Dentin Mineralization Research Articles

DPP an extracellular matrix molecule induces Wnt5a mediated signaling to promote the differentiation of adult stem cells into odontogenic lineage

Dentin phosphophoryn (DPP) an extracellular matrix protein activates Wnt signaling in DPSCs (dental pulp stem cells). Wnt/β catenin signaling is essential for tooth development but the role of DPP-mediated Wnt5a signaling in odontogenesis is not well understood. Wnt5a is typically considered as a non-canonical Wnt ligand that elicits intracellular signals through association with a specific cohort of receptors and co-receptors in a cell and context–dependent manner. In this study, DPP facilitated the interaction of Wnt5a with Frizzled 5 and LRP6 to induce nuclear translocation of β-catenin. β-catenin has several nuclear binding partners that promote the activation of Wnt target genes responsible for odontogenic differentiation. Interestingly, steady increase in the expression of Vangl2 receptor suggest planar cell polarity signaling during odontogenic differentiation. In vitro observations were further strengthened by the low expression levels of Wnt5a and β-catenin in the teeth of DSPP KO mice which exhibit impaired odontoblast differentiation and defective dentin mineralization. Together, this study suggests that the DPP-mediated Wnt5a signaling could be exploited as a therapeutic approach for the differentiation of dental pulp stem cells into functional odontoblasts and dentin regeneration.

Effect of Er:YAG Laser-Activated Irrigation with Side-Firing Spiral Endo Tip on Dentin Mineral Composition of Tooth Root Canals

Background: Treating tooth root canal systems with Er:YAG laser together with irrigants has been shown to be effective in reducing biofilms formed by Enterococcus faecalis. This study investigated whether laser-activated irrigation (LAI) with side-firing Endo tip (LiteTouch™; Light Instruments, Yokneam, Israel) affects dentin mineral composition when used with common endodontic irrigants. Methods: Root canals of extracted human teeth were treated with Er:YAG laser using a side-firing Endo tip combined with 17% ethylenediaminetetraacetic acid (EDTA) and/or 2.5% NaOCl in continuous or intermittent mode for 60 s. Dentin mineral composition (Ca, P, O) in coronal, middle, and apical regions of root canals was examined by energy dispersive X-ray spectroscopy. Results: The use of LAI with continuous EDTA resulted in the largest reduction in Ca and P levels. A final NaOCl rinse mitigated the EDTA-mediated mineral loss in all root canal regions and increased the O content. Likewise, the reduced Ca/O and Ca/P ratios caused by continuous EDTA irrigation were reversed when combined with a final NaOCl rinse. Conclusions: LAI with Er:YAG Endo tip using continuous EDTA irrigation followed by NaOCl caused minimal dentin mineral loss and can therefore be considered to be a safe treatment module for cleaning root canals.

Evaluation of hard tissue characteristics and calcifications in pulp tissue of hypomineralized permanent molars using micro-computed tomography

ObjectivesTo determine and compare pulp volume, dentin mineral density, presence of microcracks, pulp stones, and accessory canals, as well as their localizations in root regions for hypomineralized and healthy teeth. DesignThis study included 60 extracted permanent molar teeth, categorized into hypomineralized and healthy groups (n = 30 each). The hypomineralized group comprised molar teeth with limited white, yellow, or brown opacities, post-eruptive breakdown, or extensive restoration or crown damage. The healthy group included caries-free molar teeth without these characteristics. Using 3D micro-computed tomography images pulp volume, dentin mineral density, and the presence and locations of microcracks, pulp stones, and accessory canals were determined for each group. Statistical analyses were conducted using Independent T-test and Chi-square test, with significance set at p < 0.05. ResultsThere was no statistically significant difference between the groups regarding pulp volume and microcracks (p ≥ 0.05). The number of accessory canals was significantly greater in the cervical (p = 0.011; p < 0.05) and middle (p = 0.010; p < 0.05) regions of the hypomineralized teeth than healthy teeth. Dentin mineral density was statistically higher in the apical, middle, and cervical root regions (p < 0.001; p < 0.05); however, the number of pulp stones was found to be greater in the cervical regions of healthy teeth compared with those with hypomineralization (p = 0.026; p < 0.05). ConclusionThere were lower dentin mineral density measurements, a decreased number of pulp stones in the cervical region, and a greater number of accessory canals in the middle and cervical regions of hypomineralized teeth compared with healthy teeth.

