Shape‐Recoverable Anisotropic Mushroom‐Derived Scaffolds for Skeletal Muscle Regeneration

ABSTRACT Volumetric muscle loss (VML) requires scaffolds that combine anisotropic architecture, fatigue resistance, and tissue‐specific bioactivity. Here we report an injectable, shape‑recoverable scaffold engineered from decellularized Tricholoma matsutake stipes. Oxidative decellularization followed by EDC/NHS crosslinking preserved longitudinal microchannels and reinforced the chitin/β‑glucan matrix, yielding crosslinked mushroom scaffolds (CdM). To integrate muscle‑specific cues, CdM is coated with porcine muscle‑derived decellularized ECM (md‑CdM) or collagen (C‑CdM). CdM retained 97.5 ± 1.7% height after 15 cycles of 60% compressive strain and fully recovered following syringe extrusion, and these properties are preserved in the bioactive md‐CdM constructs. In vitro, md‑CdM further promoted myogenic alignment and multinucleation, lowered intracellular ROS, upregulated mechanotransduction genes (e.g., PIEZO1 , TRPV2 , RHOA/ROCK , YAP/TAZ ), and suppressed redox‑sensitive inhibitory pathways (Nrf2/Keap1, NF‑κB, TGFβ/SMAD). In a rat VML model, md‑CdM conformed to defect geometry and supported mature, peripherally nucleated myofibers, reduced fibrosis, organized microvasculature, and superior functional recovery. These findings establish mushroom‑derived hybrid scaffolds as a mechanoresponsive, redox‑buffering platform for minimally invasive VML repair and broader musculoskeletal regeneration.