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

Physiological root resorption of deciduous teeth is a normal phenomenon occurring during the developmental stages of children. Previous research has indicated the pivotal role of the inflammatory microenvironment in this process, although the specific mechanisms remain unclear. This study is aimed at elucidating the involvement of the alpha7 nicotinic acetylcholine receptors (α7 nAChR)-autophagy axis in the regulation of the inflammatory microenvironment during physiological root resorption in deciduous teeth. Samples were collected from deciduous teeth at various stages of physiological root resorption, and deciduous dental pulp stem cells (DDPSCs) were isolated and cultured during the mid-phase of root resorption. The findings revealed a substantial infiltration of the pulp of deciduous teeth at the mid-phase of root resorption, characterized by elevated expression levels of α7 nAChR and IL-1β. Significantly increased IL-1β and α7 nAChR expressions were observed in DDPSCs during the mid-phase of root resorption, with α7 nAChR demonstrating a regulatory effect on IL-1β. Moreover, evidence suggested that mechanical stress may act as a trigger, regulating autophagy and IL-1 expression via α7 nAChR. In conclusion, mechanical stress was identified as a regulator of autophagy in DDPSCs through α7 nAChR, influencing the expression of IL-1β and contributing to the formation of the inflammatory microenvironment. This mechanism plays a crucial role in the physiological root resorption of deciduous teeth. Key messages: The pulp of deciduous teeth at mid-phase of root resorption was heavily infiltrated with high expression of α7nAChR and IL-1β. α7 nAChR acts as an initiating factor to regulate IL-1β through autophagy in DDPSCs. Mechanical stress can regulate autophagy of DDPSCs through α7 nAChR and thus affect IL-1β expression and inflammatory microenvironment formation in physiological root resorption in deciduous teeth. [ABSTRACT FROM AUTHOR]

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

Dental pulp stem cells (DPSC) have shown osteogenic and bone regenerative potential. Improving the in situ bone regeneration potential of DPSC is crucial for their application as seed cells during bone defect reconstruction in clinics. This study aimed to develop DPSC-derived organoid-like microspheroids as effective seeds for bone tissue engineering applications. DPSC osteogenic microspheroids (70 μm diameter) were cultured in a polydimethylsiloxane-mold-based agarose-gel microwell-culture-system with or without cannabidiol (CBD)-treatment. Results of in vitro studies showed higher osteogenic differentiation potential of microspheroids compared with 2D-cultured-DPSC. CBD treatment further improved the osteogenic differentiation potential of microspheroids. The effect of CBD treatment in the osteogenic differentiation of microspheroids was more pronounced compared with that of CBD-treated 2D-cultured-DPSC. Microspheroids showed a higher degree of bone regeneration in nude mice calvarial bone defect compared to 2D-cultured-DPSC. CBD-treated microspheroids showed the most robust in situ bone regenerative potential compared with microspheroids or CBD-treated 2D-cultured-DPSC. According to mRNA sequencing, bioinformatic analysis, and confirmation study, the higher osteogenic potential of CBD-treated microspheroids was mainly attributed to WNT6 upregulation. Taken together, DPSC microspheroids have robust osteogenic potential and can effectively translate the effect of in vitro osteoinductive stimulation during in situ bone regeneration, indicating their application potential during bone defect reconstruction in clinics. Dental pulp stem cells-derived microspheroids have robust osteogenic potential and can effectively translate the effect of in vitro osteoinductive stimulation during in situ bone regeneration, indicating their application potential during bone defect reconstruction in the clinic. [ABSTRACT FROM AUTHOR]

Medication-related osteonecrosis of the jaw is a serious disease occurring in patients with cancer and osteoporosis, who are undergoing treatment with antiresorptive agents (ARAs) such as bisphosphonate (BP) or denosumab, an antibody targeting receptor activator of NF-κB ligand. Recently, stem cell-based therapy has been shown to be effective in preventing the development of bisphosphonate-related osteonecrosis of the jaw. However, studies on denosumab-related osteonecrosis of the jaw (DRONJ) remain limited. Here, the efficacy of treatment with dental pulp stem cell conditioned media (DPSC-CM) in preventing DRONJ in a murine model was evaluated. Local administration of DPSC-CM into the extraction socket of a mouse with DRONJ decreased the number of empty osteocyte lacunae and the prevalence of ONJ. In tissues surrounding the extraction sockets in the DPSC-CM-treated group, the expression of inflammatory cytokines was attenuated and that of osteogenesis-related molecules was enhanced compared to that in the control group. Further, the expression of Wnt signaling molecules, which had been suppressed, was improved. These findings collectively suggest that DPSC-CM prevents ONJ development in a murine DRONJ model. [ABSTRACT FROM AUTHOR]

Most of the conditions involving cartilaginous tissues are irreversible and involve degenerative processes. The aim of the present study was to fabricate a biocompatible fibrous and film scaffolds using electrospinning and casting techniques to induce chondrogenic differentiation for possible application in cartilaginous tissue regeneration. Polycaprolactone (PCL) electrospun nanofibrous scaffolds and PCL film were fabricated and incorporated with multi-walled carbon nanotubes (MWCNTs). Thereafter, coating of chondroitin sulfate (CS) on the fibrous and film structures was applied to promote chondrogenic differentiation of human dental pulp stem cells (hDPSCs). First, the morphology, hydrophilicity and mechanical properties of the scaffolds were characterized by scanning electron microscopy (SEM), spectroscopic characterization, water contact angle measurements and tensile strength testing. Subsequently, the effects of the fabricated scaffolds on stimulating the proliferation of human dental pulp stem cells (hDPSCs) and inducing their chondrogenic differentiation were evaluated via electron microscopy, flow cytometry and RT‒PCR. The results of the study demonstrated that the different forms of the fabricated PCL-MWCNTs scaffolds analyzed demonstrated biocompatibility. The nanofilm structures demonstrated a higher rate of cellular proliferation, while the nanofibrous architecture of the scaffolds supported the cellular attachment and differentiation capacity of hDPSCs and was further enhanced with CS addition. In conclusion, the results of the present investigation highlighted the significance of this combination of parameters on the viability, proliferation and chondrogenic differentiation capacity of hDPSCs seeded on PCL-MWCNT scaffolds. This approach may be applied when designing PCL-based scaffolds for future cell-based therapeutic approaches developed for chondrogenic diseases. [ABSTRACT FROM AUTHOR]

For successful root canal treatment, adequate chemomechanical instrumentation to eliminate microorganisms and pulp tissue is crucial. This study aims to assess the organic tissue dissolving activity of various irrigation solutions on bovine tooth pulp tissue. 40 extracted bovine mandibular anterior teeth (n = 10) were used for the study. Bovine pulp pieces (25 ± 5 mg) were placed in 1.5 ml Eppendorf tubes. Each tooth pulp sample was then covered with 1.5 ml of different irrigation solutions, dividing them into four groups: Group 1 with freshly prepared 5% Boric acid, Group 2 with 5% NaOCl, Group 3 with Irritrol, and Group 4 with Saline. Samples were left at room temperature for 30 min, then dried and reweighed. The efficacy of tissue dissolution ranked from highest to lowest was found to be NaOCl, Boric Acid, Irritrol, and saline (p < 0.05). It was observed that the decrease in the NaOCl group was greater than the decrease in the Irritrol and saline groups, and the decrease in the Boric acid group was significantly greater than the decrease in the saline group (p < 0.05). It also emphasizes the need for future studies to further investigate the effects of Irritrol and Boric Acid on tissue dissolution. [ABSTRACT FROM AUTHOR]

Objectives: To obtain and compare the protein profiles of supernumerary and normal permanent dental pulp tissues. Materials and methods: Dental pulp tissues were obtained from supernumerary and normal permanent teeth. Proteins were extracted and analyzed by liquid chromatography-tandem mass spectrometry (LC/MS-MS). Protein identification and quantification from MS data was performed with MaxQuant. Statistical analysis was conducted using Metaboanalyst to identify differentially expressed proteins (DEPs) (P-value < 0.05, fold-change > 2). Gene Ontology enrichment analyses were performed with gProfiler. Results: A total of 3,534 proteins were found in normal dental pulp tissue and 1,093 in supernumerary dental pulp tissue, with 174 DEPs between the two groups. This analysis revealed similar functional characteristics in terms of cellular component organization, cell differentiation, developmental process, and response to stimulus, alongside exclusive functions unique to normal permanent dental pulp tissues such as healing, vascular development and cell death. Upon examination of DEPs, these proteins were associated with the processes of wound healing and apoptosis. Conclusions: This study provides a comprehensive understanding of the protein profile of dental pulp tissue, including the first such profiling of supernumerary permanent dental pulp. There are functional differences between the proteomic profiles of supernumerary and normal permanent dental pulp tissue, despite certain biological similarities between the two groups. Differences in protein expression were identified, and the identified DEPs were linked to the healing and apoptosis processes. Clinical relevance: This discovery enhances our knowledge of supernumerary and normal permanent pulp tissue, and serves as a valuable reference for future studies on supernumerary teeth. [ABSTRACT FROM AUTHOR]