Ultrastructural evaluation of adverse effects on dentine formation from systemic fluoride application in an experimental mouse model.

Fluoride is widely used in dentistry for its caries prevention. To reduce dental caries, the optimal fluoride concentration of public water supplies in the United States is 0.7 ppm. However, excessive systemic fluoride consumption can lead to dental/enamel fluorosis. Numerous studies have explored the effects of fluoride on enamel and enamel-forming cells. However, research on systemic fluoride's impact on dentine is limited, particularly the effect of fluoride on the structure of the dentine-pulp complex. Therefore, this study aimed to identify how excessive fluoride affects dentine microstructure using an experimental mouse model. C57BL6/J male mice (6-9 weeks old) were randomized into four groups (Fluoride at 0, 50, 100, or 125 ppm in drinking water) (n = 4/group). Mice were provided water adlibitum for 6 weeks along with fluoride-free food. Thereafter, mandibular incisors were analysed. Enamel phenotypes were evaluated using light microscopy and quantitative light-induced fluorescence (QLF) to measure fluorosis levels. Dentine morphology was evaluated using micro-CT, scanning electron microscopy (SEM), SEM-EDX (energy-dispersive X-ray), microhardness test and histological imaging. Data were analysed using one-way ANOVA with Dunnett's multiple comparisons as a post hoc test and the Kruskal-Wallis test with Dunn's multiple comparisons post hoc test (p < .05). Mice treated with fluoride at 50-125 ppm developed enamel hypoplasia in their erupting incisors and micro-CT imaging revealed that fluoride 125 ppm caused external resorption of the growing incisor. Dentine mineral density, dentine volume decreased compared with the 0 ppm control, while pulp volume increased compared with the 0 ppm control group. SEM showed wider predentine layer and abnormalities in calcified matrix vesicles derived from odontoblasts in fluoride 100 and 125 ppm groups. Vickers microhardness of dentine significantly decreased in the high-dose group. Fluoride-induced dentine hypoplasia in a dose-dependent manner. Histological evaluation showed excessive fluoride 125 ppm induced micro abscess formation and inflammatory cell infiltration. Fluoride induced dentine dysplasia with a dentine microstructure resembling hypophosphatasia. High doses of systemic fluoride can cause dentine dysplasia. Both three-dimensional and microstructural analyses showed the structural, chemical and mechanical changes in the dentine and the mineralized tissue components, along with external resorption and pulp inflammation.

Assessing the impact of different Urushiol primer solvents on dentin remineralization and bond strength.

To investigate urushiol's potential as a dentin cross-linking agent, promoting remineralization of etched dentin and preventing activation of endogenous proteases causing collagen degradation within the hybrid layer. The goal is to improve bond strength and durability at the resin-dentin interface. Urushiol primers with varying concentrations were prepared using ethanol and dimethyl sulfoxide (DMSO) as solvents. Dentin from healthy molars underwent grinding and acid etching for 15s, followed by a 1min application of urushiol primer. After 14 and 28days of remineralization incubation and remineralization were usedtoassessby Attenuated Total Reflection Fourier Transform Infrared spectroscopy (ATR-FTIR), Micro-Raman spectroscopy, X-ray Diffraction (XRD), Atomic Force Microscopy (AFM), Vickers Hardness, Scanning Electron Microscopy (SEM), and Energy X-ray dispersive spectroscopy (EDS). The overall performance of urushiol primers as dentin adhesives was observed by microtensile bond strength (μTBS) testing and nanoleakage assessment. Investigated the inhibitory properties of the urushiol primers on endogenous metalloproteinases (MMPs) utilizing in situ zymography, and the cytotoxicity of the primers was tested. Based on ATR-FTIR, Raman, XRD, EM-EDS and Vickers hardness analyses, the 0.7%-Ethanol group significantly enhanced dentin mineral content and improved mechanical properties the most. Pretreatment notably increased the μTBS of restorations, promoted the stability of the mixed layer, and reduced nanoleakage and MMPs activity after 28days. The urushiol primer facilitates remineralization in demineralized dentin, enhancing remineralization in etched dentin, effectively improving the bonding interface stability, with optimal performance observed at a 0.7 wt% concentration of the urushiol primer.

Hypoxia-inducible factors in postnatal mouse molar dental pulp development: insights into expression patterns, localisation and metabolic pathways.

