In this study, chitosan-PEO blend, prepared in a 15 M acetic acid, was electrospun into nanofibers (similar to 78 nm diameter) with bead free morphology. While investigating physico-chemical parameters of blend solutions, effect of yield stress on chitosan based nanofiber fabrication was clearly evidenced. Architectural stability of nanofiber mat in aqueous medium was achieved by ionotropic cross-linking of chitosan by tripolyphosphate (TPP) ions. The TPP cross-linked nanofiber mat showed swelling up to similar to 300% in 1 hand similar to 40% degradation during 30 day study period. 3T3 fibroblast cells showed good attachment proliferation and viability on TPP treated chitosan based nanofiber mats. The results indicate non-toxic nature of TPP cross-linked chitosan based nanofibers and their potential to be explored as a tissue engineering matrix. (C) 2012 Elsevier B.V. All rights reserved.