Heterogeneity of T cells in periapical lesions and in vitro validation of the proangiogenic effect of GZMA on HUVECs.

AIM: T cells are key immunomodulatory cells in periapical lesions. This study aimed to explore the roles of T cells in chronic apical periodontitis (CAP) using single-cell RNA sequencing and to further investigate Granzyme A (GZMA) in angiogenesis regulation. METHODOLOGY: A total of five CAP samples were collected for single-cell RNA sequencing. We performed subcluster and lineage-tracing analyses for T cells. According to differential gene expression, distinct biological functions enriched in T cells of CAP were presented by gene set enrichment analysis (GSEA) and compared with healthy gingiva (data obtained from the GEO database). CellChat was used to explore potential ligand-receptor interactions between T cells and endothelial cells in CAP. The coculture of primary human umbilical vein endothelial cells (HUVECs) and Jurkat T cells, as well as the addition of GZMA recombinant protein, was used to validate the predicted pair of GZMA and coagulation factor II thrombin receptor (F2R) by RT-PCR, angiogenesis and migration assays. RESULTS: A transcriptomic atlas of 44 746 individual cells was constructed from the periapical lesions of five patients with CAP by single-cell RNA-seq, and eight cell types were identified. We identified nine subsets of T cells and deciphered the cellular heterogeneity of T cells in CAP at the functional level by subclustering and GSEA. Lineage tracing revealed a distinct lineage of T cells in CAP and predicted the transition of the T cellular state upon CAP. GSEA revealed multiple biological processes and relevant angiogenesis genes upregulated in CAP T cells. GZMA-F2R pairs were predicted by cell-cell interactions in CAP. High expression of GZMA and F2R was observed in the coculture of HUVECs and Jurkat T cells, and the proangiogenic capacity of the GZMA recombinant protein was emphasized by in vitro experiments. CONCLUSIONS: Our study provides novel insights into the heterogeneity of T cells in periapical lesions and reveals the potential role of GZMA in T cells in regulating angiogenesis in HUVECs.

A Mousepad Triboelectric-Piezoelectric Hybrid Nanogenerator (TPHNG) for Self-Powered Computer User Behavior Monitoring Sensors and Biomechanical Energy Harvesting.

Hybrid nanogenerators based on the principle of surface charging of functional films are significant in self-powering sensing and energy conversion devices due to their multiple functions and high conversion efficiency, although applications remain limited due to a lack of suitable materials and structures. Here, we investigate a triboelectric-piezoelectric hybrid nanogenerator (TPHNG) in the form of a mousepad for computer user behavior monitoring and energy harvesting. Triboelectric and piezoelectric nanogenerators with different functional films and structures work independently to detect sliding and pressing movements, and the profitable coupling between the two nanogenerators leads to enhanced device outputs/sensitivity. Different mouse operations such as clicking, scrolling, taking-up/putting-down, sliding, moving rate, and pathing can be detected by the device via distinguishable patterns of voltage ranging from 0.6 to 36 V. Based on operation recognition, human behavior monitoring is realized, with monitoring of tasks such as browsing a document and playing a computer game being successfully demonstrated. Energy harvesting from mouse sliding, patting, and bending of the device is realized with output voltages up to 37 V and power up to 48 µW while exhibiting good durability up to 20,000 cycles. This work presents a TPHNG utilizing surface charging for self-powered human behavior sensing and biomechanical energy harvesting.

The practicability of different preparation of mandibular molar restored by modified endocrown with intracanal extension: Computational analysis using finite element models.

BACKGROUND AND OBJECTIVE: Post-core-crown (PCC) and endocrown are two common restorative methods for severely damaged molars, but exhibit disadvantages. This study aimed to explore the practicability of modified endocrown with a 2 mm intracanal extension (MED) to restore defective teeth using finite element analysis (FEA). METHODS: Five groups of numerical models of mandibular molars restored by three MEDs, a PCC, and a routine endocrown after root canal treatment were devised by FEA software. We constructed 4 mm, 3 mm, and 2 mm thickness of MED restorations to restore mandibular molars that were prepared to 1 mm, 2 mm, and 3 mm from the cemento-enamel junction (CEJ). Furthermore, PCC and routine endocrown were used to compare the stress distribution with MED. Lithium disilicate glass-ceramics (EMAX) and resin nanoceramics (LU) were considered restorative materials, and a vertical load of 600 N and an oblique load of 200 N were applied to the restorations. RESULTS: In three MEDs by LU, 2 mm thickness of restoration generated the highest stress on prepared teeth, while the thickness of EMAX did not significantly influence the stress value. MED by LU generated higher stress around the CEJ, and reduced the stress on the middle and lower root compared to MEDs by EMAX, PCC by EMAX, and PCC by LU. MED by EMAX caused lower stress around the CEJ, and generated higher stress in the chamber walls after extended root canals compared with MED by LU, endocrowns by LU, and endocrowns by EMAX. There was an evident stress concentration at the last but one layer, which was a thin area of the tooth root in all restorative models. CONCLUSIONS: The use of modified endocrown may be considered an effective restorative method to restore defective mandibular molar, but suitable restorative material must be selected based on the tooth preparation method and deficiencies in the tooth structure.

