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A tricalcium-silicate-nanoparticle-containing cement (Biodentine) was developed to overcome the disadvantages of existing mineral trioxide aggregate (MTA) dental materials. This study aimed at evaluating the effects of Biodentine on the osteogenic differentiation of human periodontal ligament fibroblasts (HPLFs) in vitro and the healing of furcal perforations created experimentally in rat molars in vivo, in comparison to MTA. The in vitro studies performed the following assays: pH measurement using a pH meter, the release of calcium ions using a calcium assay kit, cell attachment and morphology using SEM, cell proliferation using a coulter counter, marker expression using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and cell mineralized deposit formation using Alizarin Red S (ARS) staining. In the in vivo studies, MTA and Biodentine were used to fill the rat molar perforations. Rat molars were processed at 7, 14 and 28 days for analysis of inflammatory processes using hematoxylin and eosin (HE) staining, immunohistochemical staining of Runx2 and tartrate-resistant acid phosphate (TRAP) staining. The results demonstrate that the nanoparticle size distribution of Biodentine is critical for osteogenic potential at an earlier stage compared to MTA. Further studies are required to elucidate the mechanism of action of Biodentine in osteogenic differentiation. [ABSTRACT FROM AUTHOR]

Odontoblasts are involved in sensory generation as sensory receptor cells and in dentin formation. We previously reported that an increase in intracellular cAMP levels by cannabinoid 1 receptor activation induces Ca2+ influx via transient receptor potential vanilloid subfamily member 1 channels in odontoblasts, indicating that intracellular cAMP/Ca2+ signal coupling is involved in dentinal pain generation and reactionary dentin formation. Here, intracellular cAMP dynamics in cultured human odontoblasts were investigated to understand the detailed expression patterns of the intracellular cAMP signaling pathway activated by the Gs protein-coupled receptor and to clarify its role in cellular functions. The presence of plasma membrane Gαs as well as prostaglandin I2 (IP), 5-hydroxytryptamine 5-HT4 (5-HT4), dopamine D1 (D1), adenosine A2A (A2A), and vasoactive intestinal polypeptide (VIP) receptor immunoreactivity was observed in human odontoblasts. In the presence of extracellular Ca2+, the application of agonists for the IP (beraprost), 5-HT4 (BIMU8), D1 (SKF83959), A2A (PSB0777), and VIP (VIP) receptors increased intracellular cAMP levels. This increase in cAMP levels was inhibited by the application of the adenylyl cyclase (AC) inhibitor SQ22536 and each receptor antagonist, dose-dependently. These results suggested that odontoblasts express Gs protein-coupled IP, 5-HT4, D1, A2A, and VIP receptors. In addition, activation of these receptors increased intracellular cAMP levels by activating AC in odontoblasts. [ABSTRACT FROM AUTHOR]

We investigated whether BMP4, FGF8, and/or WNT3a on neural crest-like cells (NCLC) derived from mouse induced pluripotent stem (miPS) cells will promote differentiation of odontoblasts-like cells. After the miPS cells matured into embryonic body (EB) cells, they were cultured in a neural induction medium to produce NCLC. As the differentiation of NCLC were confirmed by RT-qPCR, they were then disassociated and cultured with a medium containing, BMP4, FGF8, and/or WNT3a for 7 and 14 days. The effect of these stimuli on NCLC were assessed by RT-qPCR, ALP staining, and immunocytochemistry. The cultured EB cells presented a significant increase of Snai1, Slug, and Sox 10 substantiating the differentiation of NCLC. NCLC stimulated with more than two stimuli significantly increased the odontoblast markers Dmp-1, Dspp, Nestin, Alp, and Runx2 expression compared to control with no stimulus. The expression of Dmp-1 and Dspp upregulated more when FGF8 was combined with WNT3a. ALP staining was positive in groups containing BMP4 and fluorescence was observed in immunocytochemistry of the common significant groups between Dmp-1 and Dspp. After stimulation, the cell morphology demonstrated a spindle-shaped cells with long projections resembling odontoblasts. Simultaneous BMP4, FGF8, and WNT3a stimuli significantly differentiated NCLC into odontoblast-like cells. [ABSTRACT FROM AUTHOR]

Primary cultured odontoblasts rapidly lose their tissue-specific phenotype. To identify transcription factors (TF) that are important for the maintenance of the odontoblast phenotype, primary cultures of C57BL/6 J mouse dental mesenchymal cells (DMC) were isolated, and expression of TF and odontoblast marker genes in cells immediately after isolation and 2 days after culture were comprehensively evaluated and compared using RNA-sequencing (RNA-seq). The expression of odontoblast markers in mouse dental mesenchymal cells decreased rapidly after isolation. In addition, the expression of Hedgehog-related, Notch-related, and immediate- early gene (IEG)-related transcription factors significantly decreased. Forced expression of these genes in lentiviral vectors, together with fibroblast growth factor 4 (FGF4), fibroblast growth factor 9 (FGF9), and the Wnt pathway activator CHIR99021, significantly induced the expression of odontogenic marker genes. These results indicate, for the first time, that Notch signaling and early genes may be important for maintaining odontoblast cultures. Furthermore, simultaneous stimulation of FGF, Wnt, Hedgehog, Notch pathways, and IEG transcription factors cooperatively promoted the maintenance of the odontoblast phenotype. These results suggest that the Hedgehog and Notch signaling pathways may play an important role in maintaining odontoblast phenotypes, in addition to FGF and Wnt signaling. [ABSTRACT FROM AUTHOR]

