Background Mass spectrometry (MS)-based proteomics approaches have proven to be a powerful tool to discover clinical biomarkers for dental caries diagnosis and management. Saliva, acquired enamel pellicle (AEP), and dentin are three common sample medium. However, MS-based proteomics of dental caries is currently facing two major problems: (1) the analytical coverage of the dental caries proteome is relatively limited (<3000 proteins), which may hamper our understanding of the function of the key but low-abundance proteins. (2) gingival crevicular fluid (GCF) is a valuable oral fluid, but the association of GCF proteins and dental caries remains underexplored and the GCF proteomic profile in the context of dental caries has not yet been characterized. Results Based on our previously developed single-pot, solid-phase-enhanced sample-preparation (SP3)-based GCF sample preparation method, the GCF proteome of healthy control (HC) group (31 subjects, DMFT=0) and pediatric dental caries group (30 subjects, DMFT>7) were quantitatively analyzed by using a directDIA-MS method for the first time. A total of 6397 proteins were identified in all GCF samples. Compared to HC group, 75 proteins were differentially expressed (42 up-regulated, 33 down-regulated) in pediatric dental caries group. A combination of six protein biomarkers, including MUC5B, DMBT1, PIP, PIGR, TIMP1 and S100A7, showed great potential (ROC-AUC 0.944) to differentiating severe pediatric caries group from HC group. Significance and Novelty The GCF proteome for dental caries was characterized for the first time by an SP3-based directDIA-MS method. We generated the largest dataset of GCF proteome for dental caries to date and proposed a combination of six protein biomarkers to predict severe pediatric caries. The proposed protocol and findings will be an important complement and provide new insights into the diagnosis and treatment of dental caries in children.

OBJECTIVE:To apply Erbium-doped Yttrium Aluminum Garnet (Er:YAG) laser in pulpotomy for experimentally induced pulpitis in mature permanent teeth of rats, and investigate whether Er:YAG laser can improve the effect of pulpotomy in mature permanent teeth with pulpitis. METHODS:Thirty-six 3-month-old male Sprague-Dawley (SD) rats were selected, with bilateral maxillary first molars as experimental teeth. 4 rats (8 experimental teeth) were selected to construct the pulpitis model. After confirming the successful establishment of the model by this method, 32 rats were randomly divided into the control group and the experimental group, and the pulpitis models were constructed in the same way. The control group(mechanical group, 16 rats, 32 experimental teeth) received pulpotomy with conventional sterile excavators. The experimental group (laser group, 16 rats, 32 experimental teeth) received pulpotomy using Er:YAG laser. At 3, 7, 14, and 28 days post-operation, four rats from each group were randomly euthanized. Pathological changes in the dental pulp were observed using hematoxylin - eosin (HE) staining. Immunohistochemical (IHC) assessment and mean optical density (MOD) measurement were performed to evaluate the expression of interleukin-1β (IL-1β) and partitioning defective protein 3 (Par3). Inter-group differences were analyzed using the Mann-Whitney U test or the independent samples t-test. RESULTS:Histological examination by HE staining demonstrated favorable pulpal repair in the laser-treated group. The total histopathological scores were significantly lower in the laser group compared to the mechanical group at days 3 and 7 post-operation (p < 0.05). However, no statistically significant difference was observed between the two groups at days 14 and 28. IHC analysis revealed that the mean optical density (MOD) values for IL-1β were consistently lower in the laser group at all four time points (p < 0.05), while the MOD values for Par3 were consistently higher in the laser group (p < 0.05). CONCLUSION:The Er:YAG laser used during pulpotomy in mature permanent rat teeth with pulpitis preserves the remaining healthy pulp tissue, reduces IL-1β expression, and enhances Par3 expression, thereby alleviating inflammation and promoting tissue repair.

