In Situ 3D-Printing using a Bio-ink of Protein-photosensitizer Conjugates for Single-cell Manipulation

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Biotechnology

Background:

• Living tissues dynamically adapt their structure and function via microenvironmental interactions.
• In vitro manipulation of cellular functions is limited by the lack of dynamic 3D material systems.
• Controlling the spatiotemporal cellular microenvironment is crucial for biological applications.

Purpose of the Study:

• To develop a dynamic material system for precise control over the 3D cellular microenvironment.
• To demonstrate an in situ 3D-bioprinting technique for fabricating protein scaffolds.
• To investigate the potential of this system for directing cell fate.

Main Methods:

• Utilized multiphoton lithography for in situ 3D-printing.
• Developed a biocompatible bio-ink composed of protein-photosensitizer conjugates (rose bengal and bovine serum albumin).
• Leveraged singlet oxygen generation for precise protein gelation and scaffold fabrication with submicrometer resolution.

Main Results:

• Achieved submicrometer-scale precision in fabricating protein gels.
• Demonstrated improved cytocompatibility and gelation efficiency due to the properties of the protein-photosensitizer conjugates.
• Successfully microfabricated protein scaffolds and controlled single-cell behavior using 3D-bioprinting in the presence of cells.

Conclusions:

• The developed dynamic material system enables precise control over the 3D cellular microenvironment.
• This 3D-bioprinting technique offers a promising platform for advanced tissue engineering.
• The system holds potential for significant applications in drug discovery and regenerative medicine.