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Objective To investigate the effects of N-cadherin silencing on the proliferation and migration of human dental pulp stem cells (DPSCs) and to provide experimental evidence for DPSCs-based dental pulp regeneration. Methods DPSCs were transfected with N-cadherin shRNA lentivirus, and the knockdown efficiency of N-cadherin at both the mRNA and protein levels was confirmed by qRT-PCR and Western blot. The experiment included a negative control group (shRNA -NC) and an N-cadherin shRNA silencing group. Cell proliferation was detected by the CCK-8 method. Cell cycle and apoptosis were assessed by flow cytometry, and cell migration was detected using the Transwell method. Results N-cadherin shRNA significantly reduced the expression levels of N-cadherin mRNA and protein in DPSCs (P

Vital pulp therapy aims to maintain healthy pulp tissue as much as possible to improve the long-term survival of teeth. It has limited indications and uncertain curative effects. The pathological changes in inflamed pulp are the histological basis for the determination of treatment strategies and the treatment outcome; however, pulp sensitivity testing cannot reflect the actual histological status of the pulp. With the development of basic and clinical research on vital pulp therapy, the innovation of modern diagnostic and therapeutic technology and capping material, vital pulp therapy can be used as a treatment of teeth on which it was previously thought pulpectomy was necessary. Based on the evidence-based literature, this paper analyzes and summarizes the pathological changes of pulpitis and clinical research on the treatment of pulpitis. Vital pulp therapy can be a treatment for mature teeth with carious exposure and symptoms of irreversible pulpitis if comprehensive applications, including laser Doppler flowmetry, tissue oxygen monitoring, magnetic resonance imaging and microscopy, are used to determine the degree of pulp retention and if infection control and the use of biocompatible capping material are emphasized. In the future, it will be necessary to improve the success rate of vital pulp therapy for the treatment of pulpitis through research on the mechanism of pulp repair and regeneration, the precise diagnosis of pulpitis, and the development of pulp capping materials.

Objective To observe the permeability of four kinds of self-etching adhesives in aged glass-fiber-reinforced composite (GFRC). Methods After light polymerization following the manufacturers’ instructions, a total of 80 pieces of bisphenol-A-glycodal-methacrylate (Bis-GMA)+ polymethyl methacrylate (PMMA) based GFRC were randomly divided into two main groups: test group and control group, each group was then divided into four subgroups with 10 samples for each subgroup. While the test group was conducted to be aged through thermocycling at 5 ℃/55 ℃, the control group remained fresh. After the addition of a fluorescent dye (rhodamine-B-isothiocyanate), four self-etching adhesives AdperEasy One (AEO), S 3 BOND (S 3B), Tetric N-Bond Self-Etch (TNB), G-Bond (GB) were correspondently applied to the test and control groups and were light polymerized. Specimens were sectioned using hard tissue cutting and grinding system. Slices from each subgroup were observed under a confocal laser scanning microscope, the depth of dye permeation (DDP) under the surface of GFRC in each group was measured and the Results were statistically analyzed. Results The DDP of AEO was the deepest (32.58 ± 6.06) μm, and that of TNB was the shallowest (6.19 ± 1.38)μm among the four self-etching adhesive subgroups in the control group. The order of each group was AEO > GB > S 3B > TNB. The DDP of the four subgroups in the test group was significantly shallower than that in the control group (P < 0.05). The change in GB was the greatest (9.05 ± 2.35)μm/(28.93 ± 5.32)μm. In the test group, the DDP in AEO was the deepest (28.42 ± 5.32)μm, and the DDP in TNB was shallowest (1.93 ± 0.22)μm again. The order of each group was AEO > S 3B > GB > TNB. In the test group, while the layer of fluorescent dye of AEO and S 3 B could still be seen distinctly, that of TNB and GB was hard to recognize. Conclusion The self-etching adhesives of AEO and S 3 B still have good permeation effect in this kind of aged GFRC, which can help to establish a good bond between these aged GFRC and the subsequent repair of composite resin.

