Silks are naturally occurring polymers and fibroin, its filament core protein, has been shown to support stem cell differentiation in vitro, and promote tissue repair in vivo. The aim of this study is to develop a biomaterial based on silk-fibroin fibers that can be associated with mesenchymal progenitor cells from human perichondrium in vitro, in order to promote auricular reconstruction in vivo. Silk-fibroin concentrate was dissolved with formic acid solution and freeze-dried in auricular moulds. Fibroin-based material was characterized by scanning electron microscopy and by cytotoxic assays. Perichondrium mesenchymal progenitor cells were characterized by flow cytometry. They expressed the standard mesenchymal stem cell markers, and were able to differentiate into several mesenchymal lineages in vitro. This fibroin-based material is a three-dimensional fibrillar scaffold, non-woven and biocompatible, which was also well integrated with alginate and mesenchymal cells.