The synergetic effect of pulp chamber extension depth and occlusal thickness on stress distribution of molar endocrowns: a 3-dimensional finite element analysis.

The aim of this study was to evaluate the effects of butt margin, occlusal thickness and pulp chamber extension depth on stress distributions on mandibular molar endodontically treated teeth (ETT) with EMAX endocrown restoration using 3-dimensional finite element analysis (FEA). The FEA models of endocrown with flat surface or curve surface of butt margin were firstly evaluated stress distributions, and then 9 FEA models of endocrown with 1-, 2- or 3-mm pulp chamber extension depth and 1-, 2- or 3-mm occlusal thickness were generated using curve surface of butt margin. In all of FEA models, a 200 N of vertical load or horizontal load was applied, and the von Mises stress (VMS) were evaluated. The results showed that curve surface of butt margin offered more adhesive area of enamel, though VMS on the prepared teeth was similar in flat surface and curve surface models. In 9 endocrown models, 2-mm occlusal thickness showed the lowest VMS on restorations, teeth tissue and root furcations, and 2-mm extension depth displayed the lowest VMS on root furcations under vertical load. Also, 2-mm extension depth exhibited the lowest VMS on restorations and teeth tissue under horizontal load. Within the limitations of this FEA study, the results of this study could be used as an aid for dentists to better devise endocrown restorations. Graphical abstract.

Pharmacokinetic interaction between rhynchopylline and pellodendrine via CYP450 enzymes and P-gp.

CONTEXT: Rhynchopylline and pellodendrine are major extractions of commonly used Chinese medicine in gynaecology. The interaction between these two compounds could affect treatment efficiency and even result in toxicity during their co-administration in gynaecological prescription. OBJECTIVE: The pharmacokinetic interaction between rhynchopylline and pellodendrine and the potential mechanism were investigated in this study. MATERIALS AND METHODS: Sprague-Dawley rats were randomly divided into four groups to investigate the pharmacokinetic interaction between rhynchopylline (30 mg/kg) and pellodendrine (20 mg/kg) with single dose of these two drugs as the control. The transport of rhynchopylline was evaluated in the Caco-2 cell model. Additionally, the metabolic stability and the activity of corresponding CYP450 enzymes were assessed in rat liver microsomes. RESULTS: The pharmacokinetic profile of rhynchopylline was dramatically affected by pellodendrine with the increased area under the pharmacokinetic curve (3080.14 ± 454.54 vs. 1728.08 ± 220.598 µg/L*h), Cmax (395.1 ± 18.58 vs. 249.1 ± 16.20 µg/L), prolonged t1/2 (9.74 ± 2.94 vs. 4.81 ± 0.42 h) and the reduced clearance rate (from 11.39 ± 1.37 to 5.67 ± 1.42 L/h/kg). No significant changes were observed in the pharmacokinetics of pellodendrine. The transport of rhynchopylline was significantly inhibited by pellodendrine with a decreasing efflux ratio (1.43 vs. 1.79). Pellodendrine significantly inhibited the activity of CYP1A2 and CYP2C9 with IC50 values of 22.99 and 16.23 µM, which are critical enzymes responsible for the metabolism of rhynchopylline. DISCUSSION AND CONCLUSIONS: The adverse interaction between rhynchopylline and pellodendrine draws attention to the co-administration of these two herbs and provides a reference for further investigations with a broader study population.

Resistance fracture of minimally prepared endocrowns made by three types of restorative materials: a 3D finite element analysis.

A thin endocrown restoration was often applied in endodontically treated teeth with vertical bite height loss or inadequate clinical crown length. A model of mandibular molars made by endocrown restoration with 1 mm thickness and 2 mm depth of pulp chamber was constructed and imported into FEA ANSYS v18.0 software. The three CAD/CAM materials, feldspathic (Mark2), lithium disilicate (EMAX), and lava ultimate (LU), were assigned, and the five load indenters were loaded on the full occlusal (FO), occlusal center (OC), central fossa (CF), buccal groove (BG), and mesiobuccal cusp (MC) of restoration in the model. The MinPS and MaxPS of the thin endocrown were significantly higher than those of tooth tissue in five types of loads except for the LU endocrown loaded in the FO group. The smaller the contact surface of the load was, the higher MaxPS and MinPS were. MaxPS and MinPS of the MC were the highest, followed by the BG and CF in the restoration. In the stress distribution of tooth tissue, MaxPS in the LU endocrown accumulated at the external edge of enamel and was significantly higher than MaxPS in Mark2 and EMAX endocrown concentrated on the chamber wall of dentin under OC, CF and BG loads. Within the limitations of this FEA study, the LU endocrown transferred more stress to tooth tissue than Mark2 and EMAX, and the maximum principal stress on endocrown restoration and tooth tissue at the mesiobuccal cusp load was higher than that at the central fossa and buccal groove load.

