In this study, chitosan-blended poly(lactic acid) (PLA) nanofibers were fabricated via electrospinning process by using a solvent composing of chloroform, water, acetic acid, and ethanol. In particular, the introduction of ethanol made acetic acid/water and chloroform miscible. The resultant chitosan-blended PLA nanofibers were subject to characterization including scanning electron microscope (SEM), UV-visible spectroscopy, fluorescence microscope and Fourier transform infrared (FTIR) spectroscopy. The average diameter of chitosan-blended PLA nanofibers decreased with increasing chitosan content. The minimum inhibitory concentration (MIC) of chitosan-blended PLA nanofibers against Eschericia coli was 0.015 g/mL when chitosan to PLA ratio was 4 wt%. Furthermore, chitosan-blended PLA nanofibers exhibited no cytotoxicity to the L-929 cells, suggesting cytocompatibility. These results show that chitosan and PLA can be blended without using highly-toxic solvent and the electrospun chitosan-blended PLA nanofibers can potentially be employed for biomedical applications.