This work was focused on preparation and characterization of the polyurethane nanofibers containing calcium chloride (CaCl 2. prepared via electrospinning process for the bionanotechnological applications. The morphological, structural characterizations and thermal properties of the polyurethane/CaCl 2 nanofibers were determined by using scanning electron microscopy (SEM), field-emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDX), Fourier transform-infrared (FT-IR) spectroscopy and thermogravimetry (TGA). SEM images revealed that these composite nanofibers were well-oriented and had good incorporation of CaCl 2. The morphological feature of the cells attached on polyurethane/CaCl 2 nanofibers scaffold was confirmed by SEM. The in vitro cell compatibility of the polyurethane/CaCl 2 composite nanofibers was studied. This study demonstrated the non-toxic effect of electrospun polyurethane/CaCl 2 composite nanofibers. Based on this, we propose a mechanism for the cell attachment. Copyright © 2011 American Scientific Publishers.