Optimizing the decellularization process of an upper limb skeletal muscle; implications for muscle tissue engineering

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Regenerative Medicine

Background:

• Upper limb muscle defects from cancer, trauma, or congenital issues often require reconstructive surgery using flaps.
• Limitations of current flap surgeries include defect size constraints and donor site morbidity.
• Decellularized human skeletal muscle extracellular matrix (ECM) scaffolds present a potential solution for muscle reconstruction.

Purpose of the Study:

• To optimize a decellularization technique for human flexor digitorum superficialis (FDS) muscle to create viable scaffolds.
• To compare three decellularization protocols: detergent-only (DOT), detergent-enzymatic (DET), and non-detergent, non-enzymatic (NDNET).
• To evaluate scaffold quality based on DNA, glycosaminoglycan (GAG), and collagen content, alongside microstructural integrity.

Main Methods:

• Three decellularization protocols were tested on human FDS muscle.
• Qualitative assessment included histological and immunofluorescent staining.
• Quantitative assessment involved measuring DNA, GAG, and collagen content.

Main Results:

• The DOT protocol (2% SDS, 4 hours) successfully removed DNA and preserved scaffold microstructure, collagen, and GAG content.
• The DET and NDNET protocols failed to completely remove DNA after two cycles.
• DOT protocol demonstrated superior decellularization while maintaining ECM integrity.

Conclusions:

• Decellularization of skeletal muscle using a detergent-only technique is feasible for creating muscle scaffolds.
• This approach shows promise for addressing challenges in upper limb muscle reconstruction.
• Further in vivo studies are necessary to validate the efficacy of these decellularized scaffolds in skeletal tissue engineering.