To improve the electrospinnability of chitosan (CS), a series of nanofiber membrane blends comprised of CS, poly(lactic acid) (PLA), and nonionic surfactant polyoxyethylene nonylphenol ether (TX-15), were made. Uniform nanofibers with no bead-like structures were obtained from solutions of 2% TX-15 with 6% CS(50)/PLA(50). The diameter was between 200 and 300 nm. We found that with increasing TX-15 in the blend, the nanofibers displayed more hydrophilicity. Compared to CS/PLA nanofibers, the blend polymers with TX-15 had better tensile mechanical properties. Finally, all cells examined showed high levels of attachment and spreading on CS/PLA/TX-15 nanofibers with a TX-15 content of 0∼3%. Thus, the nanofibers were nontoxic. In conclusion, adding PLA and TX-15 to CS via solution-blending and electrospinning may be an effective way to toughen CS nanofibers and make them more suitable for drug delivery or tissue engineering applications. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 41098.