Odontoblast-like Cells Research Articles

Co-doped titanium dioxide nanoparticles decrease the cytotoxicity of experimental hydrogen peroxide gels for in-office tooth bleaching.

To evaluate the efficacy and cytotoxicity of experimental 6% and 35% hydrogen peroxide gels (HP6 or HP35) incorporated with titanium dioxide nanoparticles (NP) co-doped with nitrogen and fluorine and irradiated with a violet LED light (LT). Bovine enamel-dentin disks adapted to artificial pulp chambers were randomly assigned to bleaching (n = 8/group): NC (negative control), NP, HP6, HP6 + LT, HP6 + NP, HP6 + NP + LT, HP35, HP35 + LT, HP35 + NP, HP35 + NP + LT, and commercial HP35 (COM). Color (ΔE00) and whiteness index (ΔWID) changes were measured before and 14days after bleaching. The extracts (culture medium + diffused gel components) collected after the first session were applied to odontoblast-like MDPC-23 cells, which were assessed concerning their viability, oxidative stress, and morphology. The amount of HP diffused through the disks was determined. Data were analyzed by generalized linear models orKruskal Wallis Tests (α = 5%). RESULTS: HP6 + NP + LT exhibited ΔE00 and ΔWID higher than HP6 (p < 0.05) and similar to all HP35 groups. HP6 + NP + LT showed the lowest HP diffusion, and the highest cell viability (%) among bleached groups, preserving cell morphology and number of living cells similar to NC and NP. HP6 + LT, HP6 + NP, and HP6 + NP + LT exhibited the lowest cell oxidative stress among bleached groups (p < 0.05). HP35, HP35 + LT, and HP35 (COM) displayed the lowest cell viability. HP6 achieved significantly higher color and whiteness index changes when incorporated with nanoparticles and light-irradiated and caused lower cytotoxicity than HP35 gels. The nanoparticles significantly increased cell viability and reduced the hydrogen peroxide diffusion and oxidative stress, regardless of HP concentration. Incorporation of co-doped titanium dioxide nanoparticles combined with violet irradiation within the HP6 gel could promote a higher perceivable and acceptable efficacy than HP6 alone, potentially reaching the optimal esthetic outcomes rendered by HP35. This approach also holds the promise of reducing cytotoxic damages and, consequently, tooth sensitivity.

Sensory nerves drive migration of dental pulp stem cells via the CGRP-Ramp1 axis in pulp repair

Dental pulp stem cells (DPSCs) are responsible for maintaining pulp structure and function after pulp injury. DPSCs migrate directionally to the injury site before differentiating into odontoblast-like cells, which is a prerequisite and a determinant in pulp repair. Increasing evidence suggests that sensory neuron-stem cell crosstalk is critical for maintaining normal physiological functions, and sensory nerves influence stem cells mainly by neuropeptides. However, the role of sensory nerves on DPSC behaviors after pulp injury is largely unexplored. Here, we find that sensory nerves released significant amounts of calcitonin gene-related peptide (CGRP) near the injury site, acting directly on DPSCs via receptor activity modifying protein 1 (RAMP1) to promote collective migration of DPSCs to the injury site, and ultimately promoting pulp repair. Specifically, sensory denervation leads to poor pulp repair and ectopic mineralization, in parallel with that DPSCs failed to be recruited to the injury site. Furthermore, in vitro evidence shows that sensory nerve-deficient microenvironment suppressed DPSC migration prominently among all related behaviors. Mechanistically, the CGRP-Ramp1 axis between sensory neurons and DPSCs was screened by single-cell RNA-seq analysis and immunohistochemical studies confirmed that the expression of CGRP rather than Ramp1 increases substantially near the damaged site. We further demonstrated that CGRP released by sensory nerves binds the receptor Ramp1 on DPSCs to facilitate cell collective migration by an indirect co-culture system using conditioned medium from trigeminal neurons, CGRP recombinant protein and antagonists BIBN4096. The treatment with exogenous CGRP promoted the recruitment of DPSCs, and ultimately enhanced the quality of pulp repair. Targeting the sensory nerve could therefore provide a new strategy for stem cell-based pulp repair and regeneration.

Role and Molecular Mechanism of miR-586 in the Differentiation of Dental Pulp Stem Cells into Odontoblast-like Cells.