Real-Time Induction of Macrophage Apoptosis, Pyroptosis, and Necroptosis by Enterococcus faecalis OG1RF and Two Root Canal Isolated Strains.

To investigate the effects of two Enterococcus faecalis root canal isolated strains (CA1 and CA2) and of the OG1RF strain on apoptosis, pyroptosis, and necroptosis in macrophages. The virulence factors of E. faecalis CA1 and CA2 pathogenic strains were annotated in the Virulence Factors Database (VFDB). E. faecalis CA1, CA2, and OG1RF strains were used to infect RAW264.7 macrophages (MOI, 100:1). We assessed the viability of intracellular and extracellular bacteria and of macrophages at 2, 6, and 12 h post-infection. We used a live cell imaging analysis system to obtain a dynamic curve of cell death after infection by each of the three E. faecalis strains. At 6 and 12 h post-infection, we quantified the mRNA expression levels of PANoptosis-related genes and proteins by RT-qPCR and western blot, respectively. We identified ultrastructural changes in RAW264.7 cells infected with E. faecalis OG1RF using transmission electron microscopy. We found 145 and 160 virulence factors in the CA1 and CA2 strains, respectively. The extracellular CA1 strains grew faster than the CA2 and OG1RF strains, and the amount of intracellular viable bacteria in the OG1RF group was highest at 6 and 12 h post-infection. The macrophages in the CA1 infection group were the first to reach the maximum PI-positivity in the cell death time point curve. We found the expressions of mRNA expression of caspase-1, GSDMD, caspase-3, MLKL, RIPK3, NLRP3, IL-1ß and IL-18 and of proteins cleaved caspase-1, GSDMD, cleaved caspase-3 and pMIKL in the macrophages of the three infection groups to be upregulated (P<0.05). We detected ultrastructural changes of apoptosis, pyroptosis, and necroptosis in macrophages infected with E. faecalis. The three E. faecalis strains induced varying degrees of apoptosis, pyroptosis, and necroptosis that were probably associated with PANoptosis in macrophages. The E. faecalis CA1 strain exhibited faster growth and a higher real-time MOI, and it induced higher expression levels of some PANoptosis-related genes and proteins in the infected macrophages than the other strains tested.

In vitro study on the effect of leonurine hydrochloride on the enzyme activity of cytochrome P450 enzymes in human liver microsomes.

Leonurine hydrochloride (LH) is derived from an ingredient of Leonurus japonicus Houtt which is widely used for diseases in women.The influence of LH on the activity of cytochrome P450 (CYPs) enzymes was investigated in this study.The effect of LH on CYPs enzyme activities were studied using the enzyme-selective substrates phenacetin (1A2), coumarin (2A6), diclofenac (2C9), S-mephenytoin (2C19), paclitaxel (2C8), dextromethorphan (2D6), chlorzoxazone (2E1) and testosterone (3A4). The IC50 value was calculated to express the strength of inhibition. The inhibition of CYPs was fitted with competitive or non-competitive inhibition models and corresponding parameters were also obtained.LH exerted inhibitory effects on the activity of CYP1A2, 2D6, and 3A4 with the IC50 values of 18.05, 15.13, and 20.09 µM, respectively. The obtained results showed that LH inhibited the activity of CYP1A2 and CYP2D6 via competitive manners (Ki = 8.667 µM and Ki = 7.805 µM, respectively), while LH attenuated the CYP3A4 activity via a non-competitive manner (Ki = 9.507 µM). Moreover, LH showed time-dependent inhibition on CYP3A4 with the KI/Kinact value of 4.31/0.044 min-1·µM-1.The inhibition of CYP1A2, CYP2D6, and CYP3A4 by LH, demonstrated in vitro, indicated the potential herb-drug interaction. Therefore, pharmacokinetic interactions involving LH and CYP1A2 or CYP2D6 or CYP1A2 substrates are likely to occur.

Single-Cell Sequencing Unveils the Heterogeneity of Nonimmune Cells in Chronic Apical Periodontitis.