Obesity is closely linked to periodontitis development. Although semaglutide is increasingly used to manage obesity, its effects on alveolar bone repair in periodontitis are not well understood. In a mouse model of obesityrelated periodontitis, we found that obesity exacerbates alveolar bone loss in an inflammatory setting. Semaglutide treatment slightly reduced periodontal inflammation and osteoclast markers but did not improve osteogenesis or regenerate alveolar defects. Transcriptomic analysis revealed increased fatty-acid uptake but suppressed AMPK signaling and reduced fatty-acid oxidation (FAO) in inflamed alveolar bone of obese mice. In vitro, macrophage FAO was largely unaffected by inflammation or semaglutide, while FAO in bone marrow-derived mesenchymal stem cells (BMSCs) was significantly reduced, impairing osteogenic differentiation. To tackle metabolic and osteogenic issues, we created a metformin-loaded mesoporous polydopamine system (Met@MPDA) that acts as an AMPK agonist. Met@MPDA is absorbed by BMSCs, restoring cellular energy and promoting alveolar bone regeneration in periodontitis. While semaglutide provides anti-inflammatory benefits in obesity-related periodontitis, it falls short in boosting bone regeneration due to metabolic issues in BMSCs. Thus, Met@MPDA's sustained metformin delivery could enhance bone regeneration in these cases.

BackgroundThe presence of supernumerary teeth (ST) may result in varying degrees of dentition problem.PurposeThe aims of this study were to investigate the distribution and characteristics of ST in children in Shanghai, analyze the factors that have adverse effects on dentition, and provide some reference for clinicians.MethodsA total of 309 patients (427 ST) who underwent cone-beam computed tomography (CBCT) were included in this study. The age and sex of the patients, and the characteristics of the ST, such as their number, distribution, direction and effect on adjacent teeth, were retrospectively collected.ResultsThe average age of the patients was 6.3 years, and the male to female ratio was 4.15:1. ST mostly occurred in the maxillary anterior region, with the majority of cases involving one tooth. The ratio of the number of ST impacted in bone to the number of erupted ST was 14.8. The palatal side of the alveolar bone (68%) and inverted (59.5%) were associated with the greatest proportions of ST in terms of location and direction, respectively. When located on the palatal side of the dentition and in an inverted orientation, ST have minimal influence on the eruption and position of the central incisors, whereas the buccolingual ST have the most complications.ConclusionThe impact of ST in different positions and directions on the eruption of adjacent permanent teeth varies. These findings may serve as reference information for clinical decision-making on the timing of tooth extraction.

Pulpotomy is now a recommended treatment for pulpitis in primary teeth with the advancement of dental biomaterials. However, there is deficient in clinical studies which evaluate the long-term prognosis of pulpotomy in primary teeth. Thus, the study was aimed to evaluate the efficacy of iRoot BP Plus and mineral trioxide aggregate (MTA) for pulpotomy in primary molars with the aid of intraoral endoscope and to explore the influencing factors. The teeth records with iRoot BP Plus and MTA were included. The gender, age, tooth position, clinical examination, radiographs, clinical procedures, coronal restoration and follow-up information were obtained. The outcomes were analyzed by Chi-square test, Kaplan-Meier method and Cox regression (P < 0.05). The study included 82 children with 157 teeth (113 in the iRoot BP Plus group and 44 in the MTA group). The treatment procedure and the follow-up were monitored with the assistance of intraoral endoscope. The average follow-up period was 1213 days for iRoot BP Plus and 1289 days for MTA. Kaplan-Meier analysis showed a 1-year cumulative survival rate of 100

Background: Traumatic dental injuries (TDI) are considered a public health problem worldwide, especially during childhood. This study aimed to investigate parental knowledge and attitude toward TDI emergency management and associated factors among a Chinese sample. Methods: A total of 714 parents of children aged 0-14 years were recruited in this cross-sectional study. Parents were asked to fill out a questionnaire after signing an informed consent according to ethical guidelines. The questionnaire comprised two sections. One section focused on socio-demographic information, and the other was concerned about parental knowledge and attitudes toward TDI emergency management. Results: The study had an 88.2% response rate. Finally, the data of 630 participants were analyzed. Among all the respondents, 39.8% of the parents said they knew nothing about TDI emergency management; 62.9% said they knew where to seek professional help for TDI. Only 74 parents would choose fresh milk as a storage medium for avulsed permanent teeth. The interest in learning more about TDI emergency management was significantly associated with parental gender, educational level and family income. Additionally, both children's and parents' previous experience with TDI was strongly correlated with their responses to the question, "Do you know where to seek professional help for dental trauma?". Parents of children who had previously experienced TDI were more likely to believe that it is possible to reattach a fractured tooth piece due to TDI (p = 0.031). Among the 444 parents who expressed interest in learning more about the emergency management of TDI, the majority (n = 311) preferred to receive the relevant information via the Internet. Conclusions: Parental knowledge regarding TDI emergency management was inadequate. Educational programs, especially online resources, are needed to promote public health education on TDI emergency management in China.

