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Journal of Esthetic and Restorative... Mar 2023Evaluate the influence of a polymeric catalyst primer (PCP) on esthetic efficacy (EE), degradation kinetics of hydrogen peroxide (H O ), and trans-amelodentinal...
OBJECTIVE
Evaluate the influence of a polymeric catalyst primer (PCP) on esthetic efficacy (EE), degradation kinetics of hydrogen peroxide (H O ), and trans-amelodentinal cytotoxicity (TC) of bleaching gels.
MATERIALS AND METHODS
The following groups were established: G1: No treatment (NC, negative control); G2: PCP; G3: 10% H O ; G4: PCP + 10% H O ; G5: 20% H O ; G6: PCP + 20% H O ; G7: 35% H O (positive control); G8: PCP + 35% H O . To determine EE, enamel/dentin discs (E/DDs) were stained and subjected or not to bleaching protocols for 45 min. To assess TC, the E/DDs were adapted to artificial pulp chambers. The extracts (culture medium + gel components diffused through E/DDs) were applied to odontoblast-like MDPC-23 cells. The viability (VB), oxidative stress (OxS), morphology (SEM), amount of H O diffused and the production of hydroxyl radical (OH ) were assessed (two-way ANOVA/Tukey/paired Student t-test; p < 0.05).
RESULTS
The highest EE was found in G8 (p < 0.05), and G4, G6, and G7 did not differ statistically (p > 0.05). In G4, the limited H O diffusion reduced OxS and increased cell VB (p < 0.05).
CONCLUSIONS
Coating the enamel with PCP containing 10 mg/ml of manganese oxide before applying the 10% H O bleaching gel maintains the EE of conventional in-office bleaching and minimizes the toxic effects of this esthetic therapy.
CLINICAL SIGNIFICANCE
Coating the enamel with a PCP before applying the bleaching gel may potentiate the EE of the conventional in-office tooth bleaching and reduce the toxicity of this professional therapy to the dental pulp.
Topics: Humans; Tooth Bleaching; Hydrogen Peroxide; Tooth Bleaching Agents; Odontoblasts; Dental Enamel
PubMed: 36193855
DOI: 10.1111/jerd.12972 -
Journal of Cellular Biochemistry Mar 2020Cellular differentiation is caused by highly controlled modifications in the gene expression but rarely involves a change in the DNA sequence itself. Histone acetylation...
Cellular differentiation is caused by highly controlled modifications in the gene expression but rarely involves a change in the DNA sequence itself. Histone acetylation is a major epigenetic factor that adds an acetyl group to histone proteins, thus altering their interaction with DNA and nuclear proteins. Illumination of the histone acetylation during dentinogenesis is important for odontoblast differentiation and dentinogenesis. In the current study, we aimed to discover the roles and regulation of acetylation at histone 3 lysine 9 (H3K9ac) and H3K27ac during dentinogenesis. We first found that both of these modifications were enhanced during odontoblast differentiation and dentinogenesis. These modifications are dynamically catalyzed by histone acetyltransferases (HATs) and deacetylases (HDACs), among which HDAC3 was decreased while p300 increased during odontoblast differentiation. Moreover, overexpression of HDAC3 or knockdown p300 inhibited odontoblast differentiation in vitro, and inhibition of HDAC3 and p300 with trichostatin A or C646 regulated odontoblast differentiation. Taken together, the results of our present study suggest that histone acetylation is involved in dentinogenesis and coordinated expression of p300- and HDAC3-regulated odontoblast differentiation through upregulating histone acetylation.
Topics: Acetylation; Animals; Cell Differentiation; Cell Proliferation; Cells, Cultured; Dental Papilla; Dentinogenesis; E1A-Associated p300 Protein; Histone Deacetylases; Histones; Mesenchymal Stem Cells; Mice; Protein Processing, Post-Translational
PubMed: 31692090
DOI: 10.1002/jcb.29470 -
Biomimetics (Basel, Switzerland) Feb 2023The (-)-- (EGCG) metabolite is a natural polyphenol derived from green tea and is associated with antioxidant, biocompatible, and anti-inflammatory effects.
UNLABELLED
The (-)-- (EGCG) metabolite is a natural polyphenol derived from green tea and is associated with antioxidant, biocompatible, and anti-inflammatory effects.
OBJECTIVE
To evaluate the effects of EGCG to promote the odontoblast-like cells differentiated from human dental pulp stem cells (hDPSCs); the antimicrobial effects on , , and ; and improve the adhesion on enamel and dentin by shear bond strength (SBS) and the adhesive remnant index (ARI).
