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Journal of Endodontics Jan 2024Stem cell-based dental pulp regeneration has been extensively studied, mainly focusing on exploiting dental stem cells' osteogenic and angiogenic potentials. Dental stem...
Interactions of Neuronally Induced Stem Cells from Apical Papilla Spheres, Stems Cells from Apical Papilla, and Human Umbilical Vascular Endothelial Cells on Vasculogenesis and Neurogenesis.
INTRODUCTION
Stem cell-based dental pulp regeneration has been extensively studied, mainly focusing on exploiting dental stem cells' osteogenic and angiogenic potentials. Dental stem cells' neurogenic role is often overlooked. Stem cells from apical papilla (SCAPs), originating from the neural crest and capable of sphere formation, display potent neurogenic capacity. This study aimed to investigate the interactions of neuronally induced stem cells from apical papilla (iSCAP) spheres, SCAPs, and human umbilical vascular endothelial cells (HUVECs) on vasculogenesis and neurogenesis.
METHODS
SCAPs were isolated and characterized using flow cytometry and multilineage differentiation assays. SCAP monolayer culture and spheres were neuronally induced by a small molecule neural induction medium, and the neural gene expression and neurite formation at days 0, 3, and 7 were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and using phase-contrast light and fluorescence microscopy. Direct coculture or pulp-on-chip was used to investigate iSCAP sphere interaction with SCAPs and HUVECs. RT-qPCR, fluorescence microscopy, and immunostaining with β-tubulin III, alpha-smooth muscle actin, and CD31 were used to study neural gene expression, neurite formation, and neurovascular cell interactions.
RESULTS
Neural induction medium with small molecules rapidly induced SCAP differentiation toward neural-like cells. Gene expression of Nestin, β-tubulin III, microtubule-associated protein 2, neuron-specific enolase, and NeuN was higher in iSCAP spheres than in iSCAPs. iSCAP spheres formed more and longer neurites compared with iSCAPs. iSCAP sphere, HUVEC, and SCAP direct coculture significantly enhanced vessel formation along with up-regulated VEGF (P < .001) and multiple neural markers, such as Nestin (P < .01), microtubule-associated protein 2 (P < .001), S100 (P < .001), and NG2 (P < .001). iSCAP spheres, SCAPs, and HUVECs cultured in a pulp-on-chip system promoted endothelial and neural cell migration toward each other and alpha-smooth muscle actin-positive and CD31-positive cells assembling for the vascular constitution.
CONCLUSIONS
iSCAP-formed spheres interact with SCAPs and HUVECs, promoting vasculogenesis and neurogenesis.
Topics: Humans; Endothelial Cells; Dental Pulp; Nestin; Dental Papilla; Tubulin; Actins; Regeneration; Stem Cells; Cell Differentiation; Neurogenesis; Cells, Cultured; Microtubule-Associated Proteins; Osteogenesis
PubMed: 37866800
DOI: 10.1016/j.joen.2023.10.006 -
Molecules (Basel, Switzerland) Feb 2021Dental papilla cells (DPCs), precursors of odontoblasts, are considered promising seed cells for tissue engineering. Emerging evidence suggests that melatonin promotes...
Dental papilla cells (DPCs), precursors of odontoblasts, are considered promising seed cells for tissue engineering. Emerging evidence suggests that melatonin promotes odontoblastic differentiation of DPCs and affects tooth development, although the precise mechanisms remain unknown. Retinoid acid receptor-related orphan receptor α (RORα) is a nuclear receptor for melatonin that plays a critical role in cell differentiation and embryonic development. This study aimed to explore the role of RORα in odontoblastic differentiation and determine whether melatonin exerts its pro-odontogenic effect via RORα. Herein, we observed that RORα was expressed in DPCs and was significantly increased during odontoblastic differentiation in vitro and in vivo. The overexpression of RORα upregulated the expression of odontogenic markers, alkaline phosphatase (ALP) activity and mineralized nodules formation ( < 0.05). In contrast, odontoblastic differentiation of DPCs was suppressed by RORα knockdown. Moreover, we found that melatonin elevated the expression of odontogenic markers, which was accompanied by the upregulation of RORα ( < 0.001). Utilising small interfering RNA, we further demonstrated that RORα inhibition attenuated melatonin-induced odontogenic gene expression, ALP activity and matrix mineralisation ( < 0.01). Collectively, these results provide the first evidence that RORα can promote odontoblastic differentiation of DPCs and mediate the pro-odontogenic effect of melatonin.
