Embedded 3D Printing of Cryogel-Based Scaffolds.

Cryogel-based scaffolds have attracted great attention in tissue engineering due to their interconnected macroporous structures. However, three-dimensional (3D) printing of cryogels with a high degree of precision and complexity is a challenge, since the synthesis of cryogels occurs under cryogenic conditions. In this study, we demonstrated the fabrication of cryogel-based scaffolds for the first time by using an embedded printing technique. A photo-cross-linkable gelatin methacryloyl (GelMA)-based ink composition, including alginate and photoinitiator, was printed into a nanoclay-based support bath. The layer-by-layer extruded ink was held in complex and overhanging structures with the help of pre-cross-linking of alginate with Ca2+ present in the support bath. The printed 3D structures in the support bath were frozen, and then GelMA was cross-linked at a subzero temperature under UV light. The printed and cross-linked structures were successfully recovered from the support bath with an integrated shape complexity. SEM images showed the formation of a 3D printed scaffold where porous GelMA cryogel was integrated between the cross-linked alginate hydrogels. In addition, they showed excellent shape recovery under uniaxial compression cycles of up to 80% strain. In vitro studies showed that the human fibroblast cells attached to the 3D printed scaffold and displayed spread morphology with a high proliferation rate. The results revealed that the embedded 3D printing technique enables the fabrication of cytocompatible cryogel based scaffolds with desired morphology and mechanical behavior using photo-cross-linkable bioink composition. The properties of the cryogels can be modified by varying the GelMA concentration, whereby various shapes of scaffolds can be fabricated to meet the specific requirements of tissue engineering applications.

3D bioprinting of tyramine modified hydrogels under visible light for osteochondral interface.

Recent advancements in tissue engineering have demonstrated a great potential for the fabrication of three-dimensional (3D) tissue structures such as cartilage and bone. However, achieving structural integrity between different tissues and fabricating tissue interfaces are still great challenges. In this study, anin situcrosslinked hybrid, multi-material 3D bioprinting approach was used for the fabrication of hydrogel structures based on an aspiration-extrusion microcapillary method. Different cell-laden hydrogels were aspirated in the same microcapillary glass and deposited in the desired geometrical and volumetric arrangement directly from a computer model. Alginate and carboxymethyl cellulose were modified with tyramine to enhance cell bioactivity and mechanical properties of human bone marrow mesenchymal stem cells-laden bioinks. Hydrogels were prepared for extrusion by gelling in microcapillary glass utilizing anin situcrosslink approach with ruthenium (Ru) and sodium persulfate photo-initiating mechanisms under visible light. The developed bioinks were then bioprinted in precise gradient composition for cartilage-bone tissue interface using microcapillary bioprinting technique. The biofabricated constructs were co-cultured in chondrogenic/osteogenic culture media for three weeks. After cell viability and morphology evaluations of the bioprinted structures, biochemical and histological analyses, and a gene expression analysis for the bioprinted structure were carried out. Analysis of cartilage and bone formation based on cell alignment and histological evaluation indicated that mechanical cues in conjunction with chemical cues successfully induced MSC differentiation into chondrogenic and osteogenic tissues with a controlled interface.

Bisulfite-initiated crosslinking of gelatin methacryloyl hydrogels for embedded 3D bioprinting.

Recent studies on three-dimensional (3D) bioprinting of cell-laden gelatin methacryloyl (GelMA) hydrogels have provided promising outcomes for tissue engineering applications. However, the reliance on the use of photo-induced gelation processes for the bioprinting of GelMA and the lack of an alternative crosslinking process remain major challenges for the fabrication of cell-laden structures. Here, we present a novel crosslinking approach to form cell-laden GelMA hydrogel constructs through 3D embedded bioprinting without using any external irradiation that could drastically affect cell viability and functionality. This approach consists of a one-step type of crosslinking via bisulfite-initiated radical polymerization, which is combined with embedded bioprinting technology to improve the structural complexity of printed structures. By this means, complex-shaped hydrogel bio-structures with cell viability higher than 90% were successfully printed within a support bath including sodium bisulfite. This study offers an important alternative to other photo-induced gelation processes to improve the bio-fabrication of GelMA hydrogel with high cell viability.

Solvent-Free UV Polymerization of n-Octadecyl Acrylate in Butyl Rubber: A Simple Way to Produce Tough and Smart Polymeric Materials at Ambient Temperature.

One of the most fascinating challenges in recent years has been to produce mechanically robust and tough polymers with smart functions such as self-healing and shape-memory behavior. Here, we report a simple and versatile strategy for the preparation of a highly tough and highly stretchable interconnected interpenetrating polymer network (c-IPN) based on butyl rubber (IIR) and poly(n-octadecyl acrylate) (PC18A) with thermally induced healing and shape-memory functions. Solvent-free UV polymerization of n-octadecyl acrylate (C18A) at 30 ± 2 °C in the presence of IIR leads to IIR/PC18A c-IPNs with sea-island or co-continuous morphologies depending on their IIR contents. The lamellar crystals with a melting temperature Tm of 51-52 °C formed by side-by-side packed octadecyl (C18) side chains are responsible for more than 99% of effective cross-links in c-IPNs, the rest being hydrophobic associations and chemical cross-links. The c-IPNs exhibit varying stiffness (9-34 MPa), stretchability (72-740%), and a significantly higher toughness (1.9-12 MJ·m-3) than their components, which can be tuned by changing the IIR/PC18A weight ratio. The properties of c-IPNs could also be tuned by incorporating a second, noncrystallizable hydrophobic monomer, namely, lauryl methacrylate (C12M), in the melt mixture. We show that the lamellar clusters acting as sacrificial bonds break at the yield point by dissipation of energy, while the ductile amorphous continuous phase keeps the structure together, leading to the toughness improvement of c-IPNs. They exhibit a two-step healing process with >90% healing efficiency with respect to the modulus and a complete shape-recovery ratio induced by heating above Tm of alkyl crystals. The temperature-induced healing occurs via a quick step where C18 bridges form between the damaged surfaces followed by a slow step controlled by the interdiffusion of C18A segments in the bulk. We also show that the strategy developed here is suitable for a variety of rubbers and n-alkyl (meth)acrylates of various side-chain lengths.