Recombinant expression and functional characterization of human collagen III fragments

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Molecular Biology

Background:

• Collagen III (COL3) is vital for skin structure and repair.
• Full-length collagen presents stability and expression challenges.
• Microbial production offers a route to stable collagen products.

Purpose of the Study:

• To develop stable, functional collagen products using microbial expression.
• To assess collagen III fragments for mimicking full-length protein activity.
• To create enhanced biomaterials for skin regeneration and wound healing.

Main Methods:

• Extracted functional fragments (30-60 amino acids) from human COL3.
• Selected ten fragments based on melting temperature and net charge.
• Expressed fragments in Escherichia coli, achieving high titers (up to 285 mg/L).
• Assembled fragments into triple helix structures and nanofibers.
• Formulated composite hydrogels using sample-308 with chitosan, hyaluronic acid, and sodium hyaluronate.

Main Results:

• Two soluble collagen fragments were successfully expressed.
• Fragments self-assembled into triple helix structures and nanofibers.
• Composite hydrogels, particularly sample-308/SH, showed a 10-fold increase in mechanical strength.
• The sample-308/SH hydrogel significantly enhanced fibroblast (L929) adhesion, migration, and differentiation.
• Performance surpassed single-fragment hydrogels and commercial collagen I.

Conclusions:

• Microbial expression yielded bioactive COL3 fragments.
• The sample-308/SH composite hydrogel offers superior physical properties.
• This enhanced hydrogel demonstrates significant potential for wound healing applications.
• Developed a novel biomaterial solution for tissue regeneration.