Immunomodulation Through Fibroblast-Derived Extracellular Vesicles (EVs) Within 3D Polycaprolactone-Collagen Matrix

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Immunomodulation

Background:

• Extracellular vesicles (EVs) are acellular tools for immune modulation in tissue engineering.
• Human fibroblast-derived EVs show potential for therapeutic applications.
• Scaffold integrity and immune response are critical for regenerative therapies.

Purpose of the Study:

• To investigate fibroblast-derived EVs within a 3D injectable scaffold (PNCOL) for modulating macrophage behavior.
• To assess the impact of EVs on scaffold integrity under inflammatory conditions.
• To evaluate the potential of this cell-free approach for regenerative medicine.

Main Methods:

• Isolation and characterization of human fibroblast-derived EVs.
• Preparation of macrophage-laden PNCOL scaffolds with different conditions (MP, F-MP, EV-MP).
• Assessment of scaffold structure (SEM, Masson's trichrome) and immunomodulatory effects (metabolic assays, gene expression, IHC for M1/M2 markers).

Main Results:

• Fibroblast-derived EVs preserved scaffold structure and enhanced macrophage metabolic activity.
• EV-encapsulated scaffolds showed upregulation of anti-inflammatory markers (TGF-β, CD163, CCL18) and CD206.
• A phenotypic shift towards M2-like macrophages was observed in EV-encapsulated scaffolds.

Conclusions:

• Fibroblast-derived EVs in injectable PCL-collagen scaffolds offer a cell-free method to modulate inflammation.
• This strategy preserves scaffold structure and supports regenerative healing.
• The approach shows promise for immuno-instructive platforms in regenerative medicine.