To evaluate the effect of Nano-NeoMTA2 on human dental pulp stem cells (hDPSCs) differentiation and proliferation potential compared to nanohydroxyapatite. After the preparation and characterization of the nanoparticles of NeoMTA2 and hydroxyapatite, the nanoparticles were mixed prior to application. The mixed nanoparticles were added to hDPSCs harvested from third molar teeth immediately upon extraction, in addition to a negative and positive control group. Alkaline phosphatase and dentin matrix protein were used as measures for the odontogenic differentiation potential of the hDPSCs. Human DPSC count and viability of Trypan Blue and [3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide] assay were used as a measure of the proliferation potential. Nano-NeoMTA2 demonstrated significantly greater induction of hDPSC proliferation and odontogenic differentiation than nanohydroxyapatite. Both tested nano-biomaterials induce DPSC odontogenic differentiation and proliferation. Nano-NeoMTA2 stimulates odontogenic differentiation and proliferation of hDPSCs and warrants further investigation as a potential scaffold or direct pulp-capping material.

In a previous study, we demonstrated that a novel surface treatment applied to laser-melted Ti6Al4V substrates supports osteoblast-like cell adhesion, proliferation, and the activation of early osteogenic pathways. Building on these preliminary findings, the present work aimed to further investigate the ability of the same surface to promote extracellular matrix (ECM) deposition, organization, and osteogenic maturation, which are critical events for the establishment of a stable bone-implant interface in subperiosteal dental implants. Human osteoblast-like MG-63 cells were cultured on Ti6Al4V discs subjected to different surface treatments, including a proprietary surface modification (ATcs) specifically designed for subperiosteal applications. ECM formation and maturation were evaluated through scanning electron microscopy coupled with energy-dispersive spectroscopy, immunofluorescence, and semiquantitative analyses of osteogenic markers type I collagen (COL1A1), secreted protein acidic and rich in cysteine (SPARC), and dentin matrix protein 1 (DMP1) through Western blotting. The results showed that, while all tested surfaces supported cell adhesion, the ATcs surface promoted a distinct osteogenic profile characterized by enhanced DMP1 expression, organized collagen deposition, and the formation of calcium-phosphate-rich mineralized structures. Compared to surfaces that primarily stimulated cell proliferation or early matrix production, ATcs appeared to favour progression toward late-stage osteogenic maturation and matrix mineralization. Taken together, these findings extend our previous observations and indicate that this novel surface treatment not only supports osteoblast viability and early differentiation but also promotes extracellular matrix maturation, a key prerequisite for effective osseointegration. Although further in vivo studies are required, the present data provide additional biological rationale for the use of ATcs-treated Ti6Al4V surfaces in next-generation custom-made subperiosteal implant designs.

To evaluate the potential of a phosphoprotein analogue to mediate mineral deposition and enhance surface integrity in deep carious dentin lesions, compared with mineral trioxide aggregate (MTA) and Sylc air abrasion. Thirty carious dentin discs (3.5 ± 0.1 mm in thickness) were prepared and examined visually (ranging from yellow to light brown), through tactile assessment (leathery sensation), with 0-20 DIAGNOdent pen readings indicating caries-affected dentin (CAD) substrate. The specimens received three treatments: dentin matrix protein-1 (DMP-1, MCE, United States), MTA (Angelus, Brazil), and Sylc air abrasion (AquaCare, United Kingdom), and were immersed in artificial saliva (pH 7, 37°C) for 30 days. Fourier-transform infrared spectroscopy (FTIR) and contact angle measurement were performed on DMP-1. The mineral and organic content was assessed using Raman micro-spectroscopy, while cross-sectional tissue hardness was evaluated by a Vickers tester at 50 and 100 µm depth from the treated surfaces. The morphological assessment was conducted using scanning electron microscopy (SEM). One-way ANOVA followed by Tukey's multiple comparison test and paired t-test were performed for the statistical analyses. The FTIR spectra of DMP-1 displayed the amide bands associated with carbonyl and N-H stretching vibrations, in addition to the phosphate band. DMP-1 exhibited good wettability (contact angle of 27°), and enhanced the phosphate and organic content and tissue hardness of the treated CAD surfaces compared with pretreatment values (p < 0.05). Meanwhile, the MTA-treated surface demonstrated the highest phosphate content and Vickers hardness number up to 100 µm (p = 0.005). The SEM showed compact surfaces completely covered with mineral precipitation. DMP-1 provided a successful biomimetic analogue that enhanced the mineral-organic phases of mineral-depleted dentin surfaces, offering an effective treatment modality for deep carious lesions compared with MTA and Sylc air abrasion.

