Regulation of Fibroblast Spreading on Fused Silica Square Nanocone Arrays: Implications for Biomedical Applications

Abstract Developing and exploring fabrication techniques for nanostructured biomaterials that enable precise geometric control of the features for mediating cell−nanopattern interactions are essential for cutting-edge biomedical applications. Herein, we utilized a UV laser interference lithography (LIL) combined with an inductively coupled plasma (ICP) etching technique to fabricate 3D nanocones (NCs) with precisely controlled varying top geometries in square arrays on large-area fused silica substrates. The arrangement of NCs was designed via LIL, and the single parameter of the nanocone top diameter can be systematically modulated from 200 nm to 700 nm by ICP etching processing; thereby, the surface energy and wettability of the patterned surfaces were controlled accordingly. We then investigated the interactions of fibroblasts (L929) with these geometrical cues of NCs with changing adhesive area ratio (AAR) and flat substrates. Varying the AAR from 0.02 to 0.22 exerted pronounced impacts on the fibroblast morphology and directional alignment behavior of the fibroblasts. The cell circularity decreased from 0.76 to 0.35, and the aspect ratio increased from 1.66 to 4.11, the cell spreading area improved from 18.77% to 25.97%, and cell alignment shifts from a single axial orientation (0°) gradually toward a diagonal orientation (45°), finally forming dual orientations. We found that the topological nanocone-induced changes in cell morphology, spreading, and aspect ratio are predominantly governed by cytoskeletal remodeling, accompanied by adaptive nuclear morphological remodeling. The high-throughput and precise fabrication method proposed in this study, by enabling geometrically controlled tuning of the AAR, supports various morphogenetic trends in fibroblast cells, thus providing a viable platform for the design of future scaffolds.