Aim: To investigate novel diagnostic markers for pulpitis and validate by clinical samples from normal and inflamed pulp. To explore the relationship between diagnostic markers and immune cells or their phenotypes during pulp inflammation. Methodology: Two microarray datasets, GSE77459 and GSE92681, and identified differential expression genes were integrated. To understand immune features, gene functions, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Disease Ontology (DO) and ImmuneSigDB Gene Set Enrichment Analysis (GSEA) were analysed. For predictive purposes, machine learning techniques were applied to detect diagnostic markers. Immune infiltration in inflamed pulp was studied using CIBERSORT. The relationship between diagnostic markers and immune cells was investigated and validated their gene expression in clinical samples from the normal or inflamed pulp by qRT‐PCR. Finally, the correlation between one marker, secreted phosphoprotein 1 (SPP1), encoding osteopontin (OPN), and dendritic cells (DCs)/macrophages was identified via HE staining and multiplex immunohistochemistry. An in vitro inflammatory dental pulp microenvironment model of THP‐1 macrophages cocultured with dental pulp cells derived conditioned media (DPCs‐CM) to investigate OPN production and macrophage phenotypes was established. Results: Analysis revealed unique immunologic features in inflamed pulp. Three diagnostic markers for pulpitis: endothelin‐1 (EDN1), SPP1, and purine nucleoside phosphorylase (PNP), and validated them using qRT‐PCR were predicted. Multiplex immunohistochemistry demonstrated OPN co‐localized with activated DCs and M2 macrophages during pulp inflammation. In vitro experiments showed that THP‐1 macrophages produced the highest levels of OPN when stimulated with DPCs‐CM derived from the 20 μg/mL LPS pre‐conditioned group, suggesting an M2b‐like phenotype by increasing surface marker CD86 and expression of IL6, TNFα, IL10, and CCL1 but not CCL17 and MerTK. Levels of CCL1 and IL10 elevated significantly in the macrophages' supernatant from the 20 μg/mL LPS pre‐conditioned CM group. OPN was proven co‐localizing with CD86 in the inflamed pulp by immunofluorescence. Conclusions: The current findings suggest that OPN can serve as a promising biomarker for pulpitis, correlated with DCs and macrophages. OPN+ macrophages in the inflamed pulp are associated with M2b‐like phenotypes. These insights offer the potential for improved diagnosis and targeted therapy. [ABSTRACT FROM AUTHOR]

Objective To observe the stress distribution according to a model of the bucco⁃occluso⁃lingual (BOL) inlay of mandibular first molar after restoration to provide a basis for the clinical treatment of cracked tooth with BOL in⁃ lay. Methods A three⁃dimensional finite element model of mandibular first molar was established by combining micro⁃ CT scanning technology with Mimics, UG, Ansys and Midas⁃FEA software. Based on this model, a BOL inlay restoration model was established. The material parameter of inlay IPS e.max CAD was given, and a Von⁃mises stress distribution nephogram under the same loading condition was obtained. The results of the stress distribution in each model were compared. Results The stress of intact teeth is mainly concentrated in the central fissure of the occlusal surface at the crown. The stress of the cavity after BOL inlay restoration is mainly concentrated in the mesial and distal walls of the cavity, the axial⁃pulpal line angle and the gingival wall. The stress of the inlay is mainly distributed at the bottom of the inlay, axial wall and the gingival wall. Conclusion BOL inlay restoration change the stress distribution in the com⁃ plete dental model, which relieves the stress concentration in the fossa and groove of the occlusal surface and can play an active role in the treatment of cracked tooth.

Tooth defects due to dental caries, trauma, abrasion, etc., are extremely common and can be treated by di⁃ rect or indirect restoration. Compared with resin directly filling the body, an inlay can better restore the occlusal contact relationship and the adjacent surface contact relationship and has good mechanical properties. In recent years, with the development of ceramic materials and bonding systems and the popularity of chairside CAD/CAM technology, the chair⁃ side CAD/CAM porcelain inlay restoration program has been well received by doctors and patients because of its accura⁃ cy, convenience, aesthetics, hardness and stability, and this program is widely used clinically. This review covers the re⁃ search status of various aspects such as indications and contraindications for chairside CAD/CAM inlay restoration, pre⁃ restoration preparation, tooth preparation, hole type, impression taking and design, porcelain block selection, bonding, polishing, postoperative doctor􀆳s instructions, and common postoperative complications. It is expected to provide a refer⁃ ence for the clinical application of and research on chairside CAD/CAM inlay restoration technology.

Objective To investigate the short⁃term clinical effect of semiconductor laser exposure combined with total glucosides of paeony (TGP) capsules for the treatment of erosive oral lichen planus (OLP). Methods Sixty⁃four patients with erosive oral lichen planus were randomly divided into two groups: the experimental group and the control group. Patients in the control group were treated with TGP capsules, while patients in the experimental group were treat⁃ ed with TGP capsules and semiconductor laser irradiation. The clinical effects were evaluated 3 months after treatment. The data were analyzed using the SPSS 17.0 software package. Results Three months after treatment, the effective rate in the experimental group was 90.6%, which was significantly higher than that in the control group (59.4%, χ2＝ 5.62, P < 0.05). The physical condition and visual analogue scale (VAS) scores in the experimental group were 2.17 ± 1.49 and 1.25 ± 1.29, respectively. The physical condition and VAS scores in the control group were 3.55 ± 1.41 and 2.09 ± 1.24, respectively. The physical condition and VAS scores in both groups were significantly higher after treat⁃ ment than before (P < 0.05). Three months after treatment, the physical condition score (t＝3.805) and VAS score (t＝ 2.655) in the experimental group were significant higher than those in the control group (P < 0.05). Conclusion Semi⁃ conductor laser irradiation combined with TGP capsules can improve the short⁃term clinical efficacy in the treatment of erosive OLP.

