Pulp Tissue Research Articles

A novel deep learning-based pipeline architecture for pulp stone detection on panoramic radiographs.

Pulp stones are ectopic calcifications located in pulp tissue. The aim of this study is to introduce a novel method for detecting pulp stones on panoramic radiography images using a deep learning-based two-stage pipeline architecture. The first stage involved tooth localization with the YOLOv8 model, followed by pulp stone classification using ResNeXt. 375 panoramic images were included in this study, and a comprehensive set of evaluation metrics, including precision, recall, false-negative rate, false-positive rate, accuracy, and F1 score was employed to rigorously assess the performance of the proposed architecture. Despite the limited annotated training data, the proposed method achieved impressive results: an accuracy of 95.4%, precision of 97.1%, recall of 96.1%, false-negative rate of 3.9%, false-positive rate of 6.1%, and a F1 score of 96.6%, outperforming existing approaches in pulp stone detection. Unlike current studies, this approach adopted a more realistic scenario by utilizing a small dataset with few annotated samples, acknowledging the time-consuming and error-prone nature of expert labeling. The proposed system is particularly beneficial for dental students and newly graduated dentists who lack sufficient clinical experience, as it aids in the automatic detection of pulpal calcifications. To the best of our knowledge, this is the first study in the literature that propose a pipeline architecture to address the PS detection tasks on panoramic images.

ATF3 triggers M2 macrophage polarization to protect against pulp inflammation through WNT4 regulation.

Pulpitis is a common inflammatory oral disease that can lead to pulp necrosis. The aim of this study is to investigate the expression and regulatory mechanisms of ATF3, a potential therapeutic marker, in pulpitis. A mouse pulpitis model with different degrees of inflammation is established, and the expression of ATF3 in pulpitis is explored. The histological features of healthy pulp and pulpitis are analyzed by HE staining, and classical inflammatory factors are detected by immunohistochemistry (IHC). In an in vitro study, we investigate the role of ATF3 in the regulation of WNT4 transcription and explore the effects of the ATF3/WNT4 axis on the polarization of RAW264.7 macrophages, the inflammatory response and the osteogenic differentiation of human dental pulp stem/stromal cells (hDPSCs). Our results show that ATF3 is expressed at low levels in inflamed pulp tissues; overexpression of ATF3 reduces the area of pulp necrosis, decreases the level of pro-inflammatory factors, and promotes macrophage polarization toward the M2 type. Furthermore, we reveal that ATF3 binds to the WNT4 promoter region and positively regulates the expression of WNT4 and that ATF3 downregulates M1 markers and increases the expression of M2 markers by regulating WNT4 expression. In addition, ATF3 promotes the osteogenic differentiation of dental pulp stem cells. In summary, this study reveals that ATF3 promotes M2 macrophage polarization by regulating WNT4, which in turn inhibits pulpal inflammatory responses and promotes the osteogenic differentiation of dental pulp stem cells. These findings suggest that ATF3 may be a potential target for pulpitis treatment.

Efficacy of non -thermal pressure plasma versus other modalities for disinfection of primary root canals

BackgroundEndodontic treatment aims in the preservation of extremely carious primary teeth. For root canal therapy to be successful, root canals must be properly prepared and effectively irrigated .Therefore, it is necessary to select the proper root canal disinfection method to preserve the primary tooth.ObjectiveThis research was carried out to compare non-thermal pressure plasma (NTPP), diode laser, propolis, and chlorhexidine (CHX) efficacy for disinfection of deciduous anterior root canals contaminated with Enterococcus Faecalis (E. faecalis) after sterilization by gamma radiation.MethodsIn this study, forty extracted single-rooted primary anterior teeth were used. All teeth were cleaned, disinfected, and stored till use. Gaining access was provided till reaching the orifices of canals, all pulp tissue debris was removed, and root canals of all teeth were prepared. Standardized 8 mm root length was obtained through crown decronation below the cemento-enamel junction. Samples were sterilized by gamma radiation then the bacterial suspension was inoculated inside root canals. Specimens divided into four main groups; ten samples each group. Group I: Samples irrigated with chlorhexidine. Group II: Samples treated with diode laser. Group III: Samples irrigated with Ethanolic extract of propolis. Group IV: Samples treated with NTPP.ResultsA significant difference was found between values measured before and after four irrigation types (p < 0.001) for CHX, Diode Laser, NTPP and (P = 0.035) for Propolis. The highest values of colony reduction measured before and after irrigation were for NTPP (4.06 ± 0.88). Maximum reduction in colony-forming units was recorded in the NTPP group (98.79%), while the lowest reduction in colony-forming units was recorded in Propolis group (81.99%).ConclusionAll tested methods (CHX, NTPP, Diode laser and Propolis) decreased colony count, with the highest reduction noted in group treated by NTPP and the least reduction noticed in Propolis treated group.

