We have prepared carboxymethyl chitosan-alginate dialdehyde (CMCS-ADA) nanofibers via a reactive electrospinning process with the aid of polyethylene oxide (PEO). The presence of PEO delayed the gelation of CMCS and ADA, thus providing ease of use to adjust the mixing of CMCS-PEO and ADA-PEO blended solution. The mixed solution can be adjusted to come out from the needle before the gel formation or when the gel was just about to form. Defect-free CMCS-ADA-PEO nanofibers with average diameters ranging from 100nm to 900nm were obtained using water as a solvent. The in situ cross-linked CMCSADA nanofibers were then obtained following the extraction of water-soluble PEO. After immersion in phosphate-buffered saline (PBS) at a pH of 7.4 for up to 15 days, the as-spun CMCS-ADA-PEO composite nanofibers maintained structural integrity, confirming the success of the crosslinking. The PEO-extracted CMCS-ADA nanofibers promoted the adhesion, proliferation and alkaline phosphatase activity of bone marrow stromal cells. (C) 2016 Elsevier Ltd. All rights reserved.