Dental pulp stem cells (DPSCs) are a class of cells with the potential of self-replication and multi-directional differentiation, which are widely considered to have great application value. It was to investigate miR-586 in DPSCs differentiated into odontoblast-like cells. In this article, human dental pulp stem cells (hDPSCs) were used as samples, and hDPSCs were co-cultured with endothelial progenitor cells (EPCs). Furthermore, a lentiviral expression vector for the miR-586 inhibitor was established. The effect of miR-586 inhibitor expression vector on the activity of hDPSCs was detected by Cell Counting Kit-8 (CCK-8). The differentiation of hDPSCs was tested by mineralized nodule staining. The expression of miR-586 and a gene related to dental cell differentiation in the pulp was subjected to detection by real-time quantitative PCR (qRT-PCR). As against the normal hDPSCs and the empty vector, the miR-586 lentivirus expression inhibition vector could visibly raise the expression of dentin sialophosphoprotein (DSPP) in hDPSCs; and the cell proliferation activity was visibly enhanced; In addition, the mRNA expressions of dentin-matrix acidic phosphoprotein 1 (DMP-1) and alkaline phosphatase (ALP) were visibly raised in the miR-586 lentivirus expression inhibition vector (all P < 0.05). Additionally, ALP activity was significantly enhanced (P < 0.05). The number of mineralized nodules was significantly increased (P < 0.05). MiR-586 plays a key regulatory function in DPSCs differentiated into odontoblast-like cells and is associated with specific molecular mechanisms.

The Role of Mesenchymal Stem Cells in Dentistry: A Review

Mesenchymal stem cells (MSCs) have garnered significant attention in regenerative dentistry due to their remarkable ability to differentiate into various cell types and their immunomodulatory properties. This review provides a comprehensive overview of recent advancements in MSC research relevant to dentistry, focusing on their potential applications in periodontal tissue regeneration, dental pulp regeneration, and maxillofacial bone repair. Periodontal diseases, which affect the tissues surrounding and supporting the teeth, are a significant challenge in dentistry. Current treatments often involve surgical intervention and tissue grafting. MSCs have shown promise as a potential alternative for periodontal tissue regeneration, as they can differentiate into periodontal ligament cells, cementoblasts, and osteoblasts. Several preclinical and clinical studies have demonstrated the efficacy of MSC-based therapies in periodontal regeneration. Dental pulp regeneration is another area where MSCs hold promise. Damaged or infected dental pulp can lead to pulpitis or pulp necrosis, necessitating root canal treatment. MSCs have been studied for their ability to regenerate dental pulp tissue and promote pulp healing. They can differentiate into odontoblast-like cells and regenerate dentin-like tissue, making them a potential treatment option for dental pulp regeneration. In maxillofacial bone repair, MSCs have been investigated for their osteogenic differentiation potential and ability to stimulate bone regeneration. Studies have shown promising results, suggesting that MSC-based therapies could be a viable treatment option for maxillofacial bone defects. The mechanisms underlying MSC-based therapies in dentistry are not fully understood but are thought to involve a combination of paracrine effects, immunomodulation, and differentiation into specific cell types. Future research should focus on addressing these challenges and exploring novel approaches for enhancing the regenerative potential of MSCs in dentistry.

PIEZO1 Promotes Odontoblast-Mediated Reactionary Dentinogenesis via SEMA3A

Located at the interface of the dentin–pulp complex, the odontoblasts are specialized cells responsible for dentin synthesis and nociceptive signal detection in response to external stimuli. Recent studies have shown that the mechanosensitive ion channel PIEZO1 is involved in bone formation and remodeling through the influx of calcium ions, and it is abundantly expressed in odontoblasts. However, the specific role of PIEZO1 in reactionary dentinogenesis and the underlying mechanisms remain elusive. In this study, we found intense PIEZO1 expression in the plasma membrane and cytoplasm of odontoblasts in healthy human third molars, mouse mandibular molars, and human odontoblast-like cells (hOBLCs). In hOBLCs, PIEZO1 positively regulated DSPP, DMP1, and COL1A1 expression through the Ca2+/PI3K-Akt/SEMA3A signaling pathway. In addition, exogenous SEMA3A supplementation effectively reversed reduced mineralization capacity in PIEZO1-knockdown hOBLCs. In vivo, Piezo1 expression peaked at day 7 and returned to baseline at day 21 in a wild-type mice dentin injury model, with Sema3a presenting a similar expression pattern. To investigate the specific role of PIEZO1 in odontoblast-mediated reactionary dentinogenesis, mice with a conditional knockout of Piezo1 in odontoblasts were generated, and no significant differences in teeth phenotypes were observed between the control and conditional knockout (cKO) mice. Nevertheless, cKO mice exhibited reduced reactionary dentin formation and decreased Sema3a and Dsp positive staining after dentin injury, indicating impaired dental pulp repair by odontoblasts. In summary, these findings suggest that PIEZO1 enhances the mineralization capacity of hOBLCs in vitro via the Ca2+/PI3K-Akt/SEMA3A signaling pathway and contributes to reactionary dentinogenesis in vivo.