Clinical treatment difficulties and strategies for palatogingival groove

Palatogingival groove (PGG) is a common developmental deformity, which is prone to retain plaque, leading to local periodontal inflammation. Furthermore, the infection can spread to the dental pulp tissue through the PGG, eventually resulting in combined periodontal-endodontic lesions. The treatment methods include PGG treatment, periodontal treatment, endodontic treatment, etc. The abnormal anatomical structure of PGG will influence the treatment and prognosis of the affected tooth. Teeth with combined periodontal-endodontic lesions often have poor curative effect and prognosis. How to early intervene and improve its prognosis has become a hot and difficult issue for dentists in recent years. This article analyzes the challenges in the treatment of PGG and puts forward the corresponding strategies, so as to provide reference for the clinical diagnosis and treatment of PGG.

A Cerium Oxide Loaded Hyaluronic Acid Nanosystem Remits Glucose Oxidative Stress-Induced Odontoblasts Mitochondrial Apoptosis through Regulation of PGAM5 Pathway.

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.

Comparative study of pulpal response following direct pulp capping using synthesized fluorapatite and hydroxyapatite nanoparticles

ObjectiveThis 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 methodsA 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.ResultsThe 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.ConclusionMTA 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.

Microspheres of stem cells from human exfoliated deciduous teeth exhibit superior pulp regeneration capacity

ObjectivesEngineering spheroids to create three-dimensional (3D) cell cultures has gained increasing attention in recent years due to their potential advantages over traditional two-dimensional (2D) tissue culture methods. Stem cells derived from human exfoliated deciduous teeth (SHEDs) demonstrate significant potential for pulpal regeneration applications. Nevertheless, the feasibility of microsphere formation of SHEDs and its impact on pulpal regeneration remain unclear. MethodsIn this study, SHEDs were isolated, identified, and cultured in ultra-low attachment six-well plates to produce SHED microspheres. The biological properties of SHED microspheres were compared to those of traditional 2D culture using live-dead staining, Alizarin red staining, Oil-red O staining, scratch experiments, Immunofluorescence, Transmission electron microscopy scan, Western blotting, RNA sequencing, and a nude mice subcutaneous transplantation model. ResultsWe found SHED cells can form microspheres with a dense internal structure. SHED microspheres exhibited notable advantages over SHED cells in terms of biological properties, maintaining cell activity and enhancing cell differentiation, migration, and stemness in vitro. RNA-seq revealed that the SHED microspheres potentially influenced cell development, regulation of neurogenesis, skeletal system development, tissue morphogenesis singling pathway. In vivo, SHED microspheres promoted the generation of pulp tissue in dental pulp compared to traditional 2D culture. ConclusionsMicrosphereization of SHED through 3D cell culture enhances its pulp regeneration capacity, presenting a novel strategy for dental pulp regeneration and the clinical treatment of dental pulp diseases.

Clinical and radiographic evaluation of two different apexification protocols in traumatized immature permanent incisors.

