Engineered Caf1 Constructs Encoding Laminin-332 Motifs: A Novel Approach for Enhanced Cutaneous Wound Healing

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Wound Healing Research

Background:

• Laminin-332 is crucial for keratinocyte adhesion and re-epithelialization during wound healing.
• Loss of laminin-332, as seen in Junctional Epidermolysis Bullosa, impairs wound repair.
• Current laminin applications face stability and cost limitations, necessitating alternative biomaterials.

Purpose of the Study:

• To develop stable Capsular Antigen F1 (Caf1) scaffolds presenting laminin-332 motifs.
• To evaluate the therapeutic potential of these Caf1 scaffolds in promoting epithelial repair.
• To enable clinically translatable wound-healing strategies for normal and laminin-deficient skin.

Main Methods:

• Identified and ranked laminin-332-derived peptide motifs.
• Engineered selected motifs into Caf1 scaffolds and expressed them in E. coli.
• Assessed keratinocyte migration, adhesion, and signaling in vitro and in an ex vivo human skin wound model.

Main Results:

• Caf1-J3 and Caf1-J4 constructs enhanced keratinocyte migration and motility in vitro.
• These scaffolds restored adhesion and modulated signaling in laminin-deficient keratinocytes.
• Ex vivo studies demonstrated promoted re-epithelialization and homogeneous neo-epidermis formation.

Conclusions:

• Short laminin-332 motifs on Caf1 scaffolds mimic and partially restore basement membrane functions.
• Caf1-J3 and Caf1-J4 act as minimal bioactive units restoring keratinocyte adhesion and modulating wound healing pathways.
• This modular platform provides a clinically relevant strategy for chronic wounds and ECM-deficient skin, advancing regenerative therapies.