Small caliber vascular replacement (<4 mm) still remains a challenge for medical and research teams, as no available vascular substitutes (VS) are suitable for small diameter bypass. Vascular engineering proposes new models of small diameter VS but rare are those that meet the biocompatibility and mechanical criteria. In this study, we developed a new scaffold made by the combination of two natural biomacromolecules: collagen and silk fibroin. The scaffold was further cellularised with porcine smooth muscle cells. First, the behavior of cells in the collagen-fibroin constructs was verified in order to evaluate the biocompatibility of the scaffold with the cells. Then, gel mass loss and cellular attachment, morphology, spreading and viability were analysed. The results showed an excellent interaction and biocompatibility between collagen, silk fibroin fibers and cells. Thus, the collagen-fibroin construct appears to be a very attractive material for vascular tissue engineering.