Dental trauma can cause damage to the pulp tissue in immature teeth. Revascularization therapy is a possible option in the treatment of non-vital, immature permanent teeth with a history of trauma. The aim of this prospective study was to evaluate the radiographic and clinical results of immature teeth with a history of trauma treated by regenerative endodontic procedures and mineral trioxide aggregate apexification techniques. Forty-one patients aged between 7 and 12 years with traumatized immature permanent maxillary incisors were included in the study. These patients were divided into two groups: those who had previously received endodontic treatment and those who had not. Twenty-four patients who applied directly to the university clinic and had not received endodontic treatment before were included in the regenerative endodontic protocol group (Group 1). Seventeen patients who had previously undergone endodontic intervention on their relevant teeth were included in the mineral trioxide aggregate apexification group (Group 2). The patients were followed for a period of 24 months. Clinical success rates were evaluated, and pre-treatment and control radiographs were analyzed to calculate the percentage increase in root dentin width and root length. After 24 months of follow-up, positive periapical healing was detected in the radiographic findings in the majority of cases. In Group 1, a limited increase in root length and root dentin width was observed, while a narrowing in the apical opening was evident. The radiographic evaluation of two of the cases, which could be accessed 11 years later, emphasized the importance of long-term follow-up in assessing the effectiveness of the chosen methods. The revascularization method is a treatment option that has positive results in terms of root development in teeth with necrotic pulp as a result of trauma.

Small Extracellular Vesicles Derived from Lipopolysaccharide-Treated Stem Cells from the Apical Papilla Modulate Macrophage Phenotypes and Inflammatory Interactions in Pulpal and Periodontal Tissues.

Inflammation significantly influences cellular communication in the oral environment, impacting tissue repair and regeneration. This study explores the role of small extracellular vesicles (sEVs) derived from lipopolysaccharide (LPS)-treated stem cells from the apical papilla (SCAP) in modulating macrophage polarization and osteoblast differentiation. SCAPs were treated with LPS for 24 h, and sEVs from untreated (SCAP-sEVs) and LPS-treated SCAP (LPS-SCAP-sEVs) were isolated via ultracentrifugation and characterized using transmission electron microscopy, Western blot, and Tunable Resistive Pulse Sensing. LPS-SCAP-sEVs exhibited characteristic exosome morphology (~100 nm diameter) and expressed vesicular markers (CD9, CD63, CD81, and HSP70). Functional analysis revealed that LPS-SCAP-sEVs promoted M1 macrophage polarization, as evidenced by the increased pro-inflammatory cytokines (IL-6 and IL-1β) and the reduced anti-inflammatory markers (IL-10 and CD206), while impairing the M2 phenotype. Additionally, LPS-SCAP-sEVs had a minimal impact on SCAP metabolic activity or osteogenic gene expression but significantly reduced mineralization capacity in osteogenic conditions. These findings suggest that sEVs mediate the inflammatory interplay between SCAP and macrophages, skewing macrophage polarization toward a pro-inflammatory state and hindering osteoblast differentiation. Understanding this sEV-driven communication axis provides novel insights into the cellular mechanisms underlying inflammation in oral tissues and highlights potential therapeutic targets for modulating extracellular vesicle activity during acute inflammatory episodes.

Analysis of pulp histological response to pulpotomy performed with white mineral trioxide aggregate mixed with 2.25% sodium hypochlorite gel in humans: a randomized controlled clinical trial

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.

Effects of Hispidulin on the Osteo/Odontogenic and Endothelial Differentiation of Dental Pulp Stem Cells.

Background: Human dental pulp stem cells (HDPSCs) with multi-lineage differentiation potential and migration ability are required for HDPSC-based bone and dental regeneration. Hispidulin is a naturally occurring flavonoid with diverse pharmacological activities, but its effects on biological properties of HDPSCs remain unknown. Therefore, we investigated the effects of hispidulin on the differentiation potential and migration ability of HDPSCs and elucidated their underlying mechanisms. Methods: The osteo/odontogenic capacity of HDPSCs was assessed using the alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining. The migration ability of HDPSCs was evaluated using a scratch wound assay. Furthermore, the endothelial differentiation of HDPSCs was examined by using a capillary sprouting assay and by assessing CD31 expression. Results: Hispidulin significantly enhanced the osteo/odontogenic differentiation of HDPSCs with increased expression of osteo/odontogenic differentiation markers. Hispidulin increased the migration of HDPSCs, which was mediated by the upregulation of C-X-C chemokine receptor type 4 (CXCR4). The treatment of HDPSCs with hispidulin enhanced the differentiation of HDPSCs into endothelial cells, as evidenced by increased capillary sprouting and endothelial marker expression. In addition, we demonstrated that hispidulin activated the ERK1/2 signaling, and its inhibition by U0126 significantly suppressed the hispidulin-induced endothelial differentiation of HDPSCs. Conclusions: These findings demonstrate that hispidulin effectively promotes the osteo/odontogenic and endothelial differentiation, and migration of HDPSCs. These results suggest that hispidulin may have potential therapeutic applications in dental pulp regeneration and tissue engineering.

