaffolds using touch spinning of thermoplastic polycaprolactone-polyurethane elastomers and demonstrates their potential for the engineering of muscle tissue. A family of polyester-polyurethane soft copolymers based on polycaprolactone with different molecular weights and three different chain extenders such as 1,4-butanediol and polyethylene glycols with different molecular weight was synthesized. By varying the molar ratio and molecular weights between the segments of the copolymer, different physicochemical and mechanical properties were obtained. The polymers possess elastic modulus in the range of a few megapascals and good reversibility of deformation after stretching. The combination of the selected materials and fabrication methods allows several essential advantages such as biocompatibility, biodegradability, suitable mechanical properties (elasticity and softness of the fibers), high recovery ratio, and high resilience mimicking properties of the extracellular matrix of muscle tissue. Myoblasts demonstrate high viability in contact with aligned fibrous scaffolds, where they align along the fibers, allowing efficient cell patterning on top of the structures. Altogether, the importance of this approach is the fabrication of highly oriented fiber constructs that can support the proliferation and alignment of muscle cells for muscle tissue engineering applications.

Shape-Morphing Fibrous Hydrogel/Elastomer Bilayers Fabricated by a Combination of 3D Printing and Melt Electrowriting for Muscle Tissue Regeneration.

This paper reports an approach for the fabrication of shape-changing bilayered scaffolds, which allow the growth of aligned skeletal muscle cells, using a combination of 3D printing of hyaluronic acid hydrogel, melt electrowriting of thermoplastic polycaprolactone-polyurethane elastomer, and shape transformation. The combination of the selected materials and fabrication methods allows a number of important advantages such as biocompatibility, biodegradability, and suitable mechanical properties (elasticity and softness of the fibers) similar to those of important components of extracellular matrix (ECM), which allow proper cell alignment and shape transformation. Myoblasts demonstrate excellent viability on the surface of the shape-changing bilayer, where they occupy space between fibers and align along them, allowing efficient cell patterning inside folded structures. The bilayer scaffold is able to undergo a controlled shape transformation and form multilayer scroll-like structures with cells encapsulated inside. Overall, the importance of this approach is the fabrication of tubular constructs with a patterned interior that can support the proliferation and alignment of muscle cells for muscle tissue regeneration.

4D Biofabrication of fibrous artificial nerve graft for neuron regeneration.

In this paper, we describe the application of the 4D biofabrication approach for the fabrication of artificial nerve graft. Bilayer scaffolds consisting of uniaxially aligned polycaprolactone-poly(glycerol sebacate) (PCL-PGS) and randomly aligned methacrylated hyaluronic acid (HA-MA) fibers were fabricated using electrospinning and further used for the culture of PC-12 neuron cells. Tubular structures form instantly after immersion of fibrous bilayer in an aqueous buffer and the diameter of obtained tubes can be controlled by changing bilayer parameters such as the thickness of each layer, overall bilayer thickness, and medium counterion concentration. Designed scaffolds showed a self-folded scroll-like structure with high stability after four weeks of real-time degradation. The significance of this research is in the fabrication of tuneable tubular nerve guide conduits that can simplify the current existing clinical treatment of neural injuries.

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.

4D biofabrication of skeletal muscle microtissues.

Skeletal muscle is one of the most abundant tissues in the body. Although it has a relatively good regeneration capacity, it cannot heal in the case of disease or severe damage. Many current tissue engineering strategies fall short due to the complex structure of skeletal muscle. Biofabrication techniques have emerged as a popular set of methods for increasing the complexity of tissue-like constructs. In this paper, 4D biofabrication technique is introduced for fabrication of the skeletal muscle microtissues. To this end, a bilayer scaffold consisting of a layer of anisotropic methacrylated alginate fibers (AA-MA) and aligned polycaprolactone (PCL) fibers were fabricated using electrospinning and later induced to self-fold to encapsulate myoblasts. Bilayer mats undergo shape-transformation in an aqueous buffer, a process that depends on their overall thickness, the thickness of each layer and the geometry of the mat. Proper selection of these parameters allowed fabrication of scroll-like tubes encapsulating myoblasts. The myoblasts were shown to align along the axis of the anisotropic PCL fibers and further differentiated into aligned myotubes that contracted under electrical stimulation. Overall the significance of this approach is in the fabrication of hollow tubular constructs that can be further developed for the formation of a vascularized and functional muscle.