The histologic outcomes of regenerative endodontic procedures (REPs) in humans remain debated, with evidence varying between cementum-/bone-like tissue and true pulp-dentin complex regeneration. Here, we present a case of a 10-year-old girl who underwent REPs for a maxillary premolar (tooth #4) diagnosed as symptomatic irreversible pulpitis with symptomatic apical periodontitis. At the 8-month follow-up, the patient was asymptomatic with radiographic evidence of apical closure and dentinal wall thickening; the tooth was subsequently extracted due to orthodontic indications, along with the sound contralateral premolar (tooth #12) as control. Micro-computed tomography confirmed apical closure and dentinal wall thickening with heterogeneous intracanal mineralization in tooth #4. Histologic examination revealed dentin-like tissue with tubular structures, surfaced by flattened, dentin sialophosphoprotein-positive cells. Neurovascular regeneration assessed by CD31/PGP9.5 co-localization was sparse and disorganized compared to the normal control. Osteoid tissue ingrowth was present in the apical region. This case demonstrates that REPs can yield structurally disorganized pulp-dentin-like tissue, highlighting the key challenge of achieving true histologic regeneration.

Orofacial clefts, particularly cleft palate, are among the most common congenital anomalies, imposing substantial functional impairment and psychosocial burden on affected individuals. Emerging evidence implicates epigenetic modifications, especially histone lysine methylation, as critical regulators of embryonic development, yet their precise roles in palatogenesis remain poorly defined. The H3K4 methyltransferase complexes, which require the core subunit ASH2L for full enzymatic activity, govern transcriptional activation during development; however, whether ASH2L-mediated H3K4me3 deposition contributes to palatal morphogenesis is unknown. Here, we show that conditional deletion of Ash2l in Six2-positive palatal mesenchymal cells results in complete cleft palate in mice. Multi-omics analyses (RNA-seq and CUT Tag-seq) reveal that Ash2l loss causes genome-wide reduction of H3K4me3 enrichment at key developmental genes, most notably Wnt5b, leading to their transcriptional downregulation. Functionally, Ash2l deficiency impairs proliferation and osteogenic differentiation of palatal mesenchymal cells, resulting in stunted palatal shelf growth and disrupted osteogenesis. Pharmacological activation of WNT signaling downstream of Wnt5b partially rescues these cellular defects, confirming the functional relevance of this axis. These findings establish ASH2L-mediated H3K4me3 as a critical epigenetic regulator of palatal development through WNT5B modulation, and identify potential therapeutic targets for cleft palate.

Angiogenesis remains a key challenge in pulp regeneration due to the ischemic and hypoxic root canal environment. Apoptotic vesicles (ApoVs) offer unique advantages as a breakthrough strategy due to their stability in hypoxic environments, resistance to immune clearance, and high yield. However, a critical gap persists in biomaterials capable of spatiotemporal ApoVs delivery for pulp regeneration. To address this, an ApoVs-laden silk fibroin/sodium alginate (SF/SA/ApoVs) hydrogel with tunable release kinetics was fabricated. This hydrogel exhibited superior shapeability, injectability, and self-healing properties. SF/SA/ApoVs hydrogel improved angiogenesis by enhancing paracrine functions of dental pulp stem cells (DPSCs), and promoting capillary lumen formation of human umbilical vein endothelial cells via the focal adhesion signaling pathway. It also effectively created an odontogenic and neurogenic inductive micro-environment for pulp-dentin complex formation. Furthermore, in-vivo studies showed that semi-orthotopic transplantation of SF/SA/ApoVs hydrogel accelerated micro-vessels formation, and orthotopic transplantation further confirmed its capacity in generating full-length vascularized pulp tissue with aligned odontoblast-like cells and neuronal extension. Our study pioneers an ApoVs-functionalized hydrogel system with advanced angiogenic potential for promoting pulp regeneration.

