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Although silicon is a widespread constituent in dental materials, its possible influence on the formation and repair of teeth remains largely unexplored. Here, we studied the effect of two silicic acid-releasing nanomaterials, silica and bioglass, on a living model of pulp consisting of dental pulp stem cells seeded in dense type I collagen hydrogels. Silica nanoparticles and released silicic acid had little effect on cell viability and mineralization efficiency but impacted metabolic activity, delayed matrix remodeling, and led to heterogeneous cell distribution. Bioglass improved cell metabolic activity and led to a homogeneous dispersion of cells and mineral deposits within the hydrogel. These results suggest that the presence of calcium ions in bioglass is not only favorable to cell proliferation but can also counterbalance the negative effects of silicon. Both chemical and biological processes should therefore be considered when investigating the effects of silicon-containing materials on dental tissues.

The aim of the present study was to investigate the effects of the biological properties of hDPSCs exposed to Bio MTA+ & ProRoot MTA pulp capping materials on the proliferation and odontogenic differentiation of hDPSCs. Human dental pulp stem cells (hDPSCs) were isolated from impacted third molars. Extracts of Bio MTA + and ProRoot MTA were prepared at a 1:1 ratio. The effects of the extracts on hDPSCs cytotoxicity and proliferation were assessed via a CCK-8 assay. Annexin V expression was investigated to assess the effects of both materials on the induction of apoptosis. The effects of ProRoot MTA and Bio MTA + extraction media on the stemness properties of hDPSCs were assessed via real-time quantitative PCR, and the expression of odontogenic markers (RUNX2, DMP1 & DSSP) was analyzed via RT‒PCR Alizarin Red staining. Cells exposed to Bio MTA + had the greatest degree of proliferation. The results of Annexin V staining indicated that Bio MTA + caused the least amount of apoptosis. RUNX2, DMP1 and DSSP were highly expressed by Bio MTA + and indicated successful odontogenic differentiation. Compared with ProRoot MTA, Bio MTA + exhibited an exceptional level of cytocompatibility, as well as advantageous bioactivities, including the preservation of stemness and an increase in the proliferation capacity of hDPSCs. In addition, it demonstrated favorable bioactive properties by stimulating odontogenic differentiation. Bio MTA + offers significant advantages in terms of biocompatibility, bioactivity, and regenerative potential, making it an excellent choice for procedures aimed at preserving or regenerating dental pulp tissue. However, additional research is required to address the lack of in vivo validation, as replicating physiological conditions is crucial for accurately assessing clinical outcomes and comparing them with results obtained from in vitro experiments., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Ethics declarations: The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. All methods were performed in accordance with the guidelines and regulations of the Declaration of Helsinki. Informed consent was obtained from all participants. The experimental protocol was reviewed by the Ethical Committee of the Faculty of Dentistry Ain Shams University and was granted approval (FDASU-Rec ID 052329)., (© 2025. The Author(s).)

Objectives: This study investigated the in vitro effects of cannabidiol (CBD) on dental pulp cells and macrophages under pro-inflammatory conditions., Materials and Methods: Mouse dental pulp cells (OD-21) were pre-stimulated with tumor necrosis factor alpha (10 ng/mL) or left untreated, then exposed to CBD at concentrations of 0.01 µM, 0.1 µM, 1 µM, and 10 µM for 24 h and 7 days. Cell viability was assessed using the MTT assay, while gene expression related to mineralization-Dentin Sialophosphoprotein (Dspp), Dentin Matrix Protein 1 (Dmp1), Runt-related transcription factor 2 (Runx2), TNF-α (Tnf), and prostaglandin-endoperoxide synthase 2 (Ptgs2) were analyzed via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Mineralization nodule formation was evaluated using alizarin red staining. Macrophages (RAW 264.7) were stimulated with lipopolysaccharide (LPS) for 2 h before exposure to the same CBD concentrations. Data analysis included the Shapiro-Wilk normality test and comparisons using ANOVA and Tukey's post-hoc test (α = 0.05)., Results: The findings indicated that CBD did not significantly affect OD-21 cell viability, except for the 10 µM concentration after 7 days (p < 0.05). CBD treatment promoted mineralization, with significant differences observed among groups (p < 0.05). Notably, Ptgs2 expression varied between time points, while Runx2 expression was significantly reduced at 24 h (p < 0.05). In macrophages, Ptgs2 and Tnf levels were downregulated by all tested CBD concentrations (p < 0.05)., Conclusion: These results indicate that cannabidiol positively influence the biomineralization process and modulate inflammatory mediator expression., Clinical Relevance: Our research indicates that cannabidiol presents biomineralization potential within inflammatory contexts, implying its potential as a promisor bioactive substance for regenerating oral tissues by interacting with cells and tissues to induce specific responses., Competing Interests: Declaration of competing interest The authors declare that this study was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction: To prepare a gelatin methacryloyl (GelMA) hydrogel scaffold embedded with adrenomedullin (ADM) and investigate its impact and underlying mechanisms in endogenous pulp regeneration., Methods: ADM was evenly distributed within the GelMA hydrogel through a simple and conventional physical mixing technique. The scaffold underwent characterization via scanning electron microscopy, alongside assessments of swelling, degradation, and release properties. Biocompatibility was evaluated using cytoskeletal and live-dead staining techniques. The hydrogel's influence on dental pulp stem cells' proliferation, migration, and differentiation was assessed with CCK-8 assays, Transwell assays, and alizarin red and alkaline phosphatase staining. Transcriptomics provided insights into potential mechanisms. The angiogenic effects on umbilical vein endothelial cells were examined using scratch and tube formation assays. In vivo, the composite hydrogel's regenerative capacity was tested in a rat model of pulp regeneration. Statistical analysis involved Student's t-test and one-way analysis of variance, with significance set at P < .05., Results: The ADM-loaded GelMA (GelMA@ADM) hydrogel displayed a porous architecture under electron microscopy conducive to cell adhesion and demonstrated excellent biocompatibility. In vitro experiments showed that GelMA@ADM significantly boosted dental pulp stem cells' migration, proliferation, and differentiation, and enhanced the angiogenic activity of umbilical vein endothelial cells after one week of treatment. Corresponding in vivo experiments revealed that GelMA@ADM facilitated the formation of new vascularized pulp tissue after 2 weeks of treatment., Conclusions: The GelMA@ADM hydrogel effectively promotes dental pulp stem cells' proliferation and differentiation, augments vascularization by umbilical vein endothelial cells, and fosters the creation of new vascularized pulp tissue. These findings underscore the potential of GelMA@ADM hydrogel for endogenous pulp regeneration., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: Mineral trioxide aggregate (MTA) is a calcium silicate-based cement that has changed conventional dental therapeutic approaches. This study aimed to evaluate physical, chemical and biological properties of novel AGM MTA, in comparison with MTA Angelus., Methods: The samples were prepared according to the manufacturer's instructions. The initial and final setting times were measured via a Gillmore needle following the ISO 6876:2012 standard. The radiopacity of the materials was evaluated against an aluminium step wedge on the basis of the ISO 6876 and 13,116 standards. The pH changes were measured at intervals of 3, 6, 24, 72, 96 and 144 h postimmersion in Hank's solution and calcium ion release was analysed after 168 h of immersion via inductively coupled plasma optical emission spectroscopy (ICP‒OES). Moreover, the cytotoxicity was assessed through the MTT assay on human dental pulp stem cells (hDPSCs) after 24 and 72 h of exposure to the set/fresh status of various dilutions of MTA extracts, following the ISO 10993-12 standard., Results: No significant difference was found between the initial setting times of the two materials (Angelus: 11.0 ± 1.0 min, AGM: 10.3 ± 1.5 min); however, MTA Angelus demonstrated a significantly shorter final setting time. Both materials met the minimum radiopacity requirements according to the ISO 6876 standard, with MTA Angelus exhibiting greater radiopacity than AGM MTA. Both materials created an alkaline environment without presenting any differences in each time point and AGM MTA released significantly greater amounts of calcium ions. In the cytotoxicity assessment, while the diluted extracts of both materials did not elicit any cytotoxic effects, the nondiluted samples, after 72 h of exposure, as well as the 30-min set AGM MTA after 24 h of exposure, were shown to be cytotoxic., Conclusions: In conclusion, MTA Angelus presented a faster setting time and lower cytotoxicity, while AGM MTA demonstrated greater calcium ion release. However, both materials presented clinically acceptable properties and AGM MTA could be a potential alternative to MTA Angelus. However, further clinical studies are required to confirm its application., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Diabetes mellitus (DM) induced mitochondrial oxidative stress (OS) can lead to severe injury of dental pulp. The cerium oxide nanoparticles (CNP) have been proven to have excellent antioxidative activity. However, whether CNP can relieve dental pulp damage caused by DM and the underlying mechanisms remain unclear. In this study, we modified ceria with hyaluronic acid to prepare nanoceria with good biocompatibility, water solubility, and stability, namely, HACNP (hyaluronic acid cerium oxide nanoparticles). We demonstrated the protective effect of HACNP on diabetic OS-induced mitochondrial apoptosis in dental pulp-like cells. As far as the mechanism of action was concerned, glucose oxidase (GO) treatment promoted the activation of phosphoglycerate mutase family 5 (PGAM5) leading to mitochondrial abnormalities and apoptosis in an odontoblast-like cell line (mDPC6T). Knockdown or overexpression of PGAM5 further validate these results. Meanwhile, HACNP remitted GO-related toxicity via down-regulating PGAM5 expression, whereas overexpression of PGAM5 abolished the beneficial effect of HACNP. Furthermore, in the constructed animal research model of diabetic pulp injury, we also confirmed that HACNP alleviated apoptosis and mitochondrial injury of dental pulp and decreased the expression level of PGAM5 in diabetic pulp tissue. In conclusion, these results revealed that HACNP played a protective role on diabetes-associated dental pulp injury through targeting the PGAM5-mediated mitochondrial pathway, providing an idea and method for the prevention or treatment of diabetes-induced dental pulp damage.

