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Achieving Acetylcholine Receptor Clustering in Tissue-Engineered Skeletal Muscle Constructs In vitro through a
John B Scott1, Catherine L Ward2, Benjamin T Corona2
1Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Winston-SalemNC, USA; Virginia Tech - Wake Forest University School of Biomedical Engineering and Sciences, Wake Forest University Biomedical Engineering, Winston-SalemNC, USA.
Frontiers in Pharmacology
|January 27, 2017
Summary
This study developed a biomaterial construct to promote acetylcholine receptor (AChR) clustering on muscle cells, a crucial step for muscle regeneration after volumetric muscle loss (VML) injuries.
Area of Science:
- Biomaterials Science
- Muscle Tissue Engineering
- Regenerative Medicine
Background:
- Volumetric muscle loss (VML) results in permanent deficits with no effective treatments.
- Current muscle constructs lack innervation for long-term function.
Purpose of the Study:
- To develop a proof-of-concept biomaterial construct for inducing acetylcholine receptor (AChR) clustering on muscle-derived cells (MDCs) in vitro.
- To investigate strategies for enhancing AChR clustering for muscle regeneration applications.
Main Methods:
- Microspheres presenting neural agrin were embedded in fibrin hydrogels and applied to muscle cells (C2C12 or primary rat MDCs).
- AChR clustering was assessed based on microsphere-cell contact in 2D and 3D hydrogel constructs, with and without cyclic stretch.
- Agrin was presented via adsorption or covalent coupling to microspheres.
Main Results:
- Agrin-presenting microspheres induced spatially restricted AChR clustering at cell-microsphere contact sites in 2D and suspension conditions.
- Covalently coupled agrin sustained AChR clustering for over 120 hours.
- 3D fibrin constructs alone showed minimal clustering, but cyclic stretch synergistically enhanced AChR clustering with agrin-presenting microspheres.
Conclusions:
- This strategy supports the development of biomaterial constructs that mimic neuromuscular junctions for muscle tissue engineering.
- The findings represent a significant step towards improving the function of engineered muscle for treating VML injuries.
