Pulpal Inflammation Research Articles

Expression of Selected Inflammatory Mediators with Different Clinical Characteristics of Pulpal Inflammation

IntroductionAccurately diagnosing the state of dental pulp is crucial when addressing tooth pain to determine the best treatment approach. This study aimed to investigate the concentration of inflammatory mediators in the dental pulp of mature teeth that have been exposed via caries but show no signs of apical periodontitis. MethodsSamples of pulpal blood from adults with mature teeth responsive to pulp testing and have carious pulp exposures were obtained. These samples were analyzed for 12 inflammatory cytokines and other inflammatory proteins using the Luminex assay platform. Clinical factors were correlated with cytokine levels, and statistical analysis was performed to evaluate the impact of these factors on cytokine expression. ResultsOf the 36 patients that were included, 44.44% took pain medications, 33.33% had prolonged pulpal bleeding, 41.67% felt spontaneous pain, and 72.22% were diagnosed with symptomatic irreversible pulpitis. Significant correlations existed between presenting pain scores and levels of interleukin (IL)-1α, IL-6, and IL-8 (P < .05). Factors like analgesic medication intake, pain to percussion, pain to thermal testing, spontaneous pain, and nocturnal pain were significantly associated with higher levels of specific inflammatory proteins. No significant associations were observed with pain to palpation, bleeding time, or pulpal diagnosis. ConclusionsInflammatory proteins, including cytokine levels may play a critical role in characterizing pulpal inflammation. Future studies should investigate the role of these potential biomarkers in determining the diagnosis of pulpitis and the prognosis of vital pulp therapy.

Silver Diamine Fluoride Renaissance in Paediatric Dentistry: A 24-Month Retrospective and Cross-Sectional Analysis.

Background and Objectives: Silver diamine fluoride (SDF) has been incorporated into the treatment of dental caries in children, mainly in countries with high caries prevalence. In Europe, however, SDF started to gain popularity during the COVID-19 pandemic. This study aimed to investigate the efficacy of SDF and to evaluate dentists'/parents' acceptance of SDF use in paediatric patients treated in a German university setting. Materials and Methods: A retrospective analysis of all patients treated with SDF between 2017 and 2020 was carried out. Only teeth with no reported clinical/radiographic evidence of irreversible pulpal inflammation were included. The outcome measures were success, minor failures (caries progression, reversible pulpitis) and major failures (irreversible pulpitis, abscess). The treatment acceptance by dentists and the parents of SDF-treated children was cross-sectionally evaluated using questionnaires. Descriptive statistics and Kaplan-Meier survival analysis were performed. Results: A total of 93 patients (mean age 5.3 ± 2.9 years) with 455 treated teeth (418 primary/91.9%; 37 permanent/8.1%) were included and followed up for up to 24 months (19.9 ± 10.5 months). SDF was used for dental caries (98.2%) and hypersensitivity relief on MIH teeth (1.8%). Most teeth did not show any failure (total success 84.2%). A total of 5 teeth (1.1%) showed minor failures, and 67 teeth (14.7%) showed major failures (p = 0.001). Success/failure rates were not affected by patient compliance, gender, dentition, or operator (p > 0.05). In total, 30 questionnaires were collected from parents (mean age 36.8 ± 6.4 years). SDF was applied on anterior (n = 2/6.7%), posterior (n = 15/50%) and anterior/posterior teeth (n = 13/43.3%). At the 1-week follow-up, 80% of parents noticed black teeth discoloration. Treatment satisfaction was higher for posterior (95.2%) than for anterior teeth (36.4%; p < 0.001). In the 27 responses from clinicians, SDF was generally considered a viable option in paediatric dentistry (n = 23; 85%). Conclusions: SDF was found to be effective and well-accepted by parents and dentists for caries inactivation in a paediatric dentistry German university setting.

Scaffolds for Dentin-Pulp Complex Regeneration.