Objective: This study aimed to investigate and compare the histological response of rabbit dental pulp after direct pulp capping with 3 different materials: mineral trioxide aggregate (MTA), nanoparticles of fluorapatite (Nano-FA), and nanoparticles of hydroxyapatite (Nano-HA) after 4 and 6-week time intervals., Material and Methods: A total of 72 upper and lower incisor teeth from 18 rabbits were randomly categorized into 3 groups)24 incisors from six rabbits each. MTA Group: teeth were capped with MTA. Nano-FA Group: teeth were capped with fluorapatite nanoparticles. Nano-HA Group: teeth were capped with hydroxyapatite nanoparticles. Blood samples were collected to examine some antioxidant enzymes nitric oxide (NO), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione (GSH), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). After that, three rabbits from each group were euthanized after 4 and 6 weeks, respectively. Pulp tissues of all teeth in all groups were histologically observed., Results: The obtained results showed that both Nano-HA induced the formation of thick dentin bridges with irregular dentin patterns at 6 weeks, while MTA and Nano-FA induced no dentin bridge with no tubular dentin pattern. Blood examination at the two intervals revealed no significant increase or decrease in the values of NO, SOD, CAT, GPx, GSH, and TNF-α. However, there was a significant increase in p-values of IL-6 in the Nano-FA treated group compared to both MTA and Nano-HA treated groups at the two intervals. Regarding the inflammatory reaction of the dental pulp, the MTA and Nano-HA groups displayed moderate inflammation, followed by Nano-FA, which showed the highest prevalence of nonpathological inflammation. Histological results were consistent with the blood examination. After 4 weeks, the Nano-FA and Nano-HA groups showed pulp fibrosis at the operating site, but the MTA showed only granulation tissues. Plus, dilated blood vessels appeared in the Nano-FA group. After 6 weeks, MTA and Nano-FA groups showed pulp fibrosis at the operating site with the persistence of dilated blood vessels with Nano-FA. The nano-HA group showed dentin bridge formation at the operating site., Conclusion: MTA and Nano-HA could be considered favorable materials for direct pulp capping, while Nano-FA produces nonpathological inflammatory cell reactions. Moreover, the Nano-HA was the best in dentin bridge formation. Although nano-FA increased the operating site closure, it was noticed that it significantly increased IL-6 compared to MTA at the two intervals and significantly increased IL-6 compared to Nano-HA at 6 weeks, which may be manifested as some nonpathological inflammations in the Nano-FA group compared to the other groups, but it was deemed acceptable to direct pulp capping procedures., Competing Interests: Declarations. Ethics approval and consent to participate: The present investigation was conducted in adherence to the protocols established by the Research Ethics Committee of the Faculty of pharmacy, Pharos University, under ethical code 239. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Objectives: As dental pulp therapy evolves towards regenerative approaches, biomolecules such as icariin, derived from Epimedium flowers, are being evaluated for their therapeutic potential. This study investigates icariin's effectiveness in promoting odontogenic differentiation in human dental pulp stem cells (hDPSCs) in vitro and as a pulp-capping agent in vivo., Design: The study explored the effects of icariin on hDPSCs at concentrations of 10, 20, and 40 µM. Cell viability and migration assays were conducted to evaluate cytotoxicity and chemotaxis. Odontogenic differentiation was assessed using alkaline phosphatase staining and alizarin red S (ARS) staining, complemented by real-time PCR and Western blot analyses of key markers such as RUNX family transcription factor 2 (RUNX2), collagen type I alpha 1 chain (COL1A1), alkaline phosphatase (ALPL), and dentin sialophosphoprotein (DSPP). Additionally, the in vivo effects of icariin were tested in a rat maxillary molar model, where icariin-treated collagen sponges were used for direct pulp capping to evaluate its potential to induce reparative dentin formation., Results: Icariin showed no cytotoxic effects on hDPSCs at any tested concentration, enhanced migratory activity in a dose-dependent manner, and significantly increased alkaline phosphatase activity and calcium deposition. Gene and protein expression analyses revealed a dose-dependent increase in odontogenic differentiation markers in icariin-treated hDPSCs. In vivo, icariin effectively promoted reparative dentin formation in exposed rat pulp., Conclusions: Icariin enhances odontogenic differentiation of hDPSCs and has promising potential as a pulp-capping agent for vital pulp therapy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Pulpitis is inflammation of the dental pulp, often caused by bacterial infection from untreated cavities, leading to pain. The main challenge in treatment is eliminating infection while preserving tooth vitality. This study aims to address this challenge by developing a hydrogel for convenient insertion into the root canal system, securely attaching to dentin walls. An injectable hydrogel system is developed by chemically cross-linking natural polysaccharide pectin with gelatin (GPG) through reversible Schiff base reaction. The GPG system was then used to encapsulate and release drugs, such as ciprofloxacin (CIP) for infection prevention and deferoxamine (DFO) for promoting blood vessel proliferation and reducing inflammatory reactions. The GPGs absorbed significant amounts of CIP and DFO, enabling sustained release over a nearly ten-day period. When subcutaneously implanted, the GPGs formed stable gel depots, with only 50 % of the gels degrading after 3 weeks, indicating a sustained biodegradation pattern. Additionally, the GPG system demonstrated excellent antibacterial activity against both gram-negative and gram-positive bacteria. Results from in vitro scratch healing tests and in ovo chorioallantoic membrane chick model tests showed promising biocompatibility and promotion of vascular proliferation by the GPG. This study heralds a novel frontier in endodontic therapeutics, poised to potentially enable dental pulp regeneration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objectives: to assess the cytotoxicity of the following functional monomers used in dental adhesives: 10-Methacryloyloxydecyl dihydrogen phosphate (10-MDP) and glycerol phosphate dimethacrylate (GPDM), and their effect on cytokine release from human dental pulp stem cells (hDPSCs)., Methods: The hDPSCs cells were isolated from the dental pulp of extracted human third molars. The functional monomers, 10-MDP and GPDM, were diluted in dimethyl sulfoxide (DMSO) at concentrations ranging from 1 to 4 mM. Cells not exposed to the compounds served as controls. The hDPSCs were seeded into 96-well plates and incubated for 48 h. Subsequently, the cells were exposed to 10-MDP and GPDM for 24 h. Then, the culture medium was removed, the mitochondrial metabolism was evaluated using the MTT assay, while cell death analyzed by flow cytometry. Cytokine release (IL-1β, IL-6, IL-8, IL-10, TNF-α) was analyzed by the MAGPIX. The data were analyzed using one-way ANOVA and Tukey's test., Results: 10-MDP demonstrated significant toxicity to hDPSCs, reaching the IC50 at 3 mM. However, its impact on cytokine release was minimal, resulting only in IL-6 and IL-8 levels. GPDM exhibited lower toxicity, even at 4 mM, but induced an increase in IL-1β release and a reduction in IL-6, IL-8, and IL-10 levels, with no effect on TNF-α. Despite the MTT assay results indicating cytotoxicity, the cell death was low for both functional monomers., Significance: 10-MDP exhibited significant toxicity to hDPSCs, unlike GPDM, however, both monomers resulted in minimal cell death. 10-MDP had a minor impact on cytokine release, whereas GPDM demonstrated a potential to trigger an inflammatory reaction, particularly in the short term., Competing Interests: Declaration of Competing Interest The authors declare no potential conflicts of interest concerning the authorship or publication of this work. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Hyaluronic acid (HA), known for diverse properties, was investigated for its potential in dental pulp therapy. This study investigated the potential of HA in dental pulp therapy by examining the physical properties and effects of zinc oxide eugenol (ZOE) pulpotomy materials containing varying HA concentrations on rat molar teeth. In vitro tests assessed compressive strength and hardness of ZOE materials blended with HA (0.5%, 1%, 3%) and HA gels (0.54%, 0.8%). 120 samples, encompassing the control group, underwent compressive strength testing, while 60 samples were designated for hardness assessment. In vivo experiments on rat molars studied histological effects of HA-containing ZOE on dental pulp over 1 week and 1 month. Gels with HA concentrations of 0.5%, 1%, and 0.54% were used in pulpotomy on 22 rats. Each rat underwent the procedure on four teeth, with one tooth serving as a control, totaling 88 teeth subjected to the intervention. In the analyses, SPSS 22.0 was used and the significance level was set at P = 0.05. Findings showed that HA at 0.5% maintained compressive strength, but higher concentrations decreased mechanical properties significantly (P = 0.001). Histological assessments indicated better outcomes with lower HA concentrations in terms of odontoblast layer continuity (P = 0.005 at 1 month) and pulp vitality (P = 0.001 at 1 week and P = 0.018 at 1 month). The study suggests HA holds promise for pulpotomy and regenerative endodontic treatments, but further research is needed to understand long-term clinical implications., Competing Interests: Declarations. Conflict of interest: The authors have no conflicts of interest relevant to this article. Ethical approval: This study protocol was submitted to and approved by the local ethical committee (protocol code is 30.2021mar, date: 10.03.2021). All procedures involving animals were in compliance with the Guide for the Care and Use of Laboratory Animal, 8th Edition by National Research Council (US) Committee., (© 2024. The Author(s).)

Aging impairs bone marrow mesenchymal stem cell (BMSC) functions as well as associated angiogenesis which is critical for bone regeneration and repair. Hence, repairing bone defects in elderly patients poses a formidable challenge in regenerative medicine. Here, the engineered dental pulp stem cell-derived exosomes loaded with the natural derivative of adenosine Cordycepin (CY@D-exos) are fabricated by means of the intermittent ultrasonic shock, which dually rejuvenates both senescent BMSCs and endothelial cells and significantly improve bone regeneration and repair in aged animals. CY@D-exos can efficiently overcome the senescence of aged BMSCs and enhance their osteogenic differentiation by activating NRF2 signaling and maintaining heterochromatin stability. Importantly, CY@D-exos also potently overcomes the senescence of vascular endothelial cells and promotes angiogenesis. In vivo injectable gelatin methacryloyl (GelMA) hydrogels with sustained release of CY@D-exos can accelerate bone injury repair and promote new blood vessel formation in aged animals. Taken together, these results thus demonstrate that cordycepin-loaded dental pulp stem cell-derived exosomes display considerable potential to be developed as a next-generation therapeutic agent for promoting aged bone regeneration and repair., (© 2024 Wiley‐VCH GmbH.)

