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European Journal of Oral Sciences Aug 1997Human dental papilla cells were enzymatically separated from deciduous tooth germs of an 8-month-old embryo legally aborted. The second passage cells were cultured up to...
Human dental papilla cells were enzymatically separated from deciduous tooth germs of an 8-month-old embryo legally aborted. The second passage cells were cultured up to 35 days in 3 groups. The beta-GP group was cultured in the Dulbecco MEM containing ascorbic acid and beta-glycerophosphate supplemented with 15% fetal bovine serum. The Dex group was in the same medium, in addition containing dexamethasone. The control group contained none of the 3 chemicals. Mineralized nodules were formed after 15 days in the beta-GP and Dex groups. Only in the presence of ascorbic acid and organic phosphate did they mineralize. The addition of dexamethasone caused a significant increase in the number of nodules. By electron microscopy, the nodules contained needle-shaped crystals associated with a network of collagen fibrils. Calcium and phosphorus were detected by energy-dispersive X-ray microanalysis in the nodules. Furthermore, the crystalline material exhibited a pattern consistent with hydroxyapatite and dentin when examined by X-ray diffractometry. Cells showed high levels of alkaline phosphatase activity, which was increased 2-3 times in the presence of the 3 chemicals. These results indicated that human dental papilla cells have the ability to form dentin in culture. The formation of mineralized nodules by human dental papilla in vitro provides a useful model for studying the morphogenesis and differentiation of dental papilla ectomesenchyme.
Topics: Alkaline Phosphatase; Ascorbic Acid; Calcium; Cell Differentiation; Cells, Cultured; Collagen; Crystallography; Culture Media; Dental Papilla; Dentin; Dentinogenesis; Dexamethasone; Durapatite; Ectoderm; Electron Probe Microanalysis; Glucocorticoids; Glycerophosphates; Humans; Male; Mesoderm; Microscopy, Electron; Morphogenesis; Phosphorus; Tooth Calcification; Tooth Germ; Tooth, Deciduous; X-Ray Diffraction
PubMed: 9298363
DOI: 10.1111/j.1600-0722.1997.tb00247.x -
Journal of Molecular Histology Feb 2018Dental papilla cells (DPCs) belong to precursor cells differentiating to odontoblasts and play an important role in dentin formation and reproduction. This study aimed...
Dental papilla cells (DPCs) belong to precursor cells differentiating to odontoblasts and play an important role in dentin formation and reproduction. This study aimed to explore the changes and and involvement of mitochondrial respiratory function during odontogenic differentiation. Primary DPCs were obtained from first molar dental papilla of neonatal rats and cultured in odontogenic medium for 7, 14, 21 days. DPCs, which expressed mesenchymal surface markers CD29, CD44 and CD90, had the capacity for self-renewal and multipotent differentiation. Odontoblastic induction increased mineralized matrix formation in a time-dependent manner, which was accompanied by elevated alkaline phosphatase (ALP), dentin sialophosphoprotein and dentin matrix protein 1 expression at mRNA and protein levels. Notably, odontogenic medium led to an increase in adenosine-5'-triphosphate content and mitochondrial membrane potential, whereas a decrease in intercellular reactive oxygen species production and NAD/NADH ratio. Furthermore, odontogenic differentiation was significantly suppressed by treatment with rotenone, an inhibitor of mitochondrial respiratory chain. These results demonstrate that enhanced mitochondrial function is crucial for odontogenic differentiation of DPCs.
Topics: Animals; Animals, Newborn; Cell Differentiation; Cells, Cultured; Dental Papilla; Electron Transport; Mitochondria; Odontogenesis; Rats
PubMed: 29189956
DOI: 10.1007/s10735-017-9746-z -
Oral Surgery, Oral Medicine, and Oral... Dec 1991The dental pulp and its associated structures, the dentin and the cementum, are discussed. Because many of the age-related pulpal changes have components considered in... (Review)
Review
The dental pulp and its associated structures, the dentin and the cementum, are discussed. Because many of the age-related pulpal changes have components considered in several of the current theories of aging, these theories are briefly reviewed. Part 2 describes the age-related changes of the dental pulp, the dentin, and the cementum (the dental pulp complex). An attempt is made to differentiate inherent aging changes from physiologic defensive changes and pathologic irritant-induced changes. Part 3 describes the relationship of age-induced changes in the dental pulp complex to components of the current aging theories together with a unified concept of the dental pulp complex aging. Part 4 considers whether dental pulp complex aging can be used as a biomarker for generalized aging. Whether age-related changes of the dental pulp complex can be altered by interventions is discussed.