Background: Dental pulp regeneration therapy is a challenge to achieve early vascularization during treatment. Studying the regulatory mechanisms of vascular formation during human dental pulp development may provide insights for related therapies. In this study, we utilized single-cell sequencing analysis to compare the gene expression of dental pulp stem cells (DPSCs) and vascular endothelial cells (ECs) from developing and mature dental pulps. Method: Immunohistochemistry, Western blot, and real-time polymerase chain reaction (RT-PCR) were used to detect fibronectin 1 (FN1) expression and molecules, such as PI3K/AKT. Cell proliferation assay, scratch assay, tube formation assay and were used to investigate the effects of DPSCs on the vasculogenetic capability of ECs. Additionally, animal experiments involving mice were conducted. Result: The results revealed that DPSCs exist around dental pulp vasculature. FN1 expression was significantly higher in DPSCs from young permanent pulps than mature pulps, promoting HUVEC proliferation, migration, and tube formation via ITGA5 and the downstream PI3K/AKT signaling pathway. Conclusion: Our data indicate that intercellular communication between DPSCs and ECs mediated by FN1-ITGA5 signaling is crucial for vascularizationduring dental pulp development, laying an experimental foundation for future clinical studies. [ABSTRACT FROM AUTHOR]

Stem cell-based therapy is a potential alternative strategy for brain repair, with neural stem cells (NSC) presenting as the most promising candidates. Obtaining sufficient quantities of NSC for clinical applications is challenging, therefore alternative cell types, such as neural crest-derived dental pulp stem cells (DPSC), may be considered. Human DPSC possess neurogenic potential, exerting positive effects in the damaged brain through paracrine effects. However, a method for conversion of DPSC into NSC has yet to be developed. Here, overexpression of octamer-binding transcription factor 4 (OCT4) in combination with neural inductive conditions was used to reprogram human DPSC along the neural lineage. The reprogrammed DPSC demonstrated a neuronal-like phenotype, with increased expression levels of neural markers, limited capacity for sphere formation, and enhanced neuronal but not glial differentiation. Transcriptomic analysis further highlighted the expression of genes associated with neural and neuronal functions. In vivo analysis using a developmental avian model showed that implanted DPSC survived in the developing central nervous system and respond to endogenous signals, displaying neuronal phenotypes. Therefore, OCT4 enhances the neural potential of DPSC, which exhibited characteristics aligning with neuronal progenitors. This method can be used to standardise DPSC neural induction and provide an alternative source of neural cell types. [ABSTRACT FROM AUTHOR]

This in vitro study assessed peak temperature and temperature increase (ΔT) within the pulp chamber during different extended photoactivation techniques (EPT—applying similar radiant exposure values) to resin-based composites (RBCs) placed in a Class I cavity preparation in an extracted human lower third molar. A T-type thermocouple was placed in the pulp chamber and connected to a temperature analysis device (Thermes, Physitemp). The tooth was attached to an assembly simulating the in vivo environment (controlled baseline pulp chamber temperature and fluid flow). The real-time pulp chamber temperature was evaluated throughout the photoactivation (Bluephase N, Ivoclar Vivadent) of two bulk-fill RBCs: Tetric N Ceram Bulk Fill (TBF; shade: IVA; Ivoclar Vivadent); Surefill SDR flow + (SDR, shade: Universal; Dentsply Sirona), which were exposed to different curing techniques: 40 s-occlusal surface; 20 s-occlusal + 10 s-buccal + 10 s-lingual surfaces; 10 s-buccal + 10 s + lingual + 20 s-occlusal surfaces. Each EPT delivered 42.4 J/cm2. Vickers hardness (VHN) was measured on the removed, sectioned RBC restorations at the top and bottom middle areas after curing. ΔT and VHN data were analyzed using 2-way ANOVA followed by Bonferroni post-hoc test (α = 0.05). Peak temperature data were analyzed using one-way ANOVA and Dunnett's post-hoc test (α = 0.05). SDR showed higher ΔT values than TBF (p = 0.008) in some EPTs. Neither technique resulted in ΔT values greater than 5.5 °C. Both composites had acceptable bottom/top hardness ratios (greater than 80%), regardless of the photoactivation technique. The evaluated EPTs may be considered safe as a low-temperature increase was noticed within the pulp chamber. [ABSTRACT FROM AUTHOR]

The age estimation of an adult using methods accessible to the forensic routine is a goal pursued by forensic experts. Cameriere, Ferrante and Cingolani (2004) proposed the use of the pulp/tooth area ratio of canine teeth as a promising variable, but its reliability has shown conflicting results in the scientific literature. This article aimed to carry out a systematic review with meta-analysis to verify whether the pulp/tooth area ratio of canine teeth includes a variable that can be used alone to estimate dental age in adults. A systematic search was carried out in six databases using keywords related to the theme in Portuguese, English, and Spanish. The study selection process followed pre-established eligibility criteria. Assessments were carried out regarding risk of bias and publication bias of selected studies, and meta-analysis was carried out considering Pearson's correlation coefficient between pulp/tooth area ratio and chronological age as effect measure. Most selected studies showed low risk of bias; no publication bias was found when all studies were considered, and potential publication bias was found when outliers were removed. Despite the high heterogeneity among studies and the need for more research, it could be observed that the pulp/tooth area ratio has strong negative correlation with chronological age, and the pulp/tooth area ratio could be derived from both periapical radiographs and orthopantomographs. Therefore, it is suggested that there is scientific evidence that the pulp/tooth area ratio obtained from canine teeth is reliable for dental age estimation in adults. [ABSTRACT FROM AUTHOR]

The purpose of this study is to estimate chronological age and determine whether individuals were aged under or over 18 years using root pulp visibility (RPV) in cone-beam computed tomography (CBCT) images. The study included CBCT images of 699 individuals aged between 15 and 75 years. One thousand twenty-three mandibular second molar (2M) teeth were evaluated using Olze's RPV method in four stages. Descriptive statistics of the stages and the relationship between the stages and the chronological age were assessed. The distribution of the stages was analyzed according to the 18-year age threshold. There was a positive correlation between RPV stages and chronological ages in both sexes. For females and males, the mean ages of stage 0, stage 1, and stage 2, for females and males, were found as 27.21, 28.93, and 33.68 years, and 37.69, 40.9, and 44.88 years, respectively. Stage 0 and stage 1 were found both in individuals aged under and over 18 years, and stage 2 and stage 3 were not observed in individuals aged under 18 years. The presence of stage 2 and stage 3 may be an indication that an individual is aged over 18 years according to Olze's RPV age estimation method. For more reliable results, 2M teeth should be examined bilaterally in forensic science. [ABSTRACT FROM AUTHOR]

Cassava, an important subsistence crop in tropical countries, represents the third most important source of starch worldwide. Espírito Santo (Brazil) presents predominance of family farming, where table cassava is cultivated as a food crop. The objective of the study was to evaluate the genetic diversity, estimate the genetic parameters and quantify gains with the selection of traditional cassava genotypes collected in different regions of the state of Espírito Santo, using the methodology of the mixed models (REML/BLUP). A total of 106 genotypes were evaluated in three locations. Each planting date was considered an environment, totaling six environments. The evaluated traits were: shoot height (APH), total number of tuberous roots (NR); weight of commercial roots (WCR); total root weight (TWR), marketable root weight/total root ratio (MRTR); root cortex color (RCC), root pulp color (PC) and cooking time (CT). The studied cassava genotypes presented genetic diversity and selection potential. The highest heritabilities (h2 = 0.90; 0.75 and 0.75) were recorded for the traits NRT, WCR and TWR, respectively. Gains from selection were positive for all of the traits evaluated. Higher selection gains (GS = 21.77% and 20.15%) were observed for the WCR and TWR traits, considering animal and human consumption and (GS = 10.89% and 17.45%) for NRT and WCR, when intended for human consumption only. Genotypes 82, 76, 46, T3 and 2 stood out for selection purposes for animal and human consumption. Genotypes 2, 81, 69, 12 and 49 stood out for selection purposes for human food. The assessment of the diverse genotypes has uncovered a selection of superior candidates with tremendous potential for commercial crops, catering to both human and animal consumption markets. [ABSTRACT FROM AUTHOR]