Forensic age estimation is performed by assessing pulp chamber constrictions due to physiological age-related changes in dental radiographs; however, the estimated ages occasionally deviate from the actual ages. In particular, long-term steroid users tend to demonstrate pulp chamber constrictions in all teeth. Because this is uncommon among younger age groups, caution should be exercised when evaluating pulp chamber constriction. This study investigated the estimated ages of eight PSL users by applying the ratio of pulp area to total tooth area from canine radiographs. Patients in their 30-40s were examined at a dental outpatient clinic for the prevention or treatment of adverse events associated with the use of bisphosphonates to prevent steroidal osteoporosis, and radiographs were obtained. The pulp and tooth areas were measured and the estimated age was determined using regression formulas calculated from the canine teeth of the general study subjects of Japanese. The mean absolute error between the estimated ages and the chronological ages of the patients was 19.24 years for the upper canines and 17.69 years for the lower. Moreover, the root mean square error was 23.18 years for the upper canines and 20.00 years for the lower. The estimated ages were far from the actual ages of the steroid users. When estimating an unidentified individual's age, if the patient has pulp chamber constriction that is inconsistent with other forensic physical findings, this information may assist in predicting their medical background., Competing Interests: The authors report no conflicts of interest in this work., (Copyright© 2024 IOFOS This work is published and licensed by International Organization for Forensic Odonto-Stomatology.)

This study aimed to evaluate the histological success of pulpotomy in primary molars using white mineral trioxide aggregate (WMTA) mixed with 2.25% sodium hypochlorite (NaOCl) gel and to evaluate in vitro its physical and chemical properties. The study had a clinical stage and an in-vitro stage. The clinical study was conducted with 24 patients aged 8-10 years. It was a randomized controlled trial to perform a histologic evaluation of pulp response following pulpotomy. Primary first molars were randomly assigned in split mouth model to control group-WMTA + distilled water (DW) or experimental group - WMTA + NaOCl gel. Teeth were extracted after 7, 30 or 90 days and submitted to histological analysis. The second stage was an in-vitro study to evaluate the physical and chemical properties of the two materials tested. SEM and EDX analyses and pH level evaluations were performed after 24 h and 28 days. The histological findings revealed that while WMTA + NaOCl gel group showed better odontoblastic integrity (P < 0.05), WMTA + DW group had more favorable outcomes in dentin bridge formation and pulp calcification (P < 0.05). Pulp tissue hemorrhage and pulp fibrosis were similar between them (P > 0.05). Regarding materials' in vitro evaluation, the pH level indicated a higher initial pH for the WMTA + NaOCl gel group, which equalized after 28 days. SEM analysis initially showed a less homogeneous surface for WMTA + NaOCl gel, but it became similar after 28 days. EDX analysis indicated higher calcium and silicon percentages in the WMTA + NaOCl gel group initially, which increased in both groups after 28 days. Adding 2.25% NaOCl gel to WMTA enhanced odontoblastic integrity in both the short and medium term. In addition, it had a similar chemical composition, surface morphology, and alkalinity when compared to WMTA + DW mixture., Competing Interests: Declarations. Competing interests: The authors declare no competing interests. Informed consent: Informed consent was obtained from all subjects., (© 2024. The Author(s).)

Background: Propolis is a natural substance produced by honeybees that has various biological properties including, anti-inflammatory, antioxidant and antimicrobial properties. Although previous studies have evaluated the antimicrobial effects of propolis in dentistry, its effects on dental pulp stem cell (DPSC) viability, migration, and differentiation are yet not well understood. The objective of this study was to investigate the effects of Chinese propolis on viability/proliferation, migration, differentiation and cytokine expression in DPSCs., Methods: Commercially available DPSCs (Lonza) were treated with aqueous extract of propolis (AEP) or ethanolic extract of propolis (EEP), and viability/proliferation was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays and quantification of nuclear staining. DPSC differentiation into mineralizing cells was evaluated with Alizarin red staining and cell migration was assessed using Boyden Chamber Transwell inserts. Cytokine expression was measured by RT-qPCR. AEP and EEP at 0.03 and 0.1 mg/mL did not affect DPSC viability/proliferation for up to 7-days treatment., Results: Higher doses (0.33-33 mg/mL) induced a dose dependent decrease in DPSC viability/proliferation with a more prominent effect with EEP at 7 days. Neither AEP nor EEP induced DPSC differentiation into mineralizing cells, but both AEP and EEP (0.03-0.1 mg/ml) induced a dose dependent increase in DPSC migration. In addition, EEP prevents the upregulation of IL1b and IL6 but not IL8 and CCL2 in response to lipopolysaccharide stimulation. AEP has less potent anti-inflammatory effects and prevents only IL1b upregulation., Conclusion: This study provides new information about the biologic properties of ethanolic and aqueous extracts of propolis and shows that propolis, at doses that do not affect cell viability, induces DPSC migration and has anti-inflammatory properties. These data highlight the potential use of propolis as an alternative intra-canal medicament for regenerative endodontic procedures., Competing Interests: The authors declare that they have no competing interests., (© 2024 Park et al.)

This study aimed to evaluate the pulp response to F18 and cobalt-doped F18 bioglass (F18Co) in comparison with calcium hydroxide (CH) after pulp capping. The maxillary first molars of 48 rats were divided into F18, F18Co, CH, and control (no intervention) groups. The pulp was exposed, the materials were placed, and the teeth were capped. After 7 and 15 days, the animals were euthanized for pulp evaluation and interleukin (IL) expression determination. Statistical analysis was carried out using the SigmaPlot® program (Systat Software Inc., for Windows, version 12.0). The data obtained in the analyses were subjected to the non-parametric Kruskal-Wallis test, followed by Dunn's test. For all tests, statistical significance was set at p < 0.05. The CH group exhibited mild to moderate inflammation, whereas the bioglass groups displayed moderate to severe inflammation, indicating a notable difference between the control and bioglass groups. At 7 days, both the CH and most of the bioglass specimens showed moderate disorganization. On day 15, CH displayed mildto-moderate disorganization, whereas F18 and F18Co exhibited significantly more moderate-to-severe disorganization. There were no significant differences in IL-6 and IL-10 expressions between groups at 7 days, but a noteworthy increase in IL-1β was observed in both CH and F18. After 15 days, there was a greater expression of IL-6 and IL-1β in the bioglass groups. No significant IL-10 expression was observed. Bioglass performed less effectively than CH when in direct contact with the pulp tissue.

This systematic review aimed to compare the histological response of inflamed pulpodentinal complex to the hydraulic calcium silicate cements in experimental animal models of pulpitis. Articles that evaluated the histological response of inflamed pulp to mineral trioxide aggregate (MTA) in comparison with other restorative materials were selected and analysed in detail. The risk of bias assessment was conducted using SYRCLE's RoB tool. The GRADEpro tool was used to determine the overall quality of evidence. Out of the 2947 retrieved articles from databases, five articles fulfilled the inclusion criteria. MTA induced significantly more hard tissue formation compared to calcium hydroxide. The use of pulp-capping material containing fluocinolone acetonide and ASP/PLGA-ASP/ACP/PLLA-PLGA composite membrane was comparable. This systematic review could not demonstrate enhanced efficiency of capping materials compared to MTA. Future well-conducted animal studies are warranted for demonstrating the hard tissue formation abilities of pulp-capping materials with convenient inflammatory conditions., (© 2024 Australian Society of Endodontology Inc.)

Background: Ulcerative colitis is an inflammatory bowel disease (IBD) that involves inflammation of the colon lining and rectum. Although a definitive cure for IBD has not been identified, various therapeutic approaches have been proposed to mitigate the symptomatic presentation of this disease, primarily focusing on reducing inflammation. The aim of the present study was to evaluate the therapeutic potential of combining dental pulp stem cells (DPSCs) with sulfasalazine in an acetic acid-induced ulcerative colitis rat model and to assess the impact of this combination on the suppression of inflammatory cytokines and the regulation of oxidative stress in vivo., Methods: Ulcerative colitis was induced in rats through transrectal administration of 3% acetic acid. The therapeutic effect of combining DPSCs and sulfasalazine on UC was evaluated by measuring the colonic weight/length ratio and edema markers; performing histopathological investigations of colon tissue; performing immunohistochemical staining for NF-κB-P65 and IL-1β; and evaluating oxidative stress and antioxidant indices via ELISA. Moreover, the proinflammatory markers NF-κB-P65, TNF-α and TLR-4 were assessed in colon tissue via ELISA. Furthermore, qRT‒PCR was used to estimate the expression levels of the TLR-4, NF-κB-P65, and MYD88 genes in colon tissue., Results: The investigated macroscopic and microscopic signs of inflammation were markedly improved after the combined administration of sulfasalazine and DPSCs, where a noticeable improvement in histological structure, with an intact mucosal epithelium and mild inflammatory infiltration in the mucosa and submucosa, with slight hemorrhage. The administration of either DPSCs or sulfasalazine, either individually or in combination, significantly reduced ROS levels and significantly increased XOD activity. The immunohistochemical results demonstrated that the combined administration of DPSCs and sulfasalazine attenuated NFκB-p65 and IL-1β expression. Finally, the combined administration of DPSCs and sulfasalazine significantly downregulated MyD88, NF-κB and TLR4 gene expression., Conclusions: Cotreatment with DPSCs and sulfasalazine had synergistic effects on ulcerative colitis, and these effects were relieved., (© 2024. The Author(s).)

