Necroptosis in inflammatory bowel disease: A potential effective target. / ç‚Žç—‡æ€§è‚ ç— ä¸­çš„åæ­»æ€§å‡‹äº¡ï¼šä¸€ä¸ªæ½œåœ¨çš„æ²»ç–—é¶ç‚¹ï¼ˆè‹±æ–‡ï¼‰.

The morbidity of inflammatory bowel diseases (IBD) is rising rapidly but no curative therapies to prevent its recurrence. Cell death is crucial to maintaining homeostasis. Necroptosis is a newly identified programmed cell death and its roles played in IBD need to be explored. Necroptosis is mediated by receptor interacting protein kinase 1 (RIPK1), RIPK3, and mixed lineage kinase domain-like protein (MLKL), which resulted in cell swelling, plasma membrane rupture, intracellular content leaking, and eventually cell death as well as the promotion of inflammation. Studies have found that inhibiting necroptosis alleviated IBD in animal models and IBD patients with an increased level of necroptosis in inflammatory tissues, indicating that necroptosis is related to the pathogenesis of IBD. However, due to the complexity in regulation of necroptosis and the involvement of multiple functions of relevant signaling molecules, the specific mechanism remains elusive. Necroptosis may play a vital regulatory role in the pathogenesis of IBD, which provides a new idea and method for further exploring the therapeutic target of IBD.

Enhanced reparatory effect of EI1 on dental pulp via extracellular matrix remodeling by miR-181b-2-3p inhibitor.

Background/purpose: Extracellular matrix (ECM) is crucial for dental pulp repair. The aim of this paper is to investigate the ECM remodeling effect of miR-181b-2-3p (a microRNA) and to verify the reparatory effect of EI1 (an epigenetic drug) and miR-181b-2-3p inhibitor on dental pulp. Materials and methods: Levels of ECM-related factors in EI1-treated human dental pulp cells (hDPCs) were measured by qRT-PCR and Western blot. The anti-inflammation effect of EI1 was examined in Lipopolysaccharide-stimulated hDPCs. miR-181b-2-3p mimics or inhibitors were transfected into hDPCs and then the cells' functions were detected. A dual luciferase reporter assay was used to identify the targets of miR-181b-2-3p. Pulpotomy using miR-181b-2-3p antagomirs and EI1 as pulp capping materials was performed in male six-week-old Sprague-Dawley rats. Results: EI1 upregulated ECM-related genes expression in hDPCs, but failed to upregulate the collagen1A1 (COL1A1) protein level. Pro-inflammatory factors were downregulated by EI1 in Lipopolysaccharide-stimulated hDPCs. Overexpression of miR-181b-2-3p downregulated the expression of transforming growth factor-ß2 (TGF-ß2) and fibronectin type III domain-containing protein 5 precursor (FNDC5), while the inhibition had the opposite effect. Dual luciferase reporter assays demonstrated that miR-181b-2-3p targets TGF-ß2, FNDC5 and integrin alpha 4 protein (ITGA4). Compared to EI1 was used alone, EI1 combined with the inhibitor upregulated the protein levels of COL1A1, fibronectin (FN1) and TGF-ß2 in hDPCs, promoted hDPCs migration, and exhibited reparatory effects on inflamed rat pulp tissue. Conclusion: miR-181b-2-3p inhibitor could enhance the reparatory effect of EI1 via ECM remodeling in dental pulp both in vitro and in vivo.

MiR-335-3p/miR-155-5p Involved in IGFBP7-AS1-Enhanced Odontogenic Differentiation.