Mechanisms by Which miR-584-5p Regulates Osteogenic Differentiation and Inflammation in Human Dental Pulp Stem Cells.

Ferric Carboxymaltose Increases Fracture Risk in Patients and Reduces Bone Formation in Mice with Iron Deficiency Anemia.

Variants in the dentin sialophosphoprotein (DSPP) gene are associated with dentin dysplasia type II (DD-II; OMIM # 125420) and dentinogenesis imperfecta (DI) types II (OMIM # 125490) and III (OMIM # 125500). DSPP encodes a precursor protein cleaved into three dentin matrix proteins: dentin sialoprotein (DSP), dentin phosphoprotein/phosphophoryn (DPP) and dentin glycoprotein (DGP). Exon 5 contains over 200 tandem 9-base pair repeats (DSS domain), complicating sequencing with standard methods. We studied 112 individuals (42 index cases and 70 relatives) with clinical signs of DI or DD. DNA extracted from saliva was analysed using the GenoDENT next-generation sequencing panel. For inconclusive cases, long-range PCR and Oxford Nanopore Technology (ONT) long-read sequencing were used to overcome limitations in analysing the repetitive DSPP region. Pathogenic or likely pathogenic DSPP variants were identified in 41 families, including 8 known and 14 novel variants. Most were in exon 5, causing frameshifts resulting in a -1 reading-frame shift with a hydrophobic C-terminal extension and termination at a downstream stop codon. ONT sequencing enabled detection in cases where short-read methods failed. Several variants showed familial segregation and variable expressivity. This study demonstrates the value of long-read sequencing to resolve complex DSPP regions and expands the variant spectrum. The variability in clinical presentation suggests the influence of modifier factors, warranting further genotype-phenotype studies.

Dental caries demineralises the enamel and dentine of the teeth, and as infection progresses it can lead to pulpal inflammation, infection and severe pain. To determine and compare the level of mRNA expression of Toll-like receptors ((TLR)-2, TLR-4 and TLR-9), tumour necrosis factor (TNF)-α, interleukins ((IL)-1α, IL-1β, IL-4, IL-6, IL-8, IL-17 and IL-23) as well as markers of dentinogenic (dentine matrix protein (DMP)-1, dentine sialophosphoprotein (DSPP)), regulatory (nuclear factor-kappa B (NF-κB1), mitogen activated protein kinase (MAPK1)), proliferative (mitogen activated protein kinase (MKi)) and stemness (sex determining region Y-box2 (SOX2)) between non-carious and carious dental pulp tissues. This study undertook a comprehensive analysis of inflammatory markers including TLR-2, TLR-4, TLR-9, TNF-α, IL-1α, IL-1β, IL-4, IL-6, IL-8, IL-17 and IL-23, as well as markers of dentinogenic DMP-1, DSPP, NF-κB1, MAPK1, proliferative MKi and stemness SOX2 processes in healthy and carious pulp tissues using quantitative real-time reverse-transcription polymerase chain reaction. We found higher levels of TLR-2, TLR-4, IL-6, IL-8, IL-17A, IL-23A, along with NF-κB1 and MKi67 in the carious pulps (p < 0.05). The concurrent upregulation of IL-17A and IL-23A may suggest the activation of the IL-23/IL-17 signalling axis in the carious pulps, a point underreported in the literature. These findings highlight the crucial role of the immune system in pulpal inflammation and potential implications in developing targeted molecular treatments, supporting the need for further translational research.