Hypoxia is relevant to several physiological and pathological processesand this also applies for the tooth. The adaptive response to lowering oxygen concentration is mediated by hypoxia-inducible factors (HIFs). Since HIFs were shown to participate in the promotion of angiogenesis, stem cell survival, odontoblast differentiation and dentin formation, they may play a beneficial role in the tooth reparative processes. Although some data were generated in vitro, little is known about the in vivo context of HIFs in tooth development. In order to contribute to this field, the mouse mandibular first molar was used as a model.The expression and in situ localisation of HIFs were examined at postnatal (P) days P0, P7, P14, using RT-PCR and immunostaining. The expression pattern of a broad spectrum of hypoxia-related genes was monitored by customised PCR Arrays. Metabolic aspects were evaluated by determination of the lactate level and mRNA expression of the mitochondrial marker Nd1.The results show constant high mRNA expression of Hif1a, increasing expression of Hif2a, and very low expression of Hif3a during early postnatal molar development. In the examined period the localisation of HIFs in the nuclei of odontoblasts and the subodontoblastic layer identified their presence during odontoblastic differentiation. Additionally, the lower lactate level and higher expression of mitochondrial Nd1 in advanced development points to decreasing glycolysis during differentiation. Postnatal nuclear localisation of HIFs indicates a hypoxic state in specific areas of dental pulp as oxygen demands depend on physiological events such as crown and root dentin mineralization.

Novel L-(CaP-ZnP)/SA Nanocomposite Hydrogel with Dual Anti-Inflammatory and Mineralization Effects for Efficient Vital Pulp Therapy.

Vital pulp therapy (VPT) is considered a conservative treatment for preserving pulp viability in caries and trauma-induced pulpitis. However, Mineral trioxide aggregate (MTA) as the most frequently used repair material, exhibits limited efficacy under inflammatory conditions. This study introduces an innovative nanocomposite hydrogel, tailored to simultaneously target anti-inflammation and dentin mineralization, aiming to efficiently preserve vital pulp tissue. The L-(CaP-ZnP)/SA nanocomposite hydrogel was designed by combining L-Arginine modified calcium phosphate/zinc phosphate nanoparticles (L-(CaP-ZnP) NPs) with sodium alginate (SA), and was characterized with TEM, SEM, FTIR, EDX, ICP-AES, and Zeta potential. In vitro, we evaluated the cytotoxicity and anti-inflammatory properties. Human dental pulp stem cells (hDPSCs) were cultured with lipopolysaccharide (LPS) to induce an inflammatory response, and the cell odontogenic differentiation was measured and possible signaling pathways were explored by alkaline phosphatase (ALP)/alizarin red S (ARS) staining, qRT-PCR, immunofluorescence staining, and Western blotting, respectively. In vivo, a pulpitis model was utilized to explore the potential of the L-(CaP-ZnP)/SA nanocomposite hydrogel in controlling pulp inflammation and enhancing dentin mineralization by Hematoxylin and eosin (HE) staining and immunohistochemistry staining. In vitro experiments revealed that the nanocomposite hydrogel was synthesized successfully and presented desirable biocompatibility. Under inflammatory conditions, compared to MTA, the L-(CaP-ZnP)/SA nanocomposite hydrogel demonstrated superior anti-inflammatory and pro-odontogenesis effects. Furthermore, the nanocomposite hydrogel significantly augmented p38 phosphorylation, implicating the involvement of the p38 signaling pathway in pulp repair. Significantly, in a rat pulpitis model, the L-(CaP-ZnP)/SA nanocomposite hydrogel downregulated inflammatory markers while upregulating mineralization-related markers, thereby stimulating the formation of robust reparative dentin. The L-(CaP-ZnP)/SA nanocomposite hydrogel with good biocompatibility efficiently promoted inflammation resolution and enhanced dentin mineralization by activating p38 signal pathway, as a pulp-capping material, offering a promising and advanced solution for treatment of pulpitis.

Impaired Tertiary Dentin Secretion after Shallow Injury in Tgfbr2-Deficient Dental Pulp Cells Is Rescued by Extended CGRP Signaling.