MATERIAL AND METHODS
hDSPCs were isolated from pulp tissue and immunologically characterized. EEGC dose-response viability was calculated by MTT assay. Odontoblast-like cells were differentiated from hDPSCs and tested for mineral deposition activity by alizarin red, Von Kossa, and collagen/vimentin staining. Antimicrobial assays were performed in the microdilution test. Demineralization of enamel and dentin in teeth was performed, and the adhesion was conducted by incorporating EGCG in an adhesive system and testing with SBS-ARI. The data were analyzed with normalized Shapiro-Wilks test and ANOVA post hoc Tukey test.
RESULTS
The hDPSCs were positive to CD105, CD90, and vimentin and negative to CD34. EGCG (3.12 µg/mL) accelerated the differentiation of odontoblast-like cells. exhibited the highest susceptibility < < . EGCG increased ( < 0.05) the dentin adhesion, and cohesive failure was the most frequent.
CONCLUSION
(-)-- is nontoxic, promotes differentiation into odontoblast-like cells, possesses an antibacterial effect, and increases dentin adhesion.
PubMed: 36810406
DOI: 10.3390/biomimetics8010075 -
Annals of Anatomy = Anatomischer... May 2021Extracellular matrix molecules (ECMM) expression during tertiary dentinogenesis provides useful information for regenerative applications and efficacy of pulp capping... (Review)
Review
BACKGROUND
Extracellular matrix molecules (ECMM) expression during tertiary dentinogenesis provides useful information for regenerative applications and efficacy of pulp capping materials.
AIM
To identify and review the expression and roles of non-collagenous ECMM after successful direct pulp capping (DPC), following mechanical pulp exposures, via immunohistochemistry (IHC). The study addressed the question of where will successful DPC impact the IHC expression of these molecules.
DATA SOURCES
In vivo animal and human original clinical studies reporting on ECMM in relation to different follow-up periods were screened and evaluated via descriptive analysis. The electronic literature search was carried out in three databases (MEDLINE/PubMed, Web of Science, Scopus), followed by manual screening of relevant journals and cross-referencing, up to December 2018.
STUDY ELIGIBILITY CRITERIA, PARTICIPANTS, AND INTERVENTIONS
Randomized and non-randomized controlled trials, conducted in humans and animals, were selected. Histological evidence for tertiary dentine formation was a prerequisite for IHC evaluation.
STUDY APPRAISAL AND SYNTHESIS METHODS
The methodological quality of the included articles was independently assessed using the Systematic Review Centre for Laboratory animal Experimentation (SYRCLE) and the Cochrane risk of bias tool (RoB 1), respectively.
RESULTS
From a total of 1534 identified studies, 18 were included. Thirteen papers evaluated animal subjects and five studies were carried out on humans. In animals and humans, fibronectin and tenascin expressions were detected in pulp and odontoblast-like cells (OLC); dentine sialoprotein was expressed in both soft and newly-formed mineralized tissue. In animals, bone sialoprotein was early expressed, in association with OLC and predentin; the immunoreactivity for dentine sialophosphoprotein and dentine matrix protein-1 was associated with the OLC and dentine bridge; osteopontin was expressed in OLC, predentine and reparative dentine. A considerable heterogeneity was found in the methodologies of the included studies, as well as interspecies variability of results in terms of time.
CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS
Within the limited scientific evidence, all non-collagenous ECMM expressions during tertiary dentinogenesis are active and related to soft and hard tissues. There is a shortage of human studies, and future research directions should focus more on them. PROSPERO Protocol: CRD42019121304.
Topics: Animals; Dental Pulp; Dental Pulp Capping; Dentin, Secondary; Dentinogenesis; Extracellular Matrix; Humans; Odontoblasts
PubMed: 33400977
DOI: 10.1016/j.aanat.2020.151674 -
Journal of Molecular Histology Aug 2023FAM20C phosphorylates secretory proteins at S-x-E/pS motifs, and previous studies of Fam20C-dificient mice revealed that FAM20C played essential roles in bone and tooth...
