Understanding the mechanisms of vascular diseases requires elucidation of structural and functional changes in vascular smooth muscle cells (VSMCs) within the arterial wall. However, conventional long-term passage cultures on rigid, flat substrates often lead to VSMC dedifferentiation, resulting in morphological and functional changes that differ significantly from their native state. In this study, we developed a biomaterial (gelatin) cell culture substrate that replicates the mechanical microenvironment of the aortic media. The substrate was fabricated by transferring microscale rectangular grooves (5, 10, and 20 μm of groove width and 10 μm of groove depth) from polydimethylsiloxane (PDMS) mold onto a gelatin surface, followed by stabilization through glutaraldehyde crosslinking. Surface topography and elastic modulus were characterized using confocal laser scanning microscopy and atomic force microscopy (AFM), confirming groove dimension and surface elastic moduli under culture conditions. The fabricated micro-grooved gelatin substrate had sufficient groove depth to trap the cells, whose widths reflected those of the PDMS mold. VSMCs cultured on these gelatin micro-grooved substrates exhibited elongated morphologies aligned with the groove direction. Furthermore, the cell nucleus was trapped in the grooves with elongated shape, which is expected to induce differentiation. These results suggest that the developed micro grooved gelatin substrate effectively mimics key features of the in vivo arterial environment and may serve as a valuable platform for investigating the mechanobiology of vascular smooth muscle cells and the pathophysiology of vascular diseases.
ARAI et al. (Wed,) studied this question.