Biobanking of human teeth for stem cell storage: preserving stem cells for future needs

Human teeth serve as a potential reservoir for post-embryonic mesenchymal dental stem cells. Researchers have identified and isolated seven types of dental stem cells from pulp and periodontal ligament tissues. These cells have a wide range of clinical applications across the fields of medicine and dentistry due to their increased proliferative nature. Biobanking is a concept for storing human biological tissues to preserve and isolate stem cells. Until now, haematopoietic tissues have been the area of focus for biobanking facilities. Extracted human teeth may serve as a valuable resource for the isolation and preservation of dental stem cells for future therapeutic benefits. The current narrative review was planned to focus on the workflow of the teeth biobanking process, its isolation, cryopreservation, and potential therapeutic uses in medical and dental specialties. Key Words: Stem cells, Biobank, Dental stem cells.

Possible Involvement of X-Box Binding Protein-1 in the Onset of Pulpitis.

Endoplasmic reticulum (ER) stress plays important roles not only in stress avoidance, but also in cell differentiation and maturation, cell proliferation, and promotion of bone formation. This study aimed to investigate the involvement of ER stress in the onset of pulpitis. Immunohistochemical analysis was conducted on human teeth extracted for orthodontic reasons. The effects of tunicamycin (TM), an inducer of ER stress, lipopolysaccharide (LPS), and 4μ8c, an inhibitor of inositol-requiring enzyme 1 (IRE1) on cultured human dental pulp cells (hDPCs) were also examined. The expressions of two ER stress markers, X-box binding protein (XBP)-1 and binding immunoglobulin protein (BiP)/78 kDa glucose-regulated protein (GRP78), were found in the human pulp tissues of a decayed tooth that had not developed irreversible acute pulpitis, but not in an impacted tooth without inflammation in pulp tissue. Both TM and LPS increased the mRNA levels of XBP-1, interleukin (IL)-6, and IL-8, whereas TM, but not LPS, enhanced the mRNA expression of BiP/GRP78 in hDPCs. 4μ8c significantly suppressed the increased level of XBP-1 by LPS. This study is the first to demonstrate that XBP-1, in addition to inflammatory cytokines, may participate in the onset of pulpitis through IRE1. These findings provide a more comprehensive understanding of pulpitis pathogenesis through the cooperation of ER stress and inflammatory cytokines.

Cannabidiol alleviates LPS-inhibited odonto/osteogenic differentiation in human dental pulp stem cells invitro.

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. 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. 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. 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.

LL-37 regulates odontogenic differentiation of dental pulp stem cells in an inflammatory microenvironment

