A mechanobiology-driven cell-derived ECM bioink for engineering 3D glioblastoma tumor microenvironment models.

UNLABELLED: Glioblastoma (GBM) is highly aggressive and difficult to treat, partly due to the lack of in vitro models that faithfully recapitulate its biochemical and mechanobiological microenvironment. Synthetic hydrogels lack tumor-specific cues, while animal-derived dECM suffers from batch variability, limiting standardization. METHODS: Here, we describe a GBM-derived dECM bioink formulated through mechanically stimulated 3D GBM culture within GelMA/HAMA hydrogels. By controlling the matrix stiffness to match GBM tissue and applying various compressive stresses that mimic intracranial solid stress, we identified a mechanobiological activation range that maximized secretion of GBM-associated factors, including GDF15, MMP2, and MMP9. RESULTS: The resulting bioink exhibited upregulated tumor-specific biochemical signals compared to hydrogel-only controls. Micromesh-bioprinted GBM constructs fabricated from this bioink demonstrated enhanced proliferation, invasion-related gene expression, and ECM remodeling. Co-culture with endothelial cells or fibroblasts further reconstructed stromal activation, paracrine signaling, and matrix dynamics associated with GBM progression and therapeutic resistance. CONCLUSION: This strategy establishes a reproducible, bioactive GBM-specific bioink platform for physiologically relevant 3D GBM modeling and GBM-on-chip applications.