IWP-2 modulates the immunomodulatory properties of human dental pulp stem cells in vitro.

AIM: To investigate the effect of IWP-2, Wnt inhibitor, on human dental pulp stem cells (hDPSCs) responses. METHODOLOGY: hDPSCs were isolated from human dental pulp tissues. Cells were treated with 25 µM IWP-2 for 24 h, and subsequently, the gene expression profile was examined using high-throughput RNA sequencing. The mRNA expression was analysed using qPCR. The effect of IWP-2 was investigated in both normal and LPS-induced hDPSCs (inflamed hDPSCs). CD4+ T cells and CD14+ monocyte-derived macrophages were cultured with conditioned media of IWP-2 treated hDPSCs to observe the immunosuppressive property. RESULTS: RNA sequencing indicated that IWP-2 significantly downregulated several KEGG pathways, including cytokine-cytokine receptor interaction, IL-17 signalling pathway, and TNF signalling pathway. In both normal and inflamed conditions, IWP-2 markedly upregulated TGFB1 mRNA expression while the mRNA expression of pro-inflammatory cytokines, TNFA, IL1B, IFNG, and IL6, was inhibited. In the inhibition experiment, the pretreatment with p38, MAPK, or PI3K inhibitors abolished the effects of IWP-2 in LPS-induced inflammation. In terms of immune cells, IWP-2-treated-inflamed hDPSCs conditioned media attenuated T cell proliferation and regulated regulatory T cell differentiation. In addition, the migratory property of macrophage was decreased after being exposed to IWP-2-treated inflamed hDPSCs conditioned media. CONCLUSION: IWP-2 suppressed inflammatory cytokine expression in both normal and inflamed hDPSCs. Moreover, hDPSCs exerted the immunosuppressive property after IWP-2 treatment. These results suggest the role of Wnt in inflammatory responses and immunomodulation in dental pulp tissues.

Cyanoacrylate tissue adhesive versus silk sutures for mandibular third molar surgery: a systematic review and meta-analysis.

OBJECTIVE: Cyanoacrylate tissue adhesive has been presented as an alternative to sutures and several studies have compared them. The objective of this meta-analysis was to evaluate the effect of cyanoacrylate tissue adhesive on postoperative pain and swelling, following mandibular third molar surgery. MATERIALS AND METHODS: Database search was conducted in MEDLINE/PubMed and Scopus, along with extensive search in the grey literature, including randomized and non-randomized clinical trials that applied cyanoacrylate adhesive for closing mandibular third molar surgical sites and compared it with silk sutures, assessing postoperative pain and swelling. The search ended on September 22, 2023. RESULTS: Of 886 identified articles, six were included and meta-analyzed. Applying cyanoacrylate demonstrated a reduction in the overall postoperative pain (SMD = -0.57, 95% CI -1.00 to -0.15, p = 0.009). A similar outcome was noted when pain was evaluated on the first and last postoperative days, based on controlled clinical trials (SMD = -0.47, 95% CI -0.92 to -0.03, p = 0.04), and randomized trials (SMD = -0.97, 95% CI -1.31 to -0.62, p < 0.00001). Patients/sides received cyanoacrylate showed a decrease in postoperative swelling (SMD = -0.26, 95% CI -0.51 to -0.01, p = 0.04). Following the GRADE rating system, the quality of evidence on pain and swelling was judged as moderate and low, respectively. CONCLUSIONS: The use of cyanoacrylate adhesive may offer benefit in reducing postoperative pain and swelling following mandibular third molar surgery. Nevertheless, this should be further investigated, considering the low number of included reports. CLINICAL RELEVANCE: The current results could help clinicians who perform this procedure to manage postoperative pain and swelling more effectively.

6-Bromoindirubin-3'-Oxime Regulates Colony Formation, Apoptosis, and Odonto/Osteogenic Differentiation in Human Dental Pulp Stem Cells.

6-bromoindirubin-3'-oxime (BIO) is a candidate small molecule that effectively modulates Wnt signalling owing to its stable property. The present study investigated the influence of BIO on the odonto/osteogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs were treated with 200, 400, or 800 nM BIO, and the effects on hDPSC responses and osteogenic differentiation were assessed. BIO-mediated Wnt activation was confirmed by ß-catenin nuclear translocation detected by immunofluorescence staining. BIO attenuated colony formation and cell migration determined by in vitro wound-healing assay. BIO increased early apoptotic cell population evaluated using flow cytometry. For osteogenic induction, BIO promoted alkaline phosphatase (ALP) activity and mineralisation in a dose-dependent manner. ALP, RUNX2, OCN, OSX, ANKH, DMP1, and DSPP mRNA expression were significantly upregulated. The OPG/RANKL expression ratio was also increased. Further, BIO attenuated adipogenic differentiation as demonstrated by decreased lipid accumulation and adipogenic-related gene expression. Bioinformatic analysis of RNA sequencing data from the BIO-treated hDPSCs revealed that BIO modulated pathways related to autophagy and actin cytoskeleton regulation. These findings demonstrated that BIO treatment promoted hDPSC osteogenic differentiation. Therefore, this small molecule is a strong candidate as a bioactive molecule to enhance dentin repair.