A culture of cells expressing markers of mesenchymal stem cells (MSC) (CD73, CD90, CD44, CD29, and CD49b), but not hematopoietic cell markers, and capable of multilineage differentiation was isolated from the deciduous tooth pulp. Co-culturing with immature dendritic cells in the presence of LPS did not reveal an ability of the MSC to suppress the maturation of dendritic cells. On the contrary, co-culturing of MSC with monocytes in the presence of granulocyte-macrophage CSF and IL-4 led to complete suppression of monocyte differentiation into dendritic cells. However, long-term culturing of MSC from dental pulp showed that by the passage 11, they almost completely lose their suppressor ability. These results indicate that the immunological properties of MSC can change during culturing without changing their phenotypic markers. This should be taken into account when creating biomedical cell products. [ABSTRACT FROM AUTHOR]

Early diagnosis of cancer demands sensitive and accurate detection of cancer biomarkers in blood. Carbon dots (CDs) bio-functionalization with antibodies, peptides or aptamers have played significant role in cancer diagnosis and targeted cancer therapy. Herein, a biosensor for detection of cancer biomarker carcinoembryonic antigen (CEA) in blood serum has been designed using CDs bio-functionalized with HRP-conjugated CEA antibody (CUCDs@CEAAb2) as detection probe. CDs were synthesized by upscaling of cow urine, a nitrogen rich biomass waste, by hydrothermal method. Detection probe based on CDs resulted in 3.5 times higher sensitivity as compared to conventional electrochemical sandwich immunoassay. To further improve the sensor performance, hyper-branched polyethylenimine grafted poly amino aniline (PEI-g-PAANI) was used as the sensing interface, which enabled immobilization of higher amount of capture antibody. Detection of CEA in human blood serum coupled with wide linear range (0.5–50 ng/ml), good specificity, stability, reproducibility and low detection limit (10 pg/ml) signified the excellence of CUCDs based CEA immunosensor. CUCDs exhibited excitation wavelength dependent fluorescence property and showed strong blue emission under UV irradiation. MTT assay indicated that the material is not toxic towards human dental pulp stem cells (hDPSCs) and MG63 osteosarcoma cells (cell viability > 90%). The present study demonstrates a methodology for valorization of animal waste to a cost-effective carbon based functional nanomaterial for clinical detection of cancer biomarkers. [ABSTRACT FROM AUTHOR]

Leukemia inhibitory factor (LIF) has been recognized as a novel inflammatory modulator in inflammation-associated diseases. This study aimed to investigate the modulation of LIF in dental pulp inflammation. Experimental pulpitis was established in wild-type (WT) and Lif-deficient (Lif−/−) mice. Histological and immunostaining analyses were conducted to assess the role of LIF in the progression of pulpitis. Mouse macrophage cell line (RAW264.7) was treated with LPS to simulate an inflammatory environment. Exogenous LIF was added to this system to examine its modulation in macrophage inflammatory response in vitro. Primary bone marrow-derived macrophages (BMDMs) from WT and Lif−/− mice were isolated and stimulated with LPS to confirm the effect of Lif deletion on macrophage inflammatory response. Supernatants from LIF and LPS-treated human dental pulp cells (hDPCs) were collected and added to macrophages. Macrophage chemotaxis was assessed using transwell assays. The results showed an increased expression of LIF and LIFR with the progression of pulpitis, and LIFR was highly expressed in macrophages. Lif deficiency alleviated experimental pulpitis with the reduction of pro-inflammatory cytokines and macrophage infiltration. Exogenous LIF promoted inflammatory response of LPS-induced macrophages through a STAT3/p65-dependent pathway. Consistently, Lif deletion inhibited macrophage inflammatory response in vitro. Supernatants of LIF-treated hDPCs enhanced macrophage migration in LPS-induced inflammatory environment. Our findings demonstrated that LIF aggravates pulpitis by promoting macrophage inflammatory response through a STAT3/p65-dependent pathway. Furthermore, LIF plays a crucial role in driving the recruitment of macrophages to inflamed pulp tissue by promoting chemokine secretion in DPCs. [ABSTRACT FROM AUTHOR]

Reconstruction of irregular oral-maxillofacial bone defects with an inflammatory microenvironment remains a challenge, as chronic local inflammation can largely impair bone healing. Here, we used magnesium silicate nanospheres (MSNs) to load microRNA-146a-5p (miR-146a) to fabricate a nanobiomaterial, MSN+miR-146a, which showed synergistic promoting effects on the osteogenic differentiation of human dental pulp stem cells (hDPSCs). In addition, miR-146a exhibited an anti-inflammatory effect on mouse bone marrow-derived macrophages (BMMs) under lipopolysaccharide (LPS) stimulation by inhibiting the NF-κB pathway via targeting tumor necrosis factor receptor-associated factor 6 (TRAF6), and MSNs could simultaneously promote M2 polarization of BMMs. MiR-146a was also found to inhibit osteoclast formation. Finally, the dual osteogenic-promoting and immunoregulatory effects of MSN+miR-146a were further validated in a stimulated infected mouse mandibular bone defect model via delivery by a photocuring hydrogel. Collectively, the MSN+miR-146a complex revealed good potential in treating inflammatory irregular oral-maxillofacial bone defects. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) are well known for their beneficial effects, differentiation capacity and regenerative potential. Dental-derived MSCs (DSCs) are more easily accessible and have a non-invasive isolation method rather than MSCs isolated from other sources (umbilical cord, bone marrow, and adipose tissue). In addition, DSCs appear to have a relevant neuro-regenerative potential due to their neural crest origin. However, it is now known that the beneficial effects of MSCs depend, at least in part, on their secretome, referring to all the bioactive molecules (neurotrophic factors) released in the conditioned medium (CM) or in the extracellular vesicles (EVs) in particular exosomes (Exos). In this review, we described the similarities and differences between various DSCs. Our focus was on the secretome of DSCs and their applications in cell therapy for neurological disorders. For neuro-regenerative purposes, the secretome of different DSCs has been tested. Among these, the secretome of dental pulp stem cells and stem cells from human exfoliated deciduous teeth have been the most widely studied. Both CM and Exos obtained from DSCs have been shown to promote neurite outgrowth and neuroprotective effects as well as their combination with scaffold materials (to improve their functional integration in the tissue). For these reasons, the secretome obtained from DSCs in combination with scaffold materials may represent a promising tissue engineering approach for neuroprotective and neuro-regenerative treatments. [ABSTRACT FROM AUTHOR]

Hepatic fibrosis, also called cirrhosis, have wide prevalence worldwide for long yeas. Recently, many treatments for liver cirrhosis made marked progress, especially the umbilical cord-derived mesenchymal stromal cells (UCMSC) therapy. However, limited recourses and potential immune-related issues become the obstacles on UCMSC popularization in clinic. Therefore, we took dental pulp stem cells (DPSCs) into the consideration, since autologous DPSCs can be easily obtained without any ethnic or immune-related issues that heterogenous UCMSCs could encounter. We systematically compared the effects of both cell types and found that DPSCs had similar results to UCMSCs in regulating inflammation and reversing hepatic fibrosis. In our study, co-culturing T cells and PBMSCs showed that DPSCs have the ability to inhibit the proliferation of inflammatory cells and downregulate relevant inflammatory factors. In vitro and in vivo sterility tests confirmed the bio-safety of DPSCs. Moreover, the 1 year-aged mouse model demonstrated that DPSCs successfully reversed hepatic fibrosis. Overall, DPSCs demonstrated comparable effectiveness to UCMSCs in regulating inflammation and reversing hepatic fibrosis, particularly in the aged mouse model that represents middle-aged and elderly humans. Since autologous DPSCs avoid potential immune-related issues that heterogenous UCMSCs could encounter, they may be a better choice for stem cell-related therapies. [ABSTRACT FROM AUTHOR]