Topics: Animals; Cell Differentiation; Cells, Cultured; Dental Papilla; Melatonin; Nuclear Receptor Subfamily 1, Group F, Member 1; Odontoblasts; Odontogenesis; Rats, Sprague-Dawley; Up-Regulation; Rats
PubMed: 33669807
DOI: 10.3390/molecules26041098 -
Advances in Experimental Medicine and... 2018Biodegradable scaffolds are useful tools in the field of tissue engineering and regenerative medicine. The aim of this study was to test the potential of the human stem...
Biodegradable scaffolds are useful tools in the field of tissue engineering and regenerative medicine. The aim of this study was to test the potential of the human stem cells of apical papilla (SCAP) to attach, proliferate and differentiate on a polycaprolactone (PCL)-based scaffolds. SCAP were extracted from the root apical papillae of freshly extracted immature premolar teeth by using enzymatic digestion. Porous PCL scaffolds were fabricated using particle leaching method and NaCl or mannitol as porogens. SCAP of passage 3 were seeded on non-porous and porous PCL scaffolds for up to 14 days. For control, cells were cultured on glass coverslips. Picogreen DNA quantification was used to assay for cell proliferation. Cell differentiation and development of calcification nodules were examined using scanning electron microscopy and alizarin red staining. SCAP showed a comparable attachment, growth and proliferation patterns on PCL scaffolds and coverslips. Cell proliferation was enhanced on mannitol scaffolds at all time points. Calcification nodules were detected in all PCL scaffolds while it was not present on glass coverslips. These nodules were detected on NaCl-scaffolds by day 7 and on mannitol and non-porous scaffolds by day 14. In conclusion, SCAP were able to attach, proliferate and differentiate on PCL scaffolds without using any inductive media, indicating their potential application for dental tissue regeneration.
Topics: Cell Differentiation; Cell Proliferation; Cells, Cultured; Dental Papilla; Humans; Polyesters; Stem Cells; Tissue Engineering; Tissue Scaffolds
PubMed: 30357682
DOI: 10.1007/978-981-13-0947-2_3 -
PeerJ 2023Dental papilla cells (DPCs) are one of the key stem cells for tooth development, eventually forming dentin and pulp. Previous studies have reported that PER2 is...
BACKGROUND
Dental papilla cells (DPCs) are one of the key stem cells for tooth development, eventually forming dentin and pulp. Previous studies have reported that PER2 is expressed in a 24-hour oscillatory pattern in DPCs . , PER2 is highly expressed in odontoblasts (which are differentiated from DPCs). However, whether PER2 modulates the odontogenic differentiation of DPCs is uncertain. This research was to identify the function of PER2 in the odontogenic differentiation of DPCs and preliminarily explore its mechanisms.
METHODS
We monitored the expression of PER2 in DPCs differentiated . We used PER2 overexpression and knockdown studies to assess the role of PER2 in DPC differentiation and performed intracellular ATP content and reactive oxygen species (ROS) assays to further investigate the mechanism.
RESULTS
PER2 expression was considerably elevated throughout the odontoblastic differentiation of DPCs . Overexpressing boosted levels of odontogenic differentiation markers, such as dentin sialophosphoprotein (), dentin matrix protein 1 (), and alkaline phosphatase (), and enhanced mineralized nodule formation in DPCs. Conversely, the downregulation of inhibited the differentiation of DPCs. Additionally, downregulating further affected intracellular ATP content and ROS levels during DPC differentiation.
CONCLUSION
Overall, we demonstrated that PER2 positively regulates the odontogenic differentiation of DPCs, and the mechanism may be related to mitochondrial function as shown by intracellular ATP content and ROS levels.
Topics: Reactive Oxygen Species; Dental Papilla; Cell Differentiation; Odontoblasts; Adenosine Triphosphate
PubMed: 38084142
DOI: 10.7717/peerj.16489 -
Theranostics 2020Hertwig's epithelial root sheath (HERS) plays indispensable roles in tooth root development, including controlling the shape and number of roots, dentin formation, and...
