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Cell Differentiation May 1986The dental papilla is a mesenchymal cell condensation which plays an important regulatory role during tooth development. Dental papilla mesenchymes were enzymatically...
The dental papilla is a mesenchymal cell condensation which plays an important regulatory role during tooth development. Dental papilla mesenchymes were enzymatically separated from the dental epithelia from tooth germs of 17-day-old mouse embryos and disaggregated for monolayer culture. These cells were compared with gingival mesenchyme overlying the same tooth germs and with undifferentiated jaw mesenchyme from mandibles of 11-day-old embryos. The dental papilla cells were large and flat with numerous cell processes, whereas the gingival cells resembled typical spindle-shaped fibroblasts and grew to a higher cell density. Although the two mesenchymes differ in their collagen contents in vivo, no differences were detected either in the amount or type of collagen synthesized in vitro. Type I and III collagens were found in the culture media and type V collagen in the cell layer of both cell populations. The mandibular mesenchymal cells of the younger embryos resembled the dental papilla cells in morphology and growth rate. This may reflect retention of undifferentiated embryonic characteristics in the dental papilla. The successful culture of dental papilla cells now enables subsequent studies on the cellular properties related to the unique morphogenetic capabilities of these cells.
Topics: Animals; Cell Division; Collagen; Culture Techniques; Dental Papilla; Gingiva; Mesoderm; Mice; Molar; Odontogenesis; Tooth Germ
PubMed: 3708698
DOI: 10.1016/0045-6039(86)90085-0 -
In Vitro Cellular & Developmental... Sep 2013Odontogenesis is the result of the reciprocal interactions between epithelial-mesenchymal cells leading to terminally differentiated odontoblasts. This process from...
Odontogenesis is the result of the reciprocal interactions between epithelial-mesenchymal cells leading to terminally differentiated odontoblasts. This process from dental papilla mesenchymal cells to odontoblasts is regulated by a complex signaling pathway. When isolated from the developing tooth germs, odontoblasts quickly lose their potential to maintain the odontoblast-specific phenotype. Therefore, generation of an odontoblast-like cell line would be a good surrogate model for studying the dental mesenchymal cell differentiation into odontoblasts and the molecular events of dentin formation. In this study, immortalized dental papilla mesenchymal cell lines were generated from the first mouse mandibular molars at postnatal day 3 using pSV40. These transformed cells were characterized by RT-PCR, immunohistochemistry, Western blot, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iMDP-3, displayed a high proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers and demonstrated the ability to differentiate and form mineralized nodules. Furthermore, iMDP-3 cells had high transfection efficiency as well as were inducible and responded to BMP2 stimulation. We conclude that the establishment of the stable murine dental papilla mesenchymal cell line might be used for studying the mechanisms of dental cell differentiation and dentin formation.
Topics: Animals; Antigens, Viral, Tumor; Bone Morphogenetic Protein 2; Cell Differentiation; Cells, Cultured; Dental Papilla; Epithelial Cells; Gene Expression Regulation, Developmental; Humans; Mesoderm; Mice; Odontoblasts; Odontogenesis; Tooth; Transfection
PubMed: 23813243
DOI: 10.1007/s11626-013-9641-1 -
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 -
European Journal of Oral Sciences Aug 2014During tooth development, the special structure of dental follicle and dental papilla enables dental papilla cells (DPCs) and dental follicle cells (DFCs) to make...
During tooth development, the special structure of dental follicle and dental papilla enables dental papilla cells (DPCs) and dental follicle cells (DFCs) to make contact with each other. Octamer-binding transcription factor 4 (Oct-4), sex determining region Y box-2 (SOX-2), and cellular homologue of avian myelocytomatosis virus oncogene (MYC) (OSM) are associated with reprogramming and pluripotency. However, whether the expression of OSM could be activated through cell-cell communication is not known. In this study, the distribution of OSM in rat tooth germ was investigated by immunohistochemical staining. An in-vitro co-culture system of DPCs and DFCs was established. Cell proliferation, cell apoptosis, cell cycle stages, and expression of OSM were investigated by Cell Counting Kit 8 (CCK8) analysis, flow cytometry, real-time PCR, and immunohistochemical staining. We found that Oct-4 and SOX-2 were strongly expressed in tooth germ on days 7 and 9 after birth, whereas MYC was expressed only on day 9. Cell proliferation and apoptosis were inhibited, the cell cycle was arrested in the G0/G1 phase, and the propidium iodide (PI) value was downregulated. Expression of Oct-4 and SOX-2 was significantly elevated in both cell types after 3 d of co-culture, whereas expression of MYC was not significantly elevated until day 5. These results indicate that the optimized microenvironment with cell-cell communication enhanced the expression of reprogramming markers associated with reprogramming capacity in DPCs and DFCs, both in vivo and in vitro.