The transforming growth factor β (TGFβ) superfamily is a master regulator of development, adult homeostasis, and wound repair. Dysregulated TGFβ signaling can lead to cancer, fibrosis, and musculoskeletal malformations. We previously demonstrated that TGFβ receptor 2 (Tgfbr2) signaling regulates odontoblast differentiation, dentin mineralization, root elongation, and sensory innervation during tooth development. Sensory innervation also modulates the homeostasis and repair response in adult teeth. We hypothesized that Tgfbr2 regulates the neuro-pulpal responses to dentin injury. To test this, we performed a shallow dentin injury with a timed deletion of Tgfbr2 in the dental pulp mesenchyme of mice and analyzed the levels of tertiary dentin and calcitonin gene-related peptide (CGRP) axon sprouting. Microcomputed tomography imaging and histology indicated lower dentin volume in Tgfbr2cko M1s compared to WT M1s 21 days post-injury, but the volume was comparable by day 56. Immunofluorescent imaging of peptidergic afferents demonstrated that the duration of axon sprouting was longer in injured Tgfbr2cko compared to WT M1s. Thus, CGRP+ sensory afferents may provide Tgfbr2-deficient odontoblasts with compensatory signals for healing. Harnessing these neuro-pulpal signals has the potential to guide the development of treatments for enhanced dental healing and to help patients with TGFβ-related diseases.

Histological and ultrastructural evaluation of remaining dentin after selective caries removal in primary molars: Ex-vivo study

Aim: In this study, ultrastructural, chemical, and histological analyses of extracted deciduous teeth were performed to analyze the effect of two different types of glass ionomer cements used in association with selective caries removal in deep lesions of deciduous teeth. Methods: A clinical, ex vivo, longitudinal study was conducted. A total of 19 deciduous molars from 9 volunteer patients were used. The teeth were randomly restored with one of two types of glass ionomer cement following the selective caries removal technique. The groups were divided into a control group, which used Fuji IX (GC- Europe, Leuven, Belgium), and a test group, which used Maxxion R (FGM- Joinville, Santa Catarina, Brazil). After exfoliation, the teeth were stored at 4°C and subjected to scanning electron microscopy (SEM), optical microscopy (OM), and energy dispersive spectrometry (EDS). Results: EDS analysis revealed that calcium (Ca) and phosphorus (P) were present in 100% of the specimens in both the control and test groups. Fluoride was found in 60% of the control group specimens and 63% of the test group specimens. OM images showed areas of mineralized dentin in both treatment groups. Conclusions: Selective caries removal for deep lesions in deciduous teeth was shown to be a safe therapeutic protocol, demonstrating signs of dentin mineralization and control of disease progression. Keywords: Carious, Dental Caries Removal; Glass Ionomer Cement; Scanning Electron Microscopy

Low-fluoride gels supplemented with nano-sized sodium trimetaphosphate reduce dentin erosive wear in vitro

ObjectiveThe study assessed the effect of low-fluoride gels supplemented with micrometric or nano-sized sodium trimetaphosphate (TMP) on dentin erosive wear in vitro. DesignBovine dentin blocks (n = 154) were selected by surface microhardness and randomly allocated into seven groups (n = 22/group), according to the gels: Placebo; 4500 ppm F (4500F); 9000 ppm F (9000F); 5% TMP microparticulate plus 4500F (5TMPm+4500F); 2.5% TMP nanoparticulate plus 4500 F (2.5TMPn+4500F); 5% TMP nanoparticulate plus 4500F (5TMPn+4500F); and 12,300 ppm F acid gel (APF). All blocks were treated only once for 60 s and cyclically eroded (ERO, citric acid, 4 × 90 s/day) or eroded and brushed (4 × 15 s/day, five strokes/s, ERO+ABR) over five days (each subgroup n = 11). Dentin wear and integrated hardness loss in depth (ΔKHN) were determined, and the data were submitted to two-way ANOVA, followed by Tukey’s test, and Spearman’s correlation (p < 0.05). ResultsFor ERO, all gels containing 4500F supplemented with TMP significantly reduced dentin wear compared with their counterpart without TMP, reaching values similar to 9000F. For ERO+ABR, 5TMPn+ 4500F gel led to significantly lower wear than all its counterparts, reaching values similar to 9000F and APF. As for ΔKHN, all gels containing TMP promoted superior protective effects compared with 4500F, reaching values similar to 9000F and APF under both challenges. A positive correlation between dentin wear and mineral content in depth was verified. ConclusionsGels containing 4500F supplemented with TMP significantly reduced dentin erosive wear compared with pure 4500F, with additional benefit from the use of nanoparticles.