Objective: To evaluate the effect of a newly developed MTA-based material and two antibacterial-enhanced MTAs as pulp capping materials in immature permanent dental elements underwent full pulpotomy., Methods: The present animal study included 20 Wistar albino rats that, after full pulpotomy, were randomly divided into 4 groups receiving different MTA formulations as pulp capping materials: conventional MTA, Tricalcium aluminate (TCA)- free MTA, and MTA enhanced with metronidazole or doxycycline. Histopathological assessments were carried out at 7- and 28-days post-treatment to evaluate dentinal bridge formation, inflammatory reactions, pulp tissue necrosis and internal resorption., Results: Seven days post-treatment, all groups exhibited inflammation and pulp necrosis, that were minimal in Groups III and IV than Group I. Group II showed a statistically significant difference only in terms of pulp necrosis (p<0.001). At 28-days all Groups showed slight inflammation and pulp necrosis, mainly in Groups I. Dentinal bridge formation was appreciated in all samples belonging to Groups II, III and IV and in 7/10 specimens of Group I, resulting in a statistically significant difference (p≤0.001)., Conclusion: TCA-free MTA and antibiotic-enhanced MTAs showed superior performances in dentinal bridge formation and exhibited minimal pulpal necrosis than conventional MTA. The inclusion of antibiotics might contribute to create a more sterile environment that would improve the outcomes, favoring deposition of a mineralized matrix. However, further studies are needed to support these preliminary results. (EEJ-2024-10-157).

To evaluate the effects of the association of host defence peptide IDR-1002 and ciprofloxacin on human dental pulp cells (hDPSCs). hDPSCs were stimulated with ciprofloxacin and IDR-1002. Cell viability (by MTT assay), migration capacity (by scratch assay), production of inflammatory and anti-inflammatory mediators by hDPSCs (RT-PCR) and osteogenic differentiation (alizarin red staining) were evaluated. Phenotypic profile of hDPSCs demonstrated 97% for positive marked mesenchymal stem cell. Increased pulp cell migration and proliferation were observed after 24 and 48 h of exposure to IDR-1002 with ciprofloxacin. Mineral matrix formation by hDPSCs was observed of the association while its reduction was observed in the presence of peptide. After 24 h, the association between ciprofloxacin and IDR-1002 significantly downregulated TNFRSF-1, IL-1β, IL-8, IL-6 and IL-10 gene expression (p ≤ 0.0001). The association between the IDR-1002 and ciprofloxacin showed favourable immunomodulatory potential, emerging as a promising option for pulp revascularisation processes., (© 2024 Australian Society of Endodontology Inc.)

Vital pulp treatment (VPT) is crucial for preserving the health and function of the tooth in cases where the pulp tissue remains vital despite exposure. Various materials are introduced for this purpose. However, challenges such as low strength, high solubility, and tooth discoloration persist. Methylmethacrylate-based cement (MC) offers excellent sealing ability, feasibility, and mechanical properties, making it a promising alternative for VPT. Phosphate-based glass (PBG) has the potential to promote hard tissue regeneration by releasing key inducers, phosphorus (P) and calcium (Ca), for reparative odontogenesis. This study investigates PBG-integrated MC (PIMC) by characterizing its properties, assessing human dental pulp stem cell activity related to initial inflammatory adaptation and odontogenic differentiation, and evaluating hard tissue formation using an in vivo dog pulpotomy model. Results indicate that a 5% PBG-integrated MC (5PIMC) maintains the physicochemical properties of MC. Furthermore, 5PIMC demonstrates cytocompatibility, excellent expression of osteo/odontogenic markers, and resistance to inflammatory markers, significantly outperforming MC. Enhanced hard tissue formation is observed in the dental pulp of mongrel dog teeth treated with 5PIMC. These findings suggest that 5PIMC could be an optimal and suitable material for reparative odontogenesis through VPT., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Objective: To investigate the effects of quercetin (QU), hesperetin (HT), and taxifolin (TX) on human dental pulp cells (hDPCs) chronically exposed to lipopolysaccharide (LPS)., Methods: First, the cytotoxicity (alamarBlue) and bioactivity (biomineralization, Alizarin Red) of QU, HT, and TX concentrations were evaluated on healthy hDPCs. Then, the effects of non-cytotoxic and bioactive concentrations were investigated on hDPCs after previous stimulation with E. coli LPS (10 µg/mL) for 7 days. Cell culture media with and without LPS were used as positive and negative controls, respectively. Cell viability (alamarBlue), NF-κB activation (immunofluorescence), reactive oxygen species production (ROS, H2DCFDA probe), cell migration (Transwell), inflammation-related gene expression (RT-qPCR), and odontogenic differentiation (RT-qPCR and alizarin red) were evaluated (n = 8). Data were analyzed using confidence intervals and ANOVA (α = 5 %)., Results: The concentrations of 20 µM QU, 20 µM HT, and 200 µM TX reduced cell viability by more than 30 %. The 5 µM QU, 10 µM HT, and 100 µM TX concentrations were cytocompatible and stimulated biomineralization by healthy hDPCs. These concentrations were tested under the LPS challenge, and cell viability and odontogenic differentiation were significantly increased, while ROS production and inflammatory response were significantly decreased. In addition, the flavonoids significantly stimulated cell migration, reduced NF-κB activation, and increased biomineralization by LPS-challenged hDPCs compared to cells exposed to LPS alone and without any other treatment., Conclusion: Flavonoids can modulate the metabolism of hDPCs chronically exposed to LPS in vitro, stimulating cellular events compatible with stem cell-based regenerative processes., Clinical Significance: Flavonoids may be explored as adjuvant therapeutic agents during pulp capping to counteract chronic inflammatory conditions and stimulate regeneration of the dentin-pulp complex in caries-affected teeth, thereby preserving tooth vitality., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Objectives: The degradation of the restorative-dentin interface due to endogenous dentin enzymes, such as matrix metalloproteinases (MMPs), is a significant issue that accelerates the deterioration of the dentin matrix and leads to the failure of restorative treatments. Caredyne Restore (CR), a novel glass ionomer cement (GIC) with zinc ions in its formulation, represents the latest effort to mitigate this issue. This investigation aimed to evaluate the MMPs inhibitory effect, marginal integrity, and cytotoxicity of CR compared to a conventional GIC, Fuji IX (FIX)., Methods: The inhibitory effect of CR on MMPs activity was evaluated using in-situ zymography to visualize the endogenous gelatinolytic activity in the GIC-dentin interface. Additionally, CR's sealing properties were investigated using dye-assisted confocal laser microscopy (CLSM) to assess marginal leakage across the GIC-dentin interface. Both in-situ zymography and CLSM observations were conducted 1 day and seven days after pH cycling. Finally, the cytotoxicity of the GIC eluates was examined on rat dental pulp cells (RPCC2A). Assay measurements were performed after 24 and 48 h of incubation with test solutions prepared using various GIC eluate concentrations., Results: The MMPs activity of the CR specimens was significantly lower than that of FIX specimens after seven days of pH cycling. Signs of marginal leakage were also lower in the CR specimens. Comparison of CR and FIX eluates at the same concentration showed no significant difference in terms of biocompatibility., Conclusions: Caredyne restore is capable of inhibiting endogenous enzyme activity and improving sealing properties, while maintaining sufficient biocompatibility., Clinical Significance: Zinc ions released from Caredyne Restore offer a safe way to improve the quality of dental restorations and promote minimally invasive treatment, especially in lesions where the dentin matrix is susceptible to enzymatic degradation and recurrent caries., Competing Interests: Declaration of competing interest We declare that this manuscript is original and free of conflict of interest, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aim: Cannabidiol (CBD), derived from the Cannabis sativa plant, exhibits benefits in potentially alleviating a number of oral and dental pathoses, including pulpitis and periodontal diseases. This study aimed to explore the impact of CBD on several traits of human dental pulp stem cells (hDPSC), such as their proliferation, apoptosis, migration and odonto/osteogenic differentiation., Methodology: hDPSCs were harvested from human dental pulp tissues. The cells were treated with CBD at concentrations of 1.25, 2.5, 5, 10, 25 and 50 μg/mL. Cell responses in terms of cell proliferation, colony-forming unit, cell cycle progression, cell migration, apoptosis and odonto/osteogenic differentiation of hDPSCs were assessed in the normal culture condition and P. gingivalis lipopolysaccharide (LPS)-induced 'inflammatory' milieus. RNA sequencing and proteomic analysis were performed to predict target pathways impacted by CBD., Results: CBD minimally affects hDPSCs' behaviour under normal culture growth milieu in normal conditions. However, an optimal concentration of 1.25 μg/mL CBD significantly countered the harmful effects of LPS, indicated by the promoting cell proliferation and restoring the odonto/osteogenic differentiation potential of hDPSCs under LPS-treated conditions. The proteomic analysis demonstrated that several proteins involved in cell proliferation and differentiation were upregulated following CBD exposure, including CCL8, CDC42 and KFL5. RNA sequencing data indicated that CBD upregulated the Notch signalling pathway. In an inhibitory experiment, DAPT, a Notch inhibitor, reduced the effect of CBD-rescued LPS-attenuated mineralization in hDPSCs, suggesting that CBD potentially mediates Notch activation to exert its impact on odonto/osteogenic differentiation of hDPSCs., Conclusions: CBD recovers the proliferation and survival of hDPSCs following exposure to LPS. Additionally, we report that CBD-mediated Notch activation effectively restores the odonto/osteogenic differentiation ability of hDPSCs under inflamed conditions. These results underscore the potential role of CBD as a therapeutic option to enhance dentine regeneration., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Background: Dental pulp stem cells (DPSCs) are widely used in research on dental tissue regeneration and systemic disease treatment. However, the oxidative microenvironment often causes cellular senescence, leading to decreased function. Our previous study demonstrated that pleiotrophin (PTN), a secreted extracellular matrix-associated protein, could rescue the proliferative capacity and osteogenic differentiation of replicative senescent DPSCs., Objective: This study aimed to explore the influence and mechanism of PTN on dental pulp stem cells under H 2 O 2 -induced oxidative microenvironment., Materials and Methods: DPSCs isolated from human third molars were treated with 100 μm H 2 O 2 for 4 h, mimicking the oxidative microenvironment. To investigate the influence of PTN on DPSC under H 2 O 2 -induced oxidative microenvironment, 50 pg/mL PTN was added in the culture medium for 48 h. RT-qPCR, western blotting, SA-β-gal staining, intracellular ROS production and immunofluorescence staining assays were used to analyse the cellular senescence, osteogenic differentiation capacity, oxidative stress conditions and possible mechanism., Results: H 2 O 2 treatment increased the ratio of SA-β-gal-positive DPSCs and upregulated the senescence-related gene expression, including P53, P21 and P16. PTN pretreatment downregulated the ratio of SA-β-gal-positive DPSCs and the expression of these genes. Besides, PTN pretreatment partially reversed the H 2 O 2 -induced decreased osteogenic differentiation potential of DPSCs, total antioxidant capacity and Nrf2 and HO-1 mRNA expression in DPSCs. Western blotting and immunofluorescent staining results indicated that PTN pretreatment enhanced the Nrf2 nuclear translocation under oxidative stress conditions and observable higher fluorescence signals in the nucleus denoted PTN and Nrf2 colocalisation. Western blotting results showed that PTN reversed the decreased expression of p-AKT in the H 2 O 2- induced oxidative environment. However, the PI3K inhibitor LY294002 blocked the upregulated levels of total Nrf2. Immunofluorescence staining displayed that LY294002 also inhibited the nuclear translocation of Nrf2 which was enhanced under PTN pretreatment., Conclusions: This study demonstrated that PTN could prevent senescent damage induced by H 2 O 2 on DPSCs, mainly by combining with Nrf2 and enhancing its nuclear translocation., (© 2024 John Wiley & Sons Ltd.)