Introduction: Avulsion and root fracture represent the two most severe forms of traumatic dental injuries. One of the crucial goals in the treatment of traumatized is to maintain the space and site until the child reaches adulthood. Case description: A 7-year-old child experienced a root fracture of the right maxillary permanent incisor, with the crown segment being avulsed and preserved in saline for 12 hours. No pulp tissue was observed within the pulp cavity of the avulsed segment. The residual fractured root was in the alveolar bone. The root development is at Nolla stage 7. No radiographic evidence showed alveolar ridge or mandibular fractures. We performed a gingival flap surgery to expose the fractured surface of the tooth, replanted the avulsed coronal segment, and stabilized it. Subsequently, we accessed the pulp chamber, induced bleeding from the apical region below the enamel-cementum junction, and applied a filling. Nearly two years postoperatively, the tooth root continued to develop, and the replanted segment remained stable without any mobility. Discussion: Replacement resorption is a common complication following delayed replantation. However, in this particular case, the avulsed root fragment is situated near the cervical region of the tooth due to the root fracture, and the periodontal ligament (PDL) of the remaining root is healthy. Additionally, successful revascularization was achieved in the pulp cavity of the coronal segment. No replacement resorption has been observed to date. Conclusion/clinical significance: In delayed replantation, the use of revascularization may reduce ankylosis/replacement resorption and promote the establishment of a functional healing response.

BACKGROUND:Dentists are prone to a variety of occupational health problems and are at a high risk of workplace violence due to the nature of their dental practice. The aim of this study was to assess the state of occupational health and the prevalence of workplace violence among dentists in China. Additionally, this research endeavored to offer practical recommendations for Chinese dentists and relevant institutions based on the findings. METHODS:The survey was conducted based on a sample of 1109 dentists from China as respondents to an electronic questionnaire survey. This self-reported questionnaire encompassed 14 distinct types of injuries, 28 specific diseases, 11 symptoms of sub-health state, and 3 categories of workplace violence. RESULTS:In dental practice, the most common injuries are needlestick incidents and falls, each affecting 54.28% of dentists. Conditions like lumbar and cervical spondylosis, along with scapulohumeral periarthritis, are prevalent, with 53.02% and 45.99% of dentists reporting these issues, respectively. Our univariate analysis highlighted significant health disparities among dentists, influenced by gender and department. Additionally, our findings underscore the harsh reality of verbal, physical, and sexual harassment faced by Chinese dentists, which has a profound negative impact and is met with woefully inadequate protective measures. CONCLUSION AND RECOMMENDATION:Chinese dentists face significant physical and psychological stress, further exacerbated by verbal abuse, physical violence, and sexual harassment. It is imperative that hospitals, clinics, and governmental agencies step up to their responsibilities by fostering a secure work environment for Chinese dentists.

Background: Anterior crossbite is a frequently occurring clinical condition in children which can lead to Class III malocclusion. It is important to identify the recurrence predictors in children with anterior crossbite for achieving long-term stability in early orthopedic treatment. Methods: In this retrospective study, total of 36 patients with anterior crossbite in their primary dentition were enrolled. They were divided into two groups based on the subsequent relapse status in mixed dentition: the non-relapsed group and the relapsed group. The baseline characteristics for both groups were collected and compared before treatment. The cephalometric measurements were analysed at the pre-treatment and post-treatment phases. The cephalometric measurements of relapsed group were compared for the post-treatment phase and relapse phase after treatment. Results: A significant difference in participant characteristics was observed between the two groups wherein the relapsed group showed lower tongue position. The relapsed group before treatment exhibited more anteriorly and inferiorly mandible along with protrusion upper incisors. This condition did not change after the treatment. Early orthopedic treatment in both groups attained significant forward movement of maxilla and protrusive movement of upper incisors. The downward movement of hyoid bone was also noticed. In mixed dentition, patients in the relapsed group exhibited forward movement of the mandible and downward movement of hyoid bone. Conclusions: The findings indicate that the lower tongue position and specific cephalometric variables including forward growing mandible and lower positioned hyoid bone in patients with Class III growth patterns may serve as the potential predictors of relapse.

