Modular enzymatically crosslinked protein polymer hydrogels for in situ gelation

Area of Science:

• Biomaterials Science
• Polymer Chemistry
• Tissue Engineering

Background:

• Biomaterials mimicking the extracellular matrix (ECM) can guide cell fate.
• Controlling biomaterial properties is crucial for recapitulating instructive cellular signals.

Purpose of the Study:

• To develop modular protein polymer-based hydrogels using enzymatic crosslinking.
• To investigate the impact of hydrogel composition on mechanical properties and cell behavior.
• To assess the potential of these hydrogels for in situ therapies and tissue engineering.

Main Methods:

• Protein polymers with lysine or glutamine residues were engineered.
• Enzymatic crosslinking was achieved using tissue transglutaminase (tTG) or human transglutaminase (hTG).
• Particle tracking microrheology was used to determine crosslinking kinetics and mechanical properties.

Main Results:

• Rapid hydrogel formation (2 min) was achieved with tTG under physiological conditions.
• Hydrogel composition significantly influenced elastic modulus, microstructure, and swelling.
• Cultured mouse and human fibroblasts showed viability and spreading in both 2D and 3D cultures.
• Degradation by plasmin was not significantly affected by hydrogel composition.

Conclusions:

• Modular protein polymer hydrogels can be rapidly formed via enzymatic crosslinking.
• Tunable mechanical and physical properties are achievable by altering precursor composition.
• These biomaterials support cell viability and function, demonstrating potential for regenerative medicine and in situ therapies.