MicroRNA-93-5p regulates odontogenic differentiation and dentin formation via KDM6B.

BACKGROUND: Epigenetic factors influence the odontogenic differentiation of dental pulp stem cells and play indispensable roles during tooth development. Some microRNAs can epigenetically regulate other epigenetic factors like DNA methyltransferases and histone modification enzymes, functioning as epigenetic-microRNAs. In our previous study, microarray analysis suggested microRNA-93-5p (miR-93-5p) was differentially expressed during the bell stage in human tooth germ. Prediction tools indicated that miR-93-5p may target lysine-specific demethylase 6B (KDM6B). Therefore, we explored the role of miR-93-5p as an epi-miRNA in tooth development and further investigated the underlying mechanisms of miR-93-5p in regulating odontogenic differentiation and dentin formation. METHODS: The expression pattern of miR-93-5p and KDM6B of dental pulp stem cells (DPSCs) was examined during tooth development and odontogenic differentiation. Dual luciferase reporter and ChIP-qPCR assay were used to validate the target and downstream regulatory genes of miR-93-5p in human DPSCs (hDPSCs). Histological analyses and qPCR assays were conducted for investigating the effects of miR-93-5p mimic and inhibitor on odontogenic differentiation of hDPSCs. A pulpotomy rat model was further established, microCT and histological analyses were performed to explore the effects of KDM6B-overexpression and miR-93-5p inhibition on the formation of tertiary dentin. RESULTS: The expression level of miR-93-5p decreased as odontoblast differentiated, in parallel with elevated expression of histone demethylase KDM6B. In hDPSCs, miR-93-5p overexpression inhibited the odontogenic differentiation and vice versa. MiR-93-5p targeted 3' untranslated region (UTR) of KDM6B, thereby inhibiting its protein translation. Furthermore, KDM6B bound the promoter region of BMP2 to demethylate H3K27me3 marks and thus upregulated BMP2 transcription. In the rat pulpotomy model, KDM6B-overexpression or miR-93-5p inhibition suppressed H3K27me3 level in DPSCs and consequently promoted the formation of tertiary dentin. CONCLUSIONS: MiR-93-5p targets epigenetic regulator KDM6B and regulates H3K27me3 marks on BMP2 promoters, thus modulating the odontogenic differentiation of DPSCs and dentin formation.

Characteristics of pediatric mandibular condylar fractures in Southwest China: A single-center and 12-year retrospective study.

BACKGROUND/AIM: Mandibular condylar fractures in pediatric patients may exhibit distinct epidemiological characteristics attributed to their unique growth and development phase, as well as various anatomical, physiological, biomechanical, and behavioral factors that differentiate them from adults. This study aimed to investigate the demographics, injurious factors, classifications, clinical manifestations, and treatments of pediatric mandibular condylar fractures, as well as the concomitant injuries in maxillofacial and other body parts. MATERIALS AND METHODS: This retrospective study analyzed the clinical data of 189 pediatric patients with mandibular condylar fractures between 2011 and 2022. Variables investigated included age, gender, timing of onset, causes, classification of condylar fracture, concomitant injuries, clinical manifestations, and treatment modalities. RESULTS: A total of 189 patients, a higher proportion of boys compared to girls was observed, with the highest incidence rate in children aged 1-3 years. They occurred primarily in July, June, and September as well as on Saturdays and Sundays. The most prevalent cause of mandibular condylar fractures was falls from heights in 73 patients (38.62%). Pediatric patients exhibited a higher susceptibility to condylar head fractures. A significant majority (81.48%) of these fractures were accompanied by soft tissue injuries in the maxillofacial region, with the chin being particularly vulnerable to injury. In addition, 61.90% of pediatric patients experienced fractures in other areas of the maxillofacial region, with the mandibular symphysis being the most commonly affected site. Dental trauma predominantly occurred in the anterior region (44.97%). Notably, a substantial proportion (28.04%) of cases also presented with multiple systemic injuries. CONCLUSIONS: The characteristics of pediatric mandibular condylar fractures exhibit distinct features in terms of age, gender, timing of onset, etiology, location and type, the presence of concomitant maxillofacial soft/hard tissue injuries and multiple systemic injuries, as well as clinical manifestations and treatment modalities. Therefore, clinicians should pay special attention to the diagnosis and treatment of pediatric condylar fractures.