BACKGROUND:Orthodontic diagnosis and treatment planning are closely associated with the pubertal growth spurt. Previously, we developed a simplified MS-based protocol for deep quantitative analysis of human gingival crevicular fluid (GCF) proteome for skeletal maturity indicators. The purpose of this study is to perform an in-depth and comparative analysis of the GCF metaproteome at prepubertal and circumpubertal stages to aid oral health and skeletal maturity evaluation. METHODS:Based on our previously obtained and published LC-MS data, the GCF metaproteome of 45 children (24 subjects from prepubertal group and 21 subjects from circumpubertal group) were analyzed by searching against the Human Oral Microbiome Database using FragPipe software. Differentially expressed bacterial proteins between two groups were analyzed using Wilcoxon rank sum test. Differentially abundant taxa between two groups were evaluated using linear discriminant effect size (LEfSe) analysis. RESULTS:A total of 192 genera were identified in GCF. Neisseria (Neisseriales), Comamonadaceae, Burkholderiales, Proteobacteria and Betaproteobacteria were the most abundant bacterial taxa at prepubertal stage. Firmicutes, Saccharibacteria_TM7_[G-1] (Saccharibacteria TM7), Bacterium_HMT349, Bacilli and Alphaproteobacteria were the most abundant bacterial taxa of GCF microbiota at circumpubertal stage. Compared to that in the prepubertal group, enrichment of Firmicutes (Bacillus) was observed in the circumpubertal group. CONCLUSIONS:Based on our developed and already published single-pot, solid-phase enhanced sample-preparation (SP3)-based liquid chromatography (LC) - high-field asymmetric waveform ion mobility spectrometry (FAIMS)-MS protocol for deep quantitative analysis of human GCF metaproteome, we were able to generate the largest dataset of the human GCF metaproteome (14376 bacterial proteins) to date and revealed distinct microbial community at prepubertal and circumpubertal stages. The proposed protocol and findings will be useful to aid oral health and skeletal maturity evaluation for orthodontic diagnosis and treatment planning.

Dental treatments for children and adolescents have unique clinical characteristics that differ from dental care for adults in terms of children's physiology, psychology, and behavior. These differences impose specific requirements on the application of local anesthesia in pediatric dental procedures. This article presents expert consensus on the principles of local anesthesia techniques in pediatric dental therapies, including the use of common anesthetic drugs and dosage control, safety and efficacy evaluation, and prevention and management of complications. The aim is to improve the safety and quality of pediatric dental treatments and offer guidance for clinical application by dentists.

Osteonecrosis involving the permanent tooth germ in primary dentition is a rare condition that can affect dental and maxillofacial development without correct intervention. This case report presents the successful recovery from drug-induced mandibular osteonecrosis involving the permanent tooth germ in a child. A healthy 4-year-old Chinese girl visited the clinic with an unhealed gingival wound and alveolar bone exposure of the missing primary molar area after 1-day arsenic trioxide sealing during pulp therapy. Radiographic examinations indicated inflammation and sequestrum formation in the jaw. The diagnosis was mandibular osteonecrosis. The treatment plans involved sequestrectomy without extended curettage and removal of the affected permanent tooth germ with systemic antibiotic therapy. A 6-year follow-up revealed no recurrence of the lesion or complications, a gradual increase in bone density of the osteonecrosis area and the development of adjacent permanent tooth germs. A conservative treatment regimen without extended curettage may be an option for young patients with jaw osteonecrosis.

