Acoustically Responsive Nanofibrous Scaffolds with 3D Hierarchy for Tympanic Membrane Regeneration

Mesenchymal stromal cell (MSC)‐based therapies have emerged as a promising approach for repairing tympanic membrane (TM) injuries. However, the role of essential biochemical, biophysical, and biomechanical cues in guiding MSC differentiation for TM regeneration remains unclear. To address this, the current study investigates the influence of molecular, hierarchical, and acoustical stimulation on biofabricated TM scaffolds within a biomimetic environment. Relevant bioactive molecules are identified to enhance TM‐specific gene expression on electrospun meshes. Additive‐manufactured filaments are then deposited onto nanofibrous meshes to study the role of 3D hierarchy. Finally, acoustical stimulation within the human audible range is applied using a custom‐built bioreactor, partially mimicking the native tissue niche. Acousto‐vibrational analysis reveals elevated oscillatory responses at specific sound frequencies, contributing to accelerated TM wound healing. This work demonstrates that synergistically integrating bioactive agents, 3D hierarchical structures, and sound waves promotes the formation of an aligned extracellular matrix crucial for TM regeneration.