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Journal of Oral and Maxillofacial... Mar 2009This study examined the osteogenic phenotypes and mineralization of cultured human dental papilla-derived cells.
PURPOSE
This study examined the osteogenic phenotypes and mineralization of cultured human dental papilla-derived cells.
MATERIALS AND METHODS
Dental papillae were harvested from mandibles during surgical extraction of lower impacted third molars from 3 patients aged 13 to 15 years. The dental papilla-derived cells were introduced into the cell culture. After passage 3, the dental papilla-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. We examined the histochemical detection of alkaline phosphatase (ALP), the reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for ALP and osteocalcin, and von Kossa staining in the dental papilla-derived cells.
RESULTS
It was observed that ALP was strongly expressed in the earlier stage of osteoblastic differentiation, whereas osteocalcin was mainly expressed and secreted into the medium at the later stage. Von Kossa-positive mineralization nodules were first observed on day 14, which increased in number during the entire culture period.
CONCLUSIONS
These results suggest that dental papilla-derived cell have osteogenic potential and could be used as an additional source of cells for bone tissue engineering.
Topics: Adolescent; Alkaline Phosphatase; Calcification, Physiologic; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Dental Papilla; Gene Expression; Humans; Mesenchymal Stem Cells; Molar, Third; Osteoblasts; Osteocalcin; Osteogenesis; Reverse Transcriptase Polymerase Chain Reaction; Tissue Engineering
PubMed: 19231773
DOI: 10.1016/j.joms.2008.08.037 -
Dental Materials : Official Publication... Jun 2002The use of adequate target cells for cytotoxicity testing of dental restorative materials has often been experimentally assessed with respect to the clinical relevance... (Comparative Study)
Comparative Study
OBJECTIVE
The use of adequate target cells for cytotoxicity testing of dental restorative materials has often been experimentally assessed with respect to the clinical relevance of the test results. In the present study, the responses in primary bovine dental papilla-derived cells (pulp cells) were compared with those in transformed dental papilla-derived cell lines and L929 mouse fibroblasts after exposure to various dental resin compounds.
METHODS
Primary bovine dental papilla-derived cells (CPC), tCPC B (CPC cells transformed with SV40 T-antigen), tCPC E (CPC cells transformed with E6/E7 oncogen), and L929 mouse fibroblast cells were exposed to various compounds of dental resin materials for 24 h, and cytotoxicity was determined using the MTT assay. Bis-GMA, UDMA, 1,6 hexane diol dimethacrylate (HDDM), TEGDMA, HEMA, MMA, camphorquinone (CQ), bisphenol A (BPA), and glycidyl methacrylate (GMA) were tested. Concentrations leading to 50% cell survival (TC50 values) were calculated from fitted dose-response curves.
RESULTS
The simple ranking of the cytotoxic effects of the dental resin compounds in the four cell types was identical, and TC50 values determined in L929 cells here were consistent with findings by other authors using continuous cell lines. However, the concentrations of the resin compounds necessary for eliciting cytotoxic responses in the various cells were clearly different. The analyses of TC50 values of the resin compounds revealed a linear correlation between cell lines, and the overall sensitivities increased as follows: CPC=tCPC B
SIGNIFICANCE
The low sensitivities of primary cells and transformed tCPC B cells compared with the continuous L929 cell line and the transformed tCPC E cells indicates the presence of specific structural and functional properties relevant in vivo. The differences between the transformed tCPC B and tCPC E cells may indicate modifications of cellular functions caused by the different transformation processes.
Topics: Acrylates; Animals; Benzhydryl Compounds; Bisphenol A-Glycidyl Methacrylate; Cattle; Cell Line, Transformed; Composite Resins; Dental Papilla; Dental Pulp; Epoxy Compounds; L Cells; Methacrylates; Methylmethacrylate; Mice; Phenols; Polyethylene Glycols; Polymethacrylic Acids; Polyurethanes; Terpenes; Toxicity Tests
PubMed: 11992909
DOI: 10.1016/s0109-5641(01)00056-2 -
International Journal of Surgical... Jul 2007
Topics: Child; Dental Papilla; Diagnosis, Differential; Humans; Male; Mandibular Neoplasms; Myxoma; Odontogenic Tumors
PubMed: 17652537
DOI: 10.1177/1066896907302234 -
Stem Cell Research May 2014Within the field of dental tissue engineering, the establishment of adequate tissue vascularization is one of the most important burdens to overcome. As vascular access...
