Dielectrophoresis (DEP) approach was employed to achieve highly aligned multi-walled carbon nanotubes (MWCNTs) within the gelatin methacrylate (GelMA) hydrogels in a facile, rapid, inexpensive, and reproducible manner. This approach enabled us to make different CNTs alignments (e.g., vertical or horizontal alignments) within the GelMA hydrogel using different electrode designs or configurations. Anisotropically aligned GelMA-CNTs hydrogels showed considerably higher conductivity compared to randomly distributed CNTs dispersed in the GelMA hydrogel and the pristine and non-conductive GelMA hydrogel. Adding 0.3 mg/mL CNTs to the GelMA hydrogel led to a slight increase in the mechanical properties of the GelMA and made it to behave as a viscoelastic material. Therefore, it can be used as a suitable scaffold for soft tissues, such as skeletal muscle tissue. 3D microarrays of skeletal muscle myofibers were then fabricated based on the GelMA and GelMA-CNTs hydrogels and they were characterized in terms of gene expressions related to the muscle cell differentiation and contraction. Owing to high electrical conductivity of aligned GelMA-CNTs hydrogels, the engineered muscle tissues cultivated on these materials demonstrated superior maturation and functionality particularly after applying the electrical stimulation (voltage 8 V, frequency 1 Hz, and duration 10 ms for 2 days) compared to the corresponding tissues obtained on the pristine GelMA and randomly distributed CNTs within the GelMA hydrogel.
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