Topics: Aging; Biological Clocks; Cementogenesis; DNA Damage; Dental Cementum; Dental Papilla; Dental Pulp; Dental Pulp Calcification; Dentin; Dentin, Secondary; Humans; Tooth, Deciduous
PubMed: 1812456
DOI: 10.1016/0030-4220(91)90019-9 -
Shanghai Kou Qiang Yi Xue = Shanghai... Aug 2016To investigate the immunomodulatory effects of rat dental papilla cells (RDPCs) on lipopolysaccharide(LPS)-induced macrophages.
PURPOSE
To investigate the immunomodulatory effects of rat dental papilla cells (RDPCs) on lipopolysaccharide(LPS)-induced macrophages.
METHODS
Dental papilla tissues from SD rats were isolated and cultured. Cells were passaged and purified using different digesting method. After osteogenic differentiation of rat dental papilla cells, mineralized nodules were assessed by Alizarin red S staining. Oil red-O staining was used to observe the lipid after adipogenic differentiation of rat dental papilla cells. The effect of rat dental papilla cells conditioned medium (RDPC-CM) on macrophages proliferation was assessed using a cell counting kit-8 (CCK-8). LPS-stimulated macrophages were treated with RDPC-CM and the level of IL-1β, TNF-α, IL-6 and nitric oxide (NO) in the supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA) and Greiss reagent. The results were analyzed by using SPSS 13.0 software package.
RESULTS
RDPCs were found around dental papilla piece after 24h of cultures, the cells could be purified using different digesting methods. Aliarin red staining and Oil red-O staining certified the presence of mineralized nodules and lipid in rat dental papilla cells, respectively. CCK-8 assay results showed that the proliferation of macrophages was not affected by RDPC-CM. ELISA and Griess reagent assay revealed a significantly decreased level of TNF-α in the supernatants of LPS-stimulated macrophages upon RDPC-CM treatment compared with the control, but not the levels of IL-6, IL-1β and NO.
CONCLUSIONS
RDPC-CM inhibited the expression of TNF-α in LPS-treated macrophages, indicating that rat dental papilla cells may play a role in immunomodulatory effects.
Topics: Animals; Cell Differentiation; Cytokines; Dental Papilla; Enzyme-Linked Immunosorbent Assay; Interleukin-1beta; Interleukin-6; Lipopolysaccharides; Macrophages; Nitric Oxide; Osteogenesis; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha
PubMed: 27858059
DOI: No ID Found -
Zhonghua Kou Qiang Yi Xue Za Zhi =... Jun 2013To examine the expression of LIM mineralization protein-1 (LMP-1) in the apical papilla and dental pulp tissues of human immature permanent teeth and to investigate the...
OBJECTIVE
To examine the expression of LIM mineralization protein-1 (LMP-1) in the apical papilla and dental pulp tissues of human immature permanent teeth and to investigate the role of LMP-1 in the development and maturation of pulp-dentin complex.
METHODS
Forty-eight healthy premolars in need of extraction for orthodontic treatment were obtained with 24 immature permanent teeth and 24 mature permanent teeth. After extraction, the apical papilla was detached from the dental pulp in the immature permanent tooth and the dental pulp of mature permanent tooth was rapidly removed. The samples were divided into 3 groups: group 1, apical papilla of immature permanent teeth (root formed 2/3 of its full length); group 2, dental pulp tissues of immature permanent teeth; group 3, dental pulp tissues of mature permanent teeth. There were 24 samples for each group.Half of them were used for reverse transcriptien-PCR (RT-PCR) detection, and the other half were used for Western blotting detection. Band intensities were quantified using Meta Morph software 6.2.6 and subsequently normalized by dividing the band gray value of the target gene by the intensity of its corresponding β-actin. Two-sample t test was used to analyze the difference of expression intensity between group 1 and group 2 as well as group 2 and group 3 with SPSS 13.0 software package.Statistical significance was established as P < 0.05.
