Chitosan (CH), silk fibroin (SF), and hydroxyapatite (HA) were used to prepare CH/SF/HA composites and the resulting composites were electrospun into nanofibrous membrane units with gradient compositional and structural features. The optimal membrane unit was used together with CH/HA and CH/SF composites to fabricate a type of three-layer scaffold that is intended for osteochondral repair. The bottom layer of the scaffold was built with CH/HA composites and it served as a subchondral layer, the integrated nanofibrous membrane unit functioned as the middle layer for mimicking the calcified layer and the top layer was constructed using CH/SF composites for acting as a chondral layer. The nanofibrous membrane unit was found to be permeable to some molecules with limited molecular weight and was able to prevent the seeded cells from migrating cross the unit, functioning approximately like the calcified layer in the osteochondral matrix. Layered scaffolds showed abilities to promote the growth of both chondrocytes and osteoblasts that were seeded in their chondral layer and bony layer, respectively, and they were also able to support the phenotype preservation of seeded chondrocytes and the mineralization of neotissue in the bony layer. Results suggest that this type of layered scaffolds can function as an analogue of the osteochondral matrix and it has potential in osteochondral repair.
Keywords: calcified layer; chitosan; hydroxyapatite; layered scaffold; nanofibrous membrane unit; silk fibroin.