Complex tooth injuries, including dental caries, require the reestablishment of tissue architecture and functionality through the regeneration of cellular populations that enable tertiary dentin formation and the reestablishment of vascular and neural components. Among these cellular populations, the odontoblasts play a critical role, as they are responsible for dentin formation and maintenance, and can differentiate from stem cells of the dental pulp and apical papilla. We previously demonstrated that Histatin-1, a salivary peptide with wound-healing properties, enhances the mineralizing activity of primary mesenchymal cells from immature permanent teeth. Here, we demonstrate that Histatin-1 upregulates odontoblastic differentiation markers, including dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1), together with odontoblast-like cellular features. Immunohistochemistry and tissue immunofluorescence revealed increased DSPP and DMP1 expression in Histatin-1-treated apical papilla explants, and to a lesser extent, dental pulp explants. These findings were corroborated in primary cultures of these tissues, which also showed upregulation of DSPP and β-catenin. Histatin-1 further promoted primary ciliogenesis and Golgi-polarization. Moreover, Histatin-1 stimulated in vitro mineralization and cell migration in a VEGFR2-dependent manner, as confirmed by pharmacological inhibition and a VEGFR2-binding-deficient mutant of Histatin-1. Collectively, these results suggest that Histatin-1 drives odontoblastic differentiation, opening new avenues for dental regenerative medicine.

Osteopontin deficiency disturbs dentin bridge formation after direct pulp capping with mineral trioxide aggregate.

Improving phosphorus (P) utilization in broilers is crucial for reducing feed costs and environmental pollution. Bone mineralization trait is strongly associated with P utilization in poultry and is thus often used as an alternative trait for evaluating P utilization. Dentin matrix protein 1 (DMP1), an essential matrix protein for bone mineralization and P deposition, has been shown to be actively involved in P utilization in broilers, but the underlying mechanisms remain unclear. The current study aimed to investigate the possible mechanisms whereby DMP1 regulates P utilization of poultry by using gene silencing and overexpression technologies, combined with an in vitro model of primary broiler osteoblasts. The results showed that DMP1 overexpression augmented the P utilization of broiler osteoblasts, characterized by significant increases (p < 0.001) in P utilization rate, mineralization formation, alkaline phosphatase activity, and bone gla protein content. Meanwhile, DMP1 overexpression effectively (p < 0.05) activated the focal adhesion kinase (FAK) signaling, along with obvious (p < 0.01) decreases in fibroblast growth factor 23 (FGF23) expression and production. In contrast, DMP1 silencing reversed (p < 0.05) the above effects. Consistently, FAK activation promoted (p < 0.05) P utilization accompanied by remarkable (p < 0.05) decreases in FGF23 expression and production. Furthermore, gain- and loss-of-function assays demonstrated that a high level of FGF23 contributed to impaired P utilization, while a low level was beneficial. Interestingly, blocking FAK signaling not only recovered (p < 0.05) the FGF23 expression and production in DMP1 overexpressed cells but also obviously (p < 0.05) weakened their P utilization. These findings indicate that DMP1 inhibits FGF23 expression by activating FAK, thereby contributing to P utilization in broiler osteoblasts. They reveal a novel DMP1-FAK-FGF23 regulatory axis in broiler osteoblasts and provide a potential target for improving P efficiency in poultry.

Osteocytes function as central regulators of skeletal health by acting as mechanosensors that control bone remodelling mediated by osteoblasts and osteoclasts. Disrupted osteocyte function, often driven by oxidative stress and linked to ageing and osteoporosis, contributes to pathological bone remodelling. Tocotrienols (TTs), a family of vitamin E, are intensively investigated for their bone-protective effects, with mechanisms that involve reducing intracellular reactive oxygen species, enhancing antioxidant defences, and modulating signalling pathways of bone remodelling. Preliminary studies suggest that TTs exert protective and anabolic effects by influencing osteocytes, including shielding them from oxidative damage. In vivo models using ovariectomised or metabolic syndrome rats demonstrated that TT supplementation modulated key osteocyte-secreted factors, including sclerostin, dentin matrix protein 1, Dickkopf-related protein 1, fibroblast growth factor 23, and receptor activator of nuclear factor κB ligand. However, the current evidence is limited by the use of models that may not fully represent degenerative osteoporosis, restricted dose-dependent studies, and the challenge of real-time in vivo monitoring. This perspective summarises the reported effects of TTs on osteocytes' function and emphasises the critical need for future research to employ more representative animal models, advanced imaging techniques, and complex 3D co-culture or bone explant systems to accurately define the mechanism of action of TTs and their resulting functional outcomes on overall bone quality.

