Autonomously Self-Adhesive Hydrogels as Building Blocks for Additive Manufacturing.

We report a simple method of preparing autonomous and rapid self-adhesive hydrogels and their use as building blocks for additive manufacturing of functional tissue scaffolds. Dynamic cross-linking between 2-aminophenylboronic acid-functionalized hyaluronic acid and poly(vinyl alcohol) yields hydrogels that recover their mechanical integrity within 1 min after cutting or shear under both neutral and acidic pH conditions. Incorporation of this hydrogel in an interpenetrating calcium-alginate network results in an interfacially stiffer but still rapidly self-adhesive hydrogel that can be assembled into hollow perfusion channels by simple contact additive manufacturing within minutes. Such channels withstand fluid perfusion while retaining their dimensions and support endothelial cell growth and proliferation, providing a simple and modular route to produce customized cell scaffolds.

Silicon Carbide Nanoparticles as an Effective Bioadhesive to Bond Collagen Containing Composite Gel Layers for Tissue Engineering Applications.

Additive manufacturing via layer-by-layer adhesive bonding holds much promise for scalable manufacturing of tissue-like constructs, specifically scaffolds with integrated vascular networks for tissue engineering applications. However, there remains a lack of effective adhesives capable of composite layer fusion without affecting the integrity of patterned features. Here, the use of silicon carbide is introduced as an effective adhesive to achieve strong bonding (0.39 ± 0.03 kPa) between hybrid hydrogel films composed of alginate and collagen. The techniques have allowed us to fabricate multilayered, heterogeneous constructs with embedded high-resolution microchannels (150 µm-1 mm) that are precisely interspaced (500-600 µm). Hydrogel layers are effectively bonded with silicon carbide nanoparticles without blocking the hollow microchannels and high cell viability (90.61 ± 3.28%) is maintained within the scaffold. Nanosilica is also tested and found to cause clogging of smaller microchannels when used for interlayer bonding, but is successfully used to attach synthetic polymers (e.g., Tygon) to the hydrogels (32.5 ± 2.12 mN bond strength). This allows us to form inlet and outlet interconnections to the gel constructs. This ability to integrate hollow channel networks into bulk soft material structures for perfusion can be useful in 3D tissue engineering applications.

3D bioprinting of heterogeneous bi- and tri-layered hollow channels within gel scaffolds using scalable multi-axial microfluidic extrusion nozzle.

One of the primary focuses in recent years in tissue engineering has been the fabrication and integration of vascular structures into artificial tissue constructs. However, most available methodologies lack the ability to create multi-layered concentric conduits inside natural extracellular matrices (ECMs) and gels that replicate more accurately the hierarchical architecture of biological blood vessels. In this work, we present a new microfluidic nozzle design capable of multi-axial extrusion in order to 3D print and pattern bi- and tri-layered hollow channel structures. This nozzle allows, for the first time, for these structures to be embedded within layers of gels and ECMs in a fast, simple and low-cost manner. By varying flow rates (1-6 ml min-1), printspeeds (1-16 m min-1), and material concentration (25-175 mM and 1.5%-2.5% for calcium chloride and alginate, respectively) we are able to accurately determine the operational printing range as well as achieve a wide range of conduit dimensions (0.69-2.31 mm) that can be printed within a few seconds. Our scalable design allows for multi-axial extrusion and versatility in material incorporation in order to create heterogeneous structures. We demonstrate the ability to print distinct concentric layers of different cell types, namely endothelial cells and fibroblasts. By incorporating various layers of different cell-friendly materials (such as collagen and fibrin) alongside materials with high mechanical strength (i.e. alginate), we were able to increase long-term cell viability and growth without compromising the structural integrity. In this way, we can improve cellular adhesion in our biocompatible constructs as well as allow them to remain structurally sound. We are able to realize complex heterogeneous, hierarchical architectures that have strong potential for use not only in vascular tissue applications, but also in other artificially fabricated tubular or fiber-like structures such as skeletal muscle or nerve conduits.