RESULTS
As indicated by RT-PCR, LMP-1 expressed in the apical papilla, dental pulp of immature permanent teeth and the dental pulp of mature permanent teeth were 0.25 ± 0.09, 0.46 ± 0.24 and 0.31 ± 0.10 respectively. The expression intensity of LMP-1 mRNA in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t = 2.92) and that in the dental pulp tissues of human mature permanent teeth (t = 2.31) (P < 0.05). As indicated by Western blotting, LMP-1 expressed in the three groups were 0.33 ± 0.08, 0.82 ± 0.10 and 0.52 ± 0.19 respectively. The expression intensity of LMP-1 protein in the dental pulp tissues of human immature permanent teeth were significantly higher than that in the apical papilla tissues(t = 3.33) and that in the dental pulp tissues of human mature permanent teeth (t = 3.11) (P < 0.05).
CONCLUSIONS
LMP-1 were positively expressed in all the samples including apical papilla of immature permanent teeth, dental pulp of immature and mature permanent teeth at the level of both mRNA and protein, but with different intensity. LMP-1 could play an important role in the development and maturation of pulp-dentin complex.
Topics: Adaptor Proteins, Signal Transducing; Adolescent; Adult; Blotting, Western; Child; Cytoskeletal Proteins; Dental Papilla; Dental Pulp; Dentition, Permanent; Humans; LIM Domain Proteins; Maxillofacial Development; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Young Adult
PubMed: 24119999
DOI: No ID Found -
PloS One 2013Classical tooth development theory suggests that dental papilla cells (DPCs) are the precursor cells of odontoblasts, which are responsible for dentin development.... (Comparative Study)
Comparative Study
Classical tooth development theory suggests that dental papilla cells (DPCs) are the precursor cells of odontoblasts, which are responsible for dentin development. However, our previous studies have indicated that dental follicle cells (DFCs) can differentiate into odontoblasts. To further our understanding of tooth development, and the differences in dentinogenesis between DFCs and DPCs, the odontogenic differentiation of DFCs and DPCs was characterized in vitro and in vivo. DFCs and DPCs were individually combined with treated dentin matrix (TDM) before they were subcutaneously implanted into the dorsum of mice for 8 weeks. Results showed that 12 proteins were significantly differential, and phosphoserine aminotransferase 1 (PSAT1), Isoform 2 of hypoxia-inducible factor 1-alpha (HIF1A) and Isoform 1 of annexin A2 (ANXA2), were the most significantly differential proteins. These proteins are related to regulation of bone balance, angiogenesis and cell survival in an anoxic environment. Both DFCs and DPCs express odontogenic, neurogenic and peridontogenic markers. Histological examination of the harvested grafts showed that both DFCs and DPCs form pulp-dentin/cementum-periodentium-like tissues in vivo. Hence, DFCs and DPCs have similar odontogenic differentiation potential in the presence of TDM. However, differences in glucose and amino acid metabolism signal transduction and protein synthesis were observed for the two cell types. This study expands our understanding on tooth development, and provides direct evidence for the use of alternative cell sources in tooth regeneration.
Topics: Adolescent; Animals; Antigens, Surface; Biomarkers; Cell Differentiation; Cell Proliferation; Cell Separation; Cells, Cultured; Dental Papilla; Dental Sac; Dentin; Humans; Immunophenotyping; Mice; Multipotent Stem Cells; Odontogenesis; Proteomics; Real-Time Polymerase Chain Reaction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 23620822
DOI: 10.1371/journal.pone.0062332 -
Journal of Cellular Physiology Oct 2013Odontoblasts, which derive from dental papilla, are a type of terminally differentiated matrix-secreting cells. Previous studies have identified various transcription...