Background and Objectives: Regenerative dentistry aims to regenerate the pulp-dentin complex and restore those of its functions that have become compromised by pulp injury and/or inflammation. Scaffold-based techniques are a regeneration strategy that replicate a biological environment by utilizing a suitable scaffold, which is considered crucial for the successful regeneration of dental pulp. The aim of the present review is to address the main characteristics of the different scaffolds, as well as their application in dentin-pulp complex regeneration. Materials and Methods: A narrative review was conducted by two independent reviewers to answer the research question: What type of scaffolds can be used in dentin-pulp complex regeneration? An electronic search of PubMed, EMBASE and Cochrane library databases was undertaken. Keywords including "pulp-dentin regeneration scaffold" and "pulp-dentin complex regeneration" were used. To locate additional reports, reference mining of the identified papers was undertaken. Results: A wide variety of biomaterials is already available for tissue engineering and can be broadly categorized into two groups: (i) natural, and (ii) synthetic, scaffolds. Natural scaffolds often contain bioactive molecules, growth factors, and signaling cues that can positively influence cell behavior. These signaling molecules can promote specific cellular responses, such as cell proliferation and differentiation, crucial for effective tissue regeneration. Synthetic scaffolds offer flexibility in design and can be tailored to meet specific requirements, such as size, shape, and mechanical properties. Moreover, they can be functionalized with bioactive molecules, growth factors, or signaling cues to enhance their biological properties and the manufacturing process can be standardized, ensuring consistent quality for widespread clinical use. Conclusions: There is still a lack of evidence to determine the optimal scaffold composition that meets the specific requirements and complexities needed for effectively promoting dental pulp tissue engineering and achieving successful clinical outcomes.

Dental pulp cells cocultured with macrophages aggravate the inflammatory conditions stimulated by LPS

BackgroundPulp inflammation is complex interactions between different types of cells and cytokines. To mimic the interactions of different types of cells in inflamed dental pulp tissues, dental pulp cells (DPCs) were cocultured with different ratios of macrophages (THP-1) or LPS treatment.MethodsDPCs were cocultured with various ratios of THP-1, then photographed cell morphology and determined cell viability by MTT assay at preset times. Total RNA was also extracted to measure the inflammation marker-IL-6 and IL-8 expressions by RT-Q-PCR. The DPCs and THP-1 were treated with 0.01 – 1μg/ml lipopolysaccharide (LPS) and extract RNA at preset times, and detected IL-6 and IL-8 expression. DPCs were cocultured with various ratios of THP-1 with 0.1 μg/mL LPS, and detected IL-6 and IL-8 expression after 24 and 48 h. The data were analyzed by unpaired t-test or Mann-Whitney test. Differences were considered statistically significant when p < 0.05.ResultsTHP-1 and DPCs coculture models did not suppress the viability of DPCs and THP-1. Cocultured with various ratios of THP-1 could increase IL-6 and IL-8 expressions of DPCs (p = 0.0056 - p < 0.0001). The expressions of IL-6 and IL-8 were stronger in higher ratio groups (p = 0.0062 - p < 0.0001). LPS treatment also induced IL-6 and IL-8 expressions of DPCs and THP-1 (p = 0.0179 – p < 0.0001 and p = 0.0189 – p < 0.0001, separately). Under the presence of 0.1 μg/mL LPS, DPCs cocultured with THP-1 for 24 h also enhanced IL-6 and IL-8 expression (p = 0.0022). After cocultured with a higher ratio of THP-1 for 48 h, IL-6 and IL-8 expressions were even stronger in the presence of LPS (p = 0.0260).ConclusionsCoculturing dental pulp cells and macrophages under LPS treatment aggravate the inflammatory process. The responses of our models were more severe than traditional inflamed dental models and better represented what happened in the real dental pulp. Utilizing our models to explore the repair and regeneration in endodontics will be future goals.