Effects of ethylenediaminetetraacetic acid, citric acid, and etidronic acid on root dentin mineral content and bond strength of a bioceramic-based sealer: A scanning electron microscopy-energy dispersive spectroscopy study.

This study assessed the impact of chelating agents, 17% ethylenediaminetetraacetic acid (EDTA), 10% citric acid (CA), and 18% etidronic acid (HEDP), on root dentin mineral content. Scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) was applied to analyze changes, and the push-out bond strength test was used to measure dentin adhesion of Well-Root ST, a bioceramic root canal sealer. A total of 80 extracted single-rooted lower premolar teeth were included in this study and randomly divided into four groups (n=20): group 1 (17% EDTA), group 2 (10% CA), group 3 (18% HEDP), and group 4 (distilled water, control). After irrigation and drying, SEM-EDS was applied to analyze eight samples from each group at coronal, middle, and apical root regions for mineral content and SEM images. The remaining 12 samples underwent a push-out bond strength test using Well-Root ST sealer and gutta-percha. Kruskal-Wallis and Dunn's tests were used for statistical analyses. Statistically significant differences were found between groups (P<0.05). SEM-EDS showed significant differences in C, O, Ca, P, and Ca/P content, with no significant differences in Na and Mg. Push-out bond strength was significantly higher in the 17% EDTA, 10% CA, and 18% HEDP groups compared to the control group, with no significant differences between chelating agents. Chelating agents altered root dentin mineral content and improved the adhesive properties of the bioceramic sealer. These findings highlight the importance of considering the selection and use of chelating agents in the clinical practice for root canal treatment.

Efficacy of Three Remineralizing Agents on Erosion of Root Dentin by Cola Drink: An In Vitro Study

Abstract Objective The aim of the study was to investigate the effects of silver diamine fluoride (38% SDF), sodium fluoride (NaF) varnish, and casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF) on cola-eroded root dentin microhardness and mineral alteration in vitro. Materials and Methods Forty human root dentin slabs were exposed to alternating 10 cycles of cola drink and artificial saliva, repeated 3 times at 6-hour intervals. Specimens were randomly assigned to four groups: control (deionized water), 38% SDF, NaF varnish, and CPP-ACPF. All specimens underwent the second erosion process. Microhardness was measured at baseline (KHN0), pretreatment (KHN1), and posttreatment (KHN2). The mean difference of microhardness (ΔKHN2-1) was analyzed using one-way analysis of variance (ANOVA) and Tukey's post hoc tests (α = 0.05). The chemical composition and surface morphology were assessed using energy dispersive spectroscopy (EDS) and scanning electron microscope (SEM). Results All experimental groups exhibited dentinal tubule occlusion. Both 38% SDF and NaF varnish demonstrated a statistically significant increase in microhardness compared to CPP-ACPF. However, CPP-ACPF was comparable to the control group. EDS analysis showed an increase in weight percentage of fluorine in all groups. Furthermore, silver and chlorine were detected in the 38% SDF group. Conclusion All treatments enhanced eroded root dentin microhardness, with 38% SDF and NaF varnish demonstrating superior acid resistance and preventing morphological changes induced by cola re-immersion.

Effects of a Benzalkonium Chloride Surfactant-Sodium Hypochlorite Combination on Microhardness and Mineral Content of Dentin

This study aimed to investigate the impact of the combination of Benzalkonium chloride (BAC) and sodium hypochloride (NaOCl) and its application after ethylenediaminetetraacetic acid (EDTA) in root canal irrigation procedures on the microhardness and mineral content of dentin. Distal roots of mandibular third molars were embedded in auto-polymerizing acrylic resin, sectioned coronally to apically resulting in fifty-four root sections. Thirty sections underwent microhardness evaluation, and twenty-four sections were randomly chosen for mineral analysis. The microhardness assessment comprised three groups: Group 1 (2.5% NaOCl), Group 2 (2.5% NaOCl with 0.084% BAC), and Group 3 (2.5% NaOCl with 0.084 BAC post-17% EDTA). Initial and post-irrigation microhardness values were measured for each group. For mineral analysis, samples were categorized into four groups: distilled water, 2.5% NaOCl, 2.5% NaOCl with 0.084% BAC, and 17% EDTA+2.5% NaOCl with 0.084% BAC. X-ray photoelectron spectroscopy immediately measured magnesium, phosphorus, and calcium mineral contents on dentin surfaces post-irrigation. Results indicated a significant reduction in root dentin microhardness for all solutions (p&lt;0.05). However, no statistically significant difference in the percentage of reduction was observed among the groups (p&gt;0.05). XPS analysis revealed no significant disparity in dentin surface mineral content among the groups (p&gt;0.05). In conclusion, the addition of Benzalkonium chloride to sodium hypochloride, either alone or after EDTA, did not induce a significant alteration in dentin microhardness or mineral content. These findings contribute to a nuanced understanding of dental irrigation protocols and their effects on dentin properties during endodontic procedures.