Background: Aging is associated with progressive structural and functional changes in dentin, reducing its mechanical integrity and increasing vulnerability to damage. Among the most important regulators of dentin physiology are extracellular matrix proteins from the SIBLING family, including Dentin Matrix Protein 1 (DMP-1), Dentin Sialophosphoprotein (DSPP), and Osteopontin (OPN). These proteins are essential for dentin mineralization, collagen organization, and tissue remodeling. Despite their critical role, knowledge about their age-related distribution and correlation with dentin structure and morphology remains limited. Aim: To examine age-dependent changes in the expression of SIBLING proteins (DMP-1, DSPP, OPN) in human dentin and to evaluate their relationship with collagen structure and ultramorphology using polarized light microscopy (PLM), immunohistochemistry (IHC), and scanning electron microscopy (SEM). Materials and Methods: Ninety extracted human teeth were categorized into young (≤17 years), mature (18–50 years), and old (>51 years) groups. IHC was applied to detect protein distribution, PLM to assess collagen organization, and SEM to evaluate dentinal morphology. Results and Conclusions: Aging was associated with increased expression of DMP-1 and OPN and a reduction in DSPP, which is particularly evident in peritubular dentin. Older samples showed collagen disorganization, reduced birefringence, and extensive intratubular mineralization. These findings suggest that age-related alterations in SIBLING proteins contribute to structural changes in dentin, providing new insights relevant to dental care in elderly patients.

Decellularized tissue scaffolds mimic the native pulp-dentin microenvironment and support the odontogenic development of stem cells. This study investigated the dentinogenic effect of Wharton's Jelly Mesenchymal Stem Cells (WJMSCs) in decellularized human dental pulp (DHDP) with bone morphogenic protein-7 (BMP-7) at three concentrations: 0 ng/mL (control), 25 ng/mL, and 50 ng/mL. The effects of BMP-7 were evaluated by histological examination, WJMSC viability using AlamarBlue, dentinogenic gene expression by qPCR, and dentinogenic protein expression by ELISA. By day 21, all three groups exhibited cell distribution along the pore surfaces of DHDP, followed by the presence of a collagen matrix in the tissue. WJMSC viability treated with 25 ng/mL and 50 ng/mL showed a statistically significant increase on days 7, 14, and 21 compared to the control group (p < 0.05). Gene expression analysis of dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP-1), and odontogenic marker (Runx2) revealed 25 ng/mL BMP-7 resulted in significantly higher expression levels for DMP-1 and Runx2 on day 21 compared to control and 50 ng/mL BMP-7 group (p < 0.05). DSPP and DMP-1 protein expressions also showed trends similar to those of gene expressions. BMP-7 (25 ng/mL) can maintain cell viability and promote dentinogenic effects of WJMSC in the DHDP scaffold.

Photopolymerizable fish collagen peptide-GelMa composite hydrogels for dental pulp regeneration.

