In this article, different concentrations of multiwalled carbon nanotube (MWCNT) were homogeneously dispersed throughout the chitosan (CS) matrix. A simple solvent-cast method was used to fabricate chitosan films with 0.1, 0.5, and 1% of MWCNT with the average diameter around 30 nm. The CS/MWCNT films were characterized for structural, viscous and mechanical properties with optical microscopy, wide-angle X-ray diffraction, Raman spectroscopy, tensile test machine, and microindentation testing machine. Murine osteoblasts were used to examine the cell viability and attachment of the nanocomposite films at two time points. In comparison to the pure chitosan film, the mechanical properties, including the tensile modulus and strength of the films, were greatly improved by increasing the percentage of MWCNT. Furthermore, adding MWCNT up to 1% increased the viscosity of the chitosan solution by 15%. However, adding MWCNT decreased the samples ductility and transparency. In biological point of view, no toxic effect on osteoblasts was observed in the presence of different percentages of MWCNT at day 3 and day 7. This investigation suggested MWCNT could be a promising candidate for improving chitosan mechanical properties without inducing remarkable cytotoxicity on bone cells. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2704–2712, 2014.