This study aimed to evaluate and compare the effectiveness of mineral trioxide aggregate (MTA), bioceramic putty (BP) and tannic acid (TA) for experimental pulpotomy. Our in-vivo experimental study involved sample of 45 rats that were randomly divided into 4 groups: Group 1 (subdivided into negative (1-A) and positive (1-B) subgroups), Group 2 (MTA treated), Group 3 (BP treated) and Group 4 (TA treated). 4 weeks post pulpotomy, specimens were analyzed histologically, immunohistochemically using dentin sialoprotein marker, and histomorphometrically by assessing the thickness of newly formed dentin bridge. Group 1-B showed pulp necrosis without hard tissue formation. Group 2 showed moderate dentin formation while group 3 presented a thick layer of calcific barrier. Group 4 showed dentin bridge formation, however, irregular pulp calcifications and radicular pulp necrosis were seen. The thickness of newly formed dentin bridge showed a significant difference between group 1-B and group 2, 3 &4. Significant difference was found between group 2&3 and group 3&4. Dentin sialoprotein immunohistochemical expression was negative in group 1-B, mild in group 2, strong in group 3 and moderate in group 4. MTA and BP proved to be effective pulpotomy agents with BP being superior. For TA, further studies are required to explain the recorded unfavorable effects in some specimens., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Objectives: This study investigated the impact of some specific experimental calcium phosphate cements doped with different fluoride salts (FDCPCs) concentrations on the basal functions of human Dental Pulp Stem Cells (hDPSCs). Furthermore, this study also examined the migration, as well as the mineralisation through osteogenic differentiation., Methods: Experimental FDCPCs were formulated using different concentrations of calcium/sodium fluoride salts [(5 wt%: VS5F), (10 wt%: VS10F), (20 wt%: VS20F)]. A fluoride-free calcium phosphate (VS0F) was used as a control. The hDPSCs were assessed to evaluate their self-renewal and migration activity in the presence of eluates of the different FDCPCs. A viability assay in osteogenic conditions was carried out, along with the differentiation potential through Alkaline Phosphatase Activity (ALP), and Alizarin Red Staining (ARS). Moreover, the gene expression of specific markers (RUNX2, ALP, COL1α1, OCN, OPN, DSPP, MEPE, and DMP-1) was also evaluated., Results: All the tested FDCPD had no influence on cell migrations, but they caused a decrease in cell viability in osteogenic conditions when not diluted. Conversely, the eluants of VS20F showed a positive effect on stem cell differentiation. This result was corroborated through ALP activity, ARS assay. Moreover, upregulation of specific gene markers such as RUNX2, DMP-1, and DSPP was observed in hDPSCs, especially when treated with VS20F., Significance: The experimental FDCPC tested in this study exhibits a dose-dependent capacity to promote mineralisation in osteogenic environment. The FDCPC-VS20F seems to be the most promising experimental material suitable for developing of pulp-capping materials with osteogenic and bioactive properties., Competing Interests: Declaration of Competing Interest The author has no financial conflicts to disclose or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Aim: The purpose of this study was to investigate the role of calcium-sensing receptor (CaSR) in the angiogenic differentiation of lipopolysaccharide (LPS)-treated human dental pulp cells (hDPCs)., Methodology: The LPS-induced hDPCs were cultured in the medium with different combinations of CaSR agonist R568 and antagonist Calhex231. The cell proliferation, migration, and angiogenic capacity were measured by Cell Counting Kit-8 (CCK-8), scratch wound healing, and tube formation assays, respectively. Enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot were conducted to determine the gene/protein expression of CaSR, inflammatory mediators, and angiogenic-associated markers. The activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt) was assessed by western blot analysis., Results: The cell proliferation was elevated in response to R568 or Calhex231 exposure, but an enhanced cell migration was only found in cultures supplemented with Calhex231. Furthermore, R568 was found to potentiate the formation of vessel-like structure, up-regulated the protein expression of tumour necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), and stromal cell-derived factor (SDF)-1; comparable influences were also observed in R568-stimulated cells in the presence of PI3K inhibitor LY294002. In contrast, Calhex231 obviously inhibited the tube formation and VEGF protein level, whereas promoted the production of IL-6, TNF-α, and eNOS; however, in the presence of LY294002, Calhex231 showed a significant promotion on the protein expression of CaSR, VEGF, and SDF-1. In addition, R568 exhibited a promotive action on the Akt phosphorylation, which can be reversed by LY294002., Conclusions: Our results demonstrated that CaSR can regulate the angiogenic differentiation of LPS-treated hDPCs with an involvement of the PI3K/Akt signalling pathway., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objectives: The aim of this systematic review was to demonstrate the efficacy of topical application of corticosteroids in remineralization of dental pulp tissues to preserve their vitality and function., Data, Sources and Study Selection: An electronic search was performed using MEDLINE by PubMed, EMBASE, Web of Science (WOS), and Scopus databases. The inclusion criteria were in vitro studies that employed dental pulp tissue obtained from extracted healthy permanent human teeth and were subjected to topical administration of corticosteroids and evaluated tissue remineralization by performing any mineralization assay. A total of 11 studies were selected for inclusion. PRISMA guidelines were followed, and the methodological quality and risk of bias of the included studies were evaluated using the RoBDEMAT guidelines. Also, tables were designed for data extraction, including tissue mineralization and osteogenic differentiation as primary and secondary outcomes, respectively., Conclusions: Alizarin Red S (ARS) has been able to demonstrate a possible mineralizing power of corticosteroids, applied at an adequate dose. The up-regulation of Alkaline phosphatase (ALP), osteocalcin (OCN), osteopontin (OSP), sialophosphoprotein (DSPP), runt-related transcription factor 2 (RUNX2), collagen type 1 alpha 1(COL1α1) and dentin matrix protein 1 (DMP-1) induced the osteogenic/odontogenic differentiation of dental pulp stem cells (DPSCs)., Clinical Significance: Deep carious lesions treatment is still challenging in restorative dentistry. Some treatments have been focused on dental pulp tissue remineralization to maintain the function and vitality. After corticosteroids topical application, mineral deposition and osteogenic differentiation have been detected., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Background: Regenerative endodontics requires an innovative delivery system to release antibiotics/growth factors in a sequential trend. This study focuses on developing/characterizing a thermoresponsive core-shell hydrogel designed for targeted drug delivery in endodontics., Methods: The core-shell chitosan-alginate microparticles were prepared by electrospraying to deliver bone morphogenic protein-2 for 14 days and transforming growth factor-beta 1 (TGF-β1) for 7-14 days. Methylcellulose (MC) and gelatin were utilized to create the core-shell hydrogel to load a modified triple antibiotic combination (penicillin G/metronidazole/ciprofloxacin (PMC)) and growth factor-loaded microparticles in the shell and the core compartments, respectively. Morphological assessment, core-shell structural analysis, FTIR analysis, rheological analysis, swelling, and degradation rate studies were conducted for characterization. The viability of dental pulp stem cells (DPSCs) upon antibiotic exposure, antibacterial activity, and release studies of PMC and growth factors were investigated. Cellular studies (cell viability, alkaline phosphatase (ALP) activity, osteo/odontoblast gene expression (using Reverse transcription-polymerase chain reaction (RT-PCR)) and in vivo studies (inflammatory response and differentiation potential of the developed hydrogel by subcutaneous implantation in rats via histological examination) were assessed., Results: The hydrogel showed a porous microstructure with interconnected pores. Core-shell structure analysis confirmed the successful extrusion of the MC hydrogel to the surface. FTIR analysis revealed interactions between MC and gelatin. Rheological analysis indicated time-dependent gel formation, supporting thermosensitivity at 37 °C. Swelling occurred rapidly, and degradation reached 62.42% on day 45. Further, antibiotics exhibited no cytotoxicity on DPSCs. Sequential release of antibiotics and growth factors was observed for up to 5 and 14 d, respectively. The hydrogel showed antibacterial activity. DPSCs exhibited increased proliferation, ALP activity, and odontoblast gene expression. In vivo studies showed that the biocompatible drug-loaded hydrogel exhibited more mineralization than the control., Conclusions: The developed core-shell hydrogel containing PMC and growth factor-loaded core-shell microparticles provided a versatile and biocompatible platform for sequential drug delivery in regenerative endodontics. The system demonstrates promising characteristics for dentin regeneration, making it a potential candidate for clinical applications., (© 2024 The Author(s). Published by IMR Press.)

