Tailorable Biofunctionalization of Poly(acrylamide) Hydrogels via Firefly Luciferin-Bioinspired Click Ligation...

Area of Science:

• Biomaterials Science
• Cell Biology
• Organic Chemistry

Background:

• Polyacrylamide (PAM) hydrogels are widely used as extracellular matrix mimics for cell-material interaction studies.
• Conventional biofunctionalization methods for PAM hydrogels lack specificity and control over ligand density, impacting experimental reproducibility.
• There is a need for efficient, tunable, and reproducible biofunctionalization strategies for PAM hydrogels.

Purpose of the Study:

• To introduce a novel firefly luciferin-inspired click ligation method for efficient and tunable biofunctionalization of PAM hydrogels.
• To enable precise control over biomolecule density and improve the presentation of cell-adhesive cues.
• To provide a robust alternative to existing methods for engineering PAM hydrogel interfacial properties.

Main Methods:

• Synthesized a novel acrylamide-based comonomer containing cyanobenzothiazole (CBT) moieties.
• Incorporated the CBT comonomer into PAM hydrogels.
• Utilized luciferin click chemistry for rapid (minutes), mild, and light-independent biofunctionalization with N-Cys bearing biomolecules.

Main Results:

• Achieved highly efficient and homogeneous biofunctionalization of PAM hydrogels with controlled biomolecule loading.
• Demonstrated superior performance compared to the sulfo-SANPAH (SS)-based approach in terms of efficiency, homogeneity, and control.
• Observed significantly enhanced cell attachment, spreading, and proliferation due to improved presentation of cell-adhesive cues, confirmed by holotomography.

Conclusions:

• The novel luciferin click ligation provides a robust, efficient, and reproducible method for PAM hydrogel biofunctionalization.
• This approach offers precise control over ligand density while preserving bioactivity, crucial for mechanobiology applications.
• The method establishes a new standard for engineering PAM hydrogels for advanced 2D cell culture platforms and fundamental cell-material interaction studies.