Topics: Animals; Apoptosis; Cell Communication; Cell Culture Techniques; Cell Proliferation; Cellular Microenvironment; Cellular Reprogramming; Coculture Techniques; Dental Papilla; Dental Sac; G1 Phase; Octamer Transcription Factor-3; Odontogenesis; Pluripotent Stem Cells; Proto-Oncogene Proteins c-myc; Rats; Rats, Sprague-Dawley; Resting Phase, Cell Cycle; SOXB1 Transcription Factors; Time Factors; Tooth Germ
PubMed: 25039286
DOI: 10.1111/eos.12141 -
Journal of Endodontics Feb 2010The Wnt signaling pathway plays an important role in tissue development by acting on proliferation, differentiation, and cell fate decisions. Because the role of Wnt6 in...
INTRODUCTION
The Wnt signaling pathway plays an important role in tissue development by acting on proliferation, differentiation, and cell fate decisions. Because the role of Wnt6 in tooth development was still unknown, the purpose of this study was to investigate the role of Wnt6 in tooth morphogenesis and dental tissue mineralization by elucidating its effect on human dental papilla cells (hDPCs) in vitro.
METHODS
Human dental papilla cells were enzymatically separated from tooth germs. Recombinant adenovirus encoding full-length Wnt6 cDNA was constructed to overexpress Wnt6, and the biologic effects of Wnt6 on hDPCs were investigated. Wnt6-transduced changes in hDPC proliferation were examined by means of a 5-bromodeoxyuridine (BrdU) incorporation assay and cell cycle analysis. Wnt6-transduced changes in hDPC differentiation were investigated by evaluating alkaline phosphatase (ALPase) activity, by a mineralization assay, and analysis of mineralization-related gene expression including ALP, type I collagen (Col I), osteonectin (ON), osteopontin (OPN), bone sialoprotein (BSP), and dentin matrix protein-1 (DMP-1).
RESULTS
Wnt6 overexpression had no significant effect on the proliferation of hDPCs by BrdU incorporation assay and flow cytometric analysis. Wnt6 enhanced differentiation of hDPCs into functional odontoblast-like cells with up-regulated activity of ALPase and the expression of mineralization-related genes such as ALP, Col I, ON, OPN, BSP, and DMP-1. Wnt6 overexpression also promoted the mineralization of hDPCs.
CONCLUSIONS
Our findings verified that Wnt6 plays an important role in tooth development by promoting hDPC differentiation, without significant effects on hDPC proliferation.
Topics: Alkaline Phosphatase; Cell Differentiation; Cell Proliferation; Cells, Cultured; DNA, Complementary; Dental Papilla; Gene Expression Regulation, Developmental; Humans; Odontogenesis; Recombinant Proteins; Tooth Calcification; Tooth Germ; Transduction, Genetic; Wnt Proteins
PubMed: 20113781
DOI: 10.1016/j.joen.2009.09.007 -
Journal of Cellular Physiology Oct 2010Bone morphogenetic protein 2 (Bmp2) is essential for odontogensis and dentin mineralization. Generation of floxed Bmp2 dental mesenchymal cell lines is a valuable...
Bone morphogenetic protein 2 (Bmp2) is essential for odontogensis and dentin mineralization. Generation of floxed Bmp2 dental mesenchymal cell lines is a valuable application for studying the effects of Bmp2 on dental mesenchymal cell differentiation and its signaling pathways during dentinogenesis. Limitation of the primary culture of dental mesenchymal cells has led to the development of cell lines that serve as good surrogate models for the study of dental mesenchymal cell differentiation into odontoblasts and mineralization. In this study, we established and characterized immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines, which were isolated from 1st mouse mandibular molars at postnatal day 1 and immortalized with pSV40 and clonally selected. These transfected cell lines were characterized by RT-PCR, immunohistochemistry, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iBmp2-dp, displayed a higher proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers as well as demonstrated the ability to differentiate and form mineralized nodules. In addition, iBmp2-dp cells were inducible and responded to BMP2 stimulation. Thus, we for the first time described the establishment of an immortalized mouse floxed Bmp2 dental papilla mesenchyma cell line that might be used for studying the mechanisms of dental cell differentiation and dentin mineralization mediated by Bmp2 and other growth factor signaling pathways.
