A step toward 3D fibroblast organization by tuning hydrogel dimensionality

Fibroblast nematic organization is a commonly observed feature in pathologies such as keloids and fibrosis where extracellular matrix remodelling and mechanical constraints influence cell alignment. To identify key factors driving fibroblast organization, we gradually complexified the culture dimensionality, step wise transitioning from 2D to 3D. Using photopolymerized gelatin methacrylate (GelMA) with hyaluronic acid (HAMA) hydrogels, fibroblasts are 2D cultured on the hydrogel, then overlaid with an additional hydrogel layer to create a 2.5 D sandwich system that we compare to 3D culture where fibroblasts are embedded in the matrix.