Indirect Immobilised Jagged-1 Enhances Matrisome Proteins Associated with Osteogenic Differentiation of Human Dental Pulp Stem Cells: A Proteomic Study.

The indirect immobilisation of Jagged-1 (Jagged-1) promoted osteogenic differentiation of human dental pulp cells (hDPs). Furthermore, the analysis of the Reactome pathway of RNA sequencing data indicates the upregulated genes involved with the extracellular matrix (ECM). Hence, our objective was to investigate the effects of Jagged-1 on proteomic profiles of human dental pulp stem cells (hDPSC). hDPSCs were cultured on the surface coated with human IgG Fc fragment (hFc) and the surface coated with rhJagged1/Fc recombinant protein-coated surface. Cells were differentiated to the osteogenic lineage using an osteogenic differentiation medium (OM) for 14 days, and cells cultured in a growth medium were used as a control. The protein component of the cultured cells was extracted into the cytosol, membrane, nucleus, and cytoskeletal compartment. Subsequently, the proteomic analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS). Metascape gene list analysis reported that Jagged-1 stimulated the expression of the membrane trafficking protein (DOP1B), which can indirectly improve osteogenic differentiation. hDPSCs cultured on Jagged-1 surface under OM condition expressed COL27A1, MXRA5, COL7A1, and MMP16, which played an important role in osteogenic differentiation. Furthermore, common matrisome proteins of all cellular components were related to osteogenesis/osteogenic differentiation. Additionally, the gene ontology categorised by the biological process of cytosol, membrane, and cytoskeleton compartments was associated with the biomineralisation process. The gene ontology of different culture conditions in each cellular component showed several unique gene ontologies. Remarkably, the Jagged-1_OM culture condition showed the biological process related to odontogenesis in the membrane compartment. In conclusion, the Jagged-1 induces osteogenic differentiation could, mainly through the regulation of protein in the membrane compartment.

Human dental pulp stem cells derived extracellular matrix promotes mineralization via Hippo and Wnt pathways.

Extracellular matrix (ECM) is an intricate structure providing the microenvironment niche that influences stem cell differentiation. This study aimed to investigate the efficacy of decellularized ECM derived from human dental pulp stem cells (dECM_DPSCs) and gingival-derived mesenchymal stem cells (dECM_GSCs) as an inductive scaffold for osteogenic differentiation of GSCs. The proteomic analysis demonstrated that common and signature matrisome proteins from dECM_DPSCs and dECM_GSCs were related to osteogenesis/osteogenic differentiation. RNA sequencing data from GSCs reseeded on dECM_DPSCs revealed that dECM_DPSCs upregulated genes related to the Hippo and Wnt signaling pathways in GSCs. In the inhibitor experiments, results revealed that dECM_DPSCs superiorly promoted GSCs osteogenic differentiation, mainly mediated through Hippo and Wnt signaling. The present study emphasizes the promising translational application of dECM_DPSCs as a bio-scaffold rich in favorable regenerative microenvironment for tissue engineering.

The role of hemostatic agents after tooth extractions: A systematic review and meta-analysis.

BACKGROUND: Hemostatic agents are used to control bleeding after tooth extraction and have been compared with conventional measures (that is, sutures or gauze pressure) in several studies. The objective of this systematic review was to evaluate the benefits of topical hemostatic agents for controlling bleeding after tooth extractions, especially in patients receiving antithrombotic therapy. TYPES OF STUDIES REVIEWED: The authors conducted a literature search in MEDLINE (PubMed), Scopus, and the Cochrane Central Register of Controlled Trials, including prospective human randomized clinical trials in which researchers compared hemostatic agents with conventional methods and reported the time to achieve hemostasis and postoperative bleeding events. RESULTS: Seventeen articles were eligible for inclusion. Hemostatic agents resulted in a significantly shorter time to achieve hemostasis in both healthy patients and patients taking antithrombotic drugs (standardized mean difference, -1.02; 95% CI, -1.70 to -0.35; P = .003 and standardized mean difference, -2.30; 95% CI, -3.20 to -1.39; P < .00001, respectively). Significantly fewer bleeding events were noted when hemostatic agents were used (risk ratio, 0.62; 95% CI, 0.44 to 0.88; P = .007). All forms of hemostatic agents (that is, mouthrinse, gel, hemostatic plug, and gauze soaked with the agent) had better efficacy in reducing the number of postoperative bleeding events than conventional hemostasis measures, except for hemostatic sponges. However, this was based on a small number of studies in each subgroup. CONCLUSIONS: The use of hemostatic agents seemed to offer better bleeding control after tooth extractions in patients on antithrombotic drugs than conventional measures. PRACTICAL IMPLICATIONS: Findings of this systematic review may help clinicians attain more efficient hemostasis in patients requiring tooth extraction. This systematic review is registered in the PROSPERO database. The registration number is CRD42021256145.

