Embedded bio-printing has fostered significant advances toward the fabrication of soft complex tissue-like constructs, by providing a physical support that allows the freeform shape maintenance within the prescribed spatial arrangement, even under gravity force. Current supporting materials still present major drawbacks for up-scaling embedded 3D bio-printing technology towards tissue-like constructs with clinically relevant dimensions. Herein, we report a a cost-effective and widely available supporting material for embedded bio-printing consisting on a continuous pseudo-plastic matrix of xanthan-gum (XG). This natural polisaccharide exhibits peculiar rheological properties that have enabled the rapid generation of complex volumetric 3D constructs with out-of-plane features. The freedom of design within the three orthogonal axes through the independent and controlled bio-printing process opens new opportunities to produce on demand large arbitrary shapes for personalized medicine. Additionally, we have demonstrated the versatile functionality of XG as a photocurable gel reservoir to engineer perfused cell-laden hydrogel constructs, addressing other practical biomedical applications such as in vitro models and organ-on-chip platforms.
16 August 2023 in ACS Applied Materials & Interfaces
3D bioprinting is a versatile technique that allows the fabrication of living tissue analogs through the layer-by-layer deposition of cell-laden biomaterials, viz. bioinks. In this work, composite alginate hydrogel-based bioinks reinforced with curcumin-loaded particles of cellulose esters (CEpCUR) and laden with human keratinocytes (HaCaT) are developed. The addition of the CEpCUR particles, with sizes of 740 ± 147 nm, improves the rheological properties of the inks, increasing their shear stress and viscosity, while preserving the recovery rate and the mechanical and viscoelastic properties of the resulting fully cross-linked hydrogels. Moreover, the presence of these particles reduces the degradation rate of the hydrogels from 26.3 ± 0.8% (ALG) to 18.7 ± 1.3% (ALG:CEpCUR_10%) after 3 days in the culture medium. The 3D structures printed with the ALG:CEpCUR inks reveal increased printing definition and the ability to release curcumin (with nearly 70% of cumulative release after 24 h in PBS). After being laden with HaCaT cells (1.2 × 106 cells mL-1), the ALG:CEpCUR bioinks can be successfully 3D bioprinted, and the obtained living constructs show good dimensional stability and high cell viabilities at 7 days post-bioprinting (nearly 90%), confirming their great potential for application in fields like wound healing.
In this study, alginate nanocomposite hydrogel bioinks reinforced with lysozyme nanofibers (LNFs) were developed. Alginate-LNF (A-LNF) suspensions with different LNF contents (1, 5 and 10 wt.%) were prepared and pre-crosslinked with 0.5% (w/v) CaCl2 to formulate A-LNF inks. These inks exhibit proper shear-thinning behavior and good recovery properties (~90%), with the pre-crosslinking step playing a crucial role. A-LNF fully crosslinked hydrogels (with 2% (w/v) CaCl2) that mimic 3D printing scaffolds were prepared, and it was observed that the addition of LNFs improved several properties of the hydrogels, such as the morphology, swelling and degradation profiles, and mechanical properties. All formulations are also noncytotoxic towards HaCaT cells. The printing parameters and 3D scaffold model were then optimized, with A-LNF inks showing improved printability. Selected A-LNF inks (A-LNF0 and A-LNF5) were loaded with HaCaT cells (cell density 2 × 106 cells mL−1), and the cell viability within the bioprinted scaffolds was evaluated for 1, 3 and 7 days, with scaffolds printed with the A-LNF5 bioink showing the highest values for 7 days (87.99 ± 1.28%). Hence, A-LNF bioinks exhibited improved rheological performance, printability and biological properties representing a good strategy to overcome the main limitations of alginate-based bioinks.
The development of bioink formulations with suitable properties is fundamental for the progress of 3D bioprinting. The potential of cellulose, the most abundant biopolymer, in this realm has often been underestimated, relegating it essentially to a reinforcement additive of bioinks. In this work, cell-laden bioink formulations, composed exclusively of cellulose, viz., "all-cellulose bioinks", were developed by combining carboxymethyl cellulose (CMC) and nanofibrillated cellulose (NFC) in different mass proportions (90/10, 80/20, and 70/30%). The incorporation of NFC increases the printability of the inks (from Pr = 0.7 to 0.9) while maintaining their shear-thinning behavior, and increasing contents of NFC also decrease the degradation rate of the hydrogels after 7 days. The bioprinting of the cell-laden formulations, with HaCaT (keratinocyte) and ATDC5 (chondrogenic) cells, resulted in high (>80%) cell viabilities for up to 7 days, corroborating the versatility of the bioinks and their potential to originate distinct 3D living structures for biomedical applications.
The SLS additive manufacturing industry enables the development of products for diverse applications with distinct properties due to its excellent surface finish and ability to create varied part geometries, but it consumes high-performance materials with high acquisition costs. An extensive quarrying of stone leads to the accumulation of mineral residues, posing environmental hazards by contaminating soil and water when disposed of in landfills. The primary objective of the study was to incorporate mineral waste into the SLS technique and investigate the influence of its addition, along with a silane-based chemical treatment, on the mechanical performance of polymer-mineral composites (PA12-slate). Additionally, the feasibility of producing a highly loaded printed prototype, employing 50 wt% of mineral waste, was examined. Samples of PA12, PA12 blended with 50 wt% slate waste, and slate waste treated with silane underwent fabrication via selective laser sintering (SLS) and subsequent mechanical characterization, including tensile, flexural, and compressive tests. Additionally, the samples underwent accelerated aging using a QUV weathering tester, followed by mechanical characterization. The geometric accuracy, stability, and processing feasibility of these formulations were evaluated through SLS-printed composite prototypes utilizing PA12_50Sla_Si. It was found that the addition of 50% of slate to the PA12 presented mechanical properties decreasing compared to the printed PA12 only. However, an increase was verified when using silane-induced mineral bonding. The incorporation of mineral agents and silane enhanced the resistance of PA12 to aging. However, after aging, both tensile and flexural strength decreased across all printed samples. Nonetheless, this study showcased the feasibility of producing complex PA12-slate waste specimens containing up to 50 wt% of mineral waste using the SLS printing technique. Therefore, SLS presents itself as a viable means of adding value to this mineral waste.