Hydrogel Bioinks of Alginate and Curcumin-Loaded Cellulose Ester-Based Particles for the Biofabrication of Drug-Releasing Living Tissue Analogs.

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.

Nanofibrillated cellulose/gellan gum hydrogel-based bioinks for 3D bioprinting of skin cells.

The development of suitable bioinks is an important research topic in the field of three-dimensional (3D) bioprinting. Herein, novel hydrogel-based bioinks composed of nanofibrillated cellulose (NFC) and gellan gum (GG) in different NFC/GG mass proportions (90:10, 80:20, 70:30, and 60:40) were developed and characterized. The increase in the content of GG, as well as its combination with NFC, enhanced their rheological properties, increasing both storage (G') and loss (G") moduli and the G' recovery capacity of the hydrogels (from 70.05 ± 3.06 % (90:10) to 82.63 ± 1.21 % (60:40)), as well as their mechanical properties, increasing the compressive stiffness and stress from 114.02 ± 10.93 Pa (90:10) to 337.16 ± 34.03 Pa (60:40) and from 18.27 ± 1.32 kPa (90:10) to 47.17 ± 3.59 kPa (60:40), respectively. The hydrogels were non-cytotoxic against human keratinocyte cells (HaCaT), with cell viabilities above 70 % for up to 72 h. The hydrogel 60:40 was loaded with HaCaT cells (3 × 106 cells mL-1) and bioprinted. The cell viability was maintained elevated until day 7 (90 ± 3 %) after bioprinting. These results highlight that the combination of these two biopolymers was a good strategy for the development of novel hydrogel-based bioinks for extrusion 3D bioprinting applications.

Freeform 3D printing using a continuous viscoelastic supporting matrix.

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.