Preprint: Triggered sequential viral-transduction from collagen-based scaffolds for tissue regeneration

Area of Science:

• Biomaterials Science
• Regenerative Medicine
• Gene Therapy

Background:

• Chronic wounds present significant healing challenges due to dysregulated signaling.
• Traditional treatments often fail, necessitating advanced therapeutic strategies.
• Gene therapy using adeno-associated virus (AAV) is a promising approach for modulating wound healing.

Purpose of the Study:

• To develop a triggered, sequential release system for viral vectors within a tissue engineering scaffold.
• To address the stalled inflammatory phase in chronic wounds by controlling signaling pathways.
• To utilize a collagen-glycosaminoglycan (GAG) scaffold for enhanced wound healing.

Main Methods:

• Integration of two alginate pockets within a collagen-GAG scaffold to house and control AAV release.
• Development of a system for sustained diffusion of one AAV and ultrasound-triggered release of a second AAV.
• In vitro characterization using a reporter AAV and HEK293T cells, followed by demonstration with AAVs encoding angiogenesis genes.

Main Results:

• Sustained reporter gene expression over 9 days with high AAV loading in vitro.
• Ultrasound-triggered release successfully transduced cells at lower AAV loading.
• Demonstrated sequential release of AAVs encoding clinically relevant genes for angiogenesis.

Conclusions:

• The developed scaffold system enables controlled, sequential delivery of gene therapy vectors.
• This technology has potential for broad application in managing chronic wounds by restoring coordinated signaling.
• The system can be adapted to mimic various biological pathways for diverse therapeutic applications.