Use of the Hall Technique by dentists in East China: A questionnaire‐based cross‐sectional survey

AbstractBackgroundThe Hall Technique (HT) is now regarded as one of the biological management options for carious primary molars and has shown significant clinical success.AimTo investigate the perception and the use of the HT among dentists engaging in paediatric dentistry in East China.DesignThis was an electronic questionnaire‐based cross‐sectional study. A quick response code of the questionnaire via WeChat, a common communication tool in China, was sent to dentists in East China.ResultsA total of 313 dentists participated in this study. Most surveyed clinicians had heard about the HT (n = 286, 91.4%). Of them, 40.2% (n = 115) reported having used the HT. Of 67 dentists working in private clinics, 34 (50.7%) had used the HT, whereas only 37% of dentists from public hospitals had used the HT. Of 115 dentists having used the HT, 91.3% (n = 105) would consider using the HT for Class II cavitated molars; 23.5% (n = 27) would consider using the HT for Class I cavitated molars. Most dentists who had not used the HT were concerned about complications such as pulp inflammation or necrosis after applying the HT.ConclusionThe HT is well known in East China; the lack of systematic training or courses, however, may decrease its utilization.

Guanylate-binding protein 5-mediated cell-autonomous immunity suppresses inflammation in dental pulpitis: Aninvitro study.

Guanylate-binding protein 5 (GBP5) is an interferon (IFN)-inducible GTPase that plays a crucial role in the cell-autonomous immune response against microbial infections. In this study, we investigated the immunoregulatory role of GBP5 in the pathogenesis of dental pulpitis. Gene-set enrichment analysis (GSEA) was utilized to evaluate the IFN-γ signalling pathway, and the differential expression of GBP mRNA in normal versus inflamed dental pulp tissues was screened, based on Gene Expression Omnibus (GEO) datasets associated with pulpitis. Both normal pulp tissues and inflamed pulp tissues were used for experiments. The expression of IFNs and GBPs was determined by qRT-PCR. Immunoblotting and double immunofluorescence were performed to examine the cellular localization of GBP5 in dental pulp tissues. For the functional studies, IFN-γ priming or lentivirus vector-delivered shRNA was used to, respectively, overexpress or knock down endogenous GBP5 expression in human dental pulp stem cells (HDPSCs). Subsequently, LPS was used to stimulate HDPSCs (overexpressing or with knocked-down GBP5) to establish an invitro model of inflammation. qRT-PCR and ELISA were employed to examine the expression of proinflammatory cytokines (IL-6, IL-8 and IL-1β) and cyclooxygenase 2 (COX2). Every experiment has three times of biological replicates and three technical replicates, respectively. Statistical analysis was performed using the Student's t-test and one-way ANOVA, and a p-value of <.05 was considered statistically significant. GSEA analysis based on the GEO dataset revealed a significant activation of the IFN-γ signalling pathway in the human pulpitis group. Among the human GBPs evaluated, GBP5 was selectively upregulated in inflamed dental pulp tissues and predominantly expressed in dental pulp cells. Invitro experiments demonstrated that IFN-γ robustly induced the expression of GBP5 in HDPSCs. Knockdown of GBP5 expression in HDPSCs significantly amplified the LPS-induced upregulation of inflammatory mediators (IL-6, IL-8, IL-1β and COX2) both with and without IFN-γ priming. Our findings demonstrated that GBP5 partook in the pathogenesis of dental pulpitis. The involvement of GBP5 in pulpitis appeared to coordinate the regulation of inflammatory cytokines. Knockdown of GBP5 contributed to the exacerbation of LPS-mediated inflammation.

Efficient Treatment of Pulpitis via Transplantation of Human Pluripotent Stem Cell-Derived Pericytes Partially through LTBP1-Mediated T Cell Suppression.

