Matrix-over-morphogen effect depends on interaction between progenitor lineage history and mechano-chemical integration: An examination using standardized decellularized scaffold cultures

The matrix-over-morphogen effect, in which extracellular matrix architecture overrides soluble growth-factor cues during lineage specification, challenges the prevailing view that diffusible signals are the primary drivers of cell fate. Nonetheless, the reproducibility of the matrix-over-morphogen effect has remained controversial across tissue contexts. Here, we investigated the phenomenon using limb-derived decellularized scaffolds with tightly controlled collagen orientation and stiffness profiles. Murine mesenchymal progenitors (passage-matched; donor age: 10.2 ± 2.9 weeks [mean ± standard deviation]) were divided into three cohorts on the basis of their prior differentiation history. Of the 301 cultures, 171 were osteo-primed progenitors (donor age: 9.8 ± 2.7 weeks), 78 were chondro-primed progenitors (10.5 ± 3.1 weeks), and 52 were naïve fibroblastic precursors (11.1 ± 2.4 weeks). A dominant matrix effect was detected in chondro-primed progenitors (Kruskal–Wallis: p < 0.001), but was absent in osteo-primed cells (p = 0.48) and naïve precursors (p = 0.37). These findings indicate that the matrix-over-morphogen effect is contingent on developmental experience; specifically, mechanical conditioning during osteogenic training appears to dampen structural dominance. Moreover, the lack of matrix preference in naïve precursors suggests that, without lineage-specific pre-conditioning, cells may not consistently prioritize architectural information. The present data fill a critical gap in our understanding of mechano-chemical integration, particularly how varied epigenetic memories sculpt responsiveness to concurrent cues. Our results underscore the need for regenerative strategies to deliberately balance scaffold design and soluble factor regimes, especially in settings where excessive structural dominance might obscure biochemical specification.