Odontoblasts, which derive from dental papilla, are a type of terminally differentiated matrix-secreting cells. Previous studies have identified various transcription factors involved in the differentiation process of odontoblasts. We have recently found that Krüppel-like factor 4 (Klf4) was expressed in the polarizing and elongating odontoblasts, but the function of Klf4 in the differentiation of odontoblasts is still unclear. We hypothesized Klf4 promoted the differentiation of odontoblasts by up-regulating some odontoblast-related genes. In this study, we found that the expression of Klf4 increased significantly during the odontoblastic differentiation of primary mouse dental papilla cells and the mouse dental papilla cell line-mDPC6T. Overexpression of Klf4 significantly up-regulated odontoblast-related genes, such as Dmp1, Dspp, and Alp, and promoted the accumulation of mineral nodules. Knock-down of Klf4 down-regulated expression of Dmp1, Dspp, and Alp, and inhibited mineral deposition. We applied in silico analysis and identified one target gene of Klf4-Dmp1. Based on further analysis of ChIP data, EMSA and dual luciferase activity assays, we confirmed that Klf4 was able to specifically bind to the Dmp1 promoter and transactivate its expression. Furthermore, forced expression of Dmp1 in the Klf4 knock-down mDPC6T cell line significantly recovered its odontoblastic differentiation ability. Our data confirmed our hypothesis that Klf4 promotes the differentiation of odontoblasts via the up-regulation of Dmp1.
Topics: Animals; Cell Differentiation; Cell Line; Dental Papilla; Down-Regulation; Extracellular Matrix Proteins; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Mice; Odontoblasts; Promoter Regions, Genetic; Up-Regulation
PubMed: 23558921
DOI: 10.1002/jcp.24377 -
The Journal of Prosthetic Dentistry Nov 2019Polyvinylphosphonic acid (PVPA) could be used as a biomimetic remineralization analog and a matrix metalloproteinases (MMPs) inhibitor. However, studies are lacking...
STATEMENT OF PROBLEM
Polyvinylphosphonic acid (PVPA) could be used as a biomimetic remineralization analog and a matrix metalloproteinases (MMPs) inhibitor. However, studies are lacking regarding the performance of PVPA in dental bonding systems for maintaining the durability of the resin-dentin bond.
PURPOSE
The purpose of this in vitro study was to investigate the effect of PVPA on the durability of resin-dentin bonds and the viability of mouse dental papilla cell-23 (MDPC-23). The mechanical properties of resin-dentin interfaces during long-term storage were analyzed, and the potential application of PVPA as a biomimetic remineralization analog in adhesive dentistry was evaluated.
MATERIAL AND METHODS
Seventy-five extracted noncarious human third molars were collected and randomly divided into 5 groups, and then the microtensile bond strength (μTBS) data and scanning electron microscope (SEM) images were used to evaluate the preservation condition of resin-dentin bonds after 1 day, 6 months, and 1 year of storage. The cytotoxicity of PVPA was detected by cell proliferation assay and cell apoptosis assay.
RESULTS
Compared with the control and chlorhexidine (CHX) groups, the combined group (treated with both 200-μg/mL PVPA and biomimetic remineralization) had excellent bond durability. The exposed collagen fibril from the PVPA-treated groups (included 200-μg/mL and 500-μg/mL PVPA groups and a combined group) still showed integrity after 1 year of storage when compared with the control group. PVPA up to 500 μg/mL showed no cytotoxicity to MDPC-23 and did not inhibit cell growth.
CONCLUSIONS
This study offered evidence that PVPA did not result in cytotoxicity at low concentrations as an MMP inhibitor and a biomimetic remineralization analog. In addition, the application of PVPA improved bond strength and preserved collagen integrity after 1 year of in vitro storage.
Topics: Acid Etching, Dental; Animals; Dental Bonding; Dental Papilla; Dentin; Dentin-Bonding Agents; Humans; Materials Testing; Mice; Resin Cements; Surface Properties; Tensile Strength
PubMed: 31623837
DOI: 10.1016/j.prosdent.2019.08.011 -
Journal of Endodontics Mar 2014A 3-antibiotic combination (3Mix) has been widely used in regenerative endodontics. Recent studies recommend that a safe concentration of 3Mix is in the range of 0.39... (Comparative Study)
Comparative Study
INTRODUCTION
A 3-antibiotic combination (3Mix) has been widely used in regenerative endodontics. Recent studies recommend that a safe concentration of 3Mix is in the range of 0.39 μg/mL and 1 mg/mL because higher concentrations may limit tissue regeneration. The aim of this study was to determine the regenerative capacity of isolated human dental pulp cells (DPCs) and apical papilla cells (APCs) after a 7-day treatment with selected doses of 3Mix.