BackgroundInflammation often causes irreversible damage to dental pulp tissue. Dental pulp stem cells (DPSCs), which have multidirectional differentiation ability, play critical roles in the repair and regeneration of pulp tissue. However, the presence of proinflammatory factors can affect DPSCs proliferation, differentiation, migration, and other functions. LL-37 is a natural cationic polypeptide that inhibits lipopolysaccharide (LPS) activity, enhances cytokine production, and promotes the migration of stem cells. However, the potential of LL-37 in regenerative endodontics remains unknown. This study aimed to investigate the regulatory role of LL-37 in promoting the migration and odontogenic differentiation of DPSCs within an inflammatory microenvironment. These findings establish an experimental foundation for the regenerative treatment of pulpitis and provide a scientific basis for its clinical application.Materials and methodsDPSCs were isolated via enzyme digestion combined with the tissue block adhesion method and identified via flow cytometry. The impact of LL-37 on the proliferation of DPSCs was evaluated via a CCK-8 assay. The recruitment of DPSCs was assessed through a transwell assay. The mRNA expression levels of inflammatory and aging-related genes were assessed via reverse transcription‒polymerase chain reaction (RT‒PCR), western blotting, and enzyme‒linked immunosorbent assay (ELISA). The odontogenic differentiation of DPSCs was assessed through alkaline phosphatase (ALP) staining, alizarin red staining, and RT‒PCR analysis.ResultsLL-37 has the potential to enhance the migration of DPSCs. In an inflammatory microenvironment, LL-37 can suppress the expression of genes associated with inflammation and aging, such as TNF-α, IL-1β, IL-6, P21, P38 and P53. Moreover, it promotes odontogenic differentiation in DPSCs by increasing ALP activity, increasing calcium nodule formation, and increasing the expression of dentin-related genes such as DMP1, DSPP and BSP.ConclusionThese findings suggest that the polypeptide LL-37 facilitates the migration of DPSCs and plays a crucial role in resolving inflammation and promoting cell differentiation within an inflammatory microenvironment. Consequently, LL-37 has promising potential as an innovative therapeutic approach for managing inflammatory dental pulp conditions.

Dehydrated Human Amnion-Chorion Membrane as a Bioactive Scaffold for Dental Pulp Tissue Regeneration.

The dehydrated human amnion-chorion membranes (dHACMs) derived from the human placenta have emerged as a promising biomaterial for dental pulp regeneration owing to their unique biological and structural properties. The purpose of this review is to explore the potentials of dHACMs in dental pulp tissue engineering, focusing on their ability to promote cellular proliferation, differentiation, angiogenesis, and neurogenesis. dHACMs are rich in extracellular matrix proteins and growth factors such as TGF-β1, FGF2, and VEGF. They also exhibit significant anti-inflammatory and antimicrobial properties, creating an optimal environment for dental pulp regeneration. The applications of dHACMs in regenerative endodontic procedures are discussed, highlighting their ability to support the formation of dentin and well-vascularized pulp-like tissue. This review demonstrates that dHACMs hold significant potential for enhancing the success of pulp regeneration and offer a biologically based approach to preserve tooth vitality and improve tooth survival. Future research is expected to focus on conducting long-term clinical studies to establish their efficacy and safety.

The Effectiveness of Bajakah Wood Extract (Spatholobus Littoralis Hassk) From South Kalimantan against Candida Albicans ATCC 10231

Root canal treatment (RCT) is a procedure performed to remove inflamed or infected pulp tissue. Candida albicans is the most pathogenic type of fungus found in the oral cavity, accounting for 80%. Bajakah wood (Spatholobus littoralis Hassk) contains antifungal components such as phenols, flavonoids, tannins, and saponins. This research is to determine the antifungal effectiveness of Bajakah wood (Spatholobus littoralis Hassk) against Candida albicans. This research is an experimental laboratory study using the dilution method in the laboratory. A total of 20 samples of Candida albicans ATCC 10231 cultures in Sabouraud Dextrose Broth medium were tested. The treatment variations of Bajakah wood were concentrations of 50%, 75%, 100%, and 2.5% NaOCl (K+) with 5 repetitions. The results showed that the average number of fungal growths at 50% concentration was 387, at 75% concentration it was 449.4, while at 100% concentration it was 2.4, and in the positive control it was 1. Data analysis using the Kruskal-Wallistest showed significant differences (p&lt;0.05) across all treatment groups. It can be concluded that Bajakah wood at 100% concentration has the highest antifungal content and is effective in inhibiting the growth of Candida albicans.

Guidance for evaluating biomaterials' properties and biological potential for dental pulp tissue engineering and regeneration research.

