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 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.

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.

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.

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.

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.

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.