Vitamin D status in post-medieval Northern England: Insights from dental histology and enamel peptide analysis at Coach Lane, North Shields (AD 1711-1857).

The post-medieval period in Europe saw a dramatic increase in metabolic bone disease related to vitamin D deficiency (VDD). Recent paleopathological work has utilized interglobular dentin (IGD) as a proxy for poor vitamin D status during development, while enamel peptide analysis allows the identification of chromosomal sex in non-adult remains. Here we explore the relationship between sex, the presence of IGD, and macroscopic markers of VDD in an industrial era assemblage from Northeast England. 25 individuals (9 females, 9 males, 9 unknown sex) from the cemetery site at Coach Lane, North Shields (1711-1857) were selected for paleopathological analysis, histological assessment of IGD, and enamel peptide determination of chromosomal sex. Ground tooth sections from 21 individuals were of suitable quality for detection of IGD, and enamel peptide analysis confirmed the chromosomal sex of ten individuals. Sixteen individuals (76.1%) exhibited ≥1 episode of IGD. Nine of these (42.8%) exhibited >1 episode and four (19%) exhibited ≥4 episodes in regular intervals. Male sex was significantly associated with the presence of IGD (p = 0.0351; 100% males vs. 54.5% females). Females were more likely to exhibit macroscopic evidence of VDD (45.5% females vs 30% males) but this was not statistically significant. Periods of poor mineral metabolism during childhood appear much more prevalent at Coach Lane than macroscopic evidence suggests. Evidence of seasonal IGD episodes indicates that northern latitude played a major role in poor VD status in the Northeast of England. The significant association of IGD with male sex may be due to sex-related differences in dentinal mineralization or a higher risk of poor VD status in males aged <5 years. More work is needed to establish an evidence-based threshold for pathological levels of IGD before the presence of this feature can confidently be used as a biomarker for poor VD status.

Profiles of Wnt pathway gene expression during tooth morphogenesis.

Mouse and human genetic studies indicate key roles of the Wnt10a ligand in odontogenesis. Previous studies have identified effectors and regulators of the Wnt signaling pathway actively expressed during key stages of tooth morphogenesis. However, limitations in multiplexing and spatial resolution hindered a more comprehensive analysis of these signaling molecules. Here, profiling of transcriptomes using fluorescent multiplex in situ hybridization and single-cell RNA-sequencing (scRNA-seq) provide robust insight into the synchronized expression patterns of Wnt10a, Dkk1, and Sost simultaneously during tooth development. First, we identified Wnt10a transcripts restricted to the epithelium at the stage of tooth bud morphogenesis, contrasting that of Sost and Dkk1 localization to the dental mesenchyme. By embryonic day 15.5 (E15.5), a marked shift of Wnt10a expression from dental epithelium to mesenchyme was noted, while Sost and Dkk1 expression remained enriched in the mesenchyme. By postnatal day 0 (P0), co-localization patterns of Wnt10a, Dkk1, and Sost were observed in both terminally differentiating and secreting odontoblasts of molars and incisors. Interestingly, Wnt10a exhibited robust expression in fully differentiated ameloblasts at the developing cusp tip of both molars and incisors, an observation not previously noted in prior studies. At P7 and 14, after the mineralization of dentin and enamel, Wnt10a expression was limited to odontoblasts. Meanwhile, Wnt modulators showed reduced or absent signals in molars. In contrast, strong signals persisted in ameloblasts (for Wnt10a) and odontoblasts (for Wnt10a, Sost, and Dkk1) towards the proximal end of incisors, near the cervical loop. Our scRNA-seq analysis used CellChat to further contextualize Wnt pathway-mediated communication between cells by examining ligand-receptor interactions among different clusters. The co-localization pattern of Wnt10a, Dkk1, and Sost in both terminally differentiating and secreting odontoblasts of molars and incisors potentially signifies the crucial ligand-modulator interaction along the gradient of cytodifferentiation starting from each cusp tip towards the apical region. These data provide cell type-specific insight into the role of Wnt ligands and mediators during epithelial-mesenchymal interactions in odontogenesis.