BACKGROUND: The differentiation of stem cells from exfoliated deciduous teeth (SHEDs) into odontoblasts determines the regeneration of dentin-pulp complex. Non-coding RNAs (ncRNAs), including microRNA (miRNA) and long non-coding RNA (lncRNA), participate in many multiple biological processes, but the specific miRNAs involved in odontogenesis are incompletely defined. It was confirmed that lncRNA IGFBP7-AS1 could positively regulate odontogenetic differentiation in SHEDs. To investigate the downstream mechanisms of this process, miR-335-3p and miR-155-5p were found to be closely related with SHED odontogenic differentiation through whole-genome sequencing. The aim of the current study was to determine the role of miR-335-3p/miR-155-5p in IGFBP7-AS1-enhanced SHED differentiation and explore the potential mechanism of IGFBP7-AS1-mediated odontogenesis. METHODS: Putative miR-335-3p/miR-155-5p binding sites within IGFBP7-AS1 were identified by bioinformatics analysis, and the binding of miR-335-3p/miR-155-5p to these sites was confirmed by dual-luciferase reporter gene assays. The effects of miR-335-3p/miR-155-5p in odontogenesis were detected by tissue nonspecific alkaline phosphatase staining, Alizarin red staining, quantitative real-time polymerase chain reaction (qRT-PCR) analyses, and western blot testing. The molecular mechanisms of miR-335-3p/miR-155-5p involved in IGFBP7-AS1-mediated odontogenesis were analysed by qRT-PCR and western blot testing. RESULTS: Dual-luciferase reporter gene assays showed that miR-335-3p/miR-155-5p could directly bind to IGFBP7-AS1. MiR-335-3p and miR-155-5p both could down-regulate dentin sialophosphoprotein expression, and both miRNAs could inhibit IGFBP7-AS1-mediated SHED odontogenetic differentiation via suppression of the extracellular signal-regulated kinase (ERK) pathway. CONCLUSIONS: Both miR-335-3p and miR-155-5p were negative regulators to IGFBP7-AS1-enhanced odontogenic differentiation of SHED through suppression of the ERK pathway.

Willingness to Undergo Gastroscopy for Early Gastric Cancer Screening and Its Associated Factors During the COVID-19 Pandemic - A Nationwide Cross-Sectional Study in China.

Purpose: This study aimed to investigate the willingness of Chinese adults aged 40 years and older to undergo gastroscopy for gastric cancer (GC) screening during the COVID-19 pandemic in 2020. The secondary purpose was to identify factors influencing willingness to undergo gastroscopy. Methods: A cross-sectional questionnaire survey was conducted in selected cities and counties from nine provinces in China using a multi-stage sampling approach. A multivariate logistic regression model was used to determine the independent predictors of willingness to undergo gastroscopy. Results: This study included 1900 participants, and 1462 (76.95%) responded that they would undergo gastroscopy for GC screening. Participants of younger age, from the eastern region, living in an urban area, with higher educational levels, with Helicobacter pylori (H. pylori) infection, or with precancerous stomach lesions, were more willing to undergo gastroscopy. The top four reasons to reject gastroscopy were fear of pain or discomfort, worry about a possible devastating test result, no symptoms in self-feeling, and concern about the high expense. Of all those who would reject gastroscopy for GC screening, 36.76% (161/438) would be willing to accept painless gastroscopy, while 24.89% (109/438) would be willing to undergo gastroscopy screening if higher medical reimbursement rates were available. Participants considered that gastroscopy was a relatively fearful and unknown procedure, accompanied by high risks and benefits compared to all other life events. Conclusion: In general, 76.95% of participants over 40 years old were willing to undergo gastroscopy for GC screening in China during the COVID-19 pandemic. Participants' willingness to undergo GC screening increased due to medical resource constraints and increased interest in their health. Individuals with H. pylori infection are more likely to undergo gastroscopy, whereas old age individuals, those with lower educational levels, and those living in rural areas are more likely to reject gastroscopy.

Engineering core-shell chromium nanozymes with inflammation-suppressing, ROS-scavenging and antibacterial properties for pulpitis treatment.