FAM20C phosphorylates secretory proteins at S-x-E/pS motifs, and previous studies of Fam20C-dificient mice revealed that FAM20C played essential roles in bone and tooth formation. Inactivation of FAM20C in mice led to hypophosphatemia that masks direct effect of FAM20C in these tissues, and consequently the direct role of FAM20C remains unknown. Our previous study reported that osteoblast/odontoblast-specific Fam20C transgenic (Fam20C-Tg) mice had normal serum phosphate levels and that osteoblastic FAM20C-mediated phosphorylation regulated bone formation and resorption. Here, we investigated the direct role of FAM20C in dentin using Fam20C-Tg mice. The tooth of Fam20C-Tg mice contained numerous highly phosphorylated proteins, including SIBLINGs, compared to that of wild-type mice. In Fam20C-Tg mice, coronal dentin volume decreased and mineral density unchanged at early age, while the volume unchanged and the mineral density elevated at maturity. In these mice, radicular dentin volume and mineral density decreased at all ages, and histologically, the radicular dentin had wider predentin and abnormal apical-side dentin with embedded cells and argyrophilic canaliculi. Immunohistochemical analyses revealed that abnormal apical-side dentin had bone and dentin matrix properties accompanied with osteoblast-lineage cells. Further, in Fam20C-Tg mice, DSPP content which is important for dentin formation, was reduced in dentin, especially radicular dentin, which might lead to defects mainly in radicular dentin. Renal subcapsular transplantations of tooth germ revealed that newly formed radicular dentin replicated apical abnormal dentin of Fam20C-Tg mice, corroborating that FAM20C overexpression indeed caused the abnormal dentin. Our findings indicate that odontoblastic FAM20C-mediated phosphorylation in the tooth regulates dentin formation and odontoblast differentiation.
Topics: Mice; Animals; Odontoblasts; Mice, Transgenic; Tooth; Cell Differentiation; Extracellular Matrix Proteins; Dentin; Phosphoproteins; Calcium-Binding Proteins
PubMed: 37357253
DOI: 10.1007/s10735-023-10123-y -
International Endodontic Journal May 2021To investigate the role of autophagy in MTA-induced odontoblastic differentiation of human dental pulp cells (HDPCs).
AIM
To investigate the role of autophagy in MTA-induced odontoblastic differentiation of human dental pulp cells (HDPCs).
METHODOLOGY
In MTA-treated HDPCs, odontoblastic differentiation was assessed based on expression levels of dentine sialophosphoprotein (DSPP) and dentine matrix protein 1 (DMP1), alkaline phosphatase activity (ALP) activity by ALP staining and the formation of mineralized nodule by Alizarin red S staining. Expression of microtubule-associated protein 1A/1B-light chain3 (LC3), adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signalling molecules and autophagy-related genes was analysed by Western blot analysis and Acridine orange staining was used to detect autophagic lysosome. For in vivo experiments, tooth cavity preparation models on rat molars were established and the expression of proteins-related odontogenesis and autophagy markers was observed by Immunohistochemistry and Western blot analysis. Kruskal-Wallis with Dunn's multiple comparison was used for statistical analysis.
RESULTS
Mineral trioxide aggregate (MTA) promoted odontoblastic differentiation of HDPCs, accompanied by autophagy induction, including formation of autophagic lysosome and cleavage of LC3 to LC3II (P < 0.05). Conversely, inhibition of autophagy through 3MA significantly attenuated the expression level of DSPP (P < 0.05) and DMP1 (P < 0.05) as well as formation of mineralized nodules (P < 0.05), indicating the functional significance of autophagy in MTA-induced odontoblastic differentiation. Also, MTA increased the activity of AMPK (P < 0.01), whereas inhibition of AMPK by compound C downregulated DSPP (P < 0.01) and DMP1 (P < 0.05), but increased the phosphorylation of mTOR (P < 0.05), p70S6 (P < 0.01) and Unc-51-like kinases 1 (ULK1) (ser757) (P < 0.01), explaining the involvement of AMPK pathway in MTA-induced odontoblast differentiation. In vivo study, MTA treatment after tooth cavity preparation on rat molars upregulated DMP-1 and DSPP as well as autophagy-related proteins LC3II and p62, and enhanced the phosphorylation of AMPK.
CONCLUSION
MTA induced odontoblastic differentiation and mineralization by modulating autophagy with AMPK activation in HDPCs. Autophagy regulation is a new insight on regenerative endodontic therapy using MTA treatment.
Topics: Alkaline Phosphatase; Aluminum Compounds; Animals; Calcium Compounds; Cell Differentiation; Cells, Cultured; Dental Pulp; Drug Combinations; Extracellular Matrix Proteins; Humans; Odontoblasts; Oxides; Phosphoproteins; Rats; Silicates
PubMed: 33277707
DOI: 10.1111/iej.13460 -
Journal of Dentistry Jun 2022Self-assembling peptide P-4 is amphiphilic and pH-triggered, effective on repairing early enamel carious lesions and dentin remineralization. However, P-4 effects on...