Dental pulp regeneration is a complex and advancing field that requires biomaterials capable of supporting the pulp's diverse functions, including immune defense, sensory perception, vascularization, and reparative dentinogenesis. Regeneration involves orchestrating the formation of soft connective tissues, neurons, blood vessels, and mineralized structures, necessitating materials with tailored biological and mechanical properties. Numerous biomaterials have entered clinical practice, while others are being developed for tissue engineering applications. The composition and a broad range of material properties, such as surface characteristics, degradation rate, and mechanical strength, significantly influence cellular behavior and tissue outcomes. This underscores the importance of employing robust evaluation methods and ensuring precise and comprehensive reporting of findings to advance research and clinical translation. This article aims to present the biological foundations of dental pulp tissue engineering alongside potential testing methodologies and their advantages and limitations. It provides guidance for developing research protocols to evaluate the properties of biomaterials and their influences on cell and tissue behavior, supporting progress toward effective dental pulp regeneration strategies.

CRISPR-Edited DPSCs, Constitutively Expressing BDNF Enhance Dentin Regeneration in Injured Teeth.

Dental caries is one of the most common health issues worldwide arising from the complex interactions of bacteria. In response to harmful stimuli, desirable outcome for the tooth is the formation of tertiary dentin, a protective reparative process that generates new hard tissue. This reparative dentinogenesis is associated with significant inflammation, which triggers the recruitment and differentiation of dental pulp stem cells (DPSCs). Previously, we have shown that brain-derived neurotrophic factor (BDNF) and its receptor TrkB, key mediators of neural functions, are activated during the DPSC-mediated dentin regeneration process. In this study, we further define the role of inflammation in this process and apply stem cell engineering to enhance dentin regeneration in injured teeth. Our data show that TrkB expression and activation in DPSCs rapidly increase during odontogenic differentiation, further amplified by inflammatory inducers and mediators such as TNFα, LTA, and LPS. An in vivo dentin formation assessment was conducted using a mouse pulp-capping/caries model, where CRISPR-engineered DPSCs overexpressing BDNF were transplanted into inflamed pulp tissue. This transplantation significantly enhanced dentin regeneration in injured teeth. To further explore potential downstream pathways, we conducted transcriptomic profiling of TNFα-treated DPSCs, both with and without TrkB antagonist CTX-B. The results revealed significant changes in gene expression related to immune response, cytokine signaling, and extracellular matrix interactions. Taken together, our study advances our understanding of the role of BDNF in dental tissue engineering using DPSCs and identifies potential therapeutic avenues for improving dental tissue repair and regeneration strategies.

Insulin-like growth factor binding protein 7 identified in aged dental pulp by single-cell RNA sequencing.

Aging influences the regenerative and reparative functions of dental pulp, and an in-depth and complete understanding of aged dental pulp is highly important. This study aimed to explore the heterogeneity of young and aged dental pulp tissue via single-cell RNA sequencing (scRNA-seq), search novel markers of aged dental pulp, and further explore their mechanism. ScRNA-seq was employed to analyze the heterogeneity of young and aged dental pulp tissue, and immunohistochemical staining was used to detect new marker Insulin-like Growth Factor Binding Protein 7 (IGFBP7) in aged dental pulp. Differentially expressed genes (DEGs) between young and aged dental pulp tissue related with senescence-associated secretory phenotype (SASP) were validated in aging model of H2O2-induced dental pulp fibroblast (DPF). The effect of IGFBP7 on cellular senescence were validated by SA-β-Gal, γ-H2AX, and F-actin cytoskeletal staining. RNA-seq was used to analyze the mechanism of IGFBP7 alleviating senescence of H2O2-induced DPFs. A total of 32,012 cells were sequenced from 8 dental pulp samples and categorized into 8 main clusters, including fibroblasts (FB), endothelial cells, monocytes, T cells, B cells, mesenchymal stem cells, Schwann cells, and nonmyelinating ScCs. The ratio of fibroblasts was the highest, and FB1 was the largest subcluster of fibroblasts in the young group. In aged dental pulp, the ratio of fibroblasts was relatively low, and fibroblasts had more cellular communication with other cell types in fibroblast growth factor (FGF) and insulin-like growth factor (IGF) signal pathways. IGFBP7 was significantly upregulated in the aged group. Recombinant IGFBP7 reduced the senescence of H2O2-induced DPFs. These findings offer insights into the mechanisms of dental pulp aging and enhance our understanding of dental pulp at the single-cell level. Further comprehensive studies are required to clarify the exact mechanisms through which IGFBP7 influences dental pulp aging.