Calcium phosphate-based materials (CaP) are introduced as potential dental pulp capping materials for deciduous teeth. The present study investigated the influence of inorganic phosphate (P i ) on regulating stem cells isolated from human exfoliated deciduous teeth (SHED). SHEDs were treated with P i . Cell cycle progression and apoptosis were examined using flow cytometry analysis. Osteo/odontogenic and adipogenic differentiation were analyzed using alizarin red S and oil red O staining, respectively. The mRNA expression profile was investigated using a high-throughput RNA sequencing technique. P i increased the late apoptotic cell population while cell cycle progression was not altered. P i upregulated osteo/odontoblastic gene expression and enhanced calcium deposition. P i -induced mineralization was reversed by pretreatment of cells with Foscarnet, or p38 inhibitor. P i treatment inhibited adipogenic differentiation as determined by decreased PPARγ expression and reduced intracellular lipid accumulation. Bioinformatic analysis of gene expression profiles demonstrated several involved pathways, including PI3K/AKT, MAPK, EGFR, and VEGF signaling. In conclusion, P i enhanced osteo/odontogenic but inhibited adipogenic differentiation in SHED., (© 2024. The Author(s).)

This study aimed to design sustained released biodegradable calcium carbonate nanoparticles loaded with chlorhexidine (CHX-loaded NPs) and to investigate the early osteogenic differentiation and antimicrobial effects on the important bacteria involved in infections of dental implants. The microemulsion method was used to prepare the calcium carbonate nanoparticles loaded with chlorhexidine. The prepared nanoparticles were characterized using conventional methods. The release pattern determination and the biodegradation test were performed for the prepared nanoparticles. For the early osteogenic differentiation test of the prepared nanoparticles, alkaline phosphatase (ALP) activity was detected in human dental pulp stem cells (HDPSCs). The antimicrobial effects of the nanoparticles were evaluated against Escherichia coli, Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, and Pseudomonas aeruginosa. The sizes of free calcium carbonate nanoparticles and CHX-loaded NPs were 105 ± 1.63 and 118 ± 1.47 nm and their zeta potentials were - 27 and - 36, respectively. A 50% degradation of nanoparticles was achieved after 100 days. These nanoparticles showed a two-stage sustained release pattern in vitro. Microscopic images revealed that the morphology of free calcium carbonate nanoparticles primarily took on a spherical calcite form, while CHX-loaded NPs predominantly exhibited a cauliflower-like vaterite polymorph. The nanoparticles increased the activity of ALP in cells in two weeks significantly (p < 0.05). Antimicrobial and antibiofilm results showed an efficient effect of the prepared nanoparticle against the studied bacteria. Calcium carbonate nanoparticles are an efficient multifunctional vector for chlorhexidine and can be used as a bioactive antibacterial agent against various oral microorganisms to prevent implant infections., (© 2024. The Author(s).)

The regeneration of absorbed alveolar bone and reconstruction of periodontal support tissue are huge challenges in the clinical treatment of periodontitis due to the limited regenerative capacity of alveolar bone. It is essential to regulate inflammatory reaction and periodontal cell differentiation. Based on the anti-inflammatory effect of baker's yeast β-glucan (BYG) with biosafety by targeting macrophages, the BYG-based nanoparticles loading methotrexate (cBPM) were fabricated from polyethylene glycol-grafted BYG through chemical crosslinking for treatment of periodontitis. In our findings, cBPM promoted osteogenesis of human dental pulp stem cells (hDPSCs) under inflammatory microenvironment, characterized by the enhanced expression of osteogenesis-related Runx2 and activation of mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/Erk) pathway in vitro. Animal experiments further demonstrate that cBPM effectively promoted periodontal bone regeneration and achieved in a better effect of recovery indicated by 19.2 % increase in tissue volume, 7.1 % decrease in trabecular separation, and a significant increase in percent bone volume and trabecular thickness, compared with the model group. Additionally, cBPM inhibited inflammation and repaired alveolar bone by transforming macrophage phenotype from inflammatory M1 to anti-inflammatory M2. This work provides an alternative strategy for the clinical treatment of periodontitis through BYG-based delivery nanoplatform of anti-inflammatory drugs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

In the present study, nano-hydroxyapatite (n-HA) powder was extracted from carp bone waste to fabricate porous n-HA substrates by a molding and sintering process. Subsequently, the substrates were loaded with different amounts of sodium fluoride (NaF) through immersion in NaF suspensions for 10, 7.5, and 5 min. The NaF-loaded n-HA substrates were then examined for their structural and physical properties, chemical bonds, loading and release profile, pH changes, cytotoxicity, and osteogenic effect on dental pulp stem cells (DPSCs) at the level of RNA and protein expression. The results showed that the n-HA substrates were porous (> 40% porosity) and had rough surfaces. The NaF could be successfully loaded on the substrates, which was 6.43, 4.50, and 1.47 mg, respectively for n-HA substrates with immersion times of 10, 7.5, and 5 min in the NaF suspensions. It was observed that the NaF release rate was rather fast during the first 24 h in all groups (39.06%, 36.43%, and 39.57% for 10, 7.5, and 5 min, respectively), and decreased dramatically after that, indicating a slow detachment of NaF. Furthermore, the pH of the medium related to all materials was changed during the first 4 days of immersion (from 7.38 to pH of about 7.85, 7.84, 7.63, and 7.66 for C0, C5, C7.5, and C10, respectively). The pH of media associated with the C7.5, and C10 increased up to 4 days and remained relatively constant until day 14 (pH = 7.6). The results of the cytotoxicity assay rejected any toxicity of the fabricated NaF-loaded n-HA substrates on DPSCs, and the cells could adhere to their surfaces with enlarged morphology. The results showed no effect on the osteogenic differentiation at the protein level. Nevertheless, this effect was observed at the gene level., (© 2024. The Author(s).)

The cytotoxic effects of three root canal sealers with different bases on human dental pulp stem cells were assessed in this study using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. The cytotoxic effects of three root canal sealers with different bases on human dental pulp stem cells (DPSCs) were assessed in this study using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. The cytotoxicity of the sealers was tested after one, 4, and 7 d. Human dental pulp stem cell proliferation was concluded using an MTT assay. Cells not treated with sealer extract were used as controls. The absorption levels were measured using an Eliza spectrophotometer. P was set at 0.05 when the percentage of cell proliferation was matched between groups and observation times using one-way analysis of variance (ANOVA).During the second passage (P2), human dental pulp stem cells displayed a single morphological and phenotypic trait, with fibroblast morphology being the most common. There were no appreciable variations between the four groups after a day. There was a notable variation in the average percentage of cell proliferation between the groups after 4 and 7 days. The control group had the highest percentage, followed by the GuttaFlow Bioseal group, the Well Root St group, and the AH-Plus group, which had the lowest percentage. For every sealing group, after one day, the highest mean percentage of cell proliferation was recorded, followed by day four, and after day seven, the lowest mean percentage. The observation periods showed minimal cytotoxic effects of GuttaFlow Bioseal, whereas AH-Plus was the most cytotoxic to human dental pulp stem cells. The highest mean percentage of cell proliferation for all sealers was recorded on day one., (© 2024. The Author(s).)

Background: An ideal aesthetic restorative material should be attached to the tooth tissues by adhesion, have a smooth surface as possible, should not cause toxic reactions in the pulp and discoloration and microleakage. This study aims at comparatively assess the cytotoxicity of current adhesive systems on human dental pulp cells., Materials and Methods: The adequate density of human pulp cells was observed from the ready cell line. The passaging was performed and the 3rd passage cells were selected. Adhesive systems and MTA were used on the cultures. Trypan blue staining was conducted on the cells at the 1st, 2nd, 3rd days and a count of live and dead cells using a light microscope. The dead cells whose membrane integrity was impaired by staining with trypan blue and the viability rate was determined using live and dead cell numbers. Data analysis was performed using IBM SPSS Statistics 22., Results: A significant difference in vialibity rates between adhesive systems was observed on the first day. No significant statistical differences were observed on the 2nd and 3rd days (p < 0.05)., Conclusion: Futurabond M showed similar biocompatibility with MTA on human pulp cells and it can be applied in cavities with 1-1.5 mm hard tissue between pulp and dentine., (© 2024. The Author(s).)

Background: In bone tissue engineering (BTE), cell-laden scaffolds offer a promising strategy for repairing bone defects, particularly when host cell regeneration is insufficient due to age or disease. Exogenous stem cell-based BTE requires bioactive factors to activate these cells. Graphene oxide quantum dots (GOQDs), zero-dimensional derivatives of graphene oxide, have emerged as potential osteogenic nanomedicines. However, constructing biological scaffolds with GOQDs and elucidating their biological mechanisms remain critical challenges., Methods: We utilized GOQDs with a particle size of 10 nm, characterized by a surface rich in C-O-H and C-O-C functional groups. We developed a gelatin methacryloyl (GelMA) hydrogel incorporated with GOQDs-treated dental pulp stem cells (DPSCs). These constructs were transplanted into rat calvarial bone defects to estimate the effectiveness of GOQDs-induced DPSCs in repairing bone defects while also investigating the molecular mechanism underlying GOQDs-induced osteogenesis in DPSCs., Results: GOQDs at 5 μg/mL significantly enhanced the osteogenic differentiation of DPSCs without toxicity. The GOQDs-induced DPSCs showed active osteogenic potential in three-dimensional cell culture system. In vivo, transplantation of GOQDs-preactivated DPSCs/GelMA composite effectively facilitated calvarial bone regeneration. Mechanistically, GOQDs stimulated mitophagy flux through the phosphatase-and-tensin homolog-induced putative kinase 1 (PINK1)/Parkin E3 ubiquitin ligase (PRKN) pathway. Notably, inhibiting mitophagy with cyclosporin A prevented the osteogenic activity of GOQDs., Conclusion: This research presents a well-designed bionic GOQDs/DPSCs/GelMA composite scaffold and demonstrated its ability to promote bone regeneration by enhancing mitophagy. These findings highlight the significant potential of this composite for application in BTE and underscore the crucial role of mitophagy in promoting the osteogenic differentiation of GOQDs-induced stem cells., Competing Interests: The authors report no conflicts of interest in this work., (© 2024 Yan et al.)