Within the field of dental tissue engineering, the establishment of adequate tissue vascularization is one of the most important burdens to overcome. As vascular access within the tooth is restricted by the apical foramen, it is of major importance to implement effective vascularization strategies in order to recreate viable components of teeth and periodontal tissues. However, while the current regenerative approaches focus on the use of dental stem cells (DSCs), little is known about these cells and their ability to promote angiogenesis. Therefore, the present study aimed to elucidate the paracrine angiogenic properties of postnatal DSCs, in particular dental pulp stem cells (DPSCs), stem cells from the apical papilla (SCAPs) and dental follicle precursor cells (FSCs). An antibody array, together with RT-PCR and ELISA, pointed out the differential expression of pro-angiogenic as well as anti-angiogenic factors by cultured DSCs and human gingival fibroblasts (HGF-1). Despite the secretion of proliferation-promoting factors, DSCs caused no notable increase in the proliferation of human microvascular endothelial cells (HMEC-1). With regard to other aspects of the angiogenic cascade, DPSCs, SCAPs and HGF-1 significantly promoted endothelial migration in a transwell migration assay. DPSCs also had a pronounced effect on endothelial tubulogenesis, as was shown by an in vitro Matrigelâ„¢ assay. In the last part of this study, a chorioallantoic membrane assay demonstrated a sustained pro-angiogenic impact of DPSCs and SCAPs in an in vivo setting. Collectively, these data indicate a predominant pro-angiogenic influence of DPSCs and SCAPS in vitro and in vivo in comparison to FSCs, suggesting that both stem cell populations could potentially promote the vascularization of regenerated dental tissues.
Topics: Adolescent; Adult; Angiogenesis Inducing Agents; Cell Proliferation; Cells, Cultured; Dental Papilla; Dental Pulp; Female; Hepatocyte Growth Factor; Humans; In Vitro Techniques; Male; Stem Cells; Young Adult
PubMed: 24747218
DOI: 10.1016/j.scr.2014.03.008 -
Cell Proliferation Aug 2013Mesenchymal stem cells (MSCs) are a reliable resource for tissue regeneration, but their molecular mechanisms of differentiation and proliferation remain unclear; this...
OBJECTIVES
Mesenchymal stem cells (MSCs) are a reliable resource for tissue regeneration, but their molecular mechanisms of differentiation and proliferation remain unclear; this situation has restricted use of MSCs to a limited number of applications. A previous study of ours found a member of the epidermal growth factor family, epiregulin (EREG), to be involved in regulation of MSC differentiation. In the present study, we have used human dental stem cells from the apical papilla (SCAPs) to investigate the role of EREG on proliferation of MSCs.
MATERIALS AND METHODS
SCAPs were isolated from apical papillae of immature third molars. Retroviral short hairpin RNA (shRNA) was used to silence EREG gene expression, and human recombinant EREG protein was used to stimulate SCAPs. SCAP proliferation was examined using tetrazolium dye colorimetric assay/cell growth curve. Western blotting was performed to detect expressions of extracellular signal-regulated protein kinases 1 and 2 (Erk1/2), mitogen-activated protein kinases 1 and 2 (MEK1/2), protein kinase B (Akt), p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal kinase (JNK).
RESULTS
Depletion of EREG with shRNA inhibited SCAP proliferation and repressed phosphorylation of Erk1/2 and JNK. Human recombinant EREG protein promoted cell proliferation and enhanced Erk1/2, MEK and JNK phosphorylation in SCAPs. Furthermore, blocking MEK/Erk signalling with specific Erk1/2 inhibitor PD98059, or JNK signalling with specific inhibitor SP600125, abolished effects of EREG on cell proliferation.
CONCLUSION
These findings indicate that EREG could enhance cell proliferation in dental tissue-derived MSCs by activating MEK/Erk and JNK signalling pathways.