Oral diseases are usually caused by inflammation and bacterial infection. Reactive oxygen species (ROS), which come from both autologous inflammation tissue and bacterial infection, play an important role in this process. Thus, the elimination of excessive intracellular ROS can be a promising strategy for anti-inflammatory treatment. With the rapid development of nanomedicines, nanozymes, which can maintain the intracellular redox balance and protect cells against oxidative damage, have shown great application prospects in the treatment of inflammation-related diseases. However, their performance in pulpitis and their related mechanisms have yet to be explored. Herein, we prepared dozens of metallic nanoparticles with core-shell structures, and among them, chromium nanoparticles (NanoCr) were selected for their great therapeutic potential for pulpitis disease. NanoCr showed a broad antibacterial spectrum and strong anti-inflammatory function. Antibacterial assays showed that NanoCr could effectively inhibit a variety of common pathogens of oral infection. In vitro experiments offered evidence of the multienzyme activity of NanoCr and its function in suppressing ROS-induced inflammation reactions. The experimental results show that NanoCr has optimal antibacterial and anti-inflammatory properties in in vitro cell models, showing great potential for the treatment of pulpitis. Therefore, the use of NanoCr could become a new therapeutic strategy for clinical pulpitis.

Long non-coding RNA IGFBP7-AS1 accelerates the odontogenic differentiation of stem cells from human exfoliated deciduous teeth by regulating IGFBP7 expression.

Stem cells from human exfoliated deciduous teeth (SHED) are attractive seed cells for dental tissue engineering. We identified the effect of the long noncoding RNA insulin-like growth factor-binding protein 7 antisense RNA 1 (lncRNA IGFBP7-AS1) in vivo and its underlying mechanism during SHED odontogenic differentiation. IGFBP7-AS1 and insulin-like growth factor-binding protein 7 (IGFBP7) were overexpressed using lentiviruses. IGFBP7 expression was knocked down with small interfering RNA. The effect of IGFBP7-AS1 in vivo was confirmed by animal experiments. The effect of IGFBP7 on SHED odontogenic differentiation was assessed with alkaline phosphatase staining, alizarin red S staining, quantitative reverse transcription-PCR, and western blotting. The relationship between IGFBP7-AS1 and IGFBP7 was confirmed by quantitative reverse transcription-PCR and western blotting. IGFBP7-AS1 promoted SHED odontogenesis in vivo, and regulated the expression of the coding gene IGFBP7 positively. Inhibiting IGFBP7 led to suppress SHED odontogenic differentiation while IGFBP7 overexpression had the opposite effect. IGFBP7-AS1 enhanced the stability of IGFBP7. IGFBP7-AS1 promoted SHED odontogenic differentiation in vivo. The underlying mechanism may involve the enhancement of IGFBP7 stability. This may provide novel potential targets for dental tissue engineering.

Long non-coding RNA IGFBP7-AS1 promotes odontogenic differentiation of stem cells from human exfoliated deciduous teeth through autophagy: An in vitro study.

OBJECTIVE: In the present study, we aimed to investigate whether long non-coding RNA (lncRNA) insulin-like growth factor binding protein 7-antisense 1 (IGFBP7-AS1) regulates the odonto-differentiation of stem cells from human exfoliated deciduous teeth (SHED) and its underlying mechanism. DESIGN: Real-time polymerase chain reaction (PCR) and correlation analysis were used to determine the expression of IGFBP7-AS1 during odontogenesis. Alkaline phosphate staining, alizarin red S staining, and real-time PCR in vitro were performed to investigate the effects of IGFBP7-AS1 during odontogenesis. Western blot and immunostaining (with or without chloroquine treatment) were applied to detect the expression of the autophagy-related markers, microtubule-associated proteins 1A/1B light chain 3B (LC3B) and p62. The autophagy inhibitor 3-methyladenine was used to further clarify the effect of autophagy in odonto-differentiation as promoted by IGFBP7-AS1. RESULTS: The expression of lncRNA IGFBP7-AS1 is significantly upregulated during odonto-differentiation of SHED and promotes odontogenesis of SHED in vitro. IGFBP7-AS1 promotes autophagy during odontogenesis. CONCLUSIONS: IGFBP7-AS1 elicits odontogenic differentiation of SHED through autophagy. Furthermore, IGFBP7-AS1 shows promise as a gene target in the regeneration of dental hard tissue and dental-pulp complex.

Long Noncoding RNA IGFBP7-AS1 Promotes Odontogenesis of Stem Cells from Human Exfoliated Deciduous Teeth via the p38 MAPK Pathway.

