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Journal of Cellular Physiology Nov 2015Fam20c is essential for the normal mineralization of dentin and bone. The generation of odontoblast and osteoblast cell lines carrying floxed Fam20c allele can offer...
Fam20c is essential for the normal mineralization of dentin and bone. The generation of odontoblast and osteoblast cell lines carrying floxed Fam20c allele can offer valuable tools for the study of the roles of Fam20c in the mineralization of dentin and bone. The limited capability of the primary odontoblasts and osteoblasts to proliferate necessitates the development of odontoblast and osteoblast cell lines serving as substitutes for the study of differentiation and mineralization of the odontoblasts and osteoblasts. In this study, we established and characterized immortalized mouse floxed Fam20c dental papilla mesenchymal and osteoblast cell lines. The isolated primary mouse floxed Fam20c dental papilla mesenchymal cells and osteoblasts were immortalized by the infection of lentivirus containing Simian Virus 40 T-antigen (SV40 T-Ag). The immortalization of floxed Fam20c dental papilla mesenchymal cells and osteoblasts was verified by the long-term passages and genomic integration of SV40 T-Ag. The immortalized floxed Fam20c dental papilla mesenchymal and osteoblast cell lines not only proliferated at a high rate and retained the morphology of their primary counterparts, but also preserved the dentin and bone specific gene expression as the primary dental papilla mesenchymal cells and osteoblasts did. Consistently, the capability of the primary floxed Fam20c dental papilla mesenchymal cells and osteoblasts to mineralize was also inherited by the immortalized dental papilla mesenchymal and osteoblast cell lines. Thus, we have successfully generated the immortalized mouse floxed Fam20c dental papilla mesenchymal and osteoblast cell lines.
Topics: Animals; Bone Morphogenetic Protein 2; Calcification, Physiologic; Calcium-Binding Proteins; Cell Differentiation; Cell Line; Cell Proliferation; Dental Papilla; Dentin; Extracellular Matrix Proteins; Gene Expression Regulation, Developmental; Mesenchymal Stem Cells; Mice; Osteoblasts
PubMed: 25833681
DOI: 10.1002/jcp.25008 -
TGFβ3 secretion by three-dimensional cultures of human dental apical papilla mesenchymal stem cells.Journal of Tissue Engineering and... Apr 2017Mesenchymal stem cells (MSCs) can be isolated from dental tissues, such as pulp and periodontal ligament; the dental apical papilla (DAP) is a less-studied MSC source....
Mesenchymal stem cells (MSCs) can be isolated from dental tissues, such as pulp and periodontal ligament; the dental apical papilla (DAP) is a less-studied MSC source. These dental-derived MSCs are of great interest because of their potential as an accessible source for cell-based therapies and tissue-engineering (TE) approaches. Much of the interest regarding MSCs relies on the trophic-mediated repair and regenerative effects observed when they are implanted. TGFβ3 is a key growth factor involved in tissue regeneration and scarless tissue repair. We hypothesized that human DAP-derived MSCs (hSCAPs) can produce and secrete TGFβ3 in response to micro-environmental cues. For this, we encapsulated hSCAPs in different types of matrix and evaluated TGFβ3 secretion. We found that dynamic changes of cell-matrix interactions and mechanical stress that cells sense during the transition from a monolayer culture (two-dimensional, 2D) towards a three-dimensional (3D) culture condition, rather than the different chemical composition of the scaffolds, may trigger the TGFβ3 secretion, while monolayer cultures showed almost 10-fold less secretion of TGFβ3. The study of these interactions is provided as a cornerstone in designing future strategies in TE and cell therapy that are more efficient and effective for repair/regeneration of damaged tissues. Copyright © 2015 John Wiley & Sons, Ltd.
Topics: Adolescent; Adult; Antigens, CD; Cell Separation; Cells, Cultured; Dental Papilla; Gene Expression Regulation; Humans; Mesenchymal Stem Cells; Models, Biological; Principal Component Analysis; Transforming Growth Factor beta3; Young Adult
PubMed: 25690385
DOI: 10.1002/term.2004 -
General Dentistry 2017Due to improvements in esthetic dentistry, the number of patients seeking to obtain a more harmonious smile has increased. A 40-year-old woman was referred for treatment...