Comparative Evaluation of the Effect of Different Chelating Agents on Mineral Content and Erosion of Radicular Dentine: A FESEM-EDS Analysis.

Irrigating solutions play an important role in the debridement and disinfection of the root canal space, and thus, it is crucial to comprehend their effects on the composition and surface structure of radicular dentine. This study evaluated and compared the effects of 17% ethylenediaminetetraacetic acid (EDTA), 9% 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) and 0.2% chitosan on the mineral content and erosion of radicular dentine when used as a final rinse. Sixty extracted human mandibular premolar teeth were decoronated and instrumented to ProTaper size F2. After final instrumentation, the samples were randomly divided into 4 groups (n=15) according to the type of final irrigant used: Normal saline (control), 17% EDTA, 9% HEBP and 0.2% chitosan. Field emission scanning electron microscopy was used to assess the erosion of radicular dentine, and energy dispersive X- ray spectroscopy was used to quantify the radicular dentine mineral composition at the coronal, middle and apical levels of all the prepared samples after final irrigation. The one-way analysis of variance was used for intra-group and inter-group comparisons of means, the Kruskal Wallis test for intra-group and inter-group comparisons of medians and Tukey's post hoc test for pairwise comparisons. There was no significant difference in the levels of calcium (Ca), phosphorus (P) and Ca/P ratio after final rinse with 17% EDTA, 9% HEBP and 0.2% chitosan at all three root levels (p>0.05); except at the coronal level, where 0.2% chitosan caused significantly less alteration in Ca levels and Ca/P ratio than 17% EDTA and 9% HEBP respectively (p<0.05). 17% EDTA, 9% HEBP and 0.2% chitosan caused no erosion at the middle and apical levels. Meanwhile, 17% EDTA and 9% HEBP caused moderate erosion at the coronal level. Alternatives to 17% EDTA during final irrigation can be 9% HEBP and 0.2% chitosan. (EEJ-2023-06-078).

X-Linked Hypophosphatemia: Does Targeted Therapy Modify Dental Impairment?

X-linked hypophosphatemia is a rare, hereditary disorder that significant influences teeth and alveolar bone. The first clinical sign leading to the diagnosis of X-linked hypophosphatemia is often dental impairment with dental abscesses and dentin mineralization defects. Genetic analysis helped find the responsible gene and therefore opened up new ways of therapeutically managing X-linked hypophosphatemia. The human monoclonal antibody Burosumab represents a milestone in the targeted therapy of this hereditary disease by directly addressing its pathophysiology. Targeted therapy has been shown to improve skeletal impairment, pain, and phosphate metabolism. However, the influence of this new therapy on dental impairment has only been addressed in a few recent studies with varying results. Therefore, in this review, we aim to summarize the dental phenotype and analyze the different treatment modalities with a focus on dental impairment.

Glass ionomer cement with calcium-releasing particles: Effect on dentin mineral content and mechanical properties

Objectiveto evaluate the effect a glass ionomer cement (GIC) containing hydroxyapatite (HAp) or calcium silicate (CaSi) particles on mineral content and mechanical properties of demineralized dentin. Ion release and compressive strength (CS) of the cements were also evaluated. MethodsGIC (Fuji 9 Gold Label, GC), GIC+ 5%HAp and GIC+ 5%CaSi (by mass) were evaluated. Ion release was determined by induced coupled plasma optical emission spectroscopy (Ca2+/Sr2+) or ion-specific electrode (F-) (n = 3). A composite (Filtek Z250, 3 M ESPE) was used as control in remineralization tests. Demineralized dentin discs were kept in contact with materials in simulated body fluid (SBF) at 37 °C for eight weeks. Mineral:matrix ratio (MMR) was determined by ATR-FTIR spectroscopy (n = 5). Dentin hardness (H) and elastic modulus (E) were determined by nanoindentation (n = 10). CS was tested after 24 h and 7d in deionized water (n = 12). Data were analyzed by ANOVA/Tukey test (α = 0.05). ResultsCa2+ and Sr2+ release was higher for the modified materials (p < 0.05). Only GIC+ 5%HAp showed higher F- release than the control (p < 0.05). All groups showed statistically significant increases in MMR, with no differences among them after 8 weeks (p > 0.05). No differences in dentin H or E were observed among groups (p > 0.05). HAp-modified GIC showed increased initial CS, while adding CaSi had the opposite effect (p < 0.05). After 7 days, GIC+ 5%CaSi presented lower CS in relation to control and GIC+ 5%HAp (p < 0.05). SignificanceGIC modification with HAp or CaSi affected CS and increased ion release; however, none of the groups showed evidence of dentin remineralization in comparison to the negative control.