Stem cells from human exfoliated deciduous teeth (SHED) are attractive seed cells for dental tissue engineering. Epigenetics refers to heritable changes in gene expression patterns that do not alter DNA sequences. Long noncoding RNAs (lncRNAs) are one of the main methods of epigenetic regulation and participate in cell differentiation; however, little is known regarding the role of lncRNAs during SHED odontogenic differentiation. In this study, RNA sequencing (RNA-seq) was used to obtain the expression profile of lncRNAs and mRNAs during the odontogenic differentiation of SHED. The effect of IGFBP7-AS1 on odontogenic differentiation of SHED was assessed by alkaline phosphatase (ALP) staining, alizarin red S (ARS) staining, quantitative reverse transcription PCR (qRT-PCR), Western blot, and in vivo. The level of p38 and p-p38 protein expression was examined by Western blot, and the result was verified by adding the p38 inhibitor, SB203580. The expression profiles of lncRNAs and mRNAs were identified by RNA-seq analysis, which help us to further understand the mechanism in odontogenesis epigenetically. IGFBP7-AS1 expression was increased during odontogenic differentiation on days 7 and 14. The ALP staining, ARS staining, and expression of odontogenic markers were upregulated by overexpressing IGFBP7-AS1 in vitro, whereas the expression of osteogenesis markers was not significantly changed on mRNA level. The effect of IGFBP7-AS1 was also verified in vivo. IGFBP7-AS1 could further positively regulate odontogenic differentiation through the p38 MAPK pathway. This may provide novel targets for dental tissue engineering.

Fabrication and Characterization of Calcium-Phosphate Lipid System for Potential Dental Application.

Lipid has been widely studied as a vehicle and loading vector, but there have been no reports of any such related application in the dental field. The purpose of this research was to fabricate and characterize a nano-size calcium-phosphate lipid (CL) system as a potential vehicle in dental regeneration study, wherein the biocompatibility with dental pulp stem cells (DPSCs) was evaluated. The effect of CL on DPSCs proliferation was analyzed by a CCK-8 assay, and the anti-inflammatory effect was investigated by quantitative polymerase chain reaction (qPCR). Moreover, the effect of CL on odontogenic differentiation of inflamed DPSCs (iDPSCs) was studied by Alizarin red staining, tissue-non-specific alkaline phosphatase (TNAP) staining, qPCR, and western blot analyses. The results of this study showed that CL did not affect the proliferation of DPSCs, it down-regulated the inflammatory-associated markers (IL-1ß, IL-6, TNF-α, COX-2) of DPSCs treated with Escherichia coli lipopolysaccharide (LPS), and enhanced the in-vitro odontogenic differentiation potential of iDPSCs. This novel biomaterial has a broad application prospect for its bioactivity and flexible physical property, and thus represents a promising pulpal regeneration material.

Betamethasone suppresses the inflammatory response in LPS-stimulated dental pulp cells through inhibition of NF-κB.

OBJECTIVE: This study aimed to investigate the anti-inflammatory effect of betamethasone on LPS-stimulated human dental pulp stem cells (DPSCs) and its associated mechanism. The osteo-/odontogenic differentiation and osteoclast effect of betamethasone on DPSCs and stem cells from human exfoliated deciduous teeth (SHED) were evaluated. DESIGN: The proliferative effect of betamethasone on DPSCs was analyzed using a cholecystokinin octapeptide assay. The anti-inflammatory effect of betamethasone was investigated using quantitative polymerase chain reaction (qPCR) and ELISA. The anti-inflammatory mechanism was explored using qPCR, Western blot, and immunofluorescence staining. The osteo-/odontogenic differentiation and osteoclast effect of betamethasone on DPSCs and SHED were detected by qPCR. RESULTS: 1 µg L-1 betamethasone was found to have the strongest effect on DPSCs proliferation. The expression of pro-inflammatory cytokines and mediators, as well as prostaglandin E2 (PGE2) were significantly decreased following treatment with betamethasone in LPS- stimulated DPSCs. They were also decreased in response to an NF-κB inhibitor, Bay 11-7082. Betamethasone and Bay 11-7082 significantly inhibited the expression of p-p65 and promoted the nuclear exclusion of p65. Gene expression associated with osteo-/odontogenic differentiation was significantly up-regulated in betamethasone and osteogenic media (OM) treated groups. The ratio of the receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) at the mRNA level was suppressed in DPSCs and elevated in SHED. CONCLUSIONS: Betamethasone has an anti-inflammatory effect on LPS- stimulated DPSCs through a blockade of NF-κB activation and exhibits an osteo-/odonto-inductive effect on DPSCs and SHED. Although betamethasone displays an osteoclast effect on SHED.