Dental pulp pericytes are reported to have the capacity to generate odontoblasts and express multiple cytokines and chemokines that regulate the local immune microenvironment, thus participating in the repair of dental pulp injury in vivo. However, it has not yet been reported whether the transplantation of exogenous pericytes can effectively treat pulpitis, and the underlying molecular mechanism remains unknown. In this study, using a lineage-tracing mouse model, we showed that most dental pulp pericytes are derived from cranial neural crest. Then, we demonstrated that the ablation of pericytes could induce a pulpitis-like phenotype in uninfected dental pulp in mice, and we showed that the significant loss of pericytes occurs during pupal inflammation, implying that the transplantation of pericytes may help to restore dental pulp homeostasis during pulpitis. Subsequently, we successfully generated pericytes with immunomodulatory activity from human pluripotent stem cells through the intermediate stage of the cranial neural crest with a high level of efficiency. Most strikingly, for the first time we showed that, compared with the untreated pulpitis group, the transplantation of hPSC-derived pericytes could substantially inhibit vascular permeability (the extravascular deposition of fibrinogen, ** p < 0.01), alleviate pulpal inflammation (TCR+ cell infiltration, * p < 0.05), and promote the regeneration of dentin (** p < 0.01) in the mouse model of pulpitis. In addition, we discovered that the knockdown of latent transforming growth factor beta binding protein 1 (LTBP1) remarkably suppressed the immunoregulation ability of pericytes in vitro and compromised their in vivo regenerative potential in pulpitis. These results indicate that the transplantation of pericytes could efficiently rescue the aberrant phenotype of pulpal inflammation, which may be partially due to LTBP1-mediated T cell suppression.

Icariin negatively regulated lipopolysaccharide-induced inflammation and ameliorated the odontogenic activity of human dental pulp cells in vitro

Alleviating inflammation and promoting dentine regeneration is critical for the healing of pulpitis. In this study, we investigated the anti-inflammatory, angiogenesis and odontogenesis function of icariin on Human dental pulp cells (HDPCs) under inflammatory state. Furthermore, the underlying mechanisms was also evaluated. Icariin attenuated the LPS-induced pro-inflammatory marker expression, such as interleukin-1β (IL-1β), IL-6 and IL-8. The immunoblotting and immunofluorescence staining results showed that icariin suppressed the inflammatory responses mediated by the protein kinase B (Akt) and nuclear factor kappa-B (NF-κB) signaling cascades. Additionally, icariin also upregulated the expression of odontogenic and angiogenic genes and proteins (namely dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP1), anti-collagen Ⅰ (COL-Ⅰ), and vascular endothelial growth factor (VEGF) and fibroblast growth factor-1 (FGF-1)), alkaline phosphatase activity, and calcium nodule deposition in LPS-exposed HDPCs. In a word, our findings indicated that icariin attenuated pulp inflammation and promoted odontogenic and angiogenic differentiation in the inflammatory state. Icariin may be a promising vital pulp therapy agent for the regenerative treatment of the inflamed dental pulp.

Increased Purinergic Signaling in Human Dental Pulps With Inflammatory Pain is Sex-Dependent

An enhanced understanding of neurotransmitter systems contributing to pain transmission aids in drug development, while the identification of biological variables like age and sex helps in the development of personalized pain management and effective clinical trial design. This study identified enhanced expression of purinergic signaling components specifically in painful inflammation, with levels increased more in women as compared to men. Inflammatory dental pain is common and potentially debilitating; as inflammation of the dental pulp can occur with or without pain, it provides a powerful model to examine distinct pain pathways in humans. In control tissues, P2X3 and P2X2 receptors colocalized with PGP9.5-positive nerves. Expression of the ecto-nucleotidase NTPDase1 (CD39) increased with exposure to extracellular adenosine triphosphate (ATP), implying CD39 acted as a marker for sustained elevation of extracellular ATP. Both immunohistochemistry and immunoblots showed P2X2, P2X3, and CD39 increased in symptomatic pulpitis, suggesting receptors and the ATP agonist were elevated in patients with increased pain. The increased expression of P2X3 and CD39 was more frequently observed in women than men. In summary, this study identifies CD39 as a marker for chronic elevation of extracellular ATP in fixed human tissue. It supports a role for increased purinergic signaling in humans with inflammatory dental pain and suggests the contribution of purines shows sexual dimorphism. This highlights the potential for P2X antagonists to treat pain in humans and stresses the need to consider sex in clinical trials that target pain and purinergic pathways. PerspectiveThis article demonstrates an elevation of ATP-marker CD39 and of ATP receptors P2X2 and P2X3 with inflammatory pain and suggests the rise is greater in women. This highlights the potential for P2X antagonists to treat pain and stresses the consideration of sexual dimorphism in studies of purines and pain.

