In this study, the influence of degree of deacetylation ( DD) and composition on some structural and biological properties of chitosan scaffolds were examined in vitro. 3D chitosan scaffolds of 2% (w/v) and 3% (w/v) composition in different DDs i.e. 75-85% and > 85% were prepared by freeze-drying method at -80 degrees C. We noticed that > 85% deacetylated chitosan scaffolds of 2% (w/v) composition has a highly interconnected morphological structure having similar to 100 mu m pore size with 0.0917 N/mm(2) compression modulus. L929 fibroblastic cells were cultured on chitosan scaffolds in order to evaluate their biocompatibilities. Cell culture studies demonstrated that fibroblastic cell attachment and proliferation is affected by DD. The higher deacetylated chitosan scaffolds strongly supported the attachment and proliferation when compared with the lower deacetylated scaffolds. MTT assay indicated that > 85% deacetylated chitosan scaffolds of 2% (w/v) composition, having the highest specific growth rate 0.017 h(-1) of all, was found to be the most suitable for cell culture studies and a potential candidate for tissue engineering with enhanced biostability and good biocompatibility.