LncRNA CARMN facilitates odontogenic differentiation of dental pulp cells by impairing EZH2.

OBJECTIVE: This study aimed to reveal the potential role of CARMN in odontogenic differentiation of dental pulp cells (DPCs). METHODS: Laser capture microdissection was used to detect Carmn in DPCs and odontoblasts in P0 mice. After manipulating CARMN expression in odontogenic differentiation induced hDPCs, the state of odontogenic differentiation was evaluated by ALP staining, ARS, and related marker expression in qRT-PCR and western blotting. The subcutaneous transplantation of HA/ß-TCP loaded with hDPCs was performed to verify CARMN's role in promoting odontogenic differentiation in vivo. RNAplex and RIP were employed to reveal potential mechanism of CARMN in hDPCs. RESULTS: CARMN expressed more abundantly in odontoblasts than DPCs in P0 mice. CARMN expression boosted during in vitro odontogenic differentiation of hDPCs. CARMN overexpression enhanced odontogenic differentiation of hDPCs in vitro, while inhibition impaired the process. CARMN overexpression in HA/ß-TCP composites promoted more mineralized nodule formation in vivo. CARMN knockdown led to soared EZH2, while CARMN overexpression brought about EZH2 inhibition. CARMN functioned via direct interaction with EZH2. CONCLUSIONS: The results uncovered CARMN as a modulator during the odontogenic differentiation of DPCs. CARMN promoted odontogenic differentiation of DPCs by impairing EZH2.

Effects of Cold-Light Bleaching on Enamel Surface and Adhesion of Streptococcus mutans.

Tooth bleaching is becoming increasingly popular among patients with tooth staining, but the safety of bleaching agents on tooth structure has been questioned. Primarily thriving on the biofilm formation on enamel surface, Streptococcus mutans has been recognized as a major cariogenic bacterial species. The present study is aimed at investigating how cold-light bleaching would change enamel roughness and adhesion of Streptococcus mutans. Human premolars were divided into 72 enamel slices and allocated into 3 groups: (1) control, (2) cold-light bleaching with 35% hydrogen peroxide (Beyond™), and (3) 35% hydrogen peroxide (Beyond™) alone. Biofilms of Streptococcus mutans were cultivated on enamel slices in 5% CO2 (v/v) at 37°C for 1 day or 3 days. Enamel surfaces and biofilms were observed using scanning electron microscope (SEM). Atomic force microscopy (AFM) was applied to quantify the roughness of enamel surface, and the amounts of biofilms were measured by optical density of scattered biofilm and confocal laser scanning microscopy (CLSM). Cold-light bleaching significantly increased (p < 0.05) surface roughness of enamel compared to controls, but significantly inhibited (p < 0.05) adhesion of Streptococcus mutans on enamel in the bacterial cultures of both 1 day and 3 days. In conclusion, cold-light bleaching could roughen enamel surface but inhibit Streptococcus mutans adhesion at the preliminary stage after the bleaching treatment.

Effect of photobiomodulation therapy on mini-implant stability: a systematic review and meta-analysis.