Biological properties of modified bioactive glass on dental pulp cells.

Dental caries is a bacteria-caused condition classified among the most common chronic diseases worldwide. Treatment of dental caries implies the use of materials having regenerative and anti-bacterial properties, and controlling inflammation is critical for successful endodontic regeneration. OBJECTIVES: The aim of this study was to fabricate and characterize a novel composite incorporating sol-gel derived silver-doped bioactive glass (BG) in a chitosan (CS) hydrogel at a 1:1 wt ratio(Ag-BG/CS). METHODS: The effect of Ag-BG/CS on dental pulp cells (DPCs) proliferation was analyzed by CCK-8 assay, whereas the adhesion of DPCs was evaluated by confocal microscopy. The physical morphology of Ag-BG/CS was analyzed by scanning electron microscope. The anti-inflammatory effect of Ag-BG/CS was investigated by quantitative polymerase chain reaction (qPCR). Moreover, the effect of Ag-BG/CS on odontogenic differentiation of DPCs was studied by immunochemical staining, tissue-nonspecific alkaline phosphatase staining, qPCR, and western blot analyses. The antibacterial activity against dental caries key pathogenic bacteria was also evaluated. RESULTS: The results of this study showed that Ag-BG/CS did not affect the proliferation of DPCs, it down-regulated the inflammatory-associated markers (IL-1ß, IL-6, IL-8, TNF-α) of DPCs treated with Escherichia coli lipopolysaccharide (LPS) by inhibiting NF-κB pathway, and enhanced the in vitro odontogenic differentiation potential of DPCs. Furthermore, Ag-BG/CS strongly inhibited Streptococcus mutans and Lactobacillus casei growth. CONCLUSIONS: This novel biomaterial possessed antibacterial and anti-inflammatory activity, also enhanced the odontogenic differentiation potential of LPS-induced inflammatory-reacted dental pulp cells. The material introduced in this study may thus represent a suitable dental pulp-capping material for future clinical applications.

Silver-Doped Bioactive Glass/Chitosan Hydrogel with Potential Application in Dental Pulp Repair.

The major indications of a successful inflamed-pulp-capping procedure are the formation of a dense calcified dentin barrier and the preservation of healthy pulp tissue concomitant with elimination of inflammation. Our aim is to evaluate the effects of an injectable silver-doped bioactive glass/chitosan hydrogel (Ag-BG/CS), as a pulp-capping material, and explore the molecular mechanisms of Ag-BG/CS in regards to its bioactive and anti-inflammatory properties. First, the structure and component of the material were analyzed by scanning electron microscopy. Then, the downstream molecular mechanisms and anti-inflammatory effects were characterized by quantitative polymerase chain reaction (qPCR) and Western blot. Finally, a preclinical model of rat pulpitis was used to explore the potential of Ag-BG/CS in controlling pulp inflammation in vivo. The results showed that Ag-BG/CS induced stronger reparative dentin formation and enhanced preservation of vital pulp tissue when compared to the mineral trioxide aggregate (MTA) which is the currently used clinical standard. Ag-BG/CS also significantly increased the phosphorylation of p38 and ERK1/2(p42/44) of dental pulp cells, indicating that Ag-BG/CS enhanced pulpal repair through the mitogen-activated protein kinase (MAPK) pathway. This novel material may represent a superior solution for dental pulp-capping clinical scenarios with specific advantages for cases of early diffuse pulpitis in immature permanent teeth.