Three-dimensional tailor-made collagen-like proteins hydrogel for tissue engineering applications

Area of Science:

• Biomaterials Science
• Protein Engineering
• Tissue Engineering

Background:

• Collagen-like proteins (CLP) offer tunable properties as alternatives to animal collagen.
• Assembling CLPs into stable 3D hydrogels mimicking extracellular matrix is challenging.

Purpose of the Study:

• To develop stable 3D hydrogels using genetically engineered CLP variants with unnatural amino acids.
• To enhance self-assembly and stability of CLP hydrogels for biomaterial applications.

Main Methods:

• Genetic code engineering to create CLP variants (CLPhyp, CLPdopa) with hydroxyproline and dopa.
• Genipin cross-linking to induce hydrogel formation.
• Spectroscopic, rheological, and DSC analyses to characterize hydrogel properties and stability.

Main Results:

• Engineered CLPhyp and CLPdopa variants facilitated hydrogel formation with enhanced intermolecular interactions.
• Hydrogels maintained native triple-helical structure and viscoelastic properties.
• CLPhyp and CLPdopa variants increased scaffold pore size, improving cell interaction.
• CLP hydrogels exhibited enhanced stability and resistance to collagenase, particularly CLPdopa.

Conclusions:

• This study presents the first 3D biocompatible hydrogels synthesized by tailoring CLP sequences with non-natural amino acids.
• Tunable CLP hydrogels represent a novel class of synthetic protein-based biomaterials for tissue engineering.
• Engineered CLP hydrogels offer promising platforms for developing advanced biocompatible 3D biomaterials.