Jammed Micro-Flake Hydrogel for Four-Dimensional Living Cell Bioprinting.

4D bioprinting is promising to build cell-laden constructs (bioconstructs) with complex geometries and functions for tissue/organ regeneration applications. The development of hydrogel-based 4D bioinks, especially those allowing living cell printing, with easy preparation, defined composition, and controlled physical properties is critically important for 4D bioprinting. Here, a single-component jammed micro-flake hydrogel (MFH) system with heterogeneous size distribution, which differs from the conventional granular microgel, has been developed as a new cell-laden bioink for 4D bioprinting. This jammed cytocompatible MFH features scalable production and straightforward composition with shear-thinning, shear-yielding, and rapid self-healing properties. As such, it can be smoothly printed into stable 3D bioconstructs, which can be further cross-linked to form a gradient in cross-linking density when a photoinitiator and a UV absorber are incorporated. After being subject to shape morphing, a variety of complex bioconstructs with well-defined configurations and high cell viability are obtained. Based on this system, 4D cartilage-like tissue formation is demonstrated as a proof-of-concept. The establishment of this versatile new 4D bioink system may open up a number of applications in tissue engineering.

Induction of 4D spatiotemporal geometric transformations in high cell density tissues via shape changing hydrogels.

Developing and healing tissues begin as a cellular condensation. Spatiotemporal changes in tissue geometry, transformations in the spatial distribution of the cells and extracellular matrix, are essential for its evolution into a functional tissue. 4D materials, 3D materials capable of geometric changes, may have the potential to recreate the aforementioned biological phenomenon. However, most reported 4D materials are non-degradable and/or not biocompatible, which limits their application in regenerative medicine, and to date there are no systems controlling the geometry of high density cellular condensations and differentiation. Here, we describe 4D high cell density tissues based on shape-changing hydrogels. By sequential photocrosslinking of oxidized and methacrylated alginate (OMA) and methacrylated gelatin (GelMA), bi-layered hydrogels presenting controllable geometric changes without any external stimuli were fabricated. Fibroblasts and human adipose-derived stem cells (ASCs) were incorporated at concentrations up to 1.0 × 108 cells/mL to the 4D constructs, and controllable shape changes were achieved in concert with ASCs differentiated down chondrogenic and osteogenic lineages. Bioprinting of the high density cell-laden OMA and GelMA permitted the formation of more complex constructs with defined 4D geometric changes, which may further expand the promise of this approach in regenerative medicine applications.

Individual cell-only bioink and photocurable supporting medium for 3D printing and generation of engineered tissues with complex geometries.

Scaffold-free engineering of three-dimensional (3D) tissue has focused on building sophisticated structures to achieve functional constructs. Although the development of advanced manufacturing techniques such as 3D printing has brought remarkable capabilities to the field of tissue engineering, technology to create and culture individual cell only-based high-resolution tissues, without an intervening biomaterial scaffold to maintain construct shape and architecture, has been unachievable to date. In this report, we introduce a cell printing platform which addresses the aforementioned challenge and permits 3D printing and long-term culture of a living cell-only bioink lacking a biomaterial carrier for functional tissue formation. A biodegradable and photocrosslinkable microgel supporting bath serves initially as a fluid, allowing free movement of the printing nozzle for high-resolution cell extrusion, while also presenting solid-like properties to sustain the structure of the printed constructs. The printed human stem cells, which are the only component of the bioink, couple together via transmembrane adhesion proteins and differentiate down tissue-specific lineages while being cultured in a further photocrosslinked supporting bath to form bone and cartilage tissue with precisely controlled structure. Collectively, this system, which is applicable to general 3D printing strategies, is a paradigm shift for printing of scaffold-free individual cells, cellular condensations and organoids, and may have far reaching impact in the fields of regenerative medicine, drug screening, and developmental biology.

Novel processing of iron-manganese alloy-based biomaterials by inkjet 3-D printing.

The present work provides an assessment of 3-D printed iron-manganese biodegradable scaffolds as a bone scaffold material. Iron-based alloys have been investigated due to their high strength and ability to slowly corrode. Current fabrications of Fe-based materials generate raw material which must be machined into their desired form. By using inkjet 3-D printing, a technique which generates complex, customizable parts from powders mechanically milled Fe-30Mn (wt.%) powder was directly processed into scaffolds. The 3-D printed parts maintained an open porosity of 36.3% and formed a mixed phase alloy of martensitic ε and austenitic γ phases. Electrochemical corrosion tests showed the 3-D printed Fe-Mn to desirably corrode significantly more rapidly than pure iron. The scaffolds exhibited similar tensile mechanical properties to natural bone, which may reduce the risk of stress shielding. Cell viability testing of MC3T3-E1 pre-osteoblast cells seeded directly onto the Fe-Mn scaffolds using the live/dead assay and with cells cultured in the presence of the scaffolds' degradation products demonstrated good in vitro cytocompatibility compared to tissue culture plastic. Cell infiltration into the open pores of the 3-D printed scaffolds was also observed. Based on this preliminary study, we believe that 3-D printed Fe-Mn alloy is a promising material for craniofacial biomaterial applications, and represents an opportunity for other biodegradable metals to be fabricated using this unique method.