The use of solution processes—as opposed to conventional vacuum processes and vapour-phase deposition—for the fabrication of electronic devices has received considerable attention for a wide range of applications, with a view to reducing processing costs. In particular, the ability to print semiconductor devices using liquid-phase materials could prove essential for some envisaged applications, such as large-area flexible displays. Recent research in this area has largely been focused on organic semiconductors, some of which have mobilities comparable to that of amorphous silicon (a-Si); but issues of reliability remain. Solution processing of metal chalcogenide semiconductors to fabricate stable and high-performance transistors has also been reported. This class of materials is being explored as a possible substitute for silicon, given the complex and expensive manufacturing processes required to fabricate devices from the latter. However, if high-quality silicon films could be prepared by a solution process, this situation might change drastically. Here we demonstrate the solution processing of silicon thin-film transistors (TFTs) using a silane-based liquid precursor. Using this precursor, we have prepared polycrystalline silicon (poly-Si) films by both spin-coating and ink-jet printing, from which we fabricate TFTs with mobilities of 108 cm2 V-1 s-1 and 6.5 cm2 V-1 s-1, respectively. Although the processing conditions have yet to be optimized, these mobilities are already greater than those that have been achieved in solution-processed organic TFTs, and they exceed those of a-Si TFTs (≤ 1 cm2 V-1 s-1). [ABSTRACT FROM AUTHOR]

This study evaluated the microtensile bond strength (μTBS) of 1-step self-etch adhesives (1-SEAs) and 2-step self-etch adhesives (2-SEAs) to pulp chamber dentin immediately after bleaching with 2 types of common bleaching techniques. Pulp chamber dentin of bovine teeth was bleached using 30% hydrogen peroxide (H2O2) solution with quartz-tungsten-halogen light-curing unit (Group 1) and 3.5% H2O2-containing titanium dioxide (TiO2) (Pyrenees®) activated with 405-nm violet diode laser for 15 min (Group 2). Unbleached specimens were placed in distilled water for 15 min and used as controls. After treatment, dentin was bonded with resin composite using 1-SEA or 2-SEA and stored in water at 37°C for 24 h. Each specimen was sectioned and trimmed to an hourglass-shape and μTBS was measured. Fractured specimens were examined under a scanning electron microscope to determine fracture modes. All specimens in Group 1 failed before proper bonding tests. In Group 2, the μTBS of 2-SEA was significantly greater (with no failed specimens) than 1-SEA (where 21 out of 36 failed). These results indicate that 2-SEA is a better adhesive system than 1-SEA on bleached dentin. Our results also demonstrated that application of H2O2 significantly decreases bond strength of resin to dentin; however, in the case of nonvital tooth bleaching, Pyrenees® is a better alternative to the conventional 30% H2O2 bleaching. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: This study is aimed to test whether glucose-free conditions induce the activation of adenosine monophosphate-activated protein kinase (AMPK) and, to investigate association with AMPK expression and cell viability in human dental pulp cells. Design: Human dental pulp cells were initially maintained in culture medium containing glucose and the medium was subsequently changed to glucose-free medium. To evaluate the expression of AMPK, quantitative real-time RT-PCR, Western blot analysis and immunofluorescence were carried out. Cell viability was evaluated by MTT assay. Results: The expression of AMPK mRNA in glucose free conditions was 2.0–2.5 fold higher than the control at 1, 2 and 3h (P <0.01). The expression of phosphorylated-AMPK was characterized by Western blot analysis and by immunofluorescence. Compound C-pre-treated group showed a decline of both AMPK expression and cell viability, while AICAR-pre-treated group showed an increase of AMPK and maintain of cell viability at regular level. Conclusions: AMPK plays an important role on fluctuating in accordance with glucose availability and protects cell viability from glucose free condition in human dental pulp cells. [Copyright &y& Elsevier]

Abstract: Calcitonin gene-related peptides (CGRP) and adrenomedullin (ADM) belong to the calcitonin family of peptides and are structurally related. Both peptides are found in the neurons of the CNS and play a role in many neuronal functions, including the control of blood pressure. The nucleus tractus solitarius (NTS) is known to play a major role in the regulation of cardiovascular, respiratory, gustatory, hepatic and swallowing functions. Recently, hypotension and bradycardia were observed after CGRP and ADM injection in the NTS. Voltage-dependent Ca2+ channels (VDCCs) serve as crucial mediators of membrane excitability and Ca2+-dependent functions, such as neurotransmitter release, enzyme activity, and gene expression. The purpose of this study is to investigate the effects of CGRP and ADM on VDCC currents (I Ca) carried by Ba2+ (I Ba) in the NTS, using patch-clamp recording methods. Application of CGRP and ADM caused facilitation of I Ba in a concentration-dependent manner. Intracellular dialysis of the anti-Gαs-protein antibody attenuated CGRP-induced facilitation of I Ba. Intracellular dialysis of the anti-Gαi-protein antibody attenuated ADM-induced facilitation of I Ba. Pretreatment with SQ22536 (an adenylate cyclase inhibitor) and intracellular dialysis of PKI(5–24) (a protein kinase A inhibitor) attenuated CGRP-induced facilitation of I Ba. In contrast, pretreatment with PD98,059 (a mitogen-activated protein kinas inhibitor) attenuated ADM-induced facilitation of I Ba. Mainly L-type VDCCs were facilitated by both CGRP and ADM. These results indicate that CGRP facilitates L-type VDCCs via Gαs-protein involving adenylate cyclase and protein kinase A. In contrast, ADM facilitates L-type VDCCs via Gαi-protein involving mitogen-activated protein kinase in the NTS. [Copyright &y& Elsevier]