Engineering Highly Aligned and Densely Populated Cardiac Muscle Bundles via Fibrin Remodeling in 3D-Printed Anisotropic Microfibrous Lattices

Replicating the structural and functional features of native myocardium, particularly its high-density cellular alignment and efficient electrical connectivity, is essential for engineering functional cardiac tissues. Here, novel electrohydrodynamically printed InterPore microfibrous lattices with anisotropic architectures are introduced to promote high-density cellular alignment and enhanced tissue interconnectivity. The interconnected pores in the microfibrous lattice enable dynamic, cell-mediated remodeling of fibrous hydrogels, resulting in continuous, mechanically stable tissue bundles. Compared to lattices with isolated pores, the engineered aligned cardiac tissues from neonatal rat cardiomyocytes exhibit improved electrophysiological properties and synchronous contractions. Using a multiseeding strategy, an equivalent cell seeding density of 8 × 10⁷ cells mL⁻¹, facilitating the formation of multicellular, vascularized cardiac structures with maintained tissue viability and integrity, is achieved. As a demonstration, human-induced pluripotent stem cell-derived cardiac tissues are engineered with progressive maturation and functional integration over time. These findings underscore the potential of InterPore microfibrous lattices for applications in cardiac tissue engineering, drug discovery, and therapeutic development.