Topics: Adolescent; Cell Differentiation; Cell Growth Processes; Cells, Cultured; Dental Papilla; Epidermal Growth Factor; Epiregulin; Humans; MAP Kinase Signaling System; Phosphorylation; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Recombinant Proteins; Stem Cells; Young Adult; p38 Mitogen-Activated Protein Kinases
PubMed: 23829318
DOI: 10.1111/cpr.12039 -
Journal of Prosthodontics : Official... Feb 2019Soft tissue interactions with ceramic dental implants have previously been shown to have favorable esthetic outcomes. This study aimed to evaluate the papilla-crown...
PURPOSE
Soft tissue interactions with ceramic dental implants have previously been shown to have favorable esthetic outcomes. This study aimed to evaluate the papilla-crown proportion around zirconia implants in a 3-year follow-up study and the correlation between the gingival biotype and changes in papillary height.
MATERIALS AND METHODS
This was a prospective study of 39 patients with 40 single-gap implants (Straumann PURE Ceramic ZLA Implant). The papilla-crown proportion was assessed after 3 months, 1 year, and 3 years. In addition, correlations between the peri-implant biotypes and changes in papillary heights were evaluated.
RESULTS
The papilla-crown proportion improved from 35.5% after 3 months to 41.7% after 3 years. The gingival biotype was correlated very weakly to papilla height alterations. Significant papillary fill was observed in the interdental space between 3 months and 3 years (p < 0.001).
CONCLUSIONS
An ideal papilla-crown proportion of 40% around single implants was observed after 3 years. A thin or thick gingival biotype showed a very weak correlation with soft tissue alterations.
Topics: Dental Implantation, Endosseous; Dental Implants; Dental Materials; Dental Papilla; Esthetics, Dental; Female; Follow-Up Studies; Gingiva; Humans; Male; Middle Aged; Prospective Studies; Tooth Crown; Zirconium
PubMed: 29377452
DOI: 10.1111/jopr.12766 -
Biochemical and Biophysical Research... Dec 2009WNT proteins are a large family of cysteine-rich secreted molecules that are linked to both canonical and non-canonical signal pathways, and have been implicated in...
WNT proteins are a large family of cysteine-rich secreted molecules that are linked to both canonical and non-canonical signal pathways, and have been implicated in oncogenesis and tissue development. Canonical WNT proteins have been proven to play critical roles in tooth development, while little is known about the role of non-canonical WNT proteins such as WNT5A. In this study, WNT5A was localized to human dental papilla tissue and human dental papilla cells (HDPCs) cultured in vitro, using immunochemistry and RT-PCR. Recombinant adenovirus encoding full-length Wnt5a cDNA was constructed to investigate the biological role of WNT5A on HDPCs. The BrdU incorporation assay, the MTT assay and flow cytometric analysis showed that over-expression of Wnt5a strongly inhibited the proliferation of HDPCs in vitro. Wound healing and transwell migration assays indicated that over-expression of WNT5A reduced migration of HDPCs. In conclusion, our results showed that WNT5A negatively regulates both proliferation and migration of HDPCs, suggesting its important role in odontogenesis via controlling the HDPCs.
Topics: Adenoviridae; Cadaver; Cell Movement; Cell Proliferation; Cells, Cultured; Dental Papilla; Fetus; Humans; Odontogenesis; Proto-Oncogene Proteins; Wnt Proteins; Wnt-5a Protein; Wound Healing
PubMed: 19878652
DOI: 10.1016/j.bbrc.2009.10.136 -
Matrix Biology : Journal of the... May 2024The coordination between odontoblastic differentiation and directed cell migration of mesenchymal progenitors is necessary for regular dentin formation. The synthesis...
The coordination between odontoblastic differentiation and directed cell migration of mesenchymal progenitors is necessary for regular dentin formation. The synthesis and degradation of hyaluronan (HA) in the extracellular matrix create a permissive niche that directly regulates cell behaviors. However, the role and mechanisms of HA degradation in dentin formation remain unknown. In this work, we present that HA digestion promotes odontoblastic differentiation and cell migration of mouse dental papilla cells (mDPCs). Hyaluronidase 2 (HYAL2) is responsible for promoting odontoblastic differentiation through degrading HA, while hyaluronidase 1 (HYAL1) exhibits negligible effect. Silencing Hyal2 generates an extracellular environment rich in HA, which attenuates F-actin and filopodium formation and in turn inhibits cell migration of mDPCs. In addition, activating PI3K/Akt signaling significantly rescues the effects of HA accumulation on cytodifferentiation. Taken together, the results confirm the contribution of HYAL2 to HA degradation in dentinogenesis and uncover the mechanism of the HYAL2-mediated HA degradation in regulating the odontoblastic differentiation and migration of mDPCs.