Biomimetic pulp scaffolds prepared from extracellular matrix derived from stem cells from human exfoliated deciduous teeth promote pulp-dentine complex regeneration.

To evaluate the role of biomimetic pulp scaffolds derived from the extracellular matrix derived of stem cells from human exfoliated deciduous teeth (SHED-ECM-PS) in promoting pulp-dentine complex regeneration. SHED-ECM-PS was prepared through cell aggregation and decellularization techniques. Histological and immunofluorescence analyses, scanning electron microscopy, and DNA quantification assays were used to characterize the SHED-ECM-PS. Additionally, a tooth slice implantation model was established to evaluate the effects of SHED-ECM-PS on regeneration of the pulp-dentine complex invivo. Extraction medium for SHED-ECM-PS was prepared, and its effect on bone marrow mesenchymal stem cells (BMMSCs) was assessed invitro. Cell counting kit-8 and Ki-67 staining assays were performed to determine cell proliferation. The rate of apoptosis was evaluated by flow cytometry. Wound healing and transwell assays were conducted to evaluate cell migration. Alizarin red S staining was performed to examine mineralized nodule formation. Western blotting was used to detect the expression of osteogenic and odontogenic markers. The results were analysed using an independent two-tailed Student's t-test. p < .05 was considered statistically significant. SHED-ECM-PS was successfully constructed, exhibiting a striped dental pulp-like shape devoid of nuclear structures or DNA components, and rich in fibronectin, collagen I, DMP1 and DSPP. Notably, SHED-ECM-PS showed no impact on the proliferation or apoptosis of BMMSCs. Histological analysis revealed that dental pulp fibroblasts formed an interwoven mesh in the root canal, and angiogenesis was observed in the SHED-ECM-PS group. Moreover, a continuous, newly formed tubular dentine layer with polarized odontoblast-like cells was observed along the inner wall of the root canal. SHED-ECM-PS promoted the migration, polar alignment and mineralized nodule formation of BMMSCs and specifically elevated the expression levels of odontogenic markers, but not osteogenic markers, compared with the control group invitro. SHED-ECM-PS exhibited no cytotoxicity and promoted pulp-dentine complex regeneration invivo as well as cell migration and odontogenic differentiation of BMMSCs invitro. These findings provide evidence that SHED-ECM-PS, as a novel biological scaffold, has the potential to improve the outcomes of REPs.

Analysis of dentin wear and biological properties promoted by experimental inoffice desensitizing materials

BackgroundThis study aimed to evaluate dentin wear and biological performance of desensitizing materials.MethodsSeventy bovine root dentin blocks were sectioned. Half of the surface of each specimen was untreated (control) and the other half was immersed in EDTA and treated with the following desensitizing materials: placebo varnish (PLA), fluoride varnish (FLU), sodium fluoride (NaF) varnish + sodium trimetaphosphate (TMP), universal adhesive (SBU), S-PRG varnish (SPRG), biosilicate (BIOS), and amelotin solution (AMTN). After application, the specimens were submitted to an erosive-abrasive challenge and the wear analyzed by optical profilometer. Serial dilutions of extracts obtained from the culture medium containing discs impregnated with those desensitizers were applied on fibroblasts and odontoblasts-like cells cultures. Cytotoxicity and production of total protein (TP) by colorimetric assays were determined after 24 h. Data were statistically analyzed using Kruskal-Wallis, Dunn’s, One-way ANOVA and Tukey tests (p ≤ 0.05).ResultsNo dentin wear was observed only for SBU. The lowest dentin wear was observed for AMTN and TMP. Cell viability was significantly reduced after treatment with undiluted extracts of PLA, FLU, TMP and SBU in fibroblasts and TMP and SBU in odontoblast-like cells. SPRG, BIOS and AMTN were cytocompatible at all dilutions tested. Considering TP results, no statistical difference was observed among the groups and high levels for TP were observed after TMP and FLU treatments.ConclusionsUniversal adhesive system may protect dentin with opened tubules from wear after challenge. Extracts of adhesive and fluoride varnishes presented cytotoxic mainly on fibroblasts. The enamel protein may be a future alternative to treat dentin with opened tubules because it may cause low wear under erosive-abrasive challenge with low cytotoxic effects.