Objectives: This retrospective study evaluated the short-term effects of removable slow maxillary expansion (SME) on eruption patterns of labially ectopic canines in a Chinese pediatric population, comparing treated patients with untreated controls. Methods: Seventy-six patients (mean age 8.38 ± 0.88 years) underwent SME treatment for 11.04 ± 4.44 months. Canine positions were categorized as labial ectopic (TE: n = 40) or normally positioned (TN: n = 112). The TE group was stratified vertically: superior (TES; n = 15, canines above lateral incisors’ roots or adjacent to unerupted incisors) and inferior (TEI; n = 25, canines adjacent to erupted lateral incisors’ roots). Untreated controls (n = 58; mean age 8.46 ± 0.78 years) included labial ectopic (CE group; n = 32) and normal canines (CN group; n = 84), with CE further divided vertically into CES (n = 24) and CEI (n = 8). Panoramic radiographs at baseline (T0) and follow-up (T1) evaluated sector distribution, midline proximity (3c-ML: canine cusp to midline distance), vertical position (3c-OP: cusp to occlusal plane distance), and angular (3^ML: canine-midline angle). Results: SME significantly improved midline proximity (3c-ML increased) while reducing vertical height (3c-OP decreased) and angulation (3^ML reduced) in the TE group. Notably, TE patients revealed a significantly greater increase in 3c-ML compared to CE. Subgroup analysis showed that TEI canines exhibited significant improvements in all three parameters (3c-OP, 3c-ML, and 3^ML), whereas TES canines displayed minimal changes. The shifts in sector distribution were similar between the treatment and control groups. Conclusions: SME demonstrated short-term efficacy in guiding labially ectopic canines toward more favorable eruption trajectories, particularly when erupted beyond the roots of the lateral incisor. The observed positional improvements underscore SME’s potential to optimize eruption outcomes during early orthodontic intervention.

Traumatic dental injuries (TDIs) of teeth occur frequently in children and adolescents. TDIs that impact the periodontal tissues and alveolar tissue can be classified into concussion, subluxation, extrusive luxation, intrusive luxation, lateral luxation, and avulsion. In these TDIs, management of injured soft tissue, mainly periodontal ligament, and dental pulp, is crucial in maintaining the function and longevity of the injured teeth. Factors that need to be considered for management in laxation injuries include the maturation stage of the traumatic teeth, mobility, direction of displacement, distance of displacement, and whether there are alveolar fractures. In avulsion, the maturation stage of the permanent tooth, the out-socket time, storage media/condition of the avulsed tooth, and management of the PDL should also be considered. Especially, in this review, we have subdivided the immature tooth into the adolescent tooth (Nolla stage 9) and the very young tooth (Nolla stage 8 and below). This consensus paper aimed to discuss the impacts of those factors on the trauma management and prognosis of TDI to provide a streamlined guide for clinicians from clinical evaluation, diagnostic process, management plan decision, follow-up, and orthodontic treatment for tooth luxation and avulsion injuries.

Background In the regenerative endodontic procedures, scaffolds could influence the prognosis of affected teeth. Currently, there is controversy regarding the postoperative evaluation of various scaffolds for pulp regeneration. The objective of this study was to access whether other scaffolds, used alone or in combination with blood clot (BC), are more effective than BC in regenerative endodontic procedures. Methods We systematically search the PubMed, the Cochrane Central Register of Controlled Trials (CENTRAL), Embase, and Google Scholar databases. Randomized controlled trials examining the use of BC and other scaffold materials in the regenerative endodontic procedures were included. A random effects model was used for the meta-analysis. The GRADE method was used to determine the quality of the evidence. Results We screened 168 RCTs related to young permanent tooth pulp necrosis through electronic and manual retrieval. A total of 28 RCTs were related to regenerative endodontic procedures. Ultimately, 12 articles met the inclusion criteria and were included in the relevant meta-analysis. Only 2 studies were assessed to have a low risk of bias. High quality evidence indicated that there was no statistically significant difference in the success rate between the two groups (RR=0.99, 95% CI=0.96 to 1.03; 434 participants, 12 studies); low-quality evidence indicated that there was no statistically significant difference in the increase in root length or root canal wall thickness between the two groups. Medium quality evidence indicated that there was no statistically significant difference in pulp vitality testing between the two groups. Conclusions For clinical regenerative endodontic procedures, the most commonly used scaffolds include BC, PRP, and PRF. All the different scaffolds had fairly high clinical success rates, and the difference was not significant. For regenerative endodontic procedures involving young permanent teeth with pulp necrosis, clinical practitioners could choose a reasonable scaffold considering the conditions of the equipment and patients.