This study aims to experimentally compare the efficacy of different endodontic materials (iRoot BP Plus, Biodentine, MTA, Rootdent, and Trioxide) in the treatment of pulpitis and perforations on extracted tooth specimens. Additionally, the study aims to investigate the influence of iRoot BP Plus endodontic material on the regenerative processes following pulp amputation in laboratory animals. The secondary goal is to evaluate the effect of iRoot BP Plus on the restoration process in laboratory animals after pulp removal. The study presents a micropermeability analysis of the selected biomaterials performed on a sample of 50 single-rooted apical teeth in 2022. All teeth underwent endodontic treatment. Changes in molar morphology were investigated with eight laboratory animals (rabbits, 3 months old, all males) after simulated pulp removal and subsequent treatment with the iRoot BP Plus biomaterials. iRoot BP Plus appeared to be more effective in retrograde apical root filling than other biomaterials, as evidenced by its higher sealing effect. An experiment involving animal participants revealed the presence of protective adaptive mechanisms, which manifested in the form of an inflammatory process within 6 weeks after the dental pulp was removed. The connective tissue replaced the necrosis, and new capillaries began to form intensively. These dental outcomes suggest that iRoot BP Plus enables hermetical sealing in tooth restoration with good adhesion. Thus, it may have the ability to promote more active tissue regeneration after pulp removal. [ABSTRACT FROM AUTHOR]

In this study, poly (lactic-co-glycolic acid) (PLGA) microparticles loaded with cannabidiol (CBD) were synthesized (PLGA@CBD microparticles) and embedded up to 10 wt% in a chondroitin sulfate/polyvinyl alcohol hydrogel matrix. In vitro chemical, physical, and biological assays were carried out to validate the potential use of the modified hydrogels as biomaterials. The microparticles had spherical morphology and a narrow range of size distribution. CBD encapsulation efficiency was around 52%, loading was approximately 50%. Microparticle addition to the hydrogels caused minor changes in their morphology, FTIR and thermal analyses confirmed these changes. Swelling degree and total porosity were reduced in the presence of microparticles, but similar hydrophilic and degradation in phosphate buffer solution behaviors were observed by all hydrogels. Rupture force and maximum strain at rupture were higher in the modified hydrogels, whereas modulus of elasticity was similar across all materials. Viability of primary human dental pulp cells up to 21 days was generally not influenced by the addition of PLGA@CBD microparticles. The control hydrogel showed no antimicrobial activity against Staphylococcus aureus, whereas hydrogels with 5% and 10% PLGA@CBD microparticles showed inhibition zones. In conclusion, the PLGA@CBD microparticles were fabricated and successfully embedded in a hydrogel matrix. Despite the hydrophobic nature of CBD, the physicochemical and morphological properties were generally similar for the hydrogels with and without the CBD-loaded microparticles. The data reported in this study suggested that this original biomaterial loaded with CBD oil has characteristics that could enable it to be used as a scaffold for tissue/cellular regeneration. [ABSTRACT FROM AUTHOR]

Objectives: To investigate the response of pulp cells to the application of silver diamine fluoride (SDF) and potassium iodide (KI) on demineralized dentin. Materials and methods: The occlusal surfaces of human dentin discs (0.4 mm thick) with similar permeability were subjected to an artificial caries protocol, and then the discs were adapted into artificial pulp chambers. MDPC-23 cells were seeded on the healthy pulp dentin surface, while the demineralized surface was treated with SDF, KI, SDF + KI, or hydrogen peroxide (positive control—PC) (n = 8). The negative control (NC) received ultrapure water. After 24 h, cell viability (alamarBlue) and morphology (SEM) were evaluated. The extracts were then applied to new MDPC-23 cells seeded in culture plates to assess their viability and the formation of mineralized nodules (MN; Alizarin Red) after seven days. The data were analyzed using one-way analysis of variance/Tukey or Games-Howell tests (α = 5%). Results: SDF and PC significantly reduced the viability of cells seeded on discs (45.6% and 71.0%, respectively). Only cells treated with SDF or PC detached from the dentin substrate, while the remaining cells showed altered morphology. Cells in contact with extracts showed less reduction in viability, but it was still more toxic compared to NC. Only PC reduced MN deposition. SDF + KI or KI alone did not affect the cell response. Conclusions: SDF applied alone showed a mild to moderate transdentinal cytotoxic effect on pulp cells. However, the combination of SDF + KI reduced the cytotoxic effects. Both materials used alone or in combination did not affect the mineralization ability of pulp cells. Clinical relevance: Besides improving esthetic results, associating potassium iodide with silver diamine fluoride may reduce the transdentinal cytotoxic effects of this cariostatic agent on pulp cells. [ABSTRACT FROM AUTHOR]

Background: Pulpotomy is a treatment option for the preservation of pulp vitality in primary teeth with extensive caries. Propolis is a natural resinous substance with optimal antimicrobial, anti-inflammatory, and immune-regulatory properties. Thus, this study aimed to compare the cytotoxic and apoptosis-inducing effects of mineral trioxide aggregate (MTA), propolis, and MTA-propolis on immature dental pulp stem cells (IDPSCs). Methods: In this in vitro, experimental study, primary IDPSCs were exposed to propolis, MTA, and MTA-propolis for 24 and 72-h. The cytotoxicity and apoptosis-inducing effects were evaluated using the methyl thiazolyl tetrazolium (MTT) assay and flow cytometry, respectively. Data were analyzed using ANOVA and Tukey's test at 0.05 level of significance. Results: The cytotoxicity of MTA and MTA-propolis was higher than that of propolis alone at both 24/48 h. In addition, all tested concentrations showed higher biocompatibility at 72-h compared with 24-h (P < 0.0001). In the assessment of apoptosis, propolis-MTA showed higher cell viability compared with other materials (P < 0.0001). Conclusion: Propolis-MTA showed higher biocompatibility than MTA. Addition of propolis to MTA improved cell proliferation in the first 24-h. Also, the cytotoxicity of propolis was lower than other materials in the first 24-h. Thus, propolis may serve as a promising pulp capping agent given that its other properties are approved. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the prevalence of pulp stones in a Turkish paediatric cohort from the Isparta district using panoramic radiographs. Methods: Panoramic radiographs of 19,857 children and adolescents between the ages of 9 and 18 years were retrospectively analysed to determine the prevalence and distribution of pulp stones. Teeth with pulp stones were classified in terms of dentition, location, dental status and complete or incomplete root formation. Statistical analysis was carried out by applying Chi‑square and Mann–Whitney U. Univariate logistic regression analysis was applied to determine the factors affecting pulp stone status. Differences were considered as significant when p < 0.05. Results: Out of a total of 19,857 patients, pulp stones were identified in 6.5%. Among the 548,415 teeth examined, 0.96% had pulp stones. The incidence of pulp stones in girls (7.4%) was higher than in boys (5.7%) (p < 0.001). The prevalence of pulp stones increased with age and was significantly higher in molar teeth. A significant difference existed in the incidence of pulp stones according to the jaws (p < 0.001). In both jaws, no statistically significant difference was observed in terms of teeth with pulp stones being on the right and left (p = 0.834). Of molars with pulp stones, 91.8% had completed root formation. The majority of patients (37.7%) had pulp stones in four teeth. Conclusion: Knowing the incidence of pulp stones in paediatric patients by dentists, endodontists, and paediatric dentists will help prevent complications that may occur during applications by facilitating the determination of appropriate methods, especially during endodontic procedures. [ABSTRACT FROM AUTHOR]

Dentin hypersensitivity is an oral health concern affecting a large percentage of the world's adult population. Occlusion of the exposed dentinal tubules is among the treatment options available, and silver diammine fluoride (SDF) is an occluding agent used for interrupting or dampening the stimulus of the dental pulp nerves that produce pain. In addition to dentin permeability testing, the evaluation of desensitizing agents occluding dentinal tubules strongly relies on microscopic techniques, such as scanning electron microscopy (SEM). Limitations of SEM are that it provides only surface images that lack detailed information on the depth of penetration and amount of material present within the treated specimen, and it is prone to sample preparation artifacts. Here, we present high-resolution X-ray computed tomography (nano-CT) as a potential method for investigating dentin specimens with occluded tubules. We studied human dentin treated with SDF as an exemplary dentinal occlusion treatment option. We evaluated the silver deposits formed on the dentin surface region near the dentinal tubules and in the tubular regions using cross-section SEM, Energy Dispersive X-ray (EDX) analysis, and nano-CT. The resulting images obtained by SEM and nano-CT had comparable resolutions, and both techniques produced images of the tubules' occlusion. Nano-CT provided three-dimensional images adequate to quantitate tubule size and orientation in space. Moreover, it enabled clear visualization of dentinal tubules in any virtual plane and estimation of the amount and depth of occluding material. Thus, nano-CT has the potential to be a valuable technique for evaluating the occluding effects of virtually any material applied to dentinal tubules, supporting deciding between the best occluding treatment options. [ABSTRACT FROM AUTHOR]