Objective To explore methods using modified tissue enzymatic separations for culturing primary hDP⁃ SCs in vitro and further identify the cells produced. Methods Primary hDPSCs were cultured using the modified tis⁃ sue enzymatic separation method, and cells were identified by morphology, cell surface markers, and differentiation po⁃ tential and evaluated using flow cytometry and growth curves. Results The hDPSCs were successfully isolated using the modified tissue enzymatic separation method. The morphology of these cells was similar to that of fibroblasts and mesenchymal stem cells, and the growth curve was "S" ⁃type. The results of cell phenotype analysis indicated that the cells were positive for surface markers of mesenchymal stem cells, including CD29, CD44, and CD90, and negative for markers of hematopoietic stem cells, including CD34, CD45, and CD106. The cells were capable of differentiating into multiple cell types. Conclusion The modified tissue enzymatic separation method can successful be used to culture primary hDPSCs in vitro.

[Display omitted] • In vivo , Fn OMVs promoted lung metastasis in tumour-bearing mice, while chloroquine (CHQ; an autophagy inhibitor) reduced the number of pulmonary metastases resulting from intratumoral Fn OMV injection. • Fn OMVs promoted cancer cell invasion and migration in vivo , leading to changes in the expression of EMT related proteins. • RNA-seq showed that Fn OMVs activate intracellular autophagy pathways. Blocking autophagic flux with CHQ not only reduced Fn OMV-induced cancer cell migration in vitro and in vivo but also reversed the expression changes in EMT-related proteins. Metastasis is an important cause of high mortality and lethality of oral cancer. Fusobacterium nucleatum (Fn) can promote tumour metastasis. Outer membrane vesicles (OMVs) are secreted by Fn. However, the effects of Fn-derived extracellular vesicles on oral cancer metastasis and the underlying mechanisms are unclear. We aimed to determine whether and how Fn OMVs mediate oral cancer metastasis. OMVs were isolated from brain heart infusion (BHI) broth supernatant of Fn by ultracentrifugation. Tumour-bearing mice were treated with Fn OMVs to evaluate the effect of OMVs on cancer metastasis. Transwell assays were performed to determine how Fn OMVs affect cancer cell migration and invasion. The differentially expressed genes in Fn OMV-treated/untreated cancer cells were identified by RNA-seq. Transmission electron microscopy, laser confocal microscopy, and lentiviral transduction were used to detect changes in autophagic flux in cancer cells stimulated with Fn OMVs. Western blotting assay was performed to determine changes in EMT-related marker protein levels in cancer cells. Fn OMVs' effects on migration after blocking autophagic flux by autophagy inhibitors were determined by in vitro and in vivo experiments. Fn OMVs were structurally similar to vesicles. In the in vivo experiment, Fn OMVs promoted lung metastasis in tumour-bearing mice, while chloroquine (CHQ, an autophagy inhibitor) treatment reduced the number of pulmonary metastases resulting from the intratumoral Fn OMV injection. Fn OMVs promoted the migration and invasion of cancer cells in vivo , leading to altered expression levels of EMT-related proteins (E-cadherin downregulation; Vimentin/N-cadherin upregulation). RNA-seq showed that Fn OMVs activate intracellular autophagy pathways. Blocking autophagic flux with CHQ reduced in vitro and in vivo migration of cancer cells induced by Fn OMVs as well as reversed changes in EMT-related protein expression. Fn OMVs not only induced cancer metastasis but also activated autophagic flux. Blocking autophagic flux weakened Fn OMV-stimulated cancer metastasis. [ABSTRACT FROM AUTHOR]

Aim: Necroptosis participates in the pathogenesis of many inflammatory diseases, including periodontitis. Here, we aimed to investigate the role and mechanism of necroptosis inhibitors in attenuating periodontitis. Materials and Methods: The Gene Expression Omnibus (GEO) dataset GSE164241 was re‐analysed to identify the role of necroptosis in periodontitis. Gingival specimens from healthy subjects or periodontitis patients were collected to evaluate the expression level of necroptosis‐associated proteins. The therapeutic effect of necroptosis inhibitors on periodontitis was assessed in vivo and in vitro. Moreover, Transwell assays and Western blotting and siRNA transfection were used to identify the effects of necroptotic human gingival fibroblasts (hGFs) on THP‐1 macrophages. Results: Re‐analysis revealed that gingival fibroblasts (GFs) in periodontitis gingiva showed the highest area under the curve score of necroptosis. Elevated levels of necroptosis‐associated proteins were identified in GFs in periodontitis gingiva collected from patients and mice. In ligature‐induced periodontitis mice, local administration of receptor interacting protein kinase 3(RIPK3) inhibitor GSK′872 or sh‐mixed‐lineage kinase domain‐like pseudokinase (Mlkl) markedly abrogated necroptosis and rescued periodontitis. Analogously, necroptosis inhibitors alleviated the inflammatory response and release of damage‐associated molecular patterns in lipopolysaccharide‐ or LAZ (LPS + AZD'5582 + z‐VAD‐fmk, necroptosis inducer)‐induced GFs and then reduced THP‐1 cell migration and M1 polarization. Conclusions: Necroptosis in GFs aggravated gingival inflammation and alveolar bone loss. Necroptosis inhibitors attenuate this process by modulating THP‐1 macrophage migration and polarization. This study offers novel insights into the pathogenesis and potential therapeutic targets of periodontitis. [ABSTRACT FROM AUTHOR]