RESOURCES

Abstract
Four-dimensional (4D) printing is a promising technology that provides solutions for compelling needs in various fields. Most of the reported 4D printed systems are based on the temporal shape transformation of printed subjects. Induction of temporal heterogenicity in functions in addition to shape may extend the scope of 4D printing. Herein, we report a 4D printing approach using plant protein (zein) gel inspired by the amyloid fibrils formation mechanism. The printing of zein gel in a specialized layered-Carbopol supporting bath with different water concentrations in an ethanol-water mixture modulates hydrophobic and hydrogen bonding that causes temporal changes in functions. The part of the construct printed in a supporting bath with higher water content exhibits higher drug loading, faster drug release and degradation than those printed in the supporting bath with lower water content. Tri-segment conduit and butterfly-shaped construct with two asymmetrical wings are printed using this system to evaluate biomedical function as nerve conduit and drug delivery system. 4D printed conduits are also effective as a drug-eluting urethral stent in the porcine model. Overall, this study extends the concept of 4D printing beyond shape transformation and presents an approach of fabricating specialized baths for 4D printing that can also be extended to other materials to obtain 4D printed medical devices with translational potential.

Abstract
Manufacturing tubular constructs with tunable porosity can mimic the vascular structure, not only for supplying nutrients and removing metabolites to support long-term 3D cell culture but also for delivering bioactive components and drugs to tissues. There are few reports on the second purpose through 3D printing. In this study, bio-inspired tubular constructs with permeability were achieved using zein-based ink, forming structures with tunable porosity via the 3D printing technique. The parameters, e.g., zein content, with/without the addition of porogen, and drying conditions, were optimized to control the porous structure and porosity of the printed tubes. The inner wall of the resultant tube supported the adhesion of endothelial cells. A perfusion system was designed, and the penetrability of zein-based tubular constructs was demonstrated by the dialysis test. Moreover, perfusion of cell culture media and the anti-cancer drug in cell-laden hydrogels with tubular structure resulted in 3-day of 3D cell culture with a higher survival rate, and the drug was delivered to local cells around the tubular constructs, respectively. This is a new report on the preparation of 3D-printed tubular constructs using zein as the biomaterial inks with tunable porosity and porous structure, providing a general system for 3D cell culture, 3D drugs screening/pharmacokinetics in vitro, and tissue engineering.

Abstract
Bioprinting is a biofabrication technology which allows efficient and large-scale manufacture of 3D cell culture systems. However, the available biomaterials for bioinks used in bioprinting are limited by their printability and biological functionality. Fabricated constructs are often homogeneous and have limited complexity in terms of current 3D cell culture systems comprising multiple cell types. Inspired by the phenomenon that hydrogels can exchange liquids under the infiltration action, infiltration-induced suspension bioprinting (IISBP), a novel printing technique based on a hyaluronic acid (HA) suspension system to modulate the properties of the printed scaffolds by infiltration action, was described in this study. HA served as a suspension system due to its shear-thinning and self-healing rheological properties, simplicity of preparation, reusability, and ease of adjustment to osmotic pressure. Changes in osmotic pressure were able to direct the swelling or shrinkage of 3D printed gelatin methacryloyl (GelMA)-based bioinks, enabling the regulation of physical properties such as fiber diameter, micromorphology, mechanical strength, and water absorption of 3D printed scaffolds. Human umbilical vein endothelial cells (HUVEC) were applied as a cell culture model and printed within cell-laden scaffolds at high resolution and cell viability with the IISBP technique. Herein, the IISBP technique had been realized as a reliable hydrogel-based bioprinting technique, which enabled facile modulation of 3D printed hydrogel scaffolds properties, being expected to meet the scaffolds requirements of a wide range of cell culture conditions to be utilized in bioprinting applications.