Stem cells derived from the apical papilla of teeth (SCAPs) have the potential to proliferate and differentiate into osteogenic and odontoblastic lineages. However, limited information is available on the effect of osteogenic environments containing dopamine on these cells. This experimental laboratory study used SCAPs extracted from the pulp of third molar teeth of individuals aged 18years. SCAPs were treated with dopamine at doses of 1, 10, 100, 1000, and 2000μM for different treatment periods (24, 48, and 72h). No treatment was performed for the control group. Cell viability and proliferation were assessed using the MTT (Methyl Thiazolyl Tetrazolium) assay, osteogenic differentiation was evaluated using Alizarin Red S (ARS) staining, and gene expression of bone-related markers was analyzed using reverse transcription polymerase chain reaction (RT-PCR). Data analysis was performed using SPSS software (version 18), and comparisons between groups were made using t-tests and ANOVA. After 24, 48, and 72h, no significant difference was observed between the dopamine-treated and control groups in terms of SCAP viability/proliferation at low concentrations. However, a significant reduction in cell viability was observed at high dopamine concentrations (2000μM). ARS staining indicated that dopamine treatment enhanced osteogenic differentiation of SCAPs. Gene expression analysis following dopamine treatment showed an increase in osteogenic/odontogenic markers in SCAPs, including alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), and bone sialoprotein (BSP) (P < 0.05). Dopamine influenced SCAP viability in a dose- and time-dependent manner, leading to osteogenic/odontogenic differentiation and increased expression of differentiation-related genes. These findings suggest that dopamine can act as a potential bioactive molecule in regenerative endodontic therapies.

This paper explores the development of three-dimensional porous scaffolds composed of gelatin, calcium hydroxide, dentin matrix proteins, and propolis (signified as G, Ca, DP, and P, respectively). First, the scaffolds were synthesized using freeze-drying technique, then their physicochemical properties, mechanical characteristics, cytocompatibility with dental pulp stem cells, and cell adhesion, differentiation and mineralization were evaluated. The results showed that the scaffolds were porous with asperity. The surface roughness analysis identified higher roughness value for the G-Ca-P and G-Ca-DP-P scaffolds (mean Rq of 38.16 and 21.69 nm, respectively). All the scaffolds exhibited hydrophilic behavior (contact angles < 90°). However, those having propolis, calcium hydroxide and protein showed higher wettability. Furthermore, all scaffolds degraded over time with a synergistic effect between calcium hydroxide and dentin matrix proteins in making a more robust scaffold and enhancing the alkalinizing effect. The G and G-P scaffolds had elastic-plastic behavior, while G-Ca-P and G-Ca-DP-P scaffolds showed brittleness. Moreover, all scaffolds revealed no cytotoxicity (cell viability >80%). The alkaline phosphatase (ALP) concentration and matrix deposition, as measures of osteogenic differentiation, were highest for G-Ca-DP-P scaffolds, followed by G-Ca-P. Besides, the cells attached to the scaffold surfaces with a flattened morphology, and the highest cell number adhered on G-Ca-DP-P and G-Ca-P scaffolds, respectively.Altogether, the G-Ca-DP-P scaffolds acted as hydrophilic, rough, and biodegradable frameworks supporting cell viability, adhesion, differentiation and mineralization.

The dental pulp not only serves as the tooth's nutritional core but also creates a finely tuned microenvironment that is enriched with blood vessels, nerves, extracellular matrix components, and signaling molecules, all of which guide the fate of resident dental pulp stem cells (DPSCs). Trauma and microbial invasion disrupt this niche, leading to pulpitis and necrosis. Although conventional root canal treatment preserves the tooth's structure by removing infected pulp, it can increase tooth brittleness and impede root development in immature permanent teeth. Harnessing DPSCs' multipotency for pulp regeneration promises to restore the natural pulp-dentin complex in situ. Importantly, DPSCs encounter an inflammatory microenvironment composed of pathogen-associated molecular patterns, a spectrum of pro- and anti-inflammatory cytokines, diverse immune cell phenotypes, and altered matrix signals. While earlier work examined the isolated effects of mediators such as lipopolysaccharide, tumor necrosis factor-alpha, or macrophage-derived exosomes on odontogenic differentiation, this review focuses on how these mediators collectively interact in both synergistic and antagonistic ways within the inflammatory niche. We systematically delineate how these collective stimuli converge on wingless/integrated/beta-catenin, mitogen-activated protein kinase, nuclear factor kappa-B (NF-κB), and bone morphogenetic protein/Sma and Mad related protein pathways to modulate key odontogenic markers (runt-related transcription factor 2, dentin sialophosphoprotein, dentin matrix protein 1, alkaline phosphatase) and mineralization outcomes. By applying a microenvironment-centric lens, we reveal novel targets and strategies to recalibrate inflammation, steer DPSCs toward reparative odontogenesis, and ultimately enhance the efficacy of regenerative endodontic therapies.