Background Pulpotomy is a crucial method to preserve primary teeth until natural exfoliation. This study aimed to evaluate the clinical and radiographic outcomes of pulpotomy with iRoot BP Plus in primary molars and to explore the association between hemostasis time and these outcomes. Methods Primary molars that underwent iRoot BP Plus pulpotomy and were followed for at least 12 months were selected for this study. Clinical and radiographic data were collected, and the success rate was analyzed in relation to factors such as hemostasis time, tooth type, and arch type. The tests of significance used were the chi-square test, Fisher's exact test, or Kruskal-Wallis test. Statistical significance was set at P < 0.05. Results A total of 183 teeth in 106 patients were included in the analysis. The follow-up period fell into a range of 1-3 years, with a mean of 1.6 years. The clinical and radiographic success rates were 96.7% and 92.9%, respectively. The earliest time to observe the radiographic failures was half a year after the treatment, and the latest time was two years after the treatment. Among all the teeth, 130 were recorded with hemostasis time before the application of iRoot BP Plus. Compared to teeth with a hemostasis time of 5 min or less, teeth with a hemostasis time exceeding 5 min showed no significant differences in clinical and radiographic success (P = 1.000 and 0.879). Additionally, neither arch nor teeth type showed a relationship with the pulpotomy success rate (P > 0.05). Conclusions Pulpotomy using iRoot BP Plus in primary molars achieved favorable results. The hemostasis time may not significantly impact the outcomes of pulpotomy using iRoot BP Plus in primary molars.

Histone methylation assumes a crucial role in the intricate process of enamel development. Our study has illuminated the substantial prevalence of H3K4me3 distribution, spanning from the cap stage to the late bell stage of dental germs. In order to delve into the role of H3K4me3 modification in amelogenesis and unravel the underlying mechanisms, we performed a conditional knockout of Ash2l, a core subunit essential for the establishment of H3K4me3 within the dental epithelium of mice. The absence of Ash2l resulted in reduced H3K4me3 modification, subsequently leading to abnormal morphology of dental germ at the late bell stage. Notably, knockout of Ash2l resulted in a loss of polarity in ameloblasts and odontoblasts. The proliferation and apoptosis of the inner enamel epithelium cells underwent dysregulation. Moreover, there was a notable reduction in the expression of matrix-related genes, Amelx and Dspp, accompanied with impaired enamel and dentin formation. Cut&Tag-seq (cleavage under targets and tagmentation sequencing) analysis substantiated a reduction of H3K4me3 modification on Shh, Trp63, Sp6, and others in the dental epithelium of Ash2l knockout mice. Validation through real-time polymerase chain reaction, immunohistochemistry, and immunofluorescence consistently affirmed the observed downregulation of Shh and Sp6 in the dental epithelium following Ash2l knockout. Intriguingly, the expression of Trp63 isomers, DNp63 and TAp63, was perturbed in Ash2l defect dental epithelium. Furthermore, the downstream target of TAp63, P21, exhibited aberrant expression within the cervical loop of mandibular first molars and incisors. Collectively, our findings suggest that ASH2L orchestrates the regulation of crucial amelogenesis-associated genes, such as Shh, Trp63, and others, by modulating H3K4me3 modification. Loss of ASH2L and H3K4me3 can lead to aberrant differentiation, proliferation, and apoptosis of the dental epithelium by affecting the expression of Shh, Trp63, and others genes, thereby contributing to the defects of amelogenesis.