Hertwig's epithelial root sheath (HERS) plays indispensable roles in tooth root development, including controlling the shape and number of roots, dentin formation, and helping generate the cementum. Based on these characteristics, HERS cell is a potential seed cell type for tooth-related tissue regeneration. However, the application is severely limited by a lack of appropriate culture methods and small cell numbers. Here, we constructed a 3D culture method to expand functional HERS cells into spheroids, and investigated characteristics and application of dental tissue regeneration of these spheroids. HERS spheroids and HERS cells (2D monolayer culture) were compared in terms of biological characteristics (such as proliferation, self-renewal capacity, and stemness) and functions (including differentiation potential and inductive ability of dentin formation) both and . Further, transcriptome analysis was utilized to reveal the molecular mechanisms of their obvious differences. HERS spheroids showed obvious superiority in biological characteristics and functions compared to 2D monolayers of HERS cells . , HERS spheroids generated more mineralized tissue; when combined with dental papilla cells (DPCs), HERS spheroids contributed to dentin-like tissue formation. Moreover, the generation and expansion of HERS spheroids rely to some degree on the HIF-1 pathway. HERS spheroid generation is beneficial for functional HERS cell expansion and can provide a useful cell source for further tooth regeneration and mechanistic research. Notably, HIF-1 pathway plays a critical role in HERS spheroid formation and function.
Topics: Animals; Cell Differentiation; Cell Proliferation; Cell Self Renewal; Dentin; Epithelial Cells; Female; Hypoxia-Inducible Factor 1; Models, Animal; Odontogenesis; Primary Cell Culture; Rats; Regeneration; Regenerative Endodontics; Spheroids, Cellular; Stem Cells; Tooth Root
PubMed: 32642002
DOI: 10.7150/thno.44782 -
Swiss Dental Journal Sep 2021Tooth development begins in the human embryo 28 to 40 days after ovulation. Epithelial cells grow into the ectomesenchymal parts of the jaw. An epithelial protrusion...
Tooth development begins in the human embryo 28 to 40 days after ovulation. Epithelial cells grow into the ectomesenchymal parts of the jaw. An epithelial protrusion develops. Further penetration of the epithelial cells into the ectomesenchyma results in the formation of the dental papilla (Figs. 1, 2 and 3). At this point, the cells for the formation of the tooth hard substance are differentiated.
PubMed: 34472765
DOI: 10.61872/sdj-2021-09-04 -
Journal of Oral Science Oct 2022Primordial odontogenic tumor (POT) is a rare mixed odontogenic tumor composed of primitive ectomesenchyme similar to the dental papilla. The outer surface consists of...
Primordial odontogenic tumor (POT) is a rare mixed odontogenic tumor composed of primitive ectomesenchyme similar to the dental papilla. The outer surface consists of columnar/cuboidal odontogenic epithelium similar to the inner enamel epithelium, and there is no hard tissue formation. Until now, 27 cases have been reported in the English literature. This article describes the clinicopathological characteristics of one case of POT, representing the oldest patient (aged 26 years) reported to date.
Topics: Adult; Calcinosis; Epithelium; Humans; Jaw Neoplasms; Male; Odontogenic Tumors
PubMed: 36070922
DOI: 10.2334/josnusd.22-0028 -
Development, Growth & Differentiation Sep 2021As precursor cells of odontoblasts, dental papilla cells (DPCs) form the dentin-pulp complex during tooth development. Nitric oxide (NO) regulates the functions of...
As precursor cells of odontoblasts, dental papilla cells (DPCs) form the dentin-pulp complex during tooth development. Nitric oxide (NO) regulates the functions of multiple cells and organ tissues, including stem cell differentiation and bone formation. In this paper, we explored the involvement of NO in odontoblastic differentiation. We verified the expression of NO synthase (NOS) in rat odontoblasts by nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) staining and immunohistochemistry in vivo. The expression of all three NOS isoforms in rat DPCs was confirmed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), immunofluorescence, and western blotting in vitro. The expression of neuronal NOS and endothelial NOS was upregulated during the odontoblastic differentiation of DPCs. Inhibition of NOS function by NOS inhibitor l-N -monomethyl arginine (L-NMMA) resulted in reduced formation of mineralized nodules and expression of dentin sialophosphoprotein (DSPP) and dentin matrix protein (DMP1) during DPC differentiation. The NO donor S-nitroso-N-acetylpenicillamine (SNAP, 0.1, 1, 10, and 100 μM) promoted the viability of DPCs. Extracellular matrix mineralization and odontogenic markers expression were elevated by SNAP at low concentrations (0.1, 1, and 10 μM) and suppressed at high concentration (100 μM). Blocking the generation of cyclic guanosine monophosphate (cGMP) with 1H-(1,2,4)oxadiazolo-(4,3-a)quinoxalin-1-one (ODQ) abolished the positive influence of SNAP on the odontoblastic differentiation of DPCs. These findings demonstrate that NO regulates the odontoblastic differentiation of DPCs, thereby influencing dentin formation and tooth development.
