Multilayered and heterogeneous hydrogel construct printing system with crosslinking aerosol

Area of Science:

• Biotechnology
• Tissue Engineering
• Biofabrication

Background:

• Microextrusion bioprinting enables biomaterial handling but struggles with precise fabrication of complex, multilayered, and heterogeneous constructs due to bioink spreading and mixing.
• Existing methods face challenges in creating stable multi-material structures with distinct interfaces before gelation.

Purpose of the Study:

• To develop an advanced multiple-bioink printing system capable of precise fabrication of complex tissue constructs.
• To overcome the limitations of bioink spreading and mixing in creating stable, heterogeneous, and multilayered bioprinted structures.

Main Methods:

• Integration of a multibarrel nozzle for simultaneous extrusion of multiple bioinks.
• Incorporation of a nebulizer for simultaneous aerosol-based crosslinking of extruded hydrogel bioinks.
• Development of a system for precise printing of constructs with distinct interfaces and cellular compositions.

Main Results:

• The developed system demonstrated improved printing resolution and stability through aerosol-based crosslinking.
• Successfully fabricated a multilayered and heterogeneous construct using four bioinks and three cell types (breast cancer, stromal, vascular endothelial cells).
• The printed biological model allowed for the analysis of cancer cell migration and vascular network formation.

Conclusions:

• The novel multiple-bioink printing system effectively addresses challenges in fabricating complex tissue architectures.
• This technology shows significant potential for recapitulating physiological systems, including cancer microenvironments, with well-defined compartmentalized regions.
• The system is highly efficient for creating complex tissues and their environments with distinct regions for advanced research applications.