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Functional evaluation of nerve-skeletal muscle constructs engineered in vitro
Lisa M Larkin1, Jack H Van der Meulen, Robert G Dennis
1Department of Biomedical Engineering, Division of Geriatric Medicine, Muscle Mechanics Laboratory, Institute of Gerontology, University of Michigan, Ann Arbor, Michigan 48109-2007, USA. llarkin@umich.edu
In Vitro Cellular & Developmental Biology. Animal
|June 9, 2006
Summary
Engineered three-dimensional (3-D) nerve-muscle constructs exhibit spontaneous contractions and enhanced force generation. These constructs, featuring functional neuromuscular junctions, can be stimulated via neural extensions, paving the way for advanced tissue engineering applications.
Area of Science:
- Biomedical Engineering
- Neuroscience
- Muscle Physiology
Background:
- Previously engineered three-dimensional (3-D) skeletal muscle constructs exhibited fetal myosin heavy-chain (MHC) composition.
- Functional characterization of these constructs is crucial for advancing tissue engineering.
Purpose of the Study:
- To evaluate the functional characteristics of 3-D skeletal muscle constructs when cocultured with fetal nerve explants.
- To test the hypothesis that neural coculture enhances construct force and promotes adult MHC isoform expression.
Main Methods:
- Co-culturing embryonic spinal cord explants with confluent muscle cells to form 3-D nerve-muscle constructs.
- Immunohistochemical labeling to identify neural extensions and acetylcholine receptor clusters.
- Assessing spontaneous and stimulated contractions (twitch and tetanus) and MHC isoform composition.
Main Results:
- Nerve-muscle coculture resulted in constructs with neural extensions and functional acetylcholine receptor clusters at neuromuscular junctions.
- Cocultured constructs demonstrated spontaneous contractions with increased frequency and force compared to muscle-only constructs.
- Field stimulation revealed significantly greater twitch (2-fold) and tetanus (1.7-fold) forces in nerve-muscle constructs.
- Electrical stimulation of neural extensions elicited contractions, confirming functional neuromuscular transmission.
Conclusions:
- Successfully engineered functional 3-D nerve-muscle constructs with integrated neuromuscular junctions.
- Demonstrated that neural integration enhances the contractile properties of engineered muscle constructs.
- These constructs offer a promising platform for studying neuromuscular function and developing regenerative therapies.