Due to improvements in esthetic dentistry, the number of patients seeking to obtain a more harmonious smile has increased. A 40-year-old woman was referred for treatment because she was dissatisfied with the effect that diastemas and a conoid tooth had on her smile. Her maxillary anterior teeth were relatively short, resulting in a compromised esthetic relationship between height and width. Orthodontic treatment for better distribution of her teeth was recommended to the patient, but she refused, wanting faster and less expensive treatment. After diagnostic and waxed-up study casts were obtained, composite resin esthetic mock-ups were made to confirm that sufficient space was left for formation of the interdental papilla. After the patient approved the mock-ups, her anterior teeth were submitted to bleaching, definitive restoration, and additional occlusal adjustments. At the 1-year follow-up, the patient exhibited a harmonious, esthetic smile without black spaces or periodontal inflammation.
Topics: Adult; Composite Resins; Dental Papilla; Dental Restoration, Permanent; Diastema; Esthetics, Dental; Female; Humans; Smiling; Tooth Abnormalities; Tooth Bleaching
PubMed: 29099376
DOI: No ID Found -
Chang Gung Medical Journal Nov 2003Loss of the interproximal dental papilla may cause functional and, especially in the maxillary anterior region, phonetic and severe esthetic problems. The purpose of... (Clinical Trial)
Clinical Trial Randomized Controlled Trial
BACKGROUND
Loss of the interproximal dental papilla may cause functional and, especially in the maxillary anterior region, phonetic and severe esthetic problems. The purpose of this study was to investigate whether the distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth could be correlated with the presence of the interproximal papilla in Taiwanese patients.
METHODS
In total, 200 interproximal sites of maxillary anterior teeth in 45 randomly selected patients were examined. Selected subjects were adult Taiwanese with fully erupted permanent dentition. The presence of the interproximal papilla was determined visually. If there was no visible space apical to the contact area, the papilla was recorded as being present. The distance from the contact point to the crest of bone was measured on standardized periapical radiographs using a paralleling technique with a RinnXCP holder.
RESULTS
Data revealed that when the distance from the contact point to the bone crest on standardized periapical radiographs was 5 mm or less, the papillae were almost 100% present. When the distance was 6 mm, 51% of the papillae were present, and when the distance was 7 mm or greater, only 23% of the papillae were present.
CONCLUSION
The distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth is highly associated with the presence or absence of the interproximal papilla in Taiwanese patients, and is a useful guide for clinical evaluation.
Topics: Adult; Alveolar Process; Dental Occlusion; Dental Papilla; Humans; Maxilla; Radiography, Dental; Taiwan
PubMed: 14765752
DOI: No ID Found -
Journal of Dental Research 1958
Topics: Dental Papilla; Tissue Culture Techniques; Tooth Germ
PubMed: 13587798
DOI: 10.1177/00220345580370050201 -
Journal of Molecular Histology Jun 2019Interactions between the ectodermal and mesenchymal tissues are the basis of the central mechanism regulating tooth development. Based on this epithelial-mesenchymal...
Interactions between the ectodermal and mesenchymal tissues are the basis of the central mechanism regulating tooth development. Based on this epithelial-mesenchymal interaction (EMI), we demonstrated that copine-7 (CPNE7) is secreted by preameloblasts and regulates the differentiation of mesenchymal cells of dental or non-dental origin into odontoblasts. However, the precise expression patterns of CPNE7 in the stages of tooth development have not yet been elucidated. The aim of the present study was to establish the spatiotemporal expression pattern of CPNE7 during mouse tooth development. To examine the spatiotemporal expression patterns of CPNE7 during mouse tooth development, we investigate the distribution of CPNE7 in the embryonic and postnatal developing mouse tooth. Immunohistochemistry, in situ hybridization, real-time PCR, and western blot analysis are performed to investigate the CPNE7 expression pattern during tooth development of the mandibular mouse first molar. During the initiation stage (bud stage), CPNE7 protein expression is observed in the dental epithelium but not yet in the dental mesenchyme. At E18 (bell stage), expression of CPNE7 protein and mRNA is primarily observed in ectomesenchymal cells of dental papilla. At P7 (crown formation stage), CPNE7 is localized in differentiating odontoblasts but weak expression is detected in mature ameloblasts. These findings suggest that CPNE7 secreted by dental epithelium induces the differentiation of ectomesenchymal cells into preodontoblast in concert with EMI. CPNE7 is clearly expressed in differentiating odontoblasts and the odontoblast process during dentinogenesis, but is no longer expressed in fully differentiated odontoblasts. Furthermore, CPNE7 is expressed in the Hertwig's epithelial root sheath (HERS) and in the facing preodontoblasts during root dentin formation. Taken together, these results illustrate the dynamic expression of CPNE7 during tooth development and suggest its important function in entire stages of tooth development.