Electrical Stimulations Generated by P(VDF-TrFE) Films Enhance Adhesion Forces and Odontogenic Differentiation of Dental Pulp Stem Cells (DPSCs).

Biophysical and biochemical cues of biomaterials can regulate cell behaviors. Dental pulp stem cells (DPSCs) in pulp tissues can differentiate to odontoblast-like cells and secrete reparative dentin to form a barrier to protect the underlying pulp tissues and enable complete pulp healing. Promotion of the odontogenic differentiation of DPSCs is essential for dentin regeneration. The effects of the surface potentials of biomaterials on the adhesion and odontogenic differentiation of DPSCs remain unclear. Here, poly(vinylidene fluoride-trifluoro ethylene) (P(VDF-TrFE)) films with different surface potentials were prepared by the spin-coating technique and the contact poling method. The cytoskeletal organization of DPSCs grown on P(VDF-TrFE) films was studied by immunofluorescence staining. Using atomic force microscopy (AFM), the lateral detachment forces of DPSCs from P(VDF-TrFE) films were quantified. The effects of electrical stimulation generated from P(VDF-TrFE) films on odontogenic differentiation of DPSCs were evaluated in vitro and in vivo. The unpolarized, positively polarized, and negatively polarized films had surface potentials of -52.9, +902.4, and -502.2 mV, respectively. DPSCs on both negatively and positively polarized P(VDF-TrFE) films had larger cell areas and length-to-width ratios than those on the unpolarized films (P < 0.05). During the detachment of DPSCs from P(VDF-TrFE) films, the average magnitudes of the maximum detachment forces were 29.4, 72.1, and 53.9 nN for unpolarized, positively polarized, and negatively polarized groups, respectively (P < 0.05). The polarized films enhanced the mineralization activities and increased the expression levels of the odontogenic-related proteins of DPSCs compared to the unpolarized films (P < 0.05). The extracellular signal-regulated kinase (ERK) signaling pathway was involved in the odontogenic differentiation of DPSCs as induced by surface charge. In vivo, the polarized P(VDF-TrFE) films enhanced adhesion of DPSCs and promoted the odontogenic differentiation of DPSCs by electrical stimulation, demonstrating a potential application of electroactive biomaterials for reparative dentin formation in direct pulp capping.

Effects of Tenascin-C on Dental Pulp Tissue in Mice In Vivo And on The Proliferation and Differentiation of Dental Pulp Stem Cells into Odontoblasts and Calcification In Vitro

This study aimed to investigate the effects of tenascin-C (TN-C) on dental pulp tissue and on dental pulp stem cells (DPSCs). In in vivo studies, A collagen sponge with phosphate-buffered saline (PBS) for the control group or with TN-C for the experimental group was placed over the dental pulp of mice. The root pulp was excised at 7 and 21 days postoperatively and was observed microscopically using HE staining and immunohistochemistry. Inflammatory cells were found in the entire pulp tissue in the control group but no inflammatory cells were identified in the pulp tissue in the TN-C-treated experimental group. Further, nesting-positive cells at 7 days and dentin sialo phosphoprotein (DSPP)-positive cells at 21 days were seen in the experimental group. In in vitro studies, DPSCs were cultured in a medium with or without TN-C, after which the proliferation rate of DPSCs was measured mRNA expression levels were examined using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the formation of calcified nodules was investigated using alizarin red staining. There was no significant difference in the cell proliferation rate of DPSCs between the experimental and control groups during day 3 or 5, using the proliferation rate on day 1 as the baseline value. The expression of nesting mRNA on day 7 was significantly higher in the experimental group than in the control group (P&lt;0.05), but the expression of osteocalcin (OCN) mRNA was significantly higher in the control group than in the experimental group (P&lt;0.05). More calcified nodules formed and the area of calcified nodules in the control group was significantly greater than in the TN-C-treated experimental group on day 14 and day 21 (P &lt; 0.05). These results suggest that TN-C regulates inflammation during the healing process in the dental pulp and induces the differentiation of dental pulp into odontoblast-like cells. Further, TN-C promotes the early differentiation of DPSCs into odontoblast-like cells, which suggests that TN-C may further contribute to the inhibition of excessive dentin formation. From the above, TN-C is involved in differentiating dental pulp into mineralized tissue-forming cells and may have a beneficial effect in treatments that utilize this function

