Electrochemically engineered titania (TiO₂) nanopores enable tailored cellular function; however, the cellular mechanosensing mechanisms dictating the cell response and soft tissue integration are yet to be elucidated. Here, we report the fabrication of anisotropic TiO₂ nanopores with diameters of 46 and 66 nm on microrough titanium (Ti) via electrochemical anodization, towards short- and long-term guidance of human primary gingival fibroblasts (hGFs). Cells on tissue culture plates and bare Ti substrates were used as controls. Notably, we show that nanopores with a diameter of 66 nm induced more mature focal adhesions of vinculin and paxillin at the membrane, encouraged the development of actin fibers at focal adhesion sites, and led to elongated cell and nuclear shape. These topographical-driven changes were attributed to the Ras-related C3 botulinum toxin substrate 1 (Rac 1) GTPase pathway and nuclear localisation of LAMIN A/C and yes-associated protein (YAP) and associated with increased ligament differentiation with elevated expression of the ligament marker Mohawk homeobox (MKX). Study findings reveal that minor tuning of nanopore diameter is a powerful tool to explore intracellular and nuclear mechanotransduction and gain insight into the relationships between nanomaterials and mechanoresponsive cellular elements.

Anodization is a cost-effective technique to nano-engineer various metals, however, optimizations have been mostly restricted to Al and Ti, and for easy to manage polished and flat substrates, limiting industrial translation. Here, we aim at standardizing and simplifying anodization of ten metals, attempting to make metal nano-engineering more accessible. In a world-first attempt, we synthesize a standard electrolyte to fabricate controlled nanostructures on various metals, taking a close look at the influence of the substrate topography, electrolyte composition, and electrolyte aging (repeated use of same electrolyte) towards designing a standard nano-engineering strategy. Anodization of curved substrates (metal wires) with micro-roughness allows for ease of industrial translation across various applications. This study is a step closer to standardizing anodization and fabrication of controlled nanotopography on various metallic surfaces, while maintaining scalability and ease of use.