Concave/Convex Curvature of Anisotropic Grooves Differentially Alters Cellular Morphology, Adhesion, and Proliferation

Curvature is an integral part of the complex in vivo tissue architecture across various length scales. Therefore, several in vitro models with a patterned curvature in different length scales have been developed to understand the role of this in cellular behavior. At the subcellular scale, wavy patterns have been reported wherein concave and convex grooves are adjacently present. However, the independent effect of continuous subcellular concave and convex shapes has not been reported, mainly owing to the limitations in fabricating such patterns. In this study, we developed continuous concave and convex grooves on polydimethylsiloxane (PDMS) using a Dracaena sanderiana (bamboo) leaf as a template. The first (negative) replica from the abaxial side of the bamboo leaf, which imparted concave grooves on PDMS, was subsequently used as a template to fabricate a positive replica of the leaf, resulting in convex grooves of the same size and arrangement as the concave grooves. We examined the influence of the groove curvature on the morphology of bone marrow-derived human mesenchymal stem cells (BM-hMSCs) and skeletal muscle cells (C2C12). BM-hMSCs and C2C12 cells aligned on both concave and convex grooves as compared to the random orientation on a flat substrate. The significant difference was observed in the morphology of both cells, in terms of area, aspect ratio, number, and length of protrusions on concave and convex patterns. We found that the number of protrusions was also dependent on the ratio of cell to pattern length scale for convex-shaped grooves but independent of length scale for concave-shaped grooves. The proliferation of BM-hMSCs was also found to be different on concave and convex shapes. Therefore, this study shows the importance of (1) convex and concave curvatures of the subcellular length scale in cellular response, (2) dependence on the ratio of cell and curvature length scale, and (3) use of natural templates for overcoming fabrication challenges.