The study aimed to assess trials investigating the effect of PBMT on mini-implant stability. Electronic searches of seven databases and manual search were conducted up to May 2020. Randomized controlled trials and controlled clinical trials evaluating the effect of PBMT on mini-implant stability were included. The risks of bias of individual studies were performed using ROB 2.0 and ROBINS-I-tool based on different study design. Meta-analysis was conducted to compare mini-implant stability exposed to PBMT with control ones at different time points after implantation. Among the 518 records initially identified, seven studies were included in this study. Six studies investigated low-level laser therapy (LLLT) and one study evaluated light-emitting diode (LED) therapy. Two studies were eligible for meta-analysis, which showed that LLLT significantly improved mini-implant stability 60 days after initial implantation (MD - 3.01, 95% CI range [- 4.68, - 1.35], p = 0.0004). High energy density of LLLT began to show beneficial effect on mini-implant stability as early as 3 days after implantation, while the significant effect of low energy density displayed later than 30 days after insertion. LED therapy could improve mini-implant stability after 2 months post-insertion. In conclusion, PBMT appears to be beneficial in ameliorating mini-implant stability. High energy density of LLLT might exert more rapid effect than low energy density. More high-quality clinical trials are needed to further demonstrate PBMT' effects on orthodontic mini-implants.

circAKT3 positively regulates osteogenic differentiation of human dental pulp stromal cells via miR-206/CX43 axis.

BACKGROUND: Human dental pulp stromal cells (hDPSCs) are promising sources of mesenchymal stem cells (MSCs) for bone tissue regeneration. Circular RNAs (circRNAs) have been demonstrated to play critical roles in stem cell osteogenic differentiation. Herein, we aimed to investigate the role of circAKT3 during osteogenesis of hDPSCs and the underlying mechanisms of its function. METHODS: We performed circRNA sequencing to investigate the expression profiles of circular RNAs during osteogenesis of hDPSCs. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the expression pattern of circAKT3 and miR-206 in hDPSCs during osteogenesis. We knocked down circAKT3 and interfered the expression of miR-206 to verify their regulatory role in hDPSC osteogenesis. We detected hDPSCs mineralization by alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining and used dual-luciferase reporter assay to validate the direct binding between circAKT3 and miR-206. To investigate in vivo mineralization, we performed subcutaneous transplantation in nude mice and used hematoxylin and eosin, Masson's trichrome, and immunohistochemistry staining. RESULTS: Totally, 86 circRNAs were differentially expressed during hDPSC osteogenesis, in which 29 were downregulated while 57 were upregulated. circAKT3 was upregulated while miR-206 was downregulated during hDPSC osteogenesis. Knockdown of circAKT3 inhibited ALP/ARS staining and expression levels of osteogenic genes. circAKT3 directly interacted with miR-206, and the latter one suppressed osteogenesis of hDPSCs. Silencing miR-206 partially reversed the inhibitory effect of circAKT3 knockdown on osteogenesis. Connexin 43 (CX43), which positively regulates osteogenesis of stem cells, was predicted as a target of miR-206, and overexpression or knockdown of miR-206 could correspondingly decrease and increase the expression of CX43. In vivo study showed knockdown of circAKT3 suppressed the formation of mineralized nodules and expression of osteogenic proteins. CONCLUSION: During osteogenesis of hDPSCs, circAKT3 could function as a positive regulator by directly sponging miR-206 and arresting the inhibitive effect of miR-206 on CX43 expression.

Effect of clear aligners on oral health-related quality of life: A systematic review.

Clear aligners have been frequently applied in orthodontic clinic practice. However, its effect on oral health-related quality of life (OHRQoL) compared with fixed appliance treatment (FAT) remains inconclusive. This systematic review aimed to compare the impacts of clear aligner treatment (CAT) with FAT on patients' OHRQoL. Electronic searches of databases (PubMed, Cochrane Database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, Embase, Medline, two Chinese databases and six grey literature databases) were conducted up to July 2019. Randomized controlled trials, controlled clinical trials, cohort studies and cross-sectional studies comparing the impact of CAT and FAT on OHRQoL with validated instruments were included. Extraction of data and assessment of the risk of bias were conducted using ROBINS-I-tool, Newcastle-Ottawa Scale and ROB 2.0 based on study design. Of the 1112 records initially identified, 2 studies were included in this review. One study evaluated OHRQoL at the last debonding appointment, while the other made evaluation at the early stage of treatment. In the aspect of functional dimensions, both studies reported less eating disturbance in CAT patients than FAT ones. Based on currently limited information, the effect of CAT on the overall OHRQoL compared to FTA was still inconclusive. In individual dimensions, however, weak evidence supported that CAT might cause less eating disturbance than FAT. More high-quality clinical trials using validated OHRQoL instruments are needed to draw more reliable conclusions in the effect of CAT and FAT on OHRQoL.

