Modulation of 3D Bioprintability in Polysaccharide Bioink by Bioglass Nanoparticles and Multiple Metal Ions for Tissue Engineering.

BACKGROUND: Bioglasses are used in applications related to bone rehabilitation and repair. The mechanical and bioactive properties of polysaccharides like alginate and agarose can be modulated or improved using bioglass nanoparticles. Further essential metal ions used as crosslinker have the potential to supplement cultured cells for better growth and proliferation. METHOD: In this study, the alginate bioink is modulated for fabrication of tissue engineering scaffolds by extrusion-based 3D bioprinting using agarose, bioglass nanoparticles and combination of essential trace elements such as iron, zinc, and copper. Homogeneous bioink was obtained by in situ mixing and bioprinting of its components with twin screw extruder (TSE) based 3D bioprinting, and then distribution of metal ions was induced through post-printing diffusion of metal ions in the printed scaffolds. The mechanical and 3d bioprinting properties, microscopic structure, biocompatibility of the crosslinked alginate/agarose hydrogels were analyzed for different concentrations of bioglass. The adipose derived mesenchymal stem cells (ADMSC) and osteoblast cells (MC3T3) were used to evaluate this hydrogel's biological performances. RESULTS: The porosity of hydrogels significantly improves with the incorporation of the bioglass. More bioglass concentration results in improved mechanical (compressive, dynamic, and cyclic) and 3D bioprinting properties. Cell growth and extracellular matrix are also enhanced with bioglass concentration. CONCLUSION: For bioprinting of the bioinks, the advanced TSE head was attached to 3D bioprinter and in situ fabrication of cell encapsulated scaffold was obtained with optimized composition considering minimal effects on cell damage. Fabricated bioinks demonstrate a biocompatible and noncytotoxic scaffold for culturing MC3T3 and ADMSC, while bioglass controls the cellular behaviors such as cell growth and extracellular matrix formation.

Study on Bioresponsive Gelatin-Hyaluronic Acid-Genipin Hydrogel for High Cell-Density 3D Bioprinting.

The Development of bioresponsive extrudable hydrogels for 3D bioprinting is imperative to address the growing demand for scaffold design as well as efficient and reliable methods of tissue engineering and regenerative medicine. This study proposed genipin (5 mg) cross-linked gelatin (1 to 1.5 g)-hyaluronic acid (0.3 g) hydrogel bioink (20 mL) tailored for 3D bioprinting. The focus is on high cell loading and a less artificial extra-cellular matrix (ECM) effect, as well as exploring their potential applications in tissue engineering. The bioresponsiveness of these hydrogel scaffolds was successfully evaluated at 37 °C and room temperature (at pH 2.5, 7.4, and 9). The rheological and mechanical properties (more than three times) increased with the increase in gelatin content in the hydrogel; however, the hydrogel with the least amount of gelatin showed the best extrusion capability. This optimized hydrogel's high extrusion ability and post-printing shape fidelity were evident from 3D and four-axis printing of complex structures such as hollow tubes, stars, pyramids, and zigzag porous tubular (four-axis) scaffolds (printed at 90 kPa pressure, 70 mm/s speed, 22G needle, fourth axis rotation of 4 rpm). 3 million/mL MC3T3-E1 mouse osteoblast cells were used in preparing 3D bioprinted samples. The in vitro cell culture studies have been carried out in a CO2 incubator (at 37 °C, 5% CO2). In the cytocompatibility study, almost three times more cell viability was observed in 3 days compared to day 1 control, proving the non-toxicity and cell-supportiveness of these hydrogels. High cell viability and cell-to-cell interactions observed at the end of day 3 using this moderately stable hydrogel in 3D bioprinting exhibit high potential for precise cell delivery modes in tissue engineering as well as regenerative medicine.

3D bioprinting of complex tissue scaffolds with in situ homogeneously mixed alginate-chitosan-kaolin bioink using advanced portable biopen.

Control of in situ 3D bioprinting of hydrogel without toxic crosslinker is ideal for tissue regeneration by reinforcing and homogeneously distributing biocompatible reinforcing agent during fabrication of large area and complex tissue engineering scaffolds. In this study, homogeneous mixing, and simultaneous 3D bioprinting of a multicomponent bioink based on alginate (AL)-chitosan (CH), and kaolin was obtained by an advanced pen-type extruder to ensure structural and biological homogeneity during the large area tissue reconstruction. The static, dynamic and cyclic mechanical properties as well as in situ self-standing printability significantly improved with the kaolin concentration for AL-CH bioink-printed samples due to polymer-kaolin nanoclay hydrogen bonding and cross-linking with less amount of calcium ions. The Biowork pen ensures better mixing effectiveness for the kaolin-dispersed AL-CH hydrogels (evident from computational fluid dynamics study, aluminosilicate nanoclay mapping and 3D printing of complex multilayered structures) than the conventional mixing process. Two different cell lines (osteoblast and fibroblast) introduced during large area multilayered 3D bioprinting have confirmed the suitability of such multicomponent bioinks for in vitro even tissue regeneration. The effect of kaolin to promote uniform growth and proliferation of the cells throughout the bioprinted gel matrix is more significant for this advanced pen-type extruder processed samples.

Nanodiamond enhanced mechanical and biological properties of extrudable gelatin hydrogel cross-linked with tannic acid and ferrous sulphate.

BACKGROUND: The requirements for cell-encapsulated injectable and bioprintable hydrogels are extrusion ability, cell supportive micro-environment and reasonable post-printing stability for the acclimatization of the cells in the target site. Detonation nanodiamond (ND) has shown its potential to improve the mechanical and biological properties of such hydrogels. Enhancing the performance properties of natural biopolymer gelatin-based hydrogels can widen their biomedical application possibilities to various areas including drug delivery, tissue engineering and 3D bioprinting. METHOD: In this study, natural cross-linker tannic acid (TA) is used along with ferrous sulphate (FS) to optimize the swelling and disintegration of extrudable and 3D printable gelatin hydrogels. The amounts of TA and FS are restricted to improve the extrusion ability of the gels in 3D printing. Further, ND particles (detonation type) are dispersed using twin screw extrusion technology to study their effect on mechanical and biological properties of the 3D printing hydrogel. RESULTS: The improved dispersion of ND particles helps to improve compressive strength almost ten times and dynamic modulus three times using 40 mg ND (2% w/w of gelatin). The surface-functional groups of detonation ND also contributed for such improvement in mechanical properties due to higher interaction with the hydrogel matrix. The stability of the hydrogels in water was also improved to 7 days. Four times improvement of the cell growth and proliferation was observed in ND based hydrogel. CONCLUSION: The cell-supportive nature of these moderately stable and extrudable ND dispersed gelatin hydrogels makes them a good candidate for short term regenerative applications of cell-encapsulated injectable hydrogels with better mechanical properties.