Topics: Animals; Biomarkers; Bone Morphogenetic Protein 2; Calcification, Physiologic; Cell Differentiation; Cell Line; Cell Shape; Dental Papilla; Mesenchymal Stem Cells; Mice; Mice, Transgenic; Odontoblasts; Phenotype
PubMed: 20458728
DOI: 10.1002/jcp.22204 -
BioMed Research International 2019Stem cells are biological cells that can self-renew and can differentiate into multiple cell lineages. Stem cell-based therapy is emerging as a promising alternative... (Review)
Review
Stem cells are biological cells that can self-renew and can differentiate into multiple cell lineages. Stem cell-based therapy is emerging as a promising alternative therapeutic option for various disorders. Mesenchymal stem cells (MSCs) are multipotent adult stem cells that are isolated from various tissues and can be used as an alternative to embryonic stem cells. Stem cells from the apical papilla (SCAPs) are a novel population of MSCs residing in the apical papilla of immature permanent teeth. SCAPs present the characteristics of expression of MSCs markers, self-renewal, proliferation, migration, differentiation, and immunosuppression, which support the application of SCAPs in stem cell-based therapy, including the immunotherapy and the regeneration of dental tissues, bone, neural, and vascular tissues. In view of these properties and therapeutic potential, SCAPs can be considered as promising candidates for stem cell-based therapy. Thus the aim of our review was to summarize the current knowledge of SCAPs considering isolation, characterization, and multilineage differentiation. The prospects for their use in stem cell-based therapy were also discussed.
Topics: Cell Differentiation; Cell Lineage; Cell Proliferation; Dental Papilla; Humans; Mesenchymal Stem Cells; Multipotent Stem Cells; Osteogenesis; Stem Cell Transplantation
PubMed: 30834270
DOI: 10.1155/2019/6104738 -
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 -
Journal of Oral Pathology & Medicine :... Aug 2004Human telomerase reverse transcriptase (hTERT) is catalytic subunit of human telomerase.
BACKGROUND
Human telomerase reverse transcriptase (hTERT) is catalytic subunit of human telomerase.
METHODS
We studied the immortalization of a series of human dental and periodontal cells by ectopic expression of hTERT and co-expression of hTERT with human papilloma virus 16 (HPV16) or simian virus 40 (SV40). Differentiation abilities of the established cell lines were studied in terms of the mineralized matrix formation and gene expression.
RESULTS
We established immortalized gingival fibroblasts by hTERT, dental papilla and periodontal ligament cells by hTERT and HPV16, and pulp cells by hTERT and SV40. The papilla and pulp cells showed mineralization and dentin sialophosphoprotein (DSPP) expression when cultured in the presence of beta-glycerophosphate. The immortalized periodontal ligament cells did not show mineralization or DSPP expression, although expressions of alkaline phosphatase, osteopontin and osteocalcin were detected.
CONCLUSIONS
These cell lines will be useful tools for studying the repair and regeneration of dental and periodontal tissues and various diseases including odontogenic tumors.
Topics: Animals; Cell Line, Transformed; Cells, Cultured; Collagen Type I; DNA, Viral; DNA-Binding Proteins; Dental Papilla; Dental Pulp; Extracellular Matrix; Extracellular Matrix Proteins; Fibroblasts; Gene Expression; Gingiva; Humans; Mice; Mice, SCID; Osteocalcin; Osteopontin; Periodontal Ligament; Phosphoproteins; Protein Precursors; Sialoglycoproteins; Telomerase; Transfection
PubMed: 15250834
DOI: 10.1111/j.1600-0714.2004.00228.x -
International Journal of Oral and... Nov 2017A better understanding of factors that can lead to papilla formation or recession, such as the type of site where the implant was placed, is of fundamental importance to... (Review)
Review
A better understanding of factors that can lead to papilla formation or recession, such as the type of site where the implant was placed, is of fundamental importance to the aesthetic success of the rehabilitation. The aim of this study was to perform a systematic review of the literature regarding the formation or recession of papilla adjacent to implants placed in fresh, healing or healed sites. The protocol for this study was registered in the PROSPERO database (registration number CRD 42016033784). An electronic search was performed by two independent reviewers who applied the inclusion and exclusion criteria on the PubMed/MEDLINE, Scopus, and Embase databases from January 2005 up to February 2016. The initial screening yielded 1,065 articles, from which 15 were selected for a systematic review after applying the inclusion and exclusion criteria. Nine studies compared fresh and healed sites, four studies compared healing and healed sites, one study compared fresh and healing sites, and one study analysed all three sites. The majority of studies identified by this systematic review showed no difference between groups after the longer follow-up period. The sites where the implants were placed did not have a long-term influence on papilla formation or recession.
Topics: Dental Implantation, Endosseous; Dental Implants; Dental Papilla; Esthetics, Dental; Gingival Recession; Humans; Wound Healing
PubMed: 28521965
DOI: 10.1016/j.ijom.2017.04.018