Effects of direct pulp capping with recombinant human erythropoietin and/or mineral trioxide aggregate on inflamed rat dental pulp.

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

Dental Tissue Engineering by Neural Differentiation of Dental Stem Cells and Nano-systems: A Review

Background: Pulpitis is a pulpal inflammation. It generally occurs when there is inflammation within a tooth as a result of anything like grinding or decay. After dental inflammation, microcirculation and sensory nerve activity seem to play the most critical role in reducing inflammation. Therefore, researchers emphasize the study of dental nerve activity, especially in acute clinical problems in inflamed teeth and pulp regeneration. This review aims to investigate the possibility of using dental stem cells to regenerate dental nerves in order to repair dentin-pulp complexes for maintaining and restoring tooth structure and function, which nanosystems can help in this matter. Materials and Methods: In this paper, we review the literature regarding the theory of dental tissue engineering by neural differentiation of dental stem cells and nano-systems, and the comprehensive search on PubMed, Scopus, and Web of Science was conducted up to July 2022. Results: According to recent studies, dental soft and hard tissue healing also includes nerve fibers. A deeper understanding of how dental nerves are implicated in pulpitis may assist endodontic treatment. Stem cell-based treatments may be used to regenerate dental nerves to repair dentin-pulp complexes to maintain and restore tooth structure and function. Conclusion: The emphasis on dental nerve regeneration appears to be a critical stage in fostering spontaneous tooth regeneration as well as a sustainable tooth regeneration method. It is essential to further investigate dental tissue engineering by neural differentiation of dental stem cells.

Connexin43 reduces LPS-induced inflammation in hDPCs through TLR4-NF-κB pathway via hemichannels.

Connexin43 (Cx43) is involved in the inflammation of many tissue types. Dental caries is infectious disease resulting from mineralized tissue dissolution by a specific bacterial population, causing pulp inflammation. However, Cx43's role in dental pulp remains unclear. Here, we investigated the function of Cx43 during pulp inflammation. We constructed a dentin injury model in Sprague-Dawley rats to investigate changes in Cx43 expression during pulp inflammation. Cx43 was inhibited in human dental pulp cells (hDPCs) that had been stimulated with lipopolysaccharide (LPS) to investigate the effect of Cx43 on inflammatory response. Promotion of TLR4-NF-κB pathway activity and special Cx43 channel inhibitors were used to clarify the function of Cx43 in hDPCs. Dentin injury led to low-level inflammation in dental pulp. Following dentin injury, Cx43 expression initially decreased before gradually recovering to normal levels. Cx43 inhibition reduced LPS-induced expression of inflammatory cytokines and NF-κB pathway activity. Promotion of NF-κB pathway activity counteracted the effect of Cx43 in hDPCs. Furthermore, inhibition of Cx43 hemichannels reduced LPS-induced inflammatory cytokine expression. Cx43 is involved in inflammation of dental pulp, while its inhibition reduced LPS-induced inflammation in hDPCs through NF-κB pathway via blockage of hemichannels.

The association between dental caries and steroid-sensitive nephrotic syndrome in children.