The aim of this observational clinical study (OCS) was to determine the clinical anatomical diameter (CAD) in several dental groups, thus correlating them with gender, age, tooth/canal and pulpoperiradicular diagnosis. Three-hundred fifty-nine teeth/584 vital or necrotic root canals from patients of both genders and different ages composed the sample. After performing the necessary previous procedures, K-Flexofiles were used to determine the CAD. Then, the gender and age of the patients, as well as the pulpoperiradicular diagnosis of the teeth were tabulated to conduct the statistical analysis (p < 0.05). Of the 359 teeth/584 root canals evaluated, 208/342 were from women (mean age 38.85 ± 13.42 years) and 151/242 were from men (mean age 45.41 ± 14.90 years). Statistically significant differences between the CAD means of root canals from women and men were not identified (p = 0.411). The analysis of the correlation between the CAD and age also showed a total independence (p = 0.271). Teeth with pulp necrosis and asymptomatic apical periodontitis diagnosed radiographically (TPNAAPDR) had a significantly larger CAD mean than teeth with pulp necrosis and no asymptomatic apical periodontitis diagnosed radiographically (TPNNAAPDR) and teeth with vital pulp and normal apical tissues (TVPNAT) (p = 0.0297); and the last two did not differ statistically (p > 0.05). The largest CAD mean was observed in single canals of maxillary central incisors. The lowest values of this variable were identified in the mesiobuccal and mesial canals of maxillary and mandibular first molars, respectively. The CAD of the root canals was influenced only by the root canal/tooth and pulpoperiradicular diagnosis. [ABSTRACT FROM AUTHOR]

Dental pulp stem cells (DPSC) usually remain quiescent in the dental pulp tissue; however, once the dental pulp tissue is injured, DPSCs potently proliferate and migrate into the injury microenvironment and contribute to immuno-modulation and tissue repair. However, the key molecules that physiologically support the potent proliferation and migration of DPSCs have not been revealed. In this study, we searched publicly available transcriptome raw data sets, which contain comparable (i.e., equivalently cultured) DPSC and mesenchymal stem cell data. Three data sets were extracted from the Gene Expression Omnibus database and then processed and analyzed. MXRA5 was identified as the predominant DPSC-enriched gene associated with the extracellular matrix. MXRA5 is detected in human dental pulp tissues. Loss of MXRA5 drastically decreases the proliferation and migration of DSPCs, concomitantly with reduced expression of the genes associated with the cell cycle and microtubules. In addition to the known full-length isoform of MXRA5, a novel splice variant of MXRA5 was cloned in DPSCs. Recombinant MXRA5 coded by the novel splice variant potently induced the haptotaxis migration of DPSCs, which was inhibited by microtubule inhibitors. Collectively, MXRA5 is a key extracellular matrix protein in dental pulp tissue for maintaining the proliferation and migration of DPSCs. [ABSTRACT FROM AUTHOR]

We investigated the effect of femtosecond (fs) laser ablation of enamel and dentin for different pulse wavelengths: infrared (1030 nm), green (515 nm), and ultra-violet (343 nm) and for different pulse separations to determine the optimal irradiation conditions for the precise removal of dental hard tissues with the absence of structural and compositional damage. The ablation rates and efficiencies were established for all three laser wavelengths for both enamel and dentin at room temperature without using any irrigation or cooling system, and the surfaces were assessed with optical and scanning electron microscopy, optical profilometry, and Raman spectroscopy. We demonstrated that 515 nm fs irradiation provides the highest rate and efficiency for ablation, followed by infrared. Finally, we explored the temperature variations inside the dental pulp during the laser procedures for all three wavelengths and showed that the maximum increase at the optimum conditions for both infrared and green irradiations was 5.5 °C, within the acceptable limit of temperature increase during conventional dental treatments. Ultra-violet irradiation significantly increased the internal temperature of the teeth, well above the acceptable limit, and caused severe damage to tooth structures. Thus, ultra-violet is not a compatible laser wavelength for femtosecond teeth ablation. [ABSTRACT FROM AUTHOR]

The retina has restricted regeneration ability to recover injured cell layer because of reduced production of neurotrophic factors and increased inhibitory molecules against axon regrowth. A diseased retina could be regenerated by repopulating the damaged tissue with functional cell sources like mesenchymal stem cells (MSCs). The cells are able to release neurotrophic factors (NFs) to boost axonal regeneration and cell maintenance. In the current study, we comprehensively explore the potential of various types of stem cells (SCs) from oral cavity as promising therapeutic options in retinal regeneration. The oral MSCs derived from cranial neural crest cells (CNCCs) which explains their broad neural differentiation potential and secret rich NFs. They are comprised of dental pulp SCs (DPSCs), SCs from exfoliated deciduous teeth (SHED), SCs from apical papilla (SCAP), periodontal ligament-derived SCs (PDLSCs), gingival MSCs (GMSCs), and dental follicle SCs (DFSCs). The Oral MSCs are becoming a promising source of cells for cell-free or cell-based therapeutic approach to recover degenerated retinal. These cells have various mechanisms of action in retinal regeneration including cell replacement and the paracrine effect. It was demonstrated that they have more neuroprotective and neurotrophic effects on retinal cells than immediate replacement of injured cells in retina. This could be the reason that their therapeutic effects would be weakened over time. It can be concluded that neuronal and retinal regeneration through these cells is most likely due to their NFs that dramatically suppress oxidative stress, inflammation, and apoptosis. Although, oral MSCs are attractive therapeutic options for retinal injuries, more preclinical and clinical investigations are required. [ABSTRACT FROM AUTHOR]

The potential therapeutic role of the Dental Pulp Stem Cells Secretome (SECR) in a rat model of experimentally induced Temporomandibular Joint (TMJ) Osteoarthritis (OA) was evaluated. Proteomic profiling of the human SECR under specific oxygen tension (5% O2) and stimulation with Tumor Necrosis Factor-alpha (TNF-α) was performed. SECR and respective cell lysates (CL) samples were collected and subjected to SDS-PAGE, followed by LC-MS/MS analysis. The identified proteins were analyzed with Bioinformatic tools. The anti-inflammatory properties of SECR were assessed via an in vitro murine macrophages model, and were further validated in vivo, in a rat model of chemically-induced TMJ-OA by weekly recording of the head withdrawal threshold, the food intake, and the weight change, and radiographically and histologically at 4- and 8-weeks post-treatment. SECR analysis revealed the presence of 50 proteins that were enriched and/or statistically significantly upregulated compared to CL, while many of those proteins were involved in pathways related to "extracellular matrix organization" and "immune system". SECR application in vitro led to a significant downregulation on the expression of pro-inflammatory genes (MMP-13, MMP-9, MMP-3 and MCP-1), while maintaining an increased expression of IL-10 and IL-6. SECR application in vivo had a significant positive effect on all the clinical parameters, resulting in improved food intake, weight, and pain suppression. Radiographically, SECR application had a significant positive effect on trabecular bone thickness and bone density compared to the saline-treated group. Histological analysis indicated that SECR administration reduced inflammation, enhanced ECM and subchondral bone repair and regeneration, thus alleviating TMJ degeneration. [ABSTRACT FROM AUTHOR]

Recent progress in exosome based studies has revealed that they possess several advantages over cells, including "cell-free" properties, low immunogenicity and ethical controversy, high biological safety and effective action. These characteristics confer exosomes significant advantages that allow them to overcome the limitations associated with traditional "cell therapy" by circumventing the issues of immune rejection, scarcity of donor cells, heterogeneity, and ethical concerns. Identification of a complete and effective radical treatment for irreversible pulpal disease, a common clinical problem, continues to pose challenges. Although traditional root canal therapy remains the primary clinical treatment, it does not fully restore the physiological functions of pulp. Although stem cell transplantation appears to be a relatively viable treatment strategy for pulp disease, issues such as cell heterogeneity and poor regeneration effects remain problematic. Dental pulp regeneration strategies based on "cell-free" exosome therapies explored by numerous studies appear to have shown significant advantages. In particular, exosomes derived from odontogenic stem cells have demonstrated considerable potential in tooth tissue regeneration engineering, and continue to exhibit superior therapeutic effects compared to non-odontogenic stem cell-derived exosomes. However, only a few studies have comprehensively summarised their research results, particularly regarding the critical factors involved in the process. Therefore, in this study, our purpose was to review the effects exerted by odontogenic exosomes on pulp regeneration and to analyse and discus crucial factors related to this process, thereby providing scholars with a feasible and manageable new concept with respect to regeneration schemes. [ABSTRACT FROM AUTHOR]