Topics: Ameloblasts; Animals; Cell Differentiation; Dental Papilla; Dentinogenesis; Gene Expression Regulation, Developmental; Membrane Proteins; Mice; Molar; Odontoblasts; Tooth
PubMed: 30863901
DOI: 10.1007/s10735-019-09816-0 -
In Vitro Cellular & Developmental... Feb 2013Odontoblasts are a type of non-proliferating and terminally differentiated cells that play an important role in the pulpo-dentinal complex. Mouse dental papilla cells...
Odontoblasts are a type of non-proliferating and terminally differentiated cells that play an important role in the pulpo-dentinal complex. Mouse dental papilla cells (mDPCs), which can differentiate into odontoblast-like cells in vitro, have a limited life span. We combined the traditional strategy of "Cre/LoxP-based reversible immortalization" with a tamoxifen-regulated Cre recombination system to generate a tamoxifen-mediated reversibly immortalized mouse dental papilla cell line, mDPCET. mDPCs were sequentially transduced with a floxed SV40 T antigen-TK (SV40Tag-TK) and an (ERT2)Cre(ERT2)-expressing plasmid. Clonal-isolated SV40Tag- and Cre-positive cells showed modified growth characteristics and a significantly extended life span. When mDPCET cells were treated with 4-hydroxytamoxifen, (ERT2)Cre(ERT2) translocated from the cytoplasm to the nucleus and caused the excision of SV40Tag-TK, which led to the reversion of mDPCETs. After the immortalization was reversed, the cells underwent replicative senescence and transitioned into a more differentiated state. Tamoxifen-mediated reversible immortalization, therefore, allows for the expansion of primary mDPCs, leads to the production of odontoblast-like cells that retain most odontoblast-specific properties, and can represent a safe and ready-to-use method due to its simple manipulation.
Topics: Animals; Cell Differentiation; Cell Line; Dental Papilla; Genetic Vectors; Mice; Odontoblasts; Primary Cell Culture; Tamoxifen
PubMed: 23299318
DOI: 10.1007/s11626-012-9576-y -
Scandinavian Journal of Dental Research Jun 1988The mesenchymal component of the embryonic tooth, the dental papilla, shows unique cellular behavior. Only the dental papilla cells are capable of differentiating into...
The mesenchymal component of the embryonic tooth, the dental papilla, shows unique cellular behavior. Only the dental papilla cells are capable of differentiating into odontoblasts. The dental papilla also directs the morphogenesis of the tooth. Our aim is to develop suitable markers for further studies on the molecular mechanisms behind the determination and differentiation of the dental mesenchymal cells. We have produced monoclonal antibodies against this embryonal cell population. Altogether 1114 enzymatically separated mesenchymes from dissected molar tooth germs of 17-day-old mouse embryos were fixed in paraformaldehyde (PFA) and sonicated in saline. A rat was immunized and hybridomas were produced by standard methods. The supernatants were screened by immunohistology, using both frozen sections and sections of PFA-fixed paraffin-embedded mouse molar teeth. Altogether, 19 wells produced antibodies reacting with dental tissues. One monoclonal antibody recognizes a 67,000 dalton intracellular antigen enriched in secretory odontoblasts and ameloblasts. The antigen is also found in osteoblasts and chondrocytes in the developing jaw, and in some cultured murine cells. The method described here appears to be successful for generating and screening monoclonal antibodies against the dental papilla.