OBJECTIVES
Self-assembling peptide P-4 is amphiphilic and pH-triggered, effective on repairing early enamel carious lesions and dentin remineralization. However, P-4 effects on dentin biomineralization and repair ability remain unexplored. Thus, cytocompatibility and effectiveness of P-4 on inducing mineralization and migration of odontoblast-like cells (MDPC-23) were investigated.
METHODS
MDPC-23 were seeded in contact with P-4 (0.5 and 1 µg/ml), Dentin Matrix Protein 1 (DMP1 0.5 and 1 µg/ml) or Calcium hydroxide (Ca(OH) 100 µg/ml) solutions. Cell viability was verified using MTT (n = 6/group). Mineral deposition was tested using Alizarin Red (n = 4/group). Cell migration was assessed by light microscopy (n = 2/group). MTT and Alizarin Red data were compared using Kruskal-Wallis and Mann-Whitney (α=0.01).
RESULTS
P-4 (0.5 and 1 µg/ml) and DMP1 (0.5 and 1 µg/ml) resulted the highest cell viability; Ca(OH) presented the lowest. 1 µg/ml DMP1 and 1 µg/ml P-4 promoted the highest mineral deposition. Ca(OH) presented lower values of mineral deposits than DMP1 1 µg/ml (p < 0.01), but similar to P-4 1 µg/ml. P-4 and DMP1 at 0.5 µg/ml induced lesser mineral precipitation than P-4 and DMP1 at 1 µg/ml (p < 0.01), with no difference to Ca(OH). All materials stimulated cell migration, however, lower concentrations of DMP1 and P-4 demonstrated a higher migration potential.
CONCLUSION
P-4 did not affect cell viability, induces mineral deposition and MDPC-23 migration like DMP1.
CLINICAL SIGNIFICANCE
Self-assembling peptide P-4 does not affect the cell viability and induces mineral deposition comparable to native protein involved in biomineralization. Combined with its ability to bind type I collagen, P-4 is a promising bioinspired molecule that provides native-tissue conditions and foster further studies on its ability to form dentin bridges in pulp-capping strategies.
Topics: Cell Movement; Dental Enamel; Extracellular Matrix Proteins; Glycosyltransferases; Odontoblasts; Phosphoproteins
PubMed: 35460865
DOI: 10.1016/j.jdent.2022.104111 -
International Endodontic Journal Dec 2023Fat mass and obesity-associated (FTO) protein, the first discovered N6-methyladenine (m6A) demethylase, played positive roles in bone formation. In this study, the aim...
The N6-methyladenosine demethylase FTO is required for odontoblast differentiation in vitro and dentine formation in mice by promoting RUNX2 exon 5 inclusion through RBM4.
AIM
Fat mass and obesity-associated (FTO) protein, the first discovered N6-methyladenine (m6A) demethylase, played positive roles in bone formation. In this study, the aim was to investigate the function and potential mechanism of Fto in dentine formation.
METHODOLOGY
In vivo model, postnatal 12-day (PN12), 4-week-old (4 wk), 6-week-old (6 wk) healthy male C57BL/6J were randomly divided into Fto knockout (Fto ) mice and wild-type (WT) littermates according to their genotypes, with 3-5 mice in each group. The mandibles of Fto mice and WT control littermates were isolated for analysis by micro-computed tomography (micro-CT), 3-dimensional reconstruction and Haematoxylin-eosin (HE) staining. In vitro, mouse dental papilla cells (mDPCs) and human dental stem pulp cells (hDPSCs) were cultured with odontogenetic medium to evaluate differentiation capacity; expression levels of odontoblastic related genes were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). The inclusion levels of Runt-related transcription factor 2 (RUNX2) exon 5 in mDPCs and hDPSCs were detected by semiquantitative real-time polymerase chain reaction (RT-PCR). The RNA binding motif protein 4 (RBM4) m6A site was verified through m6A methylated RNA immunoprecipitation (MeRIP) and the stability of RBM4 mRNA influenced by FTO knockdown was measured by mRNA stability assay. Differences with p values < .05 were regarded as statistically significant.
RESULTS
We discovered that Fto mice showed significant dentine formation defects characterized by widened pulp cavity, enlarged pulp-tooth volume ratio, thinned dentine and pre-dentine layer of root (p < .05). Fto mDPCs and FTO-silencing hDPSCs not only exhibited insufficient mineralization ability and decreased expression levels of odontoblastic mineralization related genes (p < .05), but showed significantly reduced Runx2 exon 5 inclusion level (p < .05). FTO knockdown increased the m6A level of RBM4 and destabilized the mRNA of RBM4, thus contributing to the reduced RBM4 expression level. Moreover, Rbm4 overexpression in Fto mDPCs can partly restore Runx2 exon 5 inclusion level and the differentiation ability disrupted by Fto knockout.