Background and Objectives: This study aimed to investigate the effect of zinc phosphate (ZnP) cement, glass ionomer cement (GIC), and nano-integrated bio-ceramic (NIB) cement on mineralization when placed in contact with bone tissue-forming cells. Materials and Methods: ZnP cement, GIC, and NIB cement were divided into direct and indirect groups. A total of 72 cement pellets (24 pellets of each test sample) of 3 × 1 mm (width × height) were prepared using polytetrafluoroethylene molds. A total of 3 sample groups were demarcated using 96- cell well culture plates. In the control group, 24 wells were filled with mineralized osteoblasts and 1 µL of gingival crevicular fluid (GCF). In test group 1, to show a direct effect, 36 samples were plated with mineralized osteoblasts and 1 µL GCF for 24 h; the cells were directly exposed to cement pellets. A total of 36 samples were immersed in GCF for 24 h; later the supernatant was transferred to the mineralized osteoblasts to demonstrate an indirect effect in test group 2. To assess the mineralization, osteoblasts were stained with alizarin red and later observed under an inverted phase-contrast microscope. Data were analyzed using the statistical package for social sciences. An independent t-test compared the direct and indirect effects of the ZnP cement, GIC, NIB cement, and control groups on the mineralization of osteoblasts derived from hDPCs. Results: A statistically significant difference was observed between the ZnP cement, GIC, and NIB cement groups ( p < 0.05). ZnP cement exhibited a moderate, NIB cement the least harmful effect, and GIC showed the most harmful effect on the mineralization of osteoblast cells. Conclusions: The biocompatibility of dental luting cements is an important aspect that clinicians should consider during their selection. Nano-integrated bio-ceramic cement showed the least negative effect on the mineralization of osteoblast cells which is beneficial for the cementation of cement-retained implant prostheses. However, further studies are needed to evaluate osteoblast and osteoclast activity in vivo.

Objectives: The present study aimed to (1) investigate biocompatibility and cytotoxicity of pulp-capping materials on viability of human dental pulp stem cells (hDPSCs); (2) determine angiogenic, odontogenic, and osteogenic marker mRNA expressions; and (3) observe changes in surface morphology of the hDPSCs using scanning electron microscopy (SEM)., Methods: Impacted third molars were used to isolate the hDPSCs, which were treated with extract-release fluids of the pulp-capping materials (Harvard BioCal-Cap, NeoPUTTY MTA, TheraCal LC, and Dycal). Effects of the capping materials on cell viability were assessed using 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay and the apoptotic/necrotic cell ratios and reactive oxygen species (ROS) levels from flow cytometry. Marker expressions (alkaline phosphatase [ALP], osteocalcin [OCN], collagen type I alpha 1 [Col1A], secreted protein acidic and rich in cysteine [SPARC], osteonectin [ON], and vascular endothelial growth factor [VEGF]) were determined by quantitative reverse-transcription polymerase chain reaction. Changes in surface morphology of the hDPSCs were visualised by SEM., Results: The MTS assay results at days 1, 3, 5, and 7 indicated that Harvard BioCal-Cap, NeoPUTTY MTA, and TheraCal LC did not adversely affect cell viability when compared with the control group. According to the MTS assay results at day 14, no significant difference was found amongst Dycal, Harvard BioCal-Cap, NeoPUTTY MTA, and TheraCal LC affecting cell viability. Dycal was the only capping material that increased ROS level. High levels of VEGF expression were observed with Harvard BioCal-Cap, TheraCal LC, and NeoPUTTY MTA. NeoPUTTY MTA, and Dycal upregulated OCN expression, whereas TheraCal LC upregulated Col1A and SPARC expression. Only Dycal increased ALP expression. HDSCs were visualized in characteristic spindle morphology on SEM when treated with TheraCal LC and Harvard BioCal-Cap., Conclusions: NeoPUTTY MTA and Harvard BioCal-Cap showed suitable biocompatibility values; in particular, these pulp-capping materials were observed to support the angiogenic marker., Competing Interests: Conflict of interest None disclosed., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The use of bleaching agents to remove stains is one of the main dental procedures to improve the aesthetics of teeth. This review presents the main agents used for tooth whitening, existing clinical protocols, and the structural changes that may occur through their use. The main bleaching agents consist of hydrogen peroxide and carbamide peroxide, which are used in bleaching techniques for vital teeth. These techniques can be performed in the office by a professional or by the individual in a home en-vironment under professional guidance. Bleaching agents come in a variety of concentrations and there are over-the-counter products available on the market with lower concentrations of hydrogen peroxide. Due to the chemical characteristics of the agents, changes in the organic and inorganic content of the tooth structure can be observed. These changes are related to morphological changes characterized by in-creased permeability and surface roughness, such changes compromise the mechanical resistance of the tooth. Furthermore, bleaching agents can promote molecular changes after reaching the dental pulp, resulting in oxidative stress of pulp cells and the release of pro-inflammatory mediators. Despite the bleaching effectiveness, tooth sensitivity is considered the main side effect of use. Therefore, among the heterogeneity of protocols, those that used the bleaching agent for a prolonged time and in lower con-centrations presented more harmful effects on the tooth structure., (©The Author(s) 2024. Open Access. This article is licensed under a Creative Commons CC-BY International License.)

In response to pro-inflammatory cytokines such as interleukin (IL)-1β, dental pulp fibroblasts produce various inflammatory mediators, including IL-6, IL-8, CC chemokine ligand 20 (CCL20), and CXC chemokine ligand 10 (CXCL10), leading to the progression of pulpitis. IL-17/IL-17A (IL-17A) is a pro-inflammatory cytokine secreted by T helper (Th) 17 cells following their recruitment to inflamed sites; however, the roles of IL-17A during pulpitis remain unclear. The purpose of this study was to investigate the effect of IL-17A on IL-6, IL-8, CCL20 and CXCL10 production by human dental pulp fibroblasts (HDPFs) in vitro. IL-17A at a concentration of 100 ng/ml induced the production of 10 times more IL-8 and 4 times more CXCL10, but not IL-6 and CCL20, compared to controls. Co-stimulation of HDPFs with IL-17A and IL-1β synergistically enhanced the production of IL-6, CCL20, IL-8 and CXCL10. IL-1β increased expression of IL-17 receptor/IL-17RA (IL-17R) on HDPFs. Moreover, the cell signal pathways of p38 mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) were more potently activated by simultaneous stimulation with IL-17A and IL-1β. These findings suggest that IL-17A participates in the progression of dental pulp inflammation through the enhanced production of inflammatory mediators in HDPFs., (© 2024 Scandinavian Division of the International Association for Dental Research. Published by John Wiley & Sons Ltd.)

Objective: Drug-loaded non-resorbable polymeric nanoparticles (NPs) are proposed as an adjunctive treatment for pulp regenerative strategies. The present in vitro investigation aimed to evaluate the effectiveness of tideglusib-doped nanoparticles (TDg-NPs) in mitigating the adverse effects of bacterial lipopolysaccharide endotoxin (LPS) on the viability, morphology, migration, differentiation and mineralization potential of human dental pulp stem cells (hDPSCs)., Methods: Cell viability, proliferation, and differentiation were assessed using a MTT assay, cell migration evaluation, cell cytoskeleton staining analysis, Alizarin Red S staining and expression of the odontogenic related genes by a real-time quantitative polymerase chain reaction (RT-qPCR) were also performed. Cells were tested both with and without stimulation with LPS at various time points. One-way ANOVA and Tukey's test were employed for statistical analysis (p < 0.05)., Results: Adequate cell viability was encountered in all groups and at every tested time point (24, 48, 72 and 168 h), without differences among the groups (p > 0.05). The analysis of cell cytoskeleton showed nuclear alteration in cultures with undoped NPs after LPS stimulation. These cells exhibited an in blue diffuse and multifocal appearance. Some nuclei looked fragmented and condensed. hDPSCs after LPS stimulation but in the presence of TDg-NPs exhibited less nuclei changes. LPS induced down-regulation of Alkaline phosphatase, Osteonectin and Collagen1 gene markers, after 21d. LPS half-reduced the cells production of calcium deposits in all groups (p < 0.05), except in the group with TDg-NPs (decrease about 10 %)., Significance: LPS induced lower mineral deposition and cytoskeletal disorganization in hDPSCs. These effects were counteracted by TDg-NPs, enhancing osteogenic differentiation and mineralization., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

Objectives: The aim of this study was to assess the cytotoxicity of novel polymerization co-initiators and their effect on cytokine release from human dental pulp stem cells (hDPSCs), comparing them with commonly used co-initiators., Methods: Cells were isolated from the dental pulp of healthy human third molars. The new co-initiators, namely HDa1, HD4, HD1, and MHPTm, were evaluated and compared with the compounds dimethylaminoethyl amine benzoate (EDAB) and 2-(dimethylamino)ethyl methacrylate (DMAEMA). These compounds were diluted in dimethylsulfoxide (DMSO) at concentrations ranging from 1 to 8 mM. hDPSCs were seeded onto 96-well plates and incubated for 48 h. Subsequently, the cells were exposed to different concentrations of the co-initiators mentioned for 24 h. After this period, the culture medium was removed, and mitochondrial metabolism was evaluated using the MTT assay, while cytokine release (IL-1β, IL-6, IL-8, IL-10, TNF-α) was analyzed by the MAGPIX assay. Cells without exposure to the tested compounds served as controls. The data were analyzed using one-way ANOVA and Tukey's test., Results: The compounds showed low toxicity, with 8 mM concentration causing the most significant reduction in mitochondrial metabolism. MHPTm was the most toxic co-initiator tested (compound bearing an amine functionality). All compounds up-regulated TNF-α, IL-10, IL-6, and IL-8, with HD4 exhibiting the most pronounced increase in IL-6 and IL-8., Significance: The newly proposed co-initiators demonstrated reduced impact on mitochondrial metabolism, comparable to some traditional co-initiators. Despite their lower toxicity, HD4 increased IL-6 and IL-8 release, suggesting its potential involvement in triggering an inflammatory reaction, particularly in the short term., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: The present study aimed to evaluate the viability of human dental pulp stem cells (hDPSCs) exposed to boric acid (BA) and injectable platelet-rich fibrin (I-PRF)., Materials and Methods: hDPSCs were isolated from impacted third molars. Nine milliliters of whole blood was transferred to I-PRF tubes and centrifuged at 700 rpm for 3 minutes. A BA solution was prepared by dissolving BA in a 0.1 g/ml stock solution. The cells were divided into four groups: control, I-PRF, BA, and BA + I-PRF. Cell viability was evaluated using flow cytometry. Mineralized calcium nodules were observed using Alizarin Red staining. The data were analyzed using two-way analysis of variance and Tukey's HSD test (p<0.05)., Results: The highest percentage of viable cells was in the I-PRF group, and the lowest percentage of viable cells was in the BA group at all times. Larger calcium nodules were observed in the BA group compared to the other groups., Conclusion: The use of I-PRF with or without BA had a positive effect on cell viability. BA and I-PRF affected the formation of mineralized calcium nodules. I-PRF and BA may be used in combination because these substances minimally reduce cell viability and promote mineralized nodule formation., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024. Published by Elsevier Ltd.)