De novo regeneration of dentin pulp complex mediated by Adipose derived stem cells in an immunodeficient albino rat model (Histological, histochemical and scanning electron microscopic Study)

BackgroundDental tissue engineering is an alternative procedure for restoring damaged dental tissues. Adipose-derived stem cells are a new source of cells for regenerative endodontics in combination with scaffold materials. The descriptive data about this regenerative process is still insufficient. ObjectiveTo evaluate the regenerative potential of Adipose-derived stem cells using a self-assembling polypeptide scaffold for the dentin-pulp complex in an emptied root canal space. Material and Methods40 root segments of human single-rooted teeth were transplanted into the albino rats’ dorsal subcutaneous tissue. Root segments were divided into two groups: group I contained only a self-assembling polypeptide scaffold, and group II contained fluorescent-labeled Adipose-derived stem cells embedded in a self-assembling polypeptide scaffold. The newly formed tissues were assessed on the 60th and 90th days post-transplantation using routine histological examination, Masson trichrome staining, and scanning electron microscopy. ResultsGroup I showed granulation tissue without any signs of predentin formation or odontoblast-like cells. Group II revealed the presence of predentin tissue along the dentin margin, with arranged odontoblast-like cells. An organized connective tissue with abundant vasculature and calcific masses was observed in the pulp space. ConclusionAdipose-derived stem cells can be considered as alternative stem cells for regenerating the dentin-pulp complex. Dentin pulp complex regeneration utilizing a self-assembling polypeptide scaffold alone would not yield successful results.

Peptide KN-17-Loaded Supramolecular Hydrogel Induces the Regeneration of the Pulp-Dentin Complex.

Regenerating the pulp-dentin complex remains a decisive factor during apexification for immature permanent teeth. Peptide KN-17, which was modified based on the structure of cecropin B, could effectively interfere with bacterial growth and induce the migration of human bone marrow stromal cells (hBMSCs). This study aimed to investigate the effect of KN-17 on the tissue regeneration. To our surprise, KN-17 can significantly stimulate angiogenesis in vitro and in vivo, which may provide a guarantee for apical closure. Herein, a novel peptide/KN-17 coassembled hydrogel is developed via a heating-cooling process. Npx-FFEY/KN-17 supramolecular hydrogel can induce vessel development, stimulate odontogenic differentiation of human dental pulp stem cells (hDPSCs), and exert an antibacterial effect on Enterococcus faecalis (E. faecalis). Furthermore, coronal pulp excised rat molars are supplied with KN-17 or KN-17-loaded hydrogel and transplanted subcutaneously in BALB/c-nu mice. After 4 weeks, the hydrogel Npx-FFEY/KN-17 stimulates the formation of multiple odontoblast-like cells and dentin-like structures. Our findings demonstrate that the KN-17-loaded hydrogel can promote the regeneration of the pulp-dentin complex for continued root development.

Effects of Semiconductor Laser Irradiation on Differentiation of Human Dental Pulp Stem Cells in Co-Culture with Dentin.

This study aimed to determine the effect of photobiomodulation therapy induced by semiconductor laser irradiation on human dental pulp stem cell (hDPSC) proliferation and their differentiation into odontoblast-like cells (OLCs). The effects of various semiconductor laser irradiation conditions on hDPSCs were examined. Three groups were evaluated: a single laser irradiation at 6 h post-seeding, multiple laser irradiations up to four times every 4 days after the first dose, and a control with no laser irradiation. The cells were irradiated at 10, 30, and 150 mW using a semiconductor laser. The effect of laser irradiation on hDPSC differentiation into OLCs was also determined. Four groups were evaluated, including co-culture using basic medium and dentin discs, simple culture using OLC differentiation-inducing medium, co-culture using OLC differentiation-inducing medium and dentin discs, and control culture with basic medium. The expression of the nestin, ALP, DSPP, and DMP-1 genes was measured using real-time PCR. The multiple irradiation group irradiated at 30 mW exhibited significantly more cell proliferation than the control. The expression of nestin associated with differentiation into OLCs during each culture period tended to be lower, whereas DSPP and ALP expression was higher compared with that of the control. Multiple laser irradiations at a low power of 30 mW induced significant hDPSC proliferation and might induce differentiation into OLCs.