Pathogenesis and relapse of steroid-sensitive nephrotic syndrome (SSNS) are primarily associated with infection. Dental caries is the most common chronic progressive oral infection in children. However, clinical studies of SSNS combined with dental caries in children are rare. In our retrospective cohort study from January 2021 to June 2022, 145 children with SSNS were included in the baseline analysis and 105 in the follow-up analysis. The follow-up period was 1year. The primary study endpoints were the relapse-free period and frequently relapsing nephrotic syndrome (FRNS). Secondary endpoints included the number and triggers of relapses and concomitant medications. The median age was 5.5years, with a caries rate of 60.7%, themean DMFT/dmft was 3.86, and the caries filling rate was 1.6%. Except for the lower proportion of high household income and high parental education observed in the caries group, no statistical differences were found when comparing the other baseline data with the non-caries group. The caries group had a shorter relapse-free period and a lower 1-year cumulative relapse-free survival rate (HR = 1.90, 95% CI 1.17-3.09, P = 0.009). Univariate regression analysis showed caries associated with FRNS (OR = 2.714, 95% CI 1.021-7.219, P = 0.045), but the correlation no longer remained in the multivariate analysis. Additionally, seven cases of caries-derived pulpal periapical inflammation triggered SSNSrelapses. The caries group had more infection triggers and concomitant medication use. Dental caries and relapse of SSNS are potentially associated, but careful evaluation is needed. A higher resolution version of the Graphical abstract is available as Supplementary information.

LIF Aggravates Pulpitis by Promoting Inflammatory Response in Macrophages.

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.

MicroRNAs Function in Dental Stem Cells as a Promising Biomarker and Therapeutic Target for Dental Diseases.

Undifferentiated, highly proliferative, clonogenic, and self-renewing dental stem cells have paved the way for novel approaches to mending cleft palates, rebuilding lost jawbone and periodontal tissue, and, most significantly, recreating lost teeth. New treatment techniques may be guided by a better understanding of these cells and their potential in terms of the specificity of theregenerative response. MicroRNAs have been recognized as an essential component in stem cell biology due to their role as epigenetic regulators of the processes that determine stem cell destiny. MicroRNAs have been proven to be crucial in a wide variety of molecular and biological processes, including apoptosis, cell proliferation, migration, and necrocytosis. MicroRNAs have been recognized to control protein translation, messenger RNA stability, and transcription and have been reported to play essential roles in dental stem cell biology, including the differentiation of dental stem cells, the immunological response, apoptosis, and the inflammation of the dental pulp. Because microRNAs increase dental stem cell differentiation, they may be used in regenerative medicine to either preserve the stem cell phenotype or to aid in the development of tooth tissue. The development of novel biomarkers and therapies for dental illnesses relies heavily on progress made in our knowledge of the roles played by microRNAs in regulating dental stem cells. In this article, we discuss how dental stem cells and their associated microRNAs may be used to cure dental illness.

Vital Pulp Therapy of Young First Permanent Molars: A Retrospective Study on Radiographic Findings 24 Months Post-treatment.

With success rates comparable to that of root canal treatment, vital pulp therapy (VPT) has gained clinical interest and has been used in the management of young permanent teeth with inflamed pulps. The aim of the present study was to retrospectively evaluate the radiographic success of VPT in young first permanent molars 24 months post-treatment and correlate findings with tooth and treatment-related characteristics. Dental records of all patients with first permanent molars which received VPT in the Department of Paediatric Dentistry (National and Kapodistrian University of Athens) were retrieved. Demographic characteristics and data regarding the treatment performed were recorded. Patients' radiographs were evaluated at 6, 12 and 24 months post-treatment by two qualified paediatric dentists blinded regarding the treatment performed. Radiographic success, reasons for failure and continuation of root development were evaluated. Differences were tested using the Χ2 and Student's t-test, and possible correlations were determined by calculating the odds ratio. Overall radiographic success rate at 24 months was 77%, ranging between 50% for direct pulp capping and 92% for full pulpotomy. Differences were not statistically significant. Continuation of root development was recorded in almost 1/3 of the teeth and completion in almost 1/5. No statistically significant association was recorded between the outcome and any tooth and treatment-related variables. VPT seems to be a reliable option in the long term for the treatment of deep carious lesions in young permanent molars.

