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There is scarce information about the way that topographical cues can influence essential mesenchymal stem cell functions, such as adhesion, proliferation or differentiation to specific lineages on metallic substrates used for bone repair. Patterned substrates control cell spatial dynamics, yielding cells oriented along the surface geometry, in a process known as contact guidance [1]. In bone tissue, the alignment of cells and the extracellular matrix determines tissue-specific functions. This study aimed to investigate the interactions of human mesenchymal cells with submicron-and nano-scale grooved surfaces generated on Ti6Al4V, the metallic alloy most widely used for the fabrication of orthopedic implants. Cell adhesion, proliferation and maturation to the osteogenic and adipogenic lineages on the tested surfaces were investigated. As the Rho family of GTPases controls the assembly of focal adhesions and cytoskeleton dynamics [2], the involvement of RhoA in the process of contact guidance on the patterned surfaces was explored.