Modulation of TRPV1 on Odontoblast-like Cells Using Capsazepine-Loaded Nanogels.

The modulation of TRPV1 emerges as a promising strategy for dental pain management. This study aimed to assess TRPV1 modulation in a human odontoblast-like cell model using Capsazepine (CZP) loaded in a nanogel delivery system. Gelatin nanogels, synthesized via the emulsification-gelation technique, were characterized and loaded with the TRPV1 antagonist, CZP. HPLC determined a remarkable 67.5 ± 0.04% CZP loading efficiency, with 71.7% of nanogels falling within the 300-950 nm size range, as evidenced by light microscopy. Moreover, CZP-loaded nanogels had a low cytotoxicity. An FTIR analysis showed no adverse chemical interactions, ensuring stability and active release. When examining biological responses, TRPV1 expression and channel activity were assessed in odontoblast-like cells. On the fifth day post-treatment, cells treated with CZP-loaded nanogels exhibited an increased TRPV1 expression and a reduction in calcium fluxes after agonist stimulus (F/F0 ratio 1.18 ± 0.18), resembling the response in free CZP-treated cells (1.28 ± 0.15). A two-way analysis of variance and the Tukey's test were used to determine statistical significance (p < 0.05). This delivery system, proven to be economical and straightforward, holds promise for dental pain management and potential local use. Local administration minimizes systemic adverse effects, making it a practical solution for releasing molecules in the oral cavity.

Influence of coating dental enamel with a TiF4-loaded polymeric primer on the adverse effects caused by a bleaching gel with 35% H2O2

ObjectiveTo evaluate whether coating enamel with a polymeric primer (PPol) containing titanium tetrafluoride (TiF4) before applying a bleaching gel with 35% H2O2 (35% BG) increases esthetic efficacy, prevents changes in morphology and hardness of enamel, as well as reduces the cytotoxicity from conventional in-office bleaching. Materials and methodsStandardized enamel/dentin discs were stained and bleached for 45 min (one session) with 35% BG. Groups 2TiF4, 6TiF4, and 10TiF4 received the gel on the enamel previously coated with PPol containing 2 mg/mL, 6 mg/mL, or 10 mg/mL, respectively. No treatment or application of 35% BG directly on enamel were used as negative control (NC), and positive control (PC), respectively. UV-reflectance spectrophotometry (CIE L*a*b* system, ΔE00, and ΔWI, n = 8) determined the bleaching efficacy of treatments. Enamel microhardness (Knoop, n = 8), morphology, and composition (SEM/EDS, n = 4) were also evaluated. Enamel/dentin discs adapted to artificial pulp chambers (n = 8) were used for trans-amelodentinal cytotoxicity tests. Following the treatments, the extracts (culture medium + bleaching gel components diffused through the discs) were collected and applied to odontoblast-like MDPC-23 cells, which were assessed concerning their viability (alamarBlue, n = 8; Live/Dead, n = 4), oxidative stress (n = 8), and morphology (SEM). The amount of H2O2 in the extracts was also determined (leuco crystal violet/peroxidase, n = 8). The numerical data underwent one-criterion variance analysis (one-way ANOVA), followed by Tukey's test, at a 5% significance level. ResultsRegarding the ΔE00, no difference was observed among groups 2TiF4, 6TiF4, and PC (p > 0.05). The ΔWI was similar between groups 2TiF4 and PC (p > 0.05). The ΔWI of group 6TiF4 was superior to PC (p < 0.05), and group 10TiF4 achieved the highest ΔE00 and ΔWI values (p < 0.05). Besides limiting enamel microstructural changes compared to PC, group 10TiF4 significantly increased the hardness of this mineralized dental tissue. The highest cellular viability occurred in 10TiF4 compared to the other bleached groups (p < 0.05). Trans-amelodentinal H2O2 diffusion decreased in groups 2TiF4, 6TiF4, and 10TiF4 in comparison with PC (p < 0.05). ConclusionCoating enamel with a PPol containing TiF4 before applying a 35% BG may increase enamel microhardness and esthetic efficacy and reduce the trans-amelodentinal cytotoxicity of conventional in-office tooth bleaching. The PPol containing 10 mg/mL of TiF4 promoted the best outcomes.