To investigate the histopathology of diabetic periodontal tissues, we examined periodontal disease in Torii-Leprfa (SDT fa/fa) rats, mimicking type 2 diabetes. Forty 30-week-old male SDT fa/fa rats and age-matched Sprague-Dawley (SD) rats were fixed, and the mandibular first molars and their periodontal tissues were histochemically examined. SDT fa/fa rats exhibited epithelial downgrowth in the previous region of interradicular/interalveolar septa and periodontal spaces. Many osteoclasts and osteoblasts were present on the alveolar bone, indicating high bone turnover. Masses of bacteria were predominantly observed in the necrotic dental pulps and the cementum, where most cementocytic lacunae were empty. The cementum of the SDT fa/fa rats showed a distinct immunolocalization of osteocalcin, osteopontin, and dentin matrix protein-1 compared with the SD rats, but did not show advanced glycation end products, which were instead detected in some blood vessels and interstitial fibrous tissues associated with the down-growing epithelium. SDT fa/fa rats exhibited necrosis of dental pulp and severe periodontitis featuring markedly diminished alveolar bone because of stimulated bone resorption and formation. In addition, dead cementum may serve as bacterial nests, leading to changes in the localization of cementum matrix proteins.

Vital pulp therapy with calcium hydroxide or mineral trioxide aggregate (MTA) rapidly induces dystrophic calcification and promotes the accumulation of two members of small integrin-binding ligand N-linked glycoproteins: osteopontin (OPN) and dentin matrix protein-1 (DMP1). However, the precise relationship between these initial events and their roles in reparative dentinogenesis remain unclear. This study aimed to clarify the relationship between dystrophic calcification, OPN and DMP1 accumulation, and reparative dentin formation. Pulpotomy was performed on rat molars using MTA or zirconium oxide (ZrO2). ZrO2 was used as a control to assess pulp healing in the absence of dystrophic calcification. Pulpal responses were evaluated from 3 h to 7 days postoperatively via elemental mapping, micro-Raman spectroscopy, and histological staining. In the MTA-treated group, a calcium-rich dystrophic calcification zone containing calcite and hydroxyapatite was observed at 3 h after treatment; OPN and DMP1 accumulated under the dystrophic calcification zone by day 3; reparative dentin formed below the region of OPN and DMP1 accumulation by day 7. In contrast, these reactions did not occur in the ZrO2-treated group. These results suggest that dystrophic calcification serves as a key trigger for OPN and DMP1 accumulation and plays a pivotal role in reparative dentinogenesis.

MicroRNA (miR)-200c enhances osteogenesis, modulates inflammation, and participates in dentin development. This study was to investigate the beneficial potential of miR-200c in vital pulp therapy (VPT) by mitigating pulpitis and promoting dentin regeneration. We explored the miR-200c variations in inflamed pulp tissues from patients with symptomatic irreversible pulpitis and primary human dental pulp-derived cells (DPCs) challenged with P.g. lipopolysaccharide (Pg-LPS). We further assessed the functions of overexpression of miR-200c on odontogenic differentiation, pulpal inflammation, and dentin regeneration in vitro and in vivo. Our findings revealed a noteworthy downregulation of miR-200c expression in inflamed pulp tissues and primary human DPCs. Through the overexpression of miR-200c via transfecting plasmid DNA (pDNA), we observed a substantial downregulation of proinflammatory cytokines interleukin (IL)-6 and IL-8 in human DPCs. Furthermore, this overexpression significantly enhanced the transcript and protein levels of odontogenic differentiation markers, including Runt-related transcription factor (Runx)2, osteocalcin (OCN), dentin matrix protein (DMP)1, and dentin sialophosphoprotein (DSPP). In a rat model of pulpitis induced by Pg-LPS, we demonstrated notable benefits by local application of pDNA encoding miR-200c delivered by CaCO3-based nanoparticles to reduce pulpal inflammation and promote dentin formation. These results underscore the significant impact of locally applied miR-200c in modulating pulpal inflammation and facilitating dentin repair, showcasing its ability to improve VPT outcomes.