Collective Effect of Chemical Distribution and Micropillar Array Topography on the Migration Behavior of Human Mesenchymal Stem Cells

Abstract How cells sense and respond to ECM topographic and surface‐bound chemical cues remains unclear. In this study, engineered micropillar arrays with selectively functionalized continuous fibronectin coating (FN‑Coat) and discontinuous fibronectin stamping (FN‑Stamp) are developed, which enables a systematic analysis of microtopographical features and chemical cues that synergistically regulate migratory behavior of human bone marrow mesenchymal stem cells (hBMSCs). Cell adhesion on FN‑Stamp micropillar arrays is found to be inhibited, resulting in decreased cellular rigidity and accelerated migration. While, cells on FN‑Coat micropillar arrays form abundant filamentous pseudopodia and undergo oriented migration. Furtherly, the results demonstrate that micropillar array topography enhances the formation of cellular protrusions and promote directed cell migration, while the spatial distribution of fibronectin regulates focal adhesion expression and F‑actin organization. The greater the distribution of FN, the higher the enhancement of cell adhesion and cytoskeleton assembly, which further promotes the intracellular stress and reduces the migration speed. These findings provide insights into how cells adjust cellular mechanical properties and migration behavior in response to topographical and chemical cues.