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Traceless Photopolymerization with Non-Pulsed Red Light Enables 3D-Printable Cell-Laden Hydrogels.

Ali Eftekhari¹, Kelsey Rianne de Graaf^1,2, Ekaterina Takmakova³

¹Faculty of Engineering and Natural Sciences, Tampere University, Tampere, 33720, Finland.

Advanced Materials (Deerfield Beach, Fla.)

|May 16, 2025

Summary

A new traceless photoinitiating system using methylene blue enables colorless, red-light photocrosslinking of hydrogels. This advancement overcomes limitations of colored photoinitiators, paving the way for transparent biomaterials in tissue engineering and minimally invasive applications.

Keywords:

3D printing colorless gelatine methacrylate (GelMA)hydrogels methylene blue photopolymerization red light

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Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

Published on: September 29, 2016

Area of Science:

Biomaterials Science
Polymer Chemistry
Tissue Engineering

Background:

Red-light photocrosslinking of hydrogels offers advantages in biocompatibility and tissue penetration over UV methods.
Existing red-light systems result in colored hydrogels due to photoinitiators, limiting applications requiring optical transparency.
Photoinitiator cytotoxicity is a significant concern for biomedical applications.

Purpose of the Study:

To develop a "traceless" photoinitiating system for red-light hydrogel fabrication that eliminates residual color and photoinitiator concerns.
To demonstrate the successful polymerization of colorless, transparent hydrogels using this novel system.
To evaluate the system's suitability for 3D bioprinting and tissue engineering applications.

Main Methods:

Development of a photoinitiating system using FDA-approved methylene blue and triethanolamine.
Photopolymerization of gelatin methacrylate hydrogel using 625 nm red light irradiation.
Characterization of hydrogel properties, including color, stiffness, and biocompatibility.
Application in extrusion-based 3D bioprinting with NIH-3T3 fibroblasts and subsequent photocuring.

Main Results:

Successful fabrication of permanently colorless and transparent gelatin methacrylate hydrogels under ambient conditions.
Demonstrated light-dependent control over hydrogel stiffness.
Successful 3D bioprinting of cell-laden constructs with NIH-3T3 fibroblasts, showing cell proliferation and adhesion.
Red-light polymerization achieved through at least 5 mm of biological tissue.

Conclusions:

The developed traceless photoinitiating system enables the production of optically transparent, red-light crosslinked hydrogels.
This system addresses key limitations of existing methods, including color and cytotoxicity concerns.
The technology holds significant potential for advanced applications in tissue engineering, biosensing, and minimally invasive medical devices.