3D hierarchically aligned nanofiber scaffolds promote cell migration for tissue regeneration.

During tissue regeneration, cells are recruited from surrounding tissue to the defect site. However, when the defect site is large and morphologically complex, cell recruitment often fails to match healthy tissue morphology, resulting in a dysfunctional repair. The integration of bioscaffolds can help to direct the repair process. Here, we present a protocol that integrates electrospinning, weaving, thermal fixation and modified gas-foaming technologies to fabricate 3D hierarchically aligned nanofiber scaffolds. The scaffolds exhibit high porosity, controlled fiber alignment and diverse configurations (uniaxial, bidirectional, radial and gradient alignments), creating effective 'cell highways' for promoting collective cell migration. Applications include hemostatic materials, skin and bone regeneration, hernia repair and biomedical swabs. Both in vitro and in vivo, the highly porous and directionally arranged 3D nanofiber scaffolds markedly enhance cell migration, accelerating the reconstruction of defective tissues. This protocol resolves challenges in production scalability, facilitating the wider adoption of these scaffolds, with a procedure intended for users with expertise in biomaterials and regenerative medicine. The 3D nanofiber scaffolds require 1 d to synthesize and result in improved cell migration during in situ tissue regeneration.