Objective: This study aimed to evaluate the dental pulp responses to recombinant human erythropoietin (rhEPO) and/or mineral trioxide aggregate (MTA) in pulp capping of inflamed dental pulp in vivo., Materials and Methods: In accordance with ARRIVE guidelines, pulp inflammation was induced by exposing the maxillary first molars (n = 64) of Wistar rats (n = 32) to the oral environment for two days. The exposed pulps were randomly assigned four groups based on the pulp capping material: rhEPO, MTA, MTA + rhEPO, or an inert membrane. An additional eight rats formed the healthy control group. After four weeks, the animals were euthanized, and histological, qRT-PCR, and spectrophotometric techniques were employed to analyze the left maxillary segments, right first maxillary molars, and blood samples, respectively. Statistical significance was set at p < 0.05 and < 0.001., Results: Pulp capping with rhEPO, MTA, or MTA + rhEPO resulted in lower inflammation and higher mineralization scores compared to untreated control. MTA + rhEPO group exhibited significantly decreased expression of tumor necrosis factor-alpha, and interleukin 1-beta, while MTA group showed substantially reduced expression of interferon-gamma. Both rhEPO and MTA + rhEPO groups presented elevated dentin matrix protein 1 levels compared to untreated control. Furthermore, pulp capping with rhEPO and/or MTA led to increased transforming growth factor-beta 1 expression and reductions of pro-inflammatory/immunoregulatory cytokine ratios and prooxidative markers. Pulp capping with rhEPO also resulted in increase of systemic antioxidative stress markers., Conclusion: Capping with rhEPO or MTA + rhEPO resulted in a favorable effect that was similar or even superior to that of MTA., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Maintaining pulp vitality and function is a priority of the medicaments employed in pulp therapy to preserve tooth integrity. Aim: This study evaluated inflammatory response and reparative dentin bridge formation after direct pulp capping with two different bioceramics. Materials and Method: This was an in vivo controlled experimental study on 12 male Wistar rats. Pulpotomies were performed and the exposed pulps were capped with Biodentine or Neo MTA. After 15, 45 and 90 days, maxillary segments were obtained and prepared for histologic analysis and Micro-CT. Hounsfield Units (HU) were quantified. Results: Micro-CT analysis showed greater mineralization at 90 days with Neo MTA than with Biodentine. HU did not differ significantly (p >0.05) between molars treated with Biodentine and Neo MTA at 15 and 45 days, but at 90 days, there was statistically significant difference (p <0.05) between them. Reparative dentin was observed near the pulp exposure and canal orifice with both bioceramics. At 45 and 90 days, molars treated with Neo MTA showed mineralized tissue filling the canal orifice. Molars treated with Biodentine showed mineralized tissue and dentin bridge at the site of exposure at 45 days, and total pulp exposure coverage and mineralized entin matrix at 90 days. Conclusions: Biodentine and Neo MTA induce the formation of reparative dentin bridge after 45 days with inflammatory cell infiltrate., Competing Interests: The authors declare no potential conflicts of interest regarding the research, authorship, and/or publication of this article, (SAIO.)

Premixed calcium silicate-based materials have recently been developed and are recommended for a wide range of endodontic procedures, including vital pulp therapy. This study investigated the in vitro biocompatibility and pro-mineralization effect and in vivo reparative dentin formation of EndoSequence Root Repair Material, EndoSequence BCRRM, Bio-C Repair, and Well-pulp PT. Both fresh and set extracts had no detrimental effect on the growth of human dental pulp stem cells. The fresh extracts had a higher calcium concentration than the set extracts and induced considerably greater mineralized nodule formation. EndoSequence Root Repair Material had the longest setting time, whereas Bio-C Repair had the shortest. When these materials were applied to exposed rat molar pulps, mineralized tissue deposition was found at the exposure sites after 2 weeks. These results indicate that the premixed calcium silicate-based materials tested could have positive benefits for direct pulp capping procedures.

Objectives: This study sought to determine effects of Thai propolis extract mixed in mineral trioxide aggregate (MTA) on matrix metalloproteinase-2 (MMP-2) expression and its activity in inflamed human dental pulp cells (HDPCs)., Materials and Methods: Interleukin-1β-primed HDPCs were treated with either the eluate of MTA mixed with distilled water, of MTA mixed with 0.75 mg/ml of the propolis extract, or of Dycal®, 0.75 mg/ml of the propolis extract, or 0.2% (v/v) of chlorhexidine for 24 or 72 h. The viability of HDPCs was determined by the PrestoBlue® cytotoxic assay. HDPCs' lysates were analyzed for MMP-2 mRNA expression by RT-qPCR, while their supernatants were measured for MMP-2 activity by gelatin zymography., Results: At 24 and 72 h, a non-toxic dose of the propolis extract at 0.75 mg/ml by itself or mixed in MTA tended to reduce MMP-2 expression upregulated by MTA, while it further decreased the MMP-2 activity as compared to that of MTA mixed with distilled water. The MMP-2 activity of interleukin-1β-primed HDPCs treated with the eluate of the propolis extract mixed in MTA was significantly lower than that of interleukin-1β-primed HDPCs at 24 h (p=0.012). As a control, treatment with chlorhexidine significantly inhibited MMP-2 expression induced by MTA and MMP-2 activity enhanced by interleukin-1β (p<0.05). Treatment with Dycal® caused a significant increase in HDPC's death, resulting in a significant decrease in MMP-2 expression and activity (p<0.05)., Conclusions: MTA mixed with Thai propolis extract can reduce MMP-2 mRNA expression and activity when compared to MTA mixed with distilled water in inflamed HDPCs.

Background: Different materials have been used as wound dressings after vital pulp therapies. Some of them have limitations such as delayed setting, difficult administration, slight degree of cytotoxicity, crown discoloration and high cost. Therefore, to overcome these disadvantages, composite scaffolds have been used in regenerative dentistry. This study aims to construct and characterize the physicochemical behavior of a novel injectable alginate hydrogel loaded with different bioactive glass nanoparticles in various concentrations as a regenerative pulpotomy filling material., Methods: Alginate hydrogels were prepared by dissolving alginate powder in alcoholic distilled water containing mesoporous bioactive glass nanoparticles (MBG NPs) or boron-doped MBG NPs (BMBG NPs) at 10 and 20 wt% concentrations. The mixture was stirred and incubated overnight in a water bath at 50 0 C to ensure complete solubility. A sterile dual-syringe system was used to mix the alginate solution with 20 wt% calcium chloride solution, forming the hydrogel upon extrusion. Then, constructed hydrogel specimens from all groups were characterized by FTIR, SEM, water uptake percentage (WA%), bioactivity and ion release, and cytotoxicity. Statistical analysis was done using One-Way ANOVA test for comparisons between groups, followed by multiple pairwise comparisons using Bonferroni adjusted significance level (p < 0.05)., Results: Alginate/BMBG loaded groups exhibited remarkable increase in porosity and pore size diameter [IIB1 (168), IIB2 (183) (µm)]. Similarly, WA% increased (~ 800%) which was statistically significant (p < 0.05). Alginate/BMBG loaded groups exhibited the strongest bioactive capability displaying prominent clusters of hydroxyapatite precipitates on hydrogel surfaces. Ca/P ratio of precipitates in IIA2 and IIB1 (1.6) were like Ca/P ratio for stoichiometric pure hydroxyapatite (1.67). MTT assay data revealed that the cell viability % of human gingival fibroblast cells have declined with increasing the concentration of both powders and hydrogel extracts in all groups after 24 and 48 h but still higher than the accepted cell viability % of (˃70%)., Conclusions: The outstanding laboratory performance of the injectable alginate/BMBGNPs (20 wt%) composite hydrogel suggested it as promising candidate for pulpotomy filling material potentially enhancing dentin regeneration in clinical applications., (© 2024. The Author(s).)

Conventional pulp capping materials have limited anti-inflammatory capacity. It is necessary to develop more effective pulp capping material for the treatment of inflamed pulps. Tannic acid (TA) is a natural, water-soluble polyphenol with antimicrobial and anti-inflammatory properties. This study aimed to investigate the effects of a tannin-containing hydroxypropyl chitin hydrogel (HPCH/TA hydrogel) as an innovative pulp capping material. The physicochemical properties of the composite hydrogels were characterized. The effects of HPCH/TA hydrogel as a pulp capping material were evaluated in vitro and in vivo. The underlying mechanism of the anti-inflammatory effects of HPCH/TA hydrogel was explored. The HPCH/TA hydrogel demonstrated favorable temperature sensitivity, injectability, and antibacterial properties. In vitro, the HPCH/TA hydrogel effectively promoted the proliferation of human dental pulp cells and inhibited interleukin-1β, interleukin-6, and tumor necrosis factor-α expression, possibly by suppressing the nuclear factor kappa-B pathway. In vivo, on the fourth day after capping, the HPCH/TA hydrogel group showed lower inflammatory scores compared to the control and iRoot BP Plus (commercial pulp capping material) group. By the sixth week, complete reparative dentin formation was observed in the HPCH/TA hydrogel group, with no difference in thickness compared to the iRoot BP Plus group. Collectively, the HPCH/TA hydrogel holds promise as a bioactive pulp capping material for promoting the repair of inflamed pulp in vital pulp therapy.