METHODS
Primary human DPCs/APCs from the third passage were divided into control and experimental groups. In the control group, cells were cultured in regular complete media. In the experimental group, cells were cultured in complete media containing 0.39 μg/mL or 1 mg/mL of 3Mix for 7 days. After the treatment period, the media were changed, and the cells were further tested for proliferation and differentiation potential. For cell proliferation, a colorimetric qualification of 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide was used on days 1, 3, 5, and 7. For differentiation potential, a dentinogenic differentiation medium was added into treated cells and cultured for 7, 14, and 21 days. Results were analyzed using quantitative alizarin red S staining and real-time reverse-transcription polymerase chain reaction.
RESULTS
After 7 days of treatment, 100% cell death was discovered in the 1-mg/mL 3Mix group. The proliferative capacity of 0.39 μg/mL 3Mix-treated DPCs and APCs was significantly lower than that of untreated cells at all time points (P < .05). Mineralized nodule formation was found both in the 3Mix-treated and control groups, but it was significantly less in the 3Mix-treated groups at 7, 14, and 21 days (P < .01). Quantitative reverse-transcription polymerase chain reaction showed no statistically significant difference (95% confidence interval) in bone sialoprotein, alkaline phosphatase, and dentin matrix protein 1 gene expression in either 3Mix-treated DPCs or APCs compared with control groups.
CONCLUSIONS
One milligram per milliliter of 3Mix had strong toxicity to DPCs/APCs when applied for 7 days, whereas 0.39 μg/mL 3Mix showed no toxicity but still affected cell proliferation and mineralization potential. However, no differences in dentinogenic gene expressions were observed between the 3Mix-treated and untreated groups.
Topics: Adolescent; Adult; Alkaline Phosphatase; Anthraquinones; Anti-Bacterial Agents; Calcification, Physiologic; Cell Culture Techniques; Cell Death; Cell Differentiation; Cell Proliferation; Cells, Cultured; Ciprofloxacin; Coloring Agents; Dental Papilla; Dental Pulp; Dentinogenesis; Extracellular Matrix Proteins; Humans; Integrin-Binding Sialoprotein; Metronidazole; Minocycline; Osteogenesis; Phosphoproteins; Regeneration; Tetrazolium Salts; Thiazoles; Time Factors; Young Adult
PubMed: 24565660
DOI: 10.1016/j.joen.2013.09.027 -
Stem Cells and Development Mar 2016Recent studies have shown that secretion of bioactive factors from mesenchymal stem cells (MSCs) plays a primary role in MSC-mediated therapy; especially for bone...
Recent studies have shown that secretion of bioactive factors from mesenchymal stem cells (MSCs) plays a primary role in MSC-mediated therapy; especially for bone marrow-derived MSCs (BMSCs). MSCs from dental apical papilla (SCAPs) are involved in root development and may play a critical role in the formation of dentin and pulp. Bioactive factors secreted from SCAPs actively contribute to their environment; however, the SCAPs secretome remains unclear. To address this and gain a deeper understanding of the relevance of SCAPs secretions in a clinical setting, we used isobaric chemical tags and high-performance liquid chromatography with tandem mass spectrometry to profile the secretome of human SCAPs and then compared it to that of BMSCs. A total of 2,046 proteins were detected from the conditioned medium of SCAPs, with a false discovery rate of less than 1.0%. Included were chemokines along with angiogenic, immunomodulatory, antiapoptotic, and neuroprotective factors and extracellular matrix (ECM) proteins. The secreted levels of 151 proteins were found to differ by at least twofold when BMSCs and SCAPs were compared. Relative to BMSCs, SCAPs exhibited increased secretion of proteins that are involved in metabolic processes and transcription and lower levels of those associated with biological adhesion, developmental processes, and immune function. In addition, SCAPs secreted significantly larger amounts of chemokines and neurotrophins than BMSCs, whereas BMSCs secreted more ECM proteins and proangiogenic factors. These results may provide important clues regarding the molecular mechanisms associated with tissue regeneration and how they differ between cell sources.
Topics: Bone Marrow Cells; Cells, Cultured; Chemokines; Dental Papilla; Extracellular Matrix Proteins; Humans; Mesenchymal Stem Cells; Nerve Growth Factors; Proteome; Secretory Pathway
PubMed: 26742889
DOI: 10.1089/scd.2015.0298