Three-dimensional bioprinting is an emerging technology for fabricating living 3D constructs, and it has shown great promise in tissue engineering. Bioinks are scaffold materials mixed with cells used by 3D bioprinting to form a required cell-laden structure. In this paper, a novel bioink made of gelatin methacrylamide (GelMA) and collagen (Col) doped with tyrosinase (Ty) is presented for the 3D bioprinting of living skin tissues. Ty has the dual function of being an essential bioactive compound in the skin regeneration process and also as an enzyme to facilitate the crosslink of Col and GelMA. Further, enzyme crosslinking together with photocrosslinking can enhance the mechanical strength of the bioink. The experimental results show that the bioink is able to form stable 3D living constructs using the 3D bioprinting process. The cell culture shows that three major cell lines: human melanocytes (HEM), human keratinocytes (HaCat) and human dermal fibroblasts (HDF) exhibit high cell viabilities. The viability of these three cell lines is above 90%. The proliferation and scratching test show that Ty can enhance the proliferation of HEM, inhibit the growth and migration of HDF and not affect HaCat significantly. Animal tests show that the doped bioinks for 3D bioprinting can help form an epidermis and dermis, and thus have high potential as a skin bioink.

Hyaluronic acid (HA)-based hydrogels are widely used in biomedical applications due to their excellent biocompatibility. HA can be Ultraviolet (UV)-crosslinked by modification with methacrylic anhydride (HA-MA) and crosslinked by modification with 3,3′-dithiobis(propionylhydrazide) (DTP) (HA-SH) via click reaction. In the study presented in this paper, a 3D-bioprinted, double-crosslinked, hyaluronic-acid-based hydrogel for wound dressing was proposed. The hydrogel was produced by mixing HA-MA and HA-SH at different weight ratios. The rheological test showed that the storage modulus (G’) of the HA-SH/HA-MA hydrogel increased with the increase in the HA-MA content. The hydrogel had a high swelling ratio and a high controlled degradation rate. The in vitro degradation test showed that the hydrogel at the HA-SH/HA-MA ratio of 9:1 (S9M1) degraded by 89.91% ± 2.26% at 11 days. The rheological performance, drug release profile and the cytocompatibility of HA-SH/HA-MA hydrogels with loaded Nafcillin, which is an antibacterial drug, were evaluated. The wound dressing function of this hydrogel was evaluated by Live/Dead staining and CCK-8 assays. The foregoing results imply that the proposed HA-SH/HA-MA hydrogel has promise in wound repair applications.

In this review, we focused on a few obstacles that hinder three-dimensional (3D) bioprinting process in tissue engineering. One of the obstacles is the bioinks used to deliver cells. Hydrogels are the most widely used bioink materials; however, they are mechanically weak in nature and cannot meet the requirements for supporting structures, especially when the tissues, such as cartilage, require extracellular matrix to be mechanically strong. Secondly and more importantly, tissue regeneration is not only about building all the components in a way that mimics the structures of living tissues, but also about how to make the constructs function normally in the long term. One of the key issues is sufficient nutrient and oxygen supply to the engineered living constructs. The other is to coordinate the interplays between cells, bioactive agents and extracellular matrix in a natural way. This article reviews the approaches to improve the mechanical strength of hydrogels and their suitability for 3D bioprinting; moreover, the key issues of multiple cell lines coprinting with multiple growth factors, vascularization within engineered living constructs etc. were also reviewed.

A PLA/o-nitrobenzyl based scaffold was designed and fabricated by 3D fiber deposition to demonstrate the feasibility of photo-regulated hydrolytic degradation in vitro. It promises to approach the matched degradation with new tissues when applied in tissue engineering.

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