Topics: Animals; Cell Differentiation; Cells, Cultured; Dental Papilla; Dental Pulp; Nitric Oxide; Nitric Oxide Synthase; Odontoblasts; Rats
PubMed: 34411285
DOI: 10.1111/dgd.12745 -
Hedgehog signaling regulates dental papilla formation and tooth size during zebrafish odontogenesis.Developmental Dynamics : An Official... Apr 2015Intercellular communication by the hedgehog cell signaling pathway is necessary for tooth development throughout the vertebrates, but it remains unclear which specific...
BACKGROUND
Intercellular communication by the hedgehog cell signaling pathway is necessary for tooth development throughout the vertebrates, but it remains unclear which specific developmental signals control cell behavior at different stages of odontogenesis. To address this issue, we have manipulated hedgehog activity during zebrafish tooth development and visualized the results using confocal microscopy.
RESULTS
We first established that reporter lines for dlx2b, fli1, NF-κB, and prdm1a are markers for specific subsets of tooth germ tissues. We then blocked hedgehog signaling with cyclopamine and observed a reduction or elimination of the cranial neural crest derived dental papilla, which normally contains the cells that later give rise to dentin-producing odontoblasts. Upon further investigation, we observed that the dental papilla begins to form and then regresses in the absence of hedgehog signaling, through a mechanism unrelated to cell proliferation or apoptosis. We also found evidence of an isometric reduction in tooth size that correlates with the time of earliest hedgehog inhibition.
CONCLUSIONS
We hypothesize that these results reveal a previously uncharacterized function of hedgehog signaling during tooth morphogenesis, regulating the number of cells in the dental papilla and thereby controlling tooth size.
Topics: Animals; Apoptosis; Cell Communication; Cell Proliferation; DNA-Binding Proteins; Dental Papilla; Gene Expression Regulation, Developmental; Green Fluorescent Proteins; Hedgehog Proteins; Homeodomain Proteins; Microscopy, Fluorescence; Morphogenesis; NF-kappa B; Nuclear Proteins; Odontoblasts; Odontogenesis; Positive Regulatory Domain I-Binding Factor 1; Signal Transduction; Tooth; Tooth Germ; Transcription Factors; Veratrum Alkaloids; Zebrafish; Zebrafish Proteins
PubMed: 25645398
DOI: 10.1002/dvdy.24258 -
Journal of Esthetic and Restorative... Oct 2022To evaluate efficacy of platelet-rich fibrin (PRF) or connective tissue graft (CTG) in papilla reconstruction (PR) with the semilunar incision (SI) technique.
PURPOSE
To evaluate efficacy of platelet-rich fibrin (PRF) or connective tissue graft (CTG) in papilla reconstruction (PR) with the semilunar incision (SI) technique.
MATERIALS AND METHODS
The analysis consisted of 55 sites (27 CTG and 28 PRF) from 20 patients who underwent PR with either PRF or CTG placed in the maxillary anterior region with SI technique. Baseline (BL) and follow-up (T , first month, T , third month, T , sixth month) clinical data including periodontal evaluations (gingival index (GI), plaque index (PI), pocket depth (PD), keratinized tissue width (KTW), gingival recession), papilla-associated recordings (alveolar crest-interdental contact point [AC-IC], alveolar crest-papilla tip [AC-PT], papilla tip-interdental contact point [PT-IC], papilla height loss [PHL], interdental tissue stroke [ITS] and papilla presence index [PPI]) and patient satisfaction were analyzed.
RESULTS
CTG provided better PR outcomes. GI, PI, and PD showed a slight increase at T and then, turned to their BL levels. The other periodontal parameters showed significant improvement after both treatment modalities. No inter-group difference was found except for KTW, which was in favor of CTG.
CONCLUSION
Based on the results, CTG is recommended over PRF in PR treatment due to its superior outcomes with less recurrence risk.
CLINICAL SIGNIFICANCE
Connective tissue graft provides superior results than platelet-rich fibrin in papilla reconstruction with the semilunar incision technique.
Topics: Connective Tissue; Dental Papilla; Gingiva; Gingival Recession; Humans; Platelet-Rich Fibrin; Surgical Flaps; Treatment Outcome
PubMed: 35731089
DOI: 10.1111/jerd.12937