CONCLUSION
Thus, within the limitations of this study, the data suggest that FTO promotes odontoblastic differentiation during dentine formation by stabilizing RBM4 mRNA to promote RUNX2 exon 5 inclusion.
Topics: Animals; Humans; Male; Mice; Alpha-Ketoglutarate-Dependent Dioxygenase FTO; Cell Differentiation; Core Binding Factor Alpha 1 Subunit; Dental Pulp; Dentin; Exons; Mice, Inbred C57BL; Odontoblasts; RNA, Messenger; RNA-Binding Proteins; X-Ray Microtomography
PubMed: 37698901
DOI: 10.1111/iej.13975 -
Biochemical and Biophysical Research... Apr 2023Iroquois homeobox (Irx) genes are TALE-class homeobox genes that are evolutionarily conserved across species and have multiple critical cellular functions in fundamental...
Iroquois homeobox (Irx) genes are TALE-class homeobox genes that are evolutionarily conserved across species and have multiple critical cellular functions in fundamental tissue development processes. Previous studies have shown that Irxs genes are expressed during tooth development. However, the precise roles of genes in teeth remain unclear. Here, we demonstrated for the first time that Irx3 is an essential molecule for the proliferation and differentiation of odontoblasts. Using cDNA synthesized from postnatal day 1 (P1) tooth germs, we examined the expression of all Irx genes (Irx1-Irx6) by RT-PCR and found that all genes except Irx4 were expressed in the tooth tissue. Irx1-Irx3 a were expressed in the dental epithelial cell line M3H1 cells, while Irx3 and Irx5 were expressed in the dental mesenchymal cell line mDP cells. Only Irx3 was expressed in both undifferentiated cell lines. Immunostaining also revealed the presence of IRX3 in the dental epithelial cells and mesenchymal condensation. Inhibition of endogenous Irx3 by siRNA blocks the proliferation and differentiation of mDP cells. Wnt3a, Wnt5a, and Bmp4 are factors involved in odontoblast differentiation and were highly expressed in mDP cells by quantitative PCR analysis. Interestingly, the expression of Wnt5a (but not Wnt3a or Bmp4) was suppressed by Irx3 siRNA. These results suggest that Irx3 plays an essential role in part through the regulation of Wnt5a expression during odontoblast proliferation and differentiation.
Topics: Homeodomain Proteins; Transcription Factors; Odontoblasts; Genes, Homeobox; Cell Differentiation; Cell Proliferation
PubMed: 36773339
DOI: 10.1016/j.bbrc.2023.02.004 -
Medical Molecular Morphology Sep 2023The purpose of this study was to investigate whether fibroblast growth factor 4 (FGF4) and FGF9 are active in dentin differentiation. Dentin matrix protein 1 (Dmp1)...
The purpose of this study was to investigate whether fibroblast growth factor 4 (FGF4) and FGF9 are active in dentin differentiation. Dentin matrix protein 1 (Dmp1) -2A-Cre transgenic mice, which express the Cre-recombinase in Dmp1-expressing cells, were crossed with CAG-tdTomato mice as reporter mouse. The cell proliferation and tdTomato expressions were observed. The mesenchymal cell separated from neonatal molar tooth germ were cultured with or without FGF4, FGF9, and with or without their inhibitors ferulic acid and infigratinib (BGJ398) for 21 days. Their phenotypes were evaluated by cell count, flow cytometry, and real-time PCR. Immunohistochemistry for FGFR1, 2, and 3 expression and the expression of DMP1 were performed. FGF4 treatment of mesenchymal cells obtained promoted the expression of all odontoblast markers. FGF9 failed to enhance dentin sialophosphoprotein (Dspp) expression levels. Runt-related transcription factor 2 (Runx2) was upregulated until day 14 but was downregulated on day 21. Compared to Dmp1-negative cells, Dmp1-positive cells expressed higher levels of all odontoblast markers, except for Runx2. Simultaneous treatment with FGF4 and FGF9 had a synergistic effect on odontoblast differentiation, suggesting that they may play a role in odontoblast maturation.
Topics: Animals; Mice; Cell Differentiation; Core Binding Factor Alpha 1 Subunit; Fibroblast Growth Factor 4; Mice, Transgenic; Odontoblasts; Fibroblast Growth Factor 9
PubMed: 37012505
DOI: 10.1007/s00795-023-00351-2