A straightforward method for manufacturing stratified scaffolds with potential application in osteochondral tissue engineering is reported. The method integrates three different techniques widely used for scaffold fabrication: sponge replica method, freeze-drying and electrospinning. 45S5 Bioglass (R) was used for the fabrication of the rigid bioactive substrate intended to be in contact with bone tissue, chitosan and alginate based solutions were used for building the interface between the Bioglass (R)-based scaffold and the soft cartilage side of the construct. Finally a chitosan-based electrospun nanofibrous membrane was selected for the upper layer of the scaffold. In this design, the intermediate layer has multiple functions providing adherence between the other scaffold components, preventing delamination and acting as a soft coating for the rigid Bioglass (R) substrate. Morphological analysis, bioactivity tests in simulated body fluid (SBF), as well as mechanical and wettability tests were performed on all samples to assess the optimal stratified scaffold combination. The best results, in terms of resistance to layer delamination, preservation of the bioactivity and improvement of the mechanical properties, were obtained using a chitosan solution and the freeze gelation technique for fabrication of the intermediate layer. Key results are discussed in the context of the potential application of these novel scaffolds in osteochondral tissue engineering. (C) 2012 Elsevier B.V. All rights reserved.