Nanocelluloses are nano-scale materials with the advantages of non-toxicity, high surface area, super strength, tunable surface chemistry, biodegradability, biocompatibility, and promoting cell interaction and tissue development. Nanofibrillated cellulose (NFC), also known as cellulose nanofibers (CNF) or cellulose nanofibril, is one structural type of nanocellulosic materials. NFC has an abundance of hydroxyl groups on the fibril surfaces, which leads to strong hydrogen interactions between fibrils and with water molecules, giving viscoelasticity and shear thinning properties that are favorable for 3D bioprinting.
Amerigo Scientific offers GrowInk™ bioinks that are animal-free, biocompatible, ready-to-use and can be mixed directly with cells for 3D bioprinting applications. GrowInk™ bioinks consist primarily of NFC and water, providing a fully defined and customizable matrix. The matrix can be customized by adding molecules, such as growth factors or adhesion proteins, according to the needs of specific cell culture.
GrowInk™ bioinks mimic the in vivo environment that supports cell growth and differentiation. The stiffness of GrowInk™ can be tuned to match the requirements of specific cells. GrowInk™ is stable at room temperature and can be used with or without the cross-linking agent, depending on the application, but all enable cell printing layer-by-layer with high-precision cell positioning.
GrowInk™ bioinks have shear-thinning properties and are easy to print under force, and the gel retains shape and structure once printed. They are especially suitable for extrusion bioprinting with high printing accuracy. GrowInk™ have been used with many commercial and homemade bioprinters such as:
|GrowInk™-N Bioinks||2.5 ml; 5 ml; 10 ml||GrowInk™-N is a natural opaque bioink made of pure NFC, which can be mixed directly with cells for bioprinting applications.|
|GrowInk™-T Bioinks||2.5 ml; 5 ml; 10 ml||GrowInk™-T is a transparent, anionic NFC bioink that can be optionally cross-linked to harden the printed structures.|
|GrowInk™-ALG Bioinks||2.5 ml; 5 ml; 10 ml||GrowInk™-ALG is an NFC based bioink with alginate that can be cross-linked.|
|Property||GrowInk ™-N||GrowInk ™-T||GrowInk ™-ALG|
|Composition||Nanofibrillar cellulose||Anionic nanofibrillar cellulose||Nanofibrillar cellulose plus alginate|
|Visual appearance||Opaque gel||Transparent gel||Opaque gel|
|Endotoxins||< 5 EU/ml||< 5 EU/ml||< 5 EU/ml|
|Cell viability (HepG2)||Tested||Tested||Tested|
|Viscosity (TA Hybrid Rheometer HR-1, frequency flow sweep)||1-55 000 Pas||1-45 000 Pas||1-30 000 Pas|
|pH||6.5 ± 0.5||6.0 ± 0.5||7.0 ± 0.5|
|Storage temperature||4-22 °C||4-22 °C||4-8 °C|
The printability and cell compatibility of GrowInk™-N and GrowInk™-T with the NGB-R™ bioprinter employing a unique laser-assisted bioprinting technology were evaluated. GrowInk™-N and GrowInk™-T were easy to print by laser and were also successfully printed by pneumatic extrusion. Both bioinks remained stable over time. GrowInk™-N was slightly more viscous than GrowInk™-T, which resulted in slightly better structural stability of the printed objects.
Laser-assisted printing and extrusion printing of GrowInk™ were studied to prepare layered fibroblast/keratinocyte cell models to investigate the suitability of the bioinks for printing and tissue engineering applications. Fibroblasts were successfully laser-printed between the extrusion-printed GrowInk™-T layers, and the keratinocytes printed on top of GrowInk™-T showed correct morphology and good viability.
Figure 4. (A) Schematic representation of the 3D model printed with GrowInk™. Gel layers were printed by extrusion and the cells were printed by laser. (B) Histological characterization of the 3D printed model construct. Stained by Goldner, showing the structured morphology of keratinocytes on top of GrowInk™-T.
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