EFFECT OF ROBUSTA COFFEE BEANS AND ARABICA EXTRACT GEL AS INFLAMATION PULP MATERIAL

Background: Reversible pulpitis is a mild to moderate inflammatory pulp condition often treated with pulp capping treatments. Current study states that calcium hydroxide (CaOH2) has drawbacks for a long-time use. One of the alternative pulp capping materials is Robusta and Arabica coffee bean extract gel which contains flavonoid compounds and chlorogenic acid which have immunomodulatory properties that play role in the pulp inflammatory process by increasing the count of lymphocyte. Objective: The goal of this study is to determine and analyse the effect of Robusta coffee bean and Arabica coffee bean extract gel 95% to the of lymphocyte cells count on the 3rd and 5th day of pulp inflammation compared to placebo gel. Materials and Methods: This research is a true experimental with a post-test only control group design. This study used three treatment groups; Robusta coffee bean extract gel 95%, Arabica coffee bean extract gel 95%, and control group placebo gel. Results: Two Way Anova test results obtained p&lt;0.05, showed significant difference. Data analysis was continued with the Post Hoc Bonferroni test which showed that there was a significant difference between the 95% Robusta coffee bean extract gel group and the 95% Arabica coffee bean extract gel group with p&lt;0.05. Conclusion: The administration of Robusta coffee bean extract gel with a concentration of 95% can increase the of lymphocyte cells count in pulp inflammation on the 3rd and 5th days more effectively than the 95% Arabica coffee bean extract gel. Keywords: Arabica Coffee, Direct Pulp Capping, Inflammation, Lymphocytes, Pulpitis, Robusta Coffee

ANTIFUNGAL ACTIVITY OF RAMANIA LEAF EXTRACT (Bouea Macrophylla Griff) AGAINST Candida albicans

Background: Root canal infection is a polymicrobial infection that causes pulpitis or localized pulp inflammation. Root canal infection occurs due to the presence of pathogenic microorganisms in the root canal. One of the most common pathogenic microorganisms found in root canals is the fungus Candida albicans. Treatment that can be done to treat pulp tissue infection is endodontic treatment. Chlorhexidine gluconate 2% is one of the irrigation materials that can be used in root canal treatment. Ramania leaf extract contains compounds such as triterpenoids, phenols, flavonoids, steroids, saponins, alkaloids, and tannins that can function as antifungals. Purpose: Measuring and analyzing the antifungal activity of ramania leaf extract (Bouea Macrophylla Griff) on the growth of Candida albicans. Method: This study is a true experimental study with a post-test-only design with a control group design consisting of 7 treatment groups and repeated 4 times. The antifungal activity was assessed from the inhibition zone formed on Sabouraud Dextrose Agar media by the diffusion method. Results: Based on the Mann-Whitney test showed that there was only 1 pair of groups that did not have a significant difference, namely 100% concentration of ramania leaf extract with 2% chlorhexidine gluconate while the other groups had significant differences from each other. Conclusion: Ramania leaf extract concentrations of 6.25%, 12.5%, 25%, 50%, and 100% had antifungal activity in inhibiting the growth of the fungus Candida albicans. Keywords: Antifungal Activity, Candida albicans, Chlorhexidine gluconate 2%, Ramania Leaf Extract.