Objectives: This review aims to discuss the implications of anatomy of the root, pulp chamber, and canals on pulpotomy and revitalization procedures (RPs) as treatment alternatives to root canal treatment procedures. Methods: This narrative review was undertaken to address two main questions – why remove vital pulp tissue in teeth with complex canal anatomy when it can be preserved? And why replace the necrotic pulp in teeth with mature roots with a synthetic material when we can revitalize? This review also aims to discuss anatomical challenges with pulpotomy and revitalization procedures. Results: Maintaining the vitality of the pulp via partial or full pulpotomy procedures avoids the multiple potential challenges faced by clinicians during root canal treatment. However, carrying out pulpotomy procedures requires a meticulous understanding of the pulp chamber anatomy, which varies from tooth to tooth. Literature shows an increased interest in the application of RPs in teeth with mature roots; however, to date, the relation between the complexity of the root canal system and outcomes of RPs in necrotic multi-rooted teeth with mature roots is unclear and requires further robust comparative research and long-term follow-up. Conclusions: Whenever indicated, pulpotomy procedures are viable treatment options for vital teeth with mature roots; however, comparative, adequately powered studies with long-term follow-up are needed as a priority in this area. RPs show promising outcomes for necrotic teeth with mature roots that warrant more evidence in different tooth types with long-term follow-ups. Clinical relevance: Clinicians should be aware of the pulp chamber anatomy, which is subject to morphological changes by age or as a defensive mechanism against microbial irritation, before practicing partial and full pulpotomy procedures. RP is a promising treatment option for teeth with immature roots, but more evidence is needed for its applications in teeth with mature roots. A universal consensus and considerably more robust evidence are needed for the standardization of RPs in teeth with mature roots. [ABSTRACT FROM AUTHOR]

Vascular calcification, an ectopic calcification exacerbated by aging and renal dysfunction, is closely associated with cardiovascular disease. However, early detection indicators are limited. This study focused on dental pulp stones, ectopic calcifications found in oral tissues that are easily identifiable on dental radiographs. Our investigation explored the frequency and timing of these calcifications in different locations and their relationship to aortic calcification. In cadavers, we examined the association between the frequency of dental pulp stones and aortic calcification, revealing a significant association. Notably, dental pulp stones appeared prior to aortic calcification. Using a rat model of hyperphosphatemia, we confirmed that dental pulp stones formed earlier than calcification in the aortic arch. Interestingly, there were very few instances of aortic calcification without dental pulp stones. Additionally, we conducted cell culture experiments with vascular smooth muscle cells (SMCs) and dental pulp cells (DPCs) to explore the regulatory mechanism underlying high phosphate-mediated calcification. We found that DPCs produced calcification deposits more rapidly and exhibited a stronger augmentation of osteoblast differentiation markers compared with SMCs. In conclusion, the observation of dental pulp stones through X-ray examination during dental checkups could be a valuable method for early diagnosis of aortic calcification risk. [ABSTRACT FROM AUTHOR]

Objectives: There is a lack of studies evaluating the accuracy of the 2009 American Association of Endodontists (AAE) diagnostic criteria for diagnosing pulpal health in primary teeth. This study aimed to estimate and correlate the diagnostic accuracy of clinical diagnosis of reversible and irreversible pulpitis using the 2009 AAE criteria with histological findings in primary teeth. Methods: Eighty primary teeth that were clinically diagnosed with normal pulp (n = 10), reversible pulpitis (n = 30), irreversible pulpitis (n = 30) and pulp necrosis (n = 10) were collected. The teeth were histo-processed, and pulp tissues were diagnosed histologically as uninflamed pulp, reversible or irreversibly inflamed and necrosis based on previously proposed criteria. Results: The clinical diagnosis of pulp necrosis (sensitivity 70%, specificity 96%) and normal pulp (sensitivity 91%, specificity 100%) matched the histological diagnosis of necrosis and uninflamed pulp in 70% and 100%, respectively. The clinical diagnosis of irreversible pulpitis (sensitivity 64%, specificity 72%) matched the histological diagnosis of irreversible pulp inflammation for 47% of teeth evaluated. For the clinical diagnosis of reversible pulpitis (sensitivity: 65%, specificity: 86%), 80% matched the histological diagnosis of reversible pulp inflammation. Teeth with histologically diagnosed irreversible pulp inflammation were more likely to have lingering (OR 5.08; 95% CI 1.48–17.46, P = 0.010) and nocturnal tooth pain (OR 15.86; 95% CI 1.57–160.47, P = 0.019) when compared to teeth with reversible pulp inflammation. Using the classification and regression tree model, the presence of widened periodontal ligament space and nocturnal tooth pain were useful predictors of irreversible pulp inflammation with an accuracy of 78%. Conclusion: The 2009 AAE criteria was acceptable for primary teeth with pulp necrosis and normal pulp but poor for reversible pulpitis and irreversible pulpitis. [ABSTRACT FROM AUTHOR]

Objectives: The purpose of this study was to investigate the oxidative stress cycle consisting of reactive oxygen molecules (ROS), glutathione (GSH) and glutathione S-transferase (GST) in caries-related pulp inflammation. Methodology: Fifty-four pulp tissue samples were collected from healthy donors with the diagnosis of reversible pulpitis, symptomatic irreversible pulpitis, and healthy pulp. Twelve pulp samples from each group were homogenized and total protein, ROS, GSH, and GST were measured by spectrophotometer. The remaining 6 samples from each group were prepared for paraffin block and used for the histopathologic and immunohistochemical evaluation of oxidative stress parameters and TUNEL labeling. Data were analyzed statistically. Results: The results revealed that total protein levels significantly decreased; however, ROS levels increased in both reversible and irreversible pulpitis compared to the healthy pulp (p < 0.01). Also, as inflammation increases, GST enzyme levels decrease while GSH levels increase significantly (p < 0.05). It was found that the number of TUNEL (+) cells was increased in irreversible pulpitis samples compared to healthy and reversible pulpitis groups (p < 0.05). GSTP1 and GSH immunoreactivity were also observed in irreversible pulpitis samples. Conclusions: It has been revealed that caries-related inflammation alters the oxidative stress cycle in dental pulp tissue. The increase in GSH levels in the inflamed dental pulp due to the increase in ROS levels may improve the defensive ability of the dental pulp. Clinical Relevance: There is a relationship between oxidative stress and inflammation. Control of excessive oxidative stress in pulpitis can stimulate reparative and regenerative processes. The present findings may provide an overview of the management of oxidative stress in cases with pulpitis during regenerative treatments. [ABSTRACT FROM AUTHOR]

Vital pulp therapy (VPT) is primarily intended to preserve the vitality of pulp tissues, which have been exposed for any reason. Various materials and techniques have been proposed to improve treatment outcomes, including the use of lasers. This study aimed to review the histological results of different dental lasers including low-level lasers, carbon dioxide (CO2), erbium-doped yttrium aluminum garnet laser (Er:YAG), neodymium-doped yttrium aluminum garnet (Nd:YAG), erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) lasers, and diode lasers for VPT. This focused review included a comprehensive electronic search of Scopus, MEDLINE, Web of Science, and Google Scholar databases from 2000 to 2022 by two independent investigators. Different combinations of keywords were used, and reference mining of related papers was done. The review included studies related to histologic evaluation of laser-assisted vital pulp therapy that stated the laser parameters that were used. Articles with radiographic or clinical assessments or articles lacking necessary data were excluded. Non-English articles were excluded unless their abstract was in English and encompassed the necessary data. Most studies indicated the efficacy of lasers for reduction of inflammation, acceleration of healing, and increasing the thickness of dentinal bridge. According to the evidence, lasers used in combination with pulp capping agents are beneficial to enhance the success rate of VPT. [ABSTRACT FROM AUTHOR]

