Human DPSCs fabricate vascularized woven bone tissue: a new tool in bone tissue engineering

• DOI: 10.1042/cs20170047
• Publication Date: 2017-02-17T01:40:21Z
• Authors: Francesca Paino; Marcella La Noce; Alessandra Giuliani; Alfredo De Rosa; Serena Mazzoni; Luigi Laino; Evzen Amler; Gianpaolo Papaccio; Vincenzo Desiderio; Virginia Tirino
• Keywords: PHASE CONTRAST; Adult; 0301 basic medicine; Osteocalcin; Mice, Nude; Neovascularization, Physiologic; Cell Separation; bone differentiation; bone regeneration; bone tissue engineering; hDPSCs; holotomography; human Dental Pulp Stem Cells; human serum; phc-microCT; woven bon; woven bone; Medicine (all); Young Adult; 03 medical and health sciences; Calcification, Physiologic; Osteogenesis; PHASE CONTRAST IMAGING; Animals; Humans; Research Articles; Cells, Cultured; Dental Pulp; Cell Proliferation; Bone Transplantation; Tissue Engineering; PHASE CONTRAST MICROTOMOGRAPHY; Chemotaxis; Stem Cells; Cell Differentiation; X-Ray Microtomography; BONE MINERALIZATION; FRELON CAMERA; Bone Substitutes; bone differentiation; bone regeneration; bone tissue engineering; hDPSCs; holotomography; human Dental Pulp Stem Cells; human serum; phc-microCT; woven bon; woven bone; Adult; Animals; Bone Transplantation; Calcification, Physiologic; Cell Differentiation; Cell Proliferation; Cell Separation; Cells, Cultured; Chemotaxis; Dental Pulp; Humans; Mice, Nude; Neovascularization, Physiologic; Osteocalcin; Osteogenesis; Stem Cells; Tissue Engineering; X-Ray Microtomography; Young Adult; Bone Substitutes; Medicine (all); BONE

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.