Contributions of increased osteopontin and hypophosphatemia to dentoalveolar defects in osteomalacic Hyp mice.

X-linked hypophosphatemia (XLH) is an inherited disorder caused by inactivating mutations in the PHEX gene leading to renal phosphate wasting, rickets and osteomalacia. XLH is also associated with dentoalveolar mineralization defects in tooth enamel, dentin and cementum, and in alveolar bone, which lead to an increased prevalence of dental abscesses, periodontal disease and tooth loss. Genetic mouse experiments, and deficiencies in XLH patient therapies where treatments do not fully ameliorate mineralization defects, suggest that other pathogenic mechanisms may exist in XLH. The mineralization-inhibiting, secreted extracellular matrix phosphoprotein osteopontin (OPN, gene Spp1) is a substrate for the PHEX enzyme whereby extensive and inactivating degradation of inhibitory OPN by PHEX facilitates mineralization. Conversely, excess OPN accumulation in skeletal and dental tissues - for example in XLH where inactivating mutations in the PHEX gene limit degradation of inhibitory OPN, or as occurs in Fgf23-null mice - contributes to mineralization defects. We hypothesized that Spp1/OPN ablation in Hyp mice (a mouse model for XLH) would reduce dentoalveolar mineralization defects. Immunostaining revealed increased OPN in Hyp vs. wild-type (WT) alveolar bone, particularly in osteocyte lacunocanalicular networks where Hyp mice have characteristic hypomineralized peri-osteocytic lesions (POLs). Micro-computed tomography and histology showed that ablation of Spp1 in Hyp mice (Hyp;Spp1-/-) on a normal diet did not ameliorate bulk defects in enamel, dentin, or alveolar bone. On a high-phosphate diet, both Hyp and Hyp;Spp1-/- mice showed improved mineralization of enamel, dentin, and alveolar bone. Silver staining indicated Spp1 ablation did not improve alveolar or mandibular bone osteocyte POLs in Hyp mice; however, they were normalized by a high-phosphate diet in both Hyp and Hyp;Spp1-/- mice, although inducing increased OPN. Collectively, these data indicate that despite changes in OPN content in the dentoalveolar mineralized tissues, there exist other compensatory mineralization mechanisms that arise from knockout of Spp1/OPN in the Hyp background.

Impaired breakdown of Herwig's epithelial root sheath disturbs tooth root development.

Wnt/β-catenin signaling plays a variety of roles in both the dental epithelium and mesenchyme at most stages of tooth development. In this study, we verified the roles of Hertwig's epithelial root sheath (HERS) breakdown in tooth root development. This breakdown results in formation of epithelial cell rests of Malassez (ERM). Following induction of β-catenin stabilization in the epithelium of developing tooth at the moment of HERS breakdown, HERS failed to break down for ERM formation. HERS with stabilized β-catenin was altered into a multicellular layer enveloping elongated root dentin with higher expression of junctional proteins such as Zo-1 and E-cadherin. Importantly, this impairment of HERS breakdown led to arrest of further root elongation. In addition, the portion of root dentin enveloped by the undissociated HERS remained in a hypomineralized state. The odontoblasts showed ectopically higher expression of pyrophosphate regulators including Ank and Npp1, whereas Tnap expression was unchanged. Our data suggest that Wnt/β-catenin signaling is decreased in HERS for ERM formation during root development. Furthermore, ERM formation is important for further elongation and dentin mineralization of the tooth roots. These findings may provide new insight to understand the contribution of ERM to root formation.