Objectives: On the basis of a large sample size and a long follow-up period, the objectives of this study were to evaluate the outcomes of direct pulp capping (DPC) in mature permanent teeth with carious pulp exposure using a kind of bioaggregate putty (BP) which commercially named iRoot BP Plus (Innovative Bioceramix, Inc., Vancouver, Canada) and to analyze the potential prognostic factors. Materials and methods: The design of this research was retrospective regarding treatment procedures and prospective regarding the assessment of outcomes. The preoperative diagnosis of the teeth was either normal pulp or reversible pulpitis. Results were assessed based on clinical and radiographic examinations with at least 12 months of follow-up after DPC. No symptoms or signs, a positive response to electric pulp testing, a normal response to cold pulp testing and radiographs showing no abnormalities were considered to indicate success. Kaplan–Meier survival analysis was used to calculate the cumulative survival of teeth after DPC. Univariate and multivariate Cox proportional hazard regressions were used to analyze potential prognostic factors. Results: Three hundred thirty-four patients, including a total of 354 teeth, were available for the final clinical examination. The follow-up period ranged from 12 to 85 months, with an average of 27.0 ± 0.8 months. The total success rate was 85% (302/354), and the cumulative survival rates at 1, 2, 3, 4, and 5 years were 92%, 87%, 83%, 76%, and 72%, respectively. Univariate analysis indicated a significantly increased risk of failure in patients aged above 40 years and those treated by resident operators (P ≤ 0.01), with hazard ratios of 2.18 and 2.27, respectively. Conclusions: Under appropriate indication selection and treatment procedures, long-term success is possible in mature permanent teeth with carious pulp exposure by DPC using iRoot BP Plus. Patient age and operator experience are potential prognostic factors. Clinical relevance: Clinical data on iRoot BP Plus as a pulp capping medicament in mature permanent teeth with carious pulp exposure is lacking. This study indicated the efficacy of BP in DPC. Younger patient and sophisticated operator are beneficial for the outcome of DPC. [ABSTRACT FROM AUTHOR]

One of the most studied class of non-coding RNAs is microRNAs (miRNAs) which regulate more than 60% of human genes. A network of miRNA gene interactions participates in stem cell self-renewal, proliferation, migration, apoptosis, immunomodulation, and differentiation. Human pulp tissue-derived stem cells (PSCs) are an attractive source of dental mesenchymal stem cells (MSCs) which comprise human dental pulp stem cells (hDPSCs) obtained from the dental pulp of permanent teeth and stem cells isolated from exfoliated deciduous teeth (SHEDs) that would be a therapeutic opportunity in stomatognathic system reconstruction and repair of other damaged tissues. The regenerative capacity of hDPSCs and SHEDs is mediated by osteogenic, odontogenic, myogenic, neurogenic, angiogenic differentiation, and immunomodulatory function. Multi-lineage differentiation of PSCs can be induced or inhibited by the interaction of miRNAs with their target genes. Manipulating the expression of functional miRNAs in PSCs by mimicking miRNAs or inhibiting miRNAs emerged as a therapeutic tool in the clinical translation. However, the effectiveness and safety of miRNA-based therapeutics, besides higher stability, biocompatibility, less off-target effects, and immunologic reactions, have received particular attention. This review aimed to comprehensively overview the molecular mechanisms underlying miRNA-modified PSCs as a futuristic therapeutic option in regenerative dentistry. [ABSTRACT FROM AUTHOR]

Yes associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) are ubiquitous transcriptional co-activators that control organ development, homeostasis, and tissue regeneration. Current in vivo evidence suggests that YAP/TAZ regulates enamel knot formation during murine tooth development, and is indispensable for dental progenitor cell renewal to support constant incisor growth. Being a critical sensor for cellular mechano-transduction, YAP/TAZ lays at the center of the complex molecular network that integrates mechanical cues from the dental pulp chamber and surrounding periodontal tissue into biochemical signals, dictating in vitro cell proliferation, differentiation, stemness maintenance, and migration of dental stem cells. Moreover, YAP/TAZ-mediated cell-microenvironment interactions also display essential regulatory roles during biomaterial-guided dental tissue repair and engineering in some animal models. Here, we review recent advances in YAP/TAZ functions in tooth development, dental pulp, and periodontal physiology, as well as dental tissue regeneration. We also highlight several promising strategies that harness YAP/TAZ activation for promoting dental tissue regeneration. [ABSTRACT FROM AUTHOR]

Objectives: To evaluate the effectiveness of the 980-nm diode laser for dentinal tubule occlusion, measure the intrapulpal temperature, and investigate the dental pulp response. Materials and methods: The dentinal samples were randomly divided into G1–G7 groups: control; 980-nm laser irradiation (0.5 W, 10 s; 0.5 W, 10 s × 2; 0.8 W, 10 s; 0.8 W, 10 s × 2; 1.0 W, 10 s; 1.0 W, 10 s × 2). The dentin discs were applied for laser irradiation and analyzed by scanning electron microscopy (SEM). The intrapulpal temperature was measured on the 1.0-mm and 2.0-mm thickness samples, and then divided into G2–G7 groups according to laser irradiation. Moreover, forty Sprague Dawley rats were randomly divided into the laser-irradiated group (euthanized at 1, 7, and 14 days after irradiation) and the control group (non-irradiated). qRT-PCR, histomorphology, and immunohistochemistry analysis were employed to evaluate the response of dental pulp. Results: SEM indicated the occluding ratio of dentinal tubules in the G5 (0.8 W, 10 s × 2) and G7 (1.0 W, 10 s × 2) were significantly higher than the other groups (p < 0.05). The maximum intrapulpal temperature rises in the G5 were lower than the standard line (5.5 ℃). qRT-PCR showed that the mRNA expression level of TNF-α and HSP-70 upregulated significantly at 1 day (p < 0.05). Histomorphology and immunohistochemistry analysis showed that, compared with the control group, the inflammatory reaction was slightly higher at the 1 and 7 days (p < 0.05) and decreased to the normal levels at 14 days (p > 0.05). Conclusions: A 980-nm laser at a power of 0.8 W with 10 s × 2 defines the best treatment for dentin hypersensitivity in terms of compromise between the efficacy of the treatment and the safety of the pulp. Clinical relevance: The 980-nm laser is an effective option for treating dentin sensitivity. However, we need to ensure the safety of the pulp during laser irradiation. [ABSTRACT FROM AUTHOR]

Introduction : Usually, orthodontic movements encompass children and young adult patients, which are more susceptible to the occurrence of traumatic dental injuries. It is necessary to understand whether the effects of orthodontic movements on traumatized teeth could induce pulp necrosis. The aim of this study was to answer the following question: "Do orthodontic movements of traumatized teeth induce dental pulp necrosis?". Material and methods: Searches were performed for studies published up to May 11, 2023, in the MEDLINE/PubMed, Cochrane Library, Scopus, SciELO Citation Index, Web of Science, EMBASE, and Grey Literature Report databases, without restriction for language or year of publication. The revised Cochrane risk of bias tools for nonrandomized interventions (ROBINS-I) was used to assess the quality of the included studies. The overall quality of evidence was assessed through the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) tool. Results: Of 2671 potentially relevant studies, five were included. Four were classified as having a moderate risk of bias and one as a serious risk of bias. It was reported a higher susceptibility to pulp necrosis in teeth subjected to orthodontic movements with history of trauma involving periodontal tissues. Additionally, orthodontic movements of traumatized teeth with total pulp obliteration had an increased risk of pulp necrosis. GRADE analysis presented a moderate certainty of evidence. Conclusions: An increased risk for pulp necrosis when traumatized teeth are subjected to orthodontic movements was verified. However, this is based on evaluations performed with subjective tests. Further well-designed studies are necessary to confirm this trend. Clinical relevance: Clinicians must be aware of the possibility of pulp necrosis. However, endodontic treatment is recommended when verified signs and symptoms of pulp necrosis. [ABSTRACT FROM AUTHOR]

Dental caries has been a common health issue throughout the world, which can even lead to dental pulp and root apical inflammation eventually. Timely and effective treatment of dental caries is vital for patients to reduce pain. Traditional caries disease diagnosis methods like naked-eye detection and panoramic radiograph examinations rely on experienced doctors, which may cause misdiagnosis and high time-consuming. To this end, we propose a novel deep learning architecture called CariesNet to delineate different caries degrees from panoramic radiographs. We firstly collect a high-quality panoramic radiograph dataset with 3127 well-delineated caries lesions, including shallow caries, moderate caries, and deep caries. Then we construct CariesNet as a U-shape network with the additional full-scale axial attention module to segment these three caries types from the oral panoramic images. Moreover, we test the segmentation performance between CariesNet and other baseline methods. Experiments show that our method can achieve a mean 93.64% Dice coefficient and 93.61% accuracy in the segmentation of three different levels of caries. [ABSTRACT FROM AUTHOR]

