The effects of nanopillar and nanopit arrays on the morphology and osteogenic differentiation of adipose-derived stem cells

Abstract

Nanotopographic control of cell behavior offers great potential in designing biomimetic scaffolds for cell therapy. However, the behavior of cells on different nanotopographies is not fully understood. In this study, we investigated the effect of nanostructures on human adipose-derived stem cells (ASCs) by directly comparing nanopillar and nanopit arrays. Morphological changes, cell viability and early osteogenic differentiation of ASCs have been analyzed on the nanostructures. Nanopit arrays were found to increase cell areas and promote early osteogenic differentiation more than nanopillar arrays. Analysis of focal adhesion (FA) formation indicated a larger increase in total area as well as the number of FAs during cell spreading on nanopit arrays. The maturation of FA is related to cellular traction forces, which are known to stimulate osteogenic induction through the RhoA-ROCK pathway. We conclude that ASCs can spread more on the nanopit array than on the nanopillar array due to the presence of continuous adhesive paths on the nanopit array, which is associated with increased expression of RUNX2 as an early osteogenic marker. Our results suggest that a connected path in nanopit arrays plays a critical role in controlling stem cell behavior compared to nanopillar arrays. A comparative understanding of nanostructures can provide a guideline for designing an artificial substrate for osteogenesis and tissue engineering. © The Author(s) 2026.