Protocol for embedded 3D bioprinting of arterial models with customized contractile and metabolic functions.

3D bioprinting enables the fabrication of complex tissue structures in vitro. Here, we present a protocol for constructing multilayered arterial tissues with cellular alignment that enhances vascular smooth muscle function by modulating contractile and synthetic pathways. We describe steps for material preparation, simulation, embedded bioprinting, and biological characterization. This approach is adaptable to other tissues with cellular alignment, including skeletal and cardiac muscles. For complete details on the use and execution of this protocol, please refer to Li et al.(1).