Stem cells derived from dental/odontogenic tissue have the property of multiple differentiation and are prospective in tooth regenerative medicine and cellular and molecular studies. However, in the face of cellular senescence soon in vitro, the proliferation ability of the cells is limited, so studies are hindered to some extent. Fortunately, immortalization strategies are expected to solve the above issues. Cellular immortalization is that cells are immortalized by introducing oncogenes, human telomerase reverse transcriptase genes (hTERT), or miscellaneous immortalization genes to get unlimited proliferation. At present, a variety of immortalized stem cells from dental/odontogenic tissue has been successfully generated, such as dental pulp stem cells (DPSCs), periodontal ligament cells (PDLs), stem cells from human exfoliated deciduous teeth (SHEDs), dental papilla cells (DPCs), and tooth germ mesenchymal cells (TGMCs). This review summarized establishment and applications of immortalized stem cells from dental/odontogenic tissues and then discussed the advantages and challenges of immortalization. [ABSTRACT FROM AUTHOR]

Ischemic stroke is the major cause of death and morbidity worldwide. Stem cell treatment is at the forefront of ischemic therapeutic interventions. However, the fate of these cells following transplantation is mostly unknown. The current study examines the influence of oxidative and inflammatory pathological events associated with experimental ischemic stroke (oxygen glucose deprivation (OGD)) on the stem cell population (human Dental Pulp Stem Cells, and human Mesenchymal Stem Cells) through the involvement of the NLRP3 inflammasome. We explored the destiny of the above-mentioned stem cells in the stressed micro (-environment) and the ability of MCC950 to reverse the magnitudes. An enhanced expression of NLRP3, ASC, cleaved caspase1, active IL-1β and active IL-18 in OGD-treated DPSC and MSC was observed. The MCC950 significantly reduced NLRP3 inflammasome activation in the aforementioned cells. Further, in OGD groups, oxidative stress markers were shown to be alleviated in the stem cells under stress, which was effectively relieved by MCC950 supplementation. Interestingly, whereas OGD increased NLRP3 expression, it decreased SIRT3 levels, implying that these two processes are intertwined. In brief, we discovered that MCC950 inhibits NLRP3-mediated inflammation by inhibiting the NLRP3 inflammasome and increasing SIRT3. To conclude, according to our findings, inhibiting NLRP3 activation while enhancing SIRT3 levels with MCC950 reduces oxidative and inflammatory stress in stem cells under OGD-induced stress. These findings shed light on the causes of hDPSC and hMSC demise following transplantation and point to strategies to lessen therapeutic cell loss under ischemic-reperfusion stress. [ABSTRACT FROM AUTHOR]

The study evaluated the pulp temperature (PT) increase in Class I and V preparations when exposed to the Monet Laser (for 1 and 3 s), the PinkWave (for 3 and 10 s), the Valo Grand (for 3 and 10 s), the PowerCure, (for 3 and 10 s) and the SmartLite Pro (for 10 s). Non-retentive Class I and Class V cavities were prepared in one molar fixed in an acrylic plate and positioned in a warm water bath. The PT baseline was kept at 32 °C to simulate physiological conditions. Two T-type thermocouples were inserted through the roots into the pulp chamber in two positions: close to the pulp horn and the buccal wall close to the Class V cavity. The water flow was adjusted to 0.026 mL/min, and real-time temperature data were collected every 0.5 s. PT measurements were made with the tip of the LCU 0 and 6 mm away from the tooth surface. The radiant exitance (mW/cm2) and radiant exposure (J/cm2) were calculated. One-way ANOVA compared the effect of the pulpal flow, and ΔT values were subjected to two-way ANOVA, followed by Scheffe's post hoc tests. The Monet Laser used for 3 s and the PinkWave used for 10 s produced the greatest PT rise in the Class I cavity. The simulated pulpal flow did not influence the PT rise. Overall, cavities exposed at the 0 mm distance had higher ΔT values than groups at 6 mm distance. The placement of a rubber dam for Class V restorations may prevent positioning LCUs directly over the cavity, which may affect the rise in PT. [ABSTRACT FROM AUTHOR]

This study aimed to assess the histological changes in the maxillary sinus and its adjacent dental tissues as pulp and periodontal ligament during pregnancy and investigate the role of estrogen hormone in these changes through the detection of estrogen receptors in these tissues. Sixteen adult female rats were used and were allocated into two groups: control non-pregnant (n = 8) and pregnant (n = 8). They were sacrificed and their heads were prepared for histological and immunohistochemical examination for estrogen receptor alpha. Our results revealed that pregnant rats revealed inflammatory changes in the sinus as thick epithelial lining, loss of cilia, swollen goblet cells, intraepithelial and interstitial edema. The lamina propria demonstrated considerable infiltration of inflammatory cells, glandular hyperplasia with vacuolar degeneration, and vascular congestion. Periodontal ligament and pulp revealed hyperemia and vascular congestion. Immunohistochemical examination of estrogen receptor alpha in the maxillary sinus and adjacent dental tissues (Periodontal ligament and pulp) in pregnant rats revealed a significant increase in its expression in all examined tissues. In conclusion, there was an increase in expression of ERα in the sinus mucosa and dental tissues during pregnancy together with slight inflammatory changes in these tissues. Hence, dentists should be aware of the effect of these changes on the pregnant women avoiding teeth extraction due to misdiagnosis of dental, periodontal or sinus pain after exclusion of true pathologies. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are short, 19–23 nucleotide non-coding RNA molecules that regulate gene expression by silencing or degrading the target mRNA gene. Since their discovery in the nineties of the last century, they have emerged as key inflammatory regulators. Inflammation induces the synthesis of various miRNAs that modulate the expression of multiple molecules involved in orchestrating the inflammatory response. This review aims to provide an insight into the role of miRNAs as potential biomarkers, mediators, and potential therapeutic targets of dental pulp inflammation. A literature search was conducted using the keywords; biogenesis of microRNA, human dental pulp cells, pulpitis, and inflammation in PubMed and Scopus index databases for all the published articles dealing with the role of miRNAs in pulp inflammation in the last 10 years. According to the literature, there is a clear correlation between miRNAs and several physiological events, as well as their role as mediators of innate immune response and inflammation in dental pulp cells. Our narrative review stipulates that numerous miRNAs play a key role in modulating inflammation, delaying or enhancing cell repair, cell differentiation, and survival in dental pulp diseases. However, further studies are required for the validation of miRNAs as reliable biomarkers in dental pulp pathology and their targeted therapy. [ABSTRACT FROM AUTHOR]

Stem cell research with biological waste material is an area that holds promise to revolutionize treatment modalities and clinical practice. The interest in surgical remnants is increasing with time as research on human embryonic stem cells remains controversial due to legal and ethical issues. Perhaps, these restrictions are the motivation for the use of alternative mesenchymal stem cell (MSC) sources in the regenerative field. Stem cells (SCs) of Umbilical Cord (UC) and Dental Pulp (DP) have almost similar biological characteristics to other MSCs and can differentiate into a number of cell lineages with enormous potential future prospects. A concise critical observation of UC-MSCs and DP-MSCs is presented here reviewing articles from the last two decades along with other stem cell sources from different biological waste materials. [ABSTRACT FROM AUTHOR]

Host Defense Peptides (HDPs) have, in previous studies, been demonstrating antimicrobial, anti-inflammatory, and immunomodulatory capacity, important factors in the repair process. Knowing these characteristics, this article aims to evaluate the potential of HDPs IDR1018 and DJK-6 associated with MTA extract in the repair process of human pulp cells. Antibacterial activity of HDPs, MTA and HDPs combined with MTA in Streptococcus mutans planktonic bacteria and antibiofilm activity was evaluated. Cell toxicity was assayed with MTT and cell morphology was observed by scanning electron microscopy (SEM). Proliferation and migration of pulp cells were evaluated by trypan blue and wound healing assay. Inflammatory and mineralization related genes were evaluated by qPCR (IL-6, TNFRSF, DSPP, TGF-β). Alkaline phosphatase, phosphate quantification and alizarin red staining were also verified. The assays were performed in technical and biological triplicate (n = 9). Results were submitted for the calculation of the mean and standard deviation. Then, normality verification by Kolmogorov Smirnov test, analyzing one-way ANOVA. Analyses were considered at a 95% significance level, with a p-value < 0.05. Our study demonstrated that HDPs combined with MTA were able to reduce biofilms performed in 24 h and biofilm performed over 7 days S. mutans biofilm (p < 0.05). IDR1018 and MTA, as well as their combination, down-regulated IL-6 expression (p < 0.05). Tested materials were not cytotoxic to pulp cells. IDR1018 induced high cell proliferation and combined with MTA induced high cellular migration rates in 48 h (p < 0.05). Furthermore, the combination of IDR1018 and MTA also induced high expression levels of DSPP, ALP activity, and the production of calcification nodules. So, IDR-1018 and its combination with MTA could assist in pulp-dentine complex repair process in vitro. [ABSTRACT FROM AUTHOR]