Topics: Ameloblasts; Animals; Antibodies, Monoclonal; Cell Differentiation; Dental Papilla; Hybridomas; Mice; Odontoblasts; Rats; Tooth Germ
PubMed: 3164901
DOI: 10.1111/j.1600-0722.1988.tb01542.x -
Connective Tissue Research 1995This study describes the immunolocalization of actin, cytokeratins and vimentin during differentiation of the dental papilla in the rat. Incisors and first molars were...
This study describes the immunolocalization of actin, cytokeratins and vimentin during differentiation of the dental papilla in the rat. Incisors and first molars were sectioned from mandibles of Wistar rats from embryonic day (E)-14 to (E)-21 and weeks 1, 2, 3, 12 and 104 after birth, fixed in 90% alcohol, decalcified in EDTA, infiltrated with 5% sucrose, frozen in dry ice, and cryosectioned at 10 microns. The sections were immunolabelled using indirect immunofluorescence with a panel of monoclonal antibodies and FITC-phalloidin for F-actin localization. F-actin was present in follicular mesenchyme and odontoblast processes. Vimentin labelled dental papilla fibroblasts, differentiating, functional (secretory) and aged odontoblasts. Vimentin was uniformly localized in the cytoplasm of pre-odontoblasts but was redistributed to the apical pole of these cells during polarization. Of the cytokeratins, only cytokeratin 19 was found in differentiating odontoblasts. It was not present in dental papilla fibroblasts, functional or aged odontoblasts. These results suggest that actin and the redistribution of vimentin may be involved in odontoblast differentiation and odontoblast process formation/support and that these events may be preceded by the expression of cytokeratin 19.
Topics: Actins; Aging; Animals; Antibodies, Monoclonal; Cell Differentiation; Cytoskeletal Proteins; Cytoskeleton; Dental Papilla; Dental Pulp; Dental Sac; Embryo, Mammalian; Fluorescent Antibody Technique; Gene Expression; Keratins; Mesoderm; Odontoblasts; Rats; Rats, Wistar; Vimentin
PubMed: 7554938
DOI: 10.3109/03008209509013708 -
PloS One 2017Induced pluripotent stem cells (iPSCs) technology provides a powerful means to generate and regenerate unlimited pluripotent stem cells directly from body tissue cells....
Induced pluripotent stem cells (iPSCs) technology provides a powerful means to generate and regenerate unlimited pluripotent stem cells directly from body tissue cells. Stem cells from apical papilla (SCAP) and Dental pulp stem cells (DPSCs) are present in 'cell-rich zones' within the dental pulp region, which are capable of regenerating pulp and dentin tissues in vivo. In this study, we investigated the difference of miRNAs expression in SCAPs and DPSCs before and after the reprogramming. Using miRNA microarray, 134 and 265 differentially expressed miRNAs in DPSCs- and SCAP-iPSCs were up-regulated compared to these before reprogramming. 117 specific miRNAs with enhanced more than 2-fold were identified in both DPSCs- and SCAP-iPSCs. Among the co-regulated miRNAs, miR-19a-3p, miR-92b-3p and miR-130b-3p showed the maximum difference, which had involvement in the cell cycle, TGF beta signaling pathway and epithelial mesenchymal transition. Using qRT-PCR analysis, the expression of miR-19a-3p, miR-92b-3p and miR-130b-3p indicated substantial increases in DPSCs-iPSCs and SCAP-iPSCs. The findings suggest that miRNAs play a part in the difference between DPSCs-iPSCs and DPSCs, as well as between SCAP-iPSCs and SCAP. The variation of miRNA expression in reprogrammed dental-derived pluripotent stem cells revealed different characteristics induced by iPSC generation.
Topics: Cellular Reprogramming; Dental Papilla; Dental Pulp; Gene Expression Profiling; Humans; Induced Pluripotent Stem Cells; MicroRNAs; Young Adult
PubMed: 28542487
DOI: 10.1371/journal.pone.0177832