Engineered contractile skeletal muscle tissue on a microgrooved methacrylated gelatin substrate

Area of Science:

• Biomaterials Engineering
• Tissue Engineering
• Cell Biology

Background:

• Engineering three-dimensional (3D) tissue structures requires cells to form networks mimicking native microarchitecture.
• Microfabricated hydrogel substrates offer a platform for directing cells into biomimetic tissue architecture in vitro.

Purpose of the Study:

• To present microgrooved methacrylated gelatin hydrogels for facile and reproducible fabrication of muscle-like fibrous structures.
• To assess the impact of microgroove ridge size on myoblast orientation and myotube formation.
• To evaluate the effect of electrical stimulation on myoblast alignment and myotube development.

Main Methods:

• Fabrication of microgrooved methacrylated gelatin hydrogel substrates with 50 and 100 μm ridge sizes.
• Culturing C2C12 myoblasts on these substrates to observe alignment and multinucleated myotube formation.
• Application of electrical stimulation to assess its effects on myoblast organization and myotube characteristics.

Main Results:

• Microgroove ridge size did not significantly influence C2C12 myoblast alignment.
• Wider-ridged hydrogels (100 μm) resulted in more myotubes, with some not aligned to the groove direction.
• Electrical stimulation enhanced myoblast alignment and increased the diameter of the resulting myotubes.
• Free-standing 3D muscle sheets were successfully constructed, exhibiting contraction upon electrical stimulation.

Conclusions:

• Microgrooved methacrylated gelatin hydrogels provide a viable platform for engineering biomimetic muscle tissue.
• Engineered muscle tissues demonstrate robust contractility and potential for applications in tissue engineering, drug screening, and biorobotics.