Human dental pulp stem cells (hDPSCs) are mesenchymal stem cells that have been successfully used in human bone tissue engineering. To establish whether these cells can lead to a bone tissue ready to be grafted, we checked DPSCs for their osteogenic and angiogenic differentiation capabilities with the specific aim of obtaining a new tool for bone transplantation. Therefore, hDPSCs were specifically selected from the stromal-vascular dental pulp fraction, using appropriate markers, and cultured. Growth curves, expression of bone-related markers, calcification and angiogenesis as well as an in vivo transplantation assay were performed. We found that hDPSCs proliferate, differentiate into osteoblasts and express high levels of angiogenic genes, such as vascular endothelial growth factor and platelet-derived growth factor A. Human DPSCs, after 40 days of culture, give rise to a 3D structure resembling a woven fibrous bone. These woven bone (WB) samples were analysed using classic histology and synchrotron-based, X-ray phase-contrast microtomography and holotomography. WB showed histological and attractive physical qualities of bone with few areas of mineralization and neovessels. Such WB, when transplanted into rats, was remodelled into vascularized bone tissue. Taken together, our data lead to the assumption that WB samples, fabricated by DPSCs, constitute a noteworthy tool and do not need the use of scaffolds, and therefore they are ready for customized regeneration.

Department of Biophysics 2nd Faculty of Medicine Charles University Prague 5 Úvalu 84 150 06 Prague 5 Czech Republic

Dipartimento di Medicina Clinica e Sperimentale Università degli Studi di Foggia Plesso di Medicina Viale Luigi Pinto 1 71122 Foggia Italy

Dipartimento di Medicina Sperimentale Sezione di Biotecnologie Istologia Medica e Biologia Molecolare Università degli Studi della Campania 'L Vanvitelli' via L Armanni 5 80138 Naples Italy

Dipartimento di Scienze Cliniche Specialistiche e Odontostomatologiche Università Politecnica delle Marche Via Tronto 10 a 60020 Torrette di Ancona Ancona Italy

Dipartimento Multidisciplinare Medico chirurgico Sezione di Odontostomatologia Università degli Studi della Campania 'L Vanvitelli' Via Luigi De Crecchio 6 80138 Naples Italy

Institute of Experimental Medicine Academy of Sciences of the Czech Republic v v i Vídeňská 1083 142 20 Prague 4 Czech Republic

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