Fabrication of Dentin-Pulp-Like Organoids Using Dental-Pulp Stem Cells

Abstract

We developed a novel dentin-pulp-like organoid. It has both stem-cell and odontoblast characteristics using a mesenchymal cell lineage of human dental-pulp stem cells (hDPSCs). The mixture of hDPSCs and Matrigel was transferred into the maintenance medium (MM) and divided into four different groups according to how long they were maintained in the odontogenic differentiation medium (ODM). All organoids were harvested at 21 days and analyzed to find the optimal differentiation condition. To assess the re-fabrication of dentin-pulp-like organoid, after dissociation of the organoids, it was successfully regenerated. Additionally, its biological activity was confirmed by analyzing changes of relevant gene expression and performing a histology analysis after adding Biodentine® into the ODM. The organoid was cultured for 11 days in the ODM (ODM 11) had the most features of both stem cells and differentiated cells (odontoblasts) as confirmed by relevant gene expression and histology analyses. Micro-computed tomography and an electron microscope also showed mineralization and odontoblastic differentiation. Finally, ODM 11 demonstrated a biologically active response to Biodentine® treatment. In conclusion, for the first time, we report the fabrication of a dentin-pulp-like organoid using mesenchymal stem cells. This organoid has potential as a future therapeutic strategy for tooth regeneration.

Roles of Dental Mesenchymal Stem Cells in the Management of Immature Necrotic Permanent Teeth

“…The neurogenic potential was also confirmed with higher expression levels of neurotrophins when DPSCs were cocultured with trigeminal neurons (Jung et al, 2016;Kawase-Koga et al, 2019). DPSCs also display angiogenic potential for differentiation potential of endothelial cells and the formation of blood vessels after in vivo transplantation with HA scaffold (Jeong et al, 2020).…”

“…The neurogenic potential was also confirmed with higher expression levels of neurotrophins when DPSCs were cocultured with trigeminal neurons ( Jung et al, 2016 ; Kawase-Koga et al, 2019 ). DPSCs also display angiogenic potential for differentiation potential of endothelial cells and the formation of blood vessels after in vivo transplantation with HA scaffold ( Jeong et al, 2020 ). A clinical study shows that implanted DPSCs achieve pulp-like tissue regeneration with vasculature and innervation in the root canal of traumatized incisors ( Nakashima et al, 2017 ).…”

Dental Tissue-Derived Human Mesenchymal Stem Cells and Their Potential in Therapeutic Application

“…The higher expression level of neurotrophins like nerve growth factor (NGF) and longer axons were detected in hDPSCs cultured with a microfluidic coculture system containing trigeminal neurons. Neurospheres were also generated by hDPSCs upon specific differentiation conditions [ 33 – 35 ]. The ability of hDPSCs to differentiate into endothelial cells and their angiogenic potential have also been reported, as hDPSCs were found to secrete vascular endothelial growth factors (VEGF) and generate visible blood vessels in three-dimensional- (3D-) printed HA constructs [ 36 ].…”

“…Lastly, a novel dentin–pulp-like organoid was developed by constructs mixed with hDPSCs and Matrigel in an odontogenic differentiation medium. The organoid demonstrated a biologically active response to biodentine supplements and suggested hDPSCs as a future approach for tooth regeneration [ 33 ]. Although scientific evidence shows a positive trend to dentin regeneration using dental MSCs, especially hDPSCs in animal models, there is a lack of persuasive evidence of clinical trials up to now.…”

Ultra-High-Resolution 3D Optical Coherence Tomography Reveals Inner Structures of Human Placenta-Derived Trophoblast Organoids

“…Here, entire organoids can be imaged in detail, but fixation, labeling, and optical clearance need to be performed [17]. Further, micro-computed tomography (microCT) was established for analyzing dentin-pulp-like and retinal organoids, but again, sample fixation was required [19], [20]. Improvement of live imaging was obtained with lattice light sheet microscopy, but this technique still required fluorescence labeling [21].…”