Comparative study of stability between two different fixation systems after orthognathic surgery in mandibular prognathism skeleton.

OBJECTIVE: This study is intended to compare the skeleton stability of bioabsorbable and titanium systems after orthognathic surgery in mandibular prognathism patients. STUDY DESIGN: A Retrospective study of 28 mandibular prognathism patients who underwent BSSRO setback surgery at Chulalongkorn University. Both titanium and the bioabsorbable group would take lateral cephalometrics immediately postoperative in 1-week(T0), 3(T1), 6(T2), and 12(T3) months. These radiographs were analyzed with Dolphin imaging programTM. The vertical, horizontal, and angular indices were measured. To compare immediately postoperative and follow-up periods within the group, the Friedman difference was used, and the Man-Whitney U test was used between the two groups. RESULT: The measurements within the group presented no statistically significant differences. But this study showed that at T0-T1, there was a statistically significant difference between the two groups in the mean of Me in horizontal linear measurement. T0-T2 found differences between Me in both horizontal and vertical linear measurements, and the difference between ANB. The differences between B-point, Pog, and Me in vertical linear measurements at T0-T3 were also reported. CONCLUSION: The significant difference values were within the normal range which indicated that using the bioabsorbable system could be well maintained as well as the titanium system. STATEMENT OF CLINICAL RELEVANCE: The second operation for removing titanium plate and screw after conventional orthognathic surgery may leads patient discomforts. While a resorbable system might be the role change if the stability is place on the same level.

Reoperative genioplasty: a 10-year retrospective study.

PURPOSE: The purpose of this retrospective cohort study was to identify the causes of requiring reoperative genioplasty and determine the factors associated with reoperation. METHODS: Medical records and radiographs of patients who underwent genioplasty were reviewed. The demographic data, characteristics of operation, and treatment outcomes were gathered to analyze the causes that required reoperation. Descriptive statistics and logistic regression analysis were computed to evaluate the study. RESULTS: Of the 157 patients included, there were 12 patients (7.6%) who needed reoperation after genioplasty. Age ≤ 25 years significantly decreased the likelihood for the need for reoperative genioplasty compared with age > 35 years. However, the need for reoperative genioplasty was not directly associated with gender, simultaneous orthognathic operation, direction and amount of movement, method of fixation, or bone graft interposition. Fixation failure, esthetic problems, residual obstructive sleep apnea, and palpable step at the inferior border of the mandible were the causes that required a second operation by reposition and re-fixation with rigid fixation, recontouring, or reoperation by genioplasty. CONCLUSION: Genioplasty procedure provided a predictive result. A reoperative rate was only 7.6% and younger age decreased the risk of reoperative genioplasty.

Betaine promotes osteogenic differentiation in immortalized human dental pulp-derived cells.

OBJECTIVES: This study aimed to evaluate the effect of betaine (BET) on immortalized human dental pulp stem cell (ihDP) osteogenic differentiation. MATERIALS AND METHODS: hDPs were immortalized using SV40 T-antigen transfection. Characterization, multilineage differentiation, proliferation, cell cycle, colony-forming unit, and cellular senescence were evaluated (n = 4). The effect of BET on ihDP response was assessed (n = 4). Osteogenic differentiation was detected using ALP, ARS staining, and RT-qPCR (n = 4). To investigate the involvement of calcium signaling, the cells were pretreated with either 8-(NN-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) or thapsigargin before BET treatment (n = 6). RESULTS: ihDPs retained similar phenotypic characteristics presented in hDPs but exhibited an increase in cell proliferation and extended culture to passage 25. An increased proportion of cells in S and G2/M phases without senescence was observed in ihDPs. BET (50 mM) treatment significantly increased mineral deposition at 14 days and upregulated ALP, MSX2, BMP2, and RUNX2 expression. TMB-8 pretreatment reduced the effect of BET-induced ihDP osteogenic differentiation, whereas thapsigargin promoted osteogenic differentiation in ihDPs synergistically with BET. CONCLUSION: ihDPs showed superior proliferation ability and a longer life span, which could serve as a promising cell for regenerative dentistry. BET promoted odonto/osteogenic differentiation via intracellular calcium regulation.