Biodegradable polyurethane (PU) was synthesized based on using poly(epsilon-caprolactone) (PCL) as the soft segment. Fibers in different diameters (200-400 nm, 600-800 nm, and 1.4-1.6 mu m) were then made by electrospinning PU solution in N,N-dimethylacetamide and 2,2,2-trifluoroethanol. Human bone marrow derived mesenchymal stem cells (hMSCs) in the form of single dispersed cells or aggregates were seeded on the electrospun meshes for evaluation of cell behavior. Differentiation experiments showed that hMSC aggregates on electrospun fibers had greater differentiation capacities than single cells. Besides, nanofibers of 200-400 nm diameters significantly promoted the osteogenic and chondrogenic differentiation of hMSCs than fibers of the other diameters. The effect of substrate elasticity was further elucidated by comparing cell behaviors on the nanofibers of PCL-based PU and those of pure PCL. The more elastic PU nanofibers demonstrated more osteogenic and chondrogenic induction potential than PCL electrospun fibers. We suggested that the elastic nanofibers seeded with hMSC aggregates may be advantageous for cartilage and bone tissue engineering. (C) 2014 Elsevier B.V. All rights reserved.