Topics: Animals; Hyaluronoglucosaminidase; Mice; Cell Differentiation; Cell Movement; Hyaluronic Acid; Odontoblasts; Dental Papilla; Signal Transduction; GPI-Linked Proteins; Proto-Oncogene Proteins c-akt; Cells, Cultured; Phosphatidylinositol 3-Kinases
PubMed: 38490466
DOI: 10.1016/j.matbio.2024.03.002 -
Journal of Endodontics Jun 2008Some clinical case reports have shown that immature permanent teeth with periradicular periodontitis or abscess can undergo apexogenesis after conservative endodontic... (Review)
Review
Some clinical case reports have shown that immature permanent teeth with periradicular periodontitis or abscess can undergo apexogenesis after conservative endodontic treatment. A call for a paradigm shift and new protocol for the clinical management of these cases has been brought to attention. Concomitantly, a new population of mesenchymal stem cells residing in the apical papilla of permanent immature teeth recently has been discovered and was termed stem cells from the apical papilla (SCAP). These stem cells appear to be the source of odontoblasts that are responsible for the formation of root dentin. Conservation of these stem cells when treating immature teeth may allow continuous formation of the root to completion. This article reviews current findings on the isolation and characterization of these stem cells. The potential role of these stem cells in the following respects will be discussed: (1) their contribution in continued root maturation in endodontically treated immature teeth with periradicular periodontitis or abscess and (2) their potential utilization for pulp/dentin regeneration and bioroot engineering.
Topics: Animals; Dental Papilla; Dental Pulp; Dentin, Secondary; Dentinogenesis; Dentition, Permanent; Humans; Mesenchymal Stem Cells; Multipotent Stem Cells; Periapical Periodontitis; Periodontal Ligament; Regeneration; Root Canal Therapy; Tissue Engineering; Tissue Scaffolds; Tooth Apex
PubMed: 18498881
DOI: 10.1016/j.joen.2008.03.001 -
Biotechnology and Bioengineering Apr 2017The in vitro generation of cell-based three dimensional (3D) nerve tissue is an attractive subject to improve graft survival and integration into host tissue for neural...
The in vitro generation of cell-based three dimensional (3D) nerve tissue is an attractive subject to improve graft survival and integration into host tissue for neural tissue regeneration or to model biological events in stem cell differentiation. Although 3D organotypic culture strategies are well established for 3D nerve tissue formation of pluripotent stem cells to study underlying biology in nerve development, cell-based nerve tissues have not been developed using human postnatal stem cells with therapeutic potential. Here, we established a culture strategy for the generation of in vitro cell-based 3D nerve tissue from postnatal stem cells from apical papilla (SCAPs) of teeth, which originate from neural crest-derived ectomesenchyme cells. A stem cell population capable of differentiating into neural cell lineages was generated during the ex vivo expansion of SCAPs in the presence of EGF and bFGF, and SCAPs differentiated into neural cells, showing neural cell lineage-related molecular and gene expression profiles, morphological changes and electrophysical property under neural-inductive culture conditions. Moreover, we showed the first evidence that 3D cell-based nerve-like tissue with axons and myelin structures could be generated from SCAPs via 3D organotypic culture using an integrated bioprocess composed of polyethylene glycol (PEG) microwell-mediated cell spheroid formation and subsequent dynamic culture in a high aspect ratio vessel (HARV) bioreactor. In conclusion, the culture strategy in our study provides a novel approach to develop in vitro engineered nerve tissue using SCAPs and a foundation to study biological events in the neural differentiation of postnatal stem cells. Biotechnol. Bioeng. 2017;114: 903-914. © 2016 Wiley Periodicals, Inc.
Topics: Adolescent; Bioreactors; Cell Differentiation; Child; Dental Papilla; Humans; Molar; Nerve Tissue; Spheroids, Cellular; Stem Cells; Tissue Engineering
PubMed: 27775170
DOI: 10.1002/bit.26205