Dental Pulp Cell Transplantation Combined with Regenerative Endodontic Procedures Promotes Dentin Matrix Formation in Mature Mouse Molars.

Regenerative endodontic procedures (REPs) are promising for dental pulp tissue regeneration; however, their application in permanent teeth remains challenging. We assessed the potential combination of an REP and local dental pulp cell (DPC) transplantation in the mature molars of C57BL/6 mice with (REP + DPC group) or without (REP group) transplantation of DPCs from green fluorescent protein (GFP) transgenic mice. After 4 weeks, the regenerated tissue was evaluated by micro-computed tomography and histological analyses to detect odontoblasts, vasculogenesis, and neurogenesis. DPCs were assessed for mesenchymal and pluripotency markers. Four weeks after the REP, the molars showed no signs of periapical lesions, and both the REP and REP + DPC groups exhibited a pulp-like tissue composed of a cellular matrix with vessels surrounded by an eosin-stained acellular matrix that resembled hard tissue. However, the REP + DPC group had a broader cellular matrix and uniquely contained odontoblast-like cells co-expressing GFP. Vasculogenesis and neurogenesis were detected in both groups, with the former being more prominent in the REP + DPC group. Overall, the REP was achieved in mature mouse molars and DPC transplantation improved the outcomes by inducing the formation of odontoblast-like cells and greater vasculogenesis.

Sphingosine-1-phosphate receptor 1-mediated odontogenic differentiation of mouse apical papilla-derived stem cells

Background/purposeSphingosine-1-phosphate (S1P) exhibits receptor-mediated physiological effects by facilitating the differentiation of mesenchymal stem cells toward the osteoblast lineage. This study aimed to determine the effect of S1P on odontogenic differentiation of mouse immortalized stem cells of dental apical papilla (iSCAP) and assess the distribution of the S1P receptor 1 (S1PR1) in the apical papilla and the root canal wall of immature rat molars. Materials and methodsImmunostaining for S1PR1 was conducted at the apex of the rat mandibular first molar and within the root canal wall. The iSCAP was treated with S1P and bone morphogenetic protein (BMP)-9 (for comparison), and the expression levels of the odontogenic differentiation marker were evaluated via real-time reverse-transcriptase quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Mineralization and lipid droplet formation were evaluated via Alizarin red and Oil red O staining. ResultsS1PR1-positive cells were expressed in areas of both apical papilla and dentin-pulp interface of root canal wall. During the odontogenic differentiation of iSCAP, S1P and BMP-9 increased the expression of the differentiation marker mRNA and secreted proteins including dentin sialophosphoprotein, dentin matrix phosphoprotein 1, and matrix extracellular phosphoglycoprotein. The S1PR1 signaling pathway is involved in the action of S1P, but not that of BMP-9. S1PR1 signaling also facilitated mineralization in iSCAP and suppressed the differentiation of these cells into adipocytes. ConclusionS1P induced odontogenic differentiation of iSCAP through S1PR1. Furthermore, S1PR1-positive cells were expressed in the apical papilla of immature rat molars and in the dentin-pulp interface where odontoblast-like cells exist.

C5L2 CRISPR KO enhances dental pulp stem cell-mediated dentinogenesis via TrkB under TNFα-induced inflammation.

Background and Objectives: Dental caries is one of the most common human pathological conditions resulting from the invasion of bacteria into the dentin. Current treatment options are limited. In many cases, endodontic therapy leads to permanent pulp tissue loss. Dentin-pulp complex regeneration involves dental pulp stem cells (DPSCs) that differentiate into odontoblast-like cells under an inflammatory context. However, limited information is available on how DPSC differentiation processes are affected under inflammatory environments. We identified the crucial role of complement C5a and its receptor C5aR in the inflammation-induced odontoblastic DPSC differentiation. Methodology: Here, we further investigated the role of a second and controversial C5a receptor, C5L2, in this process and explored the underlying mechanism. Human DPSCs were examined during 7-, 10-, and 14-day odontogenic differentiation treated with TNFα, C5L2 CRISPR, and tyrosine receptor kinase B (TrkB) antagonist [cyclotraxin-B (CTX-B)]. Results: Our data demonstrate that C5L2 CRISPR knockout (KO) enhances mineralization in TNFα-stimulated differentiating DPSCs. We further confirmed that C5L2 CRISPR KO significantly enhances dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1) expression after 14-day odontoblastic DPSC differentiation, and treatment with CTX-B abolished the TNFα/C5L2 CRISPR KO-induced DSPP and DMP-1 increase, suggesting TrkB's critical role in this process. Conclusion and Key applications: Our data suggest a regulatory role of C5L2 and TrkB in the TNFα-induced odontogenic DPSC differentiation. This study may provide a useful tool to understand the mechanisms of the role of inflammation in dentinogenesis that is required for successful DPSC engineering strategies.

