Multi‐Omics Profiling Reveals Immunomodulatory and Pro‐Regenerative Effects of a Graphene Oxide–Collagen Scaffold in Massive Rotator Cuff Tears

Massive rotator cuff tears (MRCT) remain a clinical challenge, characterized by poor tendon-bone interface (TBI) healing, severe muscle degeneration, and high postoperative retear rates. Tissue engineering scaffolds offer promising alternatives, yet traditional biomaterials often lack sufficient bioactivity to orchestrate comprehensive tissue regeneration. Herein, we developed a novel graphene oxide (GO)-engineered porcine type I collagen (GO/Col) scaffold and systematically investigated its therapeutic efficacy and underlying molecular mechanisms via multi-omics analyses. Comprehensive characterization showed that GO incorporation improved scaffold stability, wettability, and biocompatibility. In vitro, the GO/Col scaffold enhanced mesenchymal stem cell adhesion and proliferation, promoted osteogenic and chondrogenic differentiation, and suppressed adipogenesis. In a macrophage model system, GO/Col was associated with a shift toward a more reparative, anti-inflammatory phenotype. Using a clinically relevant chronic MRCT rat model, we observed that GO/Col scaffolds significantly improved motor function, biomechanical properties, and tendon-bone regeneration, while inhibiting muscle fibrosis and fatty infiltration. Mechanistically, integrated transcriptomics, proteomics, and mass cytometry analyses revealed GO-mediated modulation of critical signaling pathways involved in immune regulation, stem cell differentiation, and tissue regeneration. Notably, GO activated pro-osteogenic/chondrogenic pathways and anti-inflammatory signatures, while downregulating adipogenic and pro-inflammatory pathways. Collectively, these findings support the potential of GO/Col scaffolds as a bioactive tissue-engineering strategy for chronic MRCT repair.