Insight into the Role of Long Non-coding RNAs During Osteogenesis in Mesenchymal Stem Cells.

BACKGROUND: Long non-coding RNAs (LncRNAs) are non-protein coding transcripts longer than 200 nucleotides in length. Instead of being "transcriptional noise", lncRNAs are emerging as a key modulator in various biological processes and disease development. Mesenchymal stem cells can be isolated from various adult tissues, such as bone marrow and dental tissues. The differentiation processes into multiple lineages, such as osteogenic differentiation, are precisely orchestrated by molecular signals in both genetic and epigenetic ways. Recently, several lines of evidence suggested the role of lncRNAs participating in cell differentiation through the regulation of gene transcriptions. And the involvement of lncRNAs may be associated with initiation and progression of mesenchymal stem cell-related diseases. OBJECTIVE: We aimed at addressing the role of lncRNAs in the regulation of osteogenesis of mesenchymal stem cells derived from bone marrow and dental tissues, and discussing the potential utility of lncRNAs as biomarkers and therapeutic targets for mesenchymal stem cell-related diseases. RESULTS: Numerous lncRNAs were differentially expressed during osteogenesis or odontogenesis of mesenchymal stem cells, and some of them were confirmed to be able to regulate the differentiation processes through the modifications of chromatin, transcriptional and post-transcriptional processes. LncRNAs were also associated with some diseases related with pathologic differentiation of mesenchymal stem cells. CONCLUSION: LncRNAs involve in the osteogenic differentiation of bone marrow and dental tissuederived mesenchymal stem cells, and they could become promising therapeutic targets and prognosis parameters. However, the mechanisms of the role of lncRNAs are still enigmatic and require further investigation.

Nicotine is a risk factor for dental caries: An in vivo study.

BACKGROUND/PURPOSE: Streptococcus mutans is an important pathogen in the development of dental caries. Many studies have focused on the relationship between nicotine and S. mutans in vitro. The aim of this study was to investigate the effect of nicotine on the growth of S. mutans and its cariogenic potential in vivo. MATERIALS AND METHODS: Sixteen male Specific-pathogen-free Wistar rats were divided into 2 groups (nicotine-treated and nicotine-untreated group) and infected with S. mutans. The S. mutans suspension was treated with 1 mg/mL nicotine in the nicotine-treated group. The Keyes method was used to evaluate sulcal caries of rats, and dental plaque on molar teeth was observed by scanning electron microscopy (SEM). RESULTS: Incidence of sulcal caries was higher in nicotine-treated group compared to nicotine-untreated group (42.7 ±â€¯1.7 vs 37.3 ±â€¯4.9, P = 0.009). Severity of caries increased with nicotine treatment. The slightly dentinal caries scores and moderate dentinal caries scores were higher in the presence of nicotine (P < 0.001). Increased number of S. mutans cells attached to dental surface was observed under SEM in the nicotine-treated group. CONCLUSION: Nicotine would promote the attachment of S. mutans to dental surface, and further increase the incidence and severity of dental caries. Therefore, nicotine might be a risk factor for smoking-induced caries.

[A review of the effect of tooth bleaching agents on oral microbes].

Tooth bleaching agents contain powerful oxidizing agents, which serve as the main part of bleaching agents because of its release of effective bleaching component. It has been a hot topic whether tooth bleaching agents exert negative influence on oral health. In order to provide train of thoughts and reference for further clinical researches and treatments, this review paper focuses on bleaching agents' effects on the growth of oral microbes and the formation of biofilms.