Regeneration of dental pulp/dentine complex with a three-dimensional and scaffold-free stem-cell sheet-derived pellet.
Dental pulp/dentine complex regeneration is indispensable to the construction of biotissue-engineered tooth roots and represents a promising approach to therapy for irreversible pulpitis. We used a tissue-engineering method based on odontogenic stem cells to design a three-dimensional (3D) and scaffold-free stem-cell sheet-derived pellet (CSDP) with the necessary physical and biological properties. Stem cells were isolated and identified and stem cells from root apical papilla (SCAPs)-based CSDPs were then fabricated and examined. Compact cell aggregates containing a high proportion of extracellular matrix (ECM) components were observed, and the CSDP culture time was prolonged. The expression of alkaline phosphatase (ALP), dentine sialoprotein (DSPP), bone sialoprotein (BSP) and runt-related gene 2 (RUNX2) mRNA was higher in CSDPs than in cell sheets (CSs), indicating that CSDPs have greater odonto/osteogenic potential. To further investigate this hypothesis, CSDPs and CSs were inserted into human treated dentine matrix fragments (hTDMFs) and transplanted into the subcutaneous space in the backs of immunodeficient mice, where they were cultured in vivo for 6 weeks. The root space with CSDPs was filled entirely with a dental pulp-like tissue with well-established vascularity, and a continuous layer of dentine-like tissue was deposited onto the existing dentine. A layer of odontoblast-like cells was found to express DSPP, ALP and BSP, and human mitochondria lined the surface of the newly formed dentine-like tissue. These results clearly indicate that SCAP-CSDPs with a mount of endogenous ECM have a strong capacity to form a heterotopic dental pulp/dentine complex in empty root canals; this method can be used in the fabrication of bioengineered dental roots and also provides an alternative treatment approach for pulp disease.
Research and Development Centre for Tissue Engineering, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.|Department of Oral and Maxillofacial Surgery, College of Stomatology, Jiamusi University, Jiamusi, Hei Longjiang, People's Republic of China.
Sijia
S
Na
Research and Development Centre for Tissue Engineering, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.|Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
Hao
H
Zhang
Department of Oral and Maxillofacial Surgery, College of Stomatology, Jiamusi University, Jiamusi, Hei Longjiang, People's Republic of China.
Fang
F
Huang
Department of Oral and Maxillofacial Surgery, College of Stomatology, Jiamusi University, Jiamusi, Hei Longjiang, People's Republic of China.
Weiqi
W
Wang
Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
Yin
Y
Ding
Department of Oral and Maxillofacial Surgery, College of Stomatology, Jiamusi University, Jiamusi, Hei Longjiang, People's Republic of China.
Dechao
D
Li
Research and Development Centre for Tissue Engineering, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.|Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi'an, Shaanxi, People's Republic of China.
Yan
Y
Jin
D000818:Animals; D002454:Cell Differentiation; D002469:Cell Separation; D003782:Dental Pulp / Q000502:physiology*; D003804:Dentin / Q000502:physiology*; D006801:Humans; D051379:Mice; D012038:Regeneration; D013234:Stem Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
cell sheet; cell sheet-derived pellet; dental pulp; dentine; irreversible pulpitis; stem cells from apical papilla; tissue regeneration