As a part of regenerative medicine, biomaterials are largely used in this field of nanotechnology and tissue engineering research. We have recently developed a new scaffold using electrospun nanofibers of Poly (ε-caprolactone), PCL which is able to mimic the collagen extracellular matrix of cells. The aim of this study was to engineer a biological and implantable structure leading the regeneration of the tooth-bone unit. For this aim, we have cultured mouse osteoblasts embedded in a collagen gel on the nanofibrous membrane and coupled this structure with an embryonic dental germ before implantation. To follow bone and tooth regeneration, we have performed RT-PCR, histology and immunofluorescence analysis. We showed here that this leaving implantable structure represents an accurate strategy for bone-tooth unit regeneration. We report here the first demonstration of bone-tooth unit regeneration by using a strategy based on a synthetic nanostructured membrane. This electrospun membrane is manufactured by using an FDA approved polymer, PCL and functionalized with osteoblasts before incorporation of the tooth germs at ED14 (the first lower molars) to generate bone-tooth unit in vivo after implantation in mice. Our technology represents an excellent platform on which other sophisticated products could be based.