Catalysis-based approaches with biopolymers and violet LED to improve in-office dental bleaching.

This in vitro experimental investigation aimed to evaluate the impact of the combined application of a nanofiber scaffold (NS), a polymeric catalyst primer (PCP) containing 10mg/mL of heme peroxidase enzyme, and violet LED (LEDv) on the esthetic efficacy (EE), trans-amelodentinal cytotoxicity (TC), and procedural duration of conventional in-office bleaching therapy. To achieve this, 96 standardized enamel/dentin discs were individually placed in artificial pulp chambers. A 35% hydrogen peroxide (H2O2) bleaching gel was administered for 45, 30, or 15min to the enamel, either previously coated with NS + PCP or left uncoated, followed by irradiation with LEDv for 15min or no irradiation. The established groups were as follows: G1, negative control (no treatment); G2, 35% H2O2/45min; G3, NS + PCP + LEDv; G4, NS + PCP + 35%H2O2/45min + LEDv; G5, NS + PCP + 35%H2O2/30min + LEDv; and G6, NS + PCP + 35%H2O2/15min + LEDv. Extracts (culture medium + gel components diffused through the discs) were collected and applied to odontoblast-like MDPC-23 cells. EE (ΔE00 and ΔWI) and TC were assessed using ANOVA/Tukey analysis (p < 0.05). The EE analysis revealed no statistical differences between G6 and G2 (p > 0.05). Cells in G6 exhibited higher viability and lower oxidative stress compared to other bleached groups (p < 0.05). In conclusion, employing NS + PCP + LEDv to catalyze a 35%H2O2 bleaching gel applied for 15min to the enamel resulted in successful esthetic improvements and reduced the cytotoxicity commonly linked with traditional in-office bleaching procedures.

Fusobacterium nucleatum exacerbates the progression of pulpitis by regulating the STING-dependent pathway.

Bacterial infection is the main cause of pulpitis. However, whether a dominant bacteria can promote the progression of pulpitis and its underlying mechanism remains unclear. We provided a comprehensive assessment of the microbiota alteration in pulpitis using 16S rRNA sequencing. Fusobacterium nucleatum was the most enriched in pulpitis and played a pathogenic role accelerating pulpitis progression in rat pulpitis model. After odontoblast-like cells cocultured with F. nucleatum, the stimulator of interferon genes (STING) pathway and autophagy were activation. There was a float of STING expression during F. nucleatum stimulation. STING was degraded by autophagy at the early stage. At the late stage, F.nucleatum stimulated mitochondrial Reactive Oxygen Species (ROS) production, mitochondrial dysfunction and then mtDNA escape into cytosol. mtDNA, which escaped into cytosol, caused more cytosolic mtDNA binds to cyclic GMP-AMP synthase (cGAS). The release of IFN-β was dramatically reduced when mtDNA-cGAS-STING pathway inhibited. STING-/- mice showed milder periapical bone loss and lower serum IFN-β levels compared with wildtype mice after 28 days F. nucleatum-infected pulpitis model establishment. Our data demonstrated that F. nucleatum exacerbated the progression of pulpitis, which was mediated by the STING-dependent pathway.

Physiologic dentin regeneration: its past, present, and future perspectives.

Regenerative dentistry has rapidly progressed since the advancement of stem cell biology and material science. However, more emphasis has been placed on the success of tissue formation than on how well the newly generated tissue retains the original structure and function. Once dentin is lost, tertiary dentinogenesis can be induced by new odontoblastic differentiation or re-activation of existing odontoblasts. The characteristic morphology of odontoblasts generates the tubular nature of dentin, which is a reservoir of fluid, ions, and a number of growth factors, and protects the inner pulp tissue. Therefore, understanding the dynamic but delicate process of new dentin formation by odontoblasts, or odontoblast-like cells, following dentinal defects is crucial. In this regard, various efforts have been conducted to identify novel molecules and materials that can promote the regeneration of dentin with strength and longevity. In this review, we focus on recent progress in dentin regeneration research with biological molecules identified, and discuss its potential in future clinical applications.