Regeneration of dental pulp/dentine complex with a three-dimensional and scaffold-free stem-cell sheet-derived pellet

Abstract

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.

Three-Dimensional Printed Multiphase Scaffolds for Regeneration of Periodontium Complex

“…[4][5][6][7] The field of tooth regeneration has grown robustly. [8][9][10][11][12][13][14][15][16][17][18][19][20] The ambitious goal to regenerate an entire tooth, including the enamel, has encountered several barriers, such as the unavailability of patient compatible, postnatal stem/progenitor cells, and lack of strategy to derive functional ameloblasts. 12 A less ambitious and pragmatic goal to regenerate mineralized tooth roots has gained substantial momentum.…”

Exposure to a continuous low dose of tetrachlorodibenzo-p-dioxin impairs the development of the tooth root in lactational rats and alters the function of apical papilla-derived stem cells

“…Referring to the previous studies, 18,19 the apical papilla tissue from the immature roots of the first molars were removed on PND6 and minced into approximately 1-mm 3 fragments. The SCAPs were purified from the apical papilla using the Percoll Runx2, and alkaline phosphatase, suggesting that the AHR pathway mediated the effects of TCDD.…”

“…The methods about root dentine generation in vivo was performed as previously described. 18 A total of 2 Â 10 6 SCAPs were seeded into 5-cm dishes without thermoresponsive surfaces. When 90% confluent the cells were cultured in basic medium with 10 pM TCDD or DMSO for 10 days until a sheet of cells was formed.…”

The Scope of Stem Cells in Periodontal Regeneration

“…Also more recently cell sheet fragments and cell sheet pellets have been developed to increase the efficacy of the cells transplanted especially in cases where the target sit is too small for the entire cell sheet [93,94]. Further Co-culturing and micro-patterning of different types of cells are under trials for creation of more tissue-like materials which would give better results than single cell-sheets [95].…”