Chronic apical periodontitis (CAP) is a unique dynamic interaction between microbial invasions and host defense mechanisms, resulting in infiltration of immune cells, bone absorption, and periapical granuloma formation. To help to understand periapical tissue pathophysiology, we constituted a single-cell atlas for 26,737 high-quality cells from inflammatory periapical tissue and uncovered the complex cellular landscape. The eight types of cells, including nonimmune cells and immune cells, were identified in the periapical tissue of CAP. Considering the key roles of nonimmune cells in CAP, we emphasized osteo-like cells, basal/stromal cells, endothelial cells, and epithelial cells, and discovered their diversity and heterogeneity. The temporal profiling of genomic alterations from common CAP to typical periapical granuloma provided predictions for transcription factors and biological processes. Our study presented potential clues that the shift of inflammatory cytokines, chemokines, proteases, and growth factors initiated polymorphic cell differentiation, lymphangiogenesis, and angiogenesis during CAP.

Enhanced performances of triboelectric nanogenerators by filling hierarchical flower-like TiO2 particles into polymethyl methacrylate film.

In this study, a flower-like TiO2 filled polymethyl methacrylate (PMMA) composite is presented as a positive tribo-material to produce an excellent-performance triboelectric nanogenerator (TENG). By working in conjunction with polydimethylsiloxane (PDMS), the flat-surface PDMS/PMMA-flower TiO2 TENG generates a voltage of 1200 V, a current of 139 mA m-2 and an output power of 34.85 W m-2, showing significant enhancement compared with its counterpart utilizing neat PMMA as the positive tribo-material under the same operating conditions, whose voltage is 620 V, current is 78 mA m-2 and output power is 13.89 W m-2, respectively. The performance of the TENG is highly dependent on filler loadings of TiO2 flower particles in PMMA composites with an optimal filler loading of 40 wt% with the highest performances. The flower TiO2 is vital to the enhanced performances of the TENG, which is due to the modified surface, the tailored dielectric constant and the space charge polarization. The TENG is capable of powering 600 light emitting diodes, a calculator and a digit display, and applied in self-powered electrophoretic deposition of oxide films. This work demonstrates a facile, low-cost approach for obtaining high-performance TENGs utilizing a PMMA-flower TiO2 composite as the positive tribo-material for applications in sustainable power systems.

A Reconfigurable energy-resolving method for a layered edge-on detector.

Currently, most of the x-ray spectral detectors can extract signals in a set number of energy bins, that inevitably reduces the dynamic range and energy resolution of the imaging system. Inspired by the idea of dynamic thresholding, we previously proposed a pixel architecture and an energy-resolving method for layered edge-on detector. However, the complicated energy exchange mechanism of x-rays in the detector that ultimately affects the practical applications of the layered detectors had not been previously considered. In this study, we modify the energy-depositing model of x-ray photons and propose a reconfigurable energy-resolving method to improve the spectral performance of a layered energy integrating detector. We analyze the errors associated with the energy-resolving process and present our numerical simulation results obtained with energy bins and dynamically changed detection layers to demonstrate the utility and reliability of the proposed method.

Inhibitor of DNA binding 1 (Id1) induces differentiation and proliferation of mouse embryonic carcinoma P19CL6 cells.

The inhibitor of DNA binding (Id) family of genes encodes negative regulators of basic helix-loop-helix transcription factors and has been implicated in such diverse cellular processes as differentiation, proliferation, apoptosis and migration. Id knockout mouse embryos display multiple cardiac defects but the specific role of Id1 in cardiac differentiation is unclear. In the present study, we investigated the function of Id1 in DMSO-induced P19CL6 cells, a widely-accepted cell model of cardiac differentiation. We found that Id1 was upregulated during the cardiac differentiation of P19CL6 cells. The expression of cardiac specific marker genes, Gata4, α-MHC and ISL1, was upregulated in P19CL6 cells stably transfected with Id1 (P19CL6-Id1) during cardiac differentiation. The overexpression of Id1 reduced the number of cells in G1 phase and increased the cell population in G2, M and S phases, while knockdown of Id1 increased the number of cells in G1 phase from 48.6 ± 2.51% to 62.2 ± 1.52% at day 0 of cardiac induction, and from 52.5 ± 3.41% to 63.7 ± 1.02% at day 3 after cardiac induction, indicating that Id1 promoted proliferation of P19CL6 cells. Luciferase assays showed that the activity of TOP flash was higher in P19CL6-Id1 cells than wildtype P19CL6 cells, while Id1 expression was also upregulated in P19CL6 cells treated with Wnt3a or LiCl. This indicates that there may be positive feedback between Id1 and Wnt signaling which plays an important role in cardiac differentiation.