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

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.