Human Dental Pulp Stem Cells Are Subjected to Metabolic Reprogramming and Repressed Proliferation and Migration by the Sympathetic Nervous System via α1B-Adrenergic Receptor

IntroductionHuman dental pulp stem cells (hDPSCs) reside in specialized microenvironments in the dental pulp, termed “niches,” which are composed of diverse cellular components including nerves. Sensory nerves can positively regulate the expansion and differentiation of pulp cells, while the biological effects of the sympathetic nervous system (SNS) on hDPSCs remain elusive. This study is devoted to investigating the effects and underlying mechanisms of the SNS on the proliferation and migration of hDPSCs. MethodsThe distribution of sympathetic nerve fibers in human dental pulp was examined by immunofluorescence staining of tyrosine hydroxylase. The concentration of norepinephrine in healthy and carious human dental pulp tissues was detected using enzyme-linked immunosorbent assay. RNA-sequencing was applied to identify the dominant sympathetic neurotransmitter receptor in hDPSCs. Seahorse metabolic assay, adenosine triphosphate assay, lactate assay, and mitochondrial DNA copy number were performed to determine the level of glycometabolism. Transwell assay, wound healing assay, 5-ethynyl-2'-deoxyuridine staining assay, cell cycle assay, and Cell Counting Kit-8 assay were conducted to analyze the migratory and proliferative capacities of hDPSCs. ResultsSprouting of sympathetic nerve fibers and an increased concentration of norepinephrine were observed in inflammatory pulp tissues. Sympathetic nerve fibers were mainly distributed along blood vessels, and aldehyde dehydrogenase 1–positive hDPSCs resided in close proximity to neurovascular bundles. ADRA1B was identified as the major sympathetic neurotransmitter receptor expressed in hDPSCs, and its expression was enhanced in inflammatory pulp tissues. In addition, the SNS inhibited the proliferation and migration of hDPSCs through metabolic reprogramming via ADRA1B and its crosstalk with serine-threonine kinase and p38 mitogen-activated protein kinase signaling pathways. ConclusionsThis study demonstrates that the SNS can shift the metabolism of hDPSCs from oxidative phosphorylation to anaerobic glycolysis via ADRA1B and its crosstalk with serine-threonine kinase and p38 mitogen-activated protein kinase signaling pathways, thereby inhibiting the proliferative and migratory abilities of hDPSCs. This metabolic shift may facilitate the maintenance of the quiescent state of hDPSCs.

IPO7 promotes lipopolysaccharide-induced inflammatory responses in human dental pulp cells via p38 MAPK and NF-κB signaling pathways

Pulpitis is a chronic inflammatory process that greatly affects the physical, mental health and life quality of patients. Human dental pulp cells (hDPCs) are essential components of dental pulp tissue and play a significant role in pulpitis. Lipopolysaccharide (LPS) is an initiator of pulpitis and can induce the production of inflammatory cytokines in hDPCs by activating p38 MAPK and NF-κB signaling pathways. Importin7 (IPO7), a member of the importin-β family, is widely expressed in many tissues. Previous studies have shown that IPO7 mediated nuclear translocation of p-p38 after stimulation, and IPO7 homologous protein IPO8 participated in human dental pulp inflammation. This research aims to investigate whether IPO7 is involved in pulpitis and explore its underlying mechanisms. In the current study, we found the expression of IPO7 was increased in pulpitis tissue. In vitro, hDPCs treated with LPS to mimic the inflammatory environment, the expression of IPO7 was increased. Knockdown of IPO7 significantly inhibited the production of inflammatory cytokines and suppressed the p38 MAPK and NF-κB signaling pathways. Activating the p38 MAPK and NF-κB signaling pathways by the p38 activator and p65 activator reversed the inflammatory responses. IPO7 interacted with p-p38 under LPS stimulation in hDPCs. In addition, the increased binding between IPO7 and p-p38 is associated with the decreased binding ability of IPO7 to Sirt2. In conclusion, we found that IPO7 was highly expressed in pulpitis and played a vital role in modulating human dental pulp inflammation.