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Tumor cells undergo an epithelial-mesenchymal transition (EMT) accompanied by a reduction in elasticity to initiate metastasis. However, in vivo, tumor cells typically exhibit partial EMT rather than fully EMT. Whether cell mechanics can accurately identify the status of partial EMT, especially the dynamic process, remains unclear. To elucidate the relationship between cell mechanics and partial EMT, we employed scanning ion conductance microscopy (SICM) to analyze the dynamic changes in cell mechanics during the TGFß-induced partial EMT of HCT116 colon cancer cells. Cells undergoing partial EMT, characterized by increased expression of EMT transcription factors, Snai1 and Zeb1, and EMT-related genes, Fn1 and MMP9, while retaining the expression of the epithelial markers E-cadherin (E-cad) and EpCAM, did not exhibit significant changes in cell morphology, suggesting that morphological changes alone were inadequate for identifying partial EMT status. However, cell elasticity markedly decreased in partial EMT cells, and this reduction was reversed with the reversible transition of partial EMT. These findings suggest a strong correlation between cell mechanics and the dynamic process of partial EMT, indicating that cell mechanics could serve as a valuable label-free marker for identifying the status of partial EMT while preserving the physiological characteristics of tumor cells.
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Bio-inspired in nature, using nanomaterials to fabricate the vivid bionic structural color and intelligent stimulus responsive interface as smart skin or optical devices are widely concerned and remain a huge challenge. Here, the bionic flexible film is designed and fabricated with chiral nanointerface and tunable hydrophilic-hydrophobic by the ultrasonic energy perturbation strategy and crosslinking of the cellulose nanocrystals (CNC). An intelligent nanointerface with adjustable hydrophilic and hydrophobic properties is constructed by the supramolecular assembly using a smart ionic liquid molecule. The bionic flexible film possessed the variable hydrophilic-hydrophobic, stimulus responsive, and robust iridescent structural color. The reflective wavelength and the helical pitch of the film can be easily modulated through the ultrasonic energy perturbation strategy. The bionic flexible film by covalent cross-linking has excellent robustness, good elasticity and flexibility. The tunable brilliant structural color of the chiral nanointerface is attributed to the surface charge change of the CNC photonic crystal, which is disturbed by ultrasonic energy perturbation. The bionic flexible film with tunable structure color has intelligent hydrophilic and hydrophobic stimulus response properties. The chiral bionic materials have potential applications in smart skin, optical devices, bionic materials, robots, anti-counterfeiting, colorful displays, and stealth materials.
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Hypoxia was proved to enhance the angiogenesis of stem cells. However, the mechanism of the angiogenic potential in hypoxia-pretreated dental pulp stem cells (DPSCs) is poorly understood. We previously confirmed that hypoxia enhances the angiogenic potential of DPSC-derived exosomes with upregulation of lysyl oxidase-like 2 (LOXL2). Therefore, our study aimed to illuminate whether these exosomes promote angiogenesis via transfer of LOXL2. Exosomes were generated from hypoxia-pretreated DPSCs (Hypo-Exos) stably silencing LOXL2 after lentiviral transfection and characterized with transmission electron microscopy, nanosight and Western blot. The efficiency of silencing was verified using quantitative real-time PCR (qRT-PCR) and Western blot. CCK-8, scratch and transwell assays were conducted to explore the effects of LOXL2 silencing on DPSCs proliferation and migration. Human umbilical vein endothelial cells (HUVECs) were co-incubated with exosomes to assess the migration and angiogenic capacity through transwell and matrigel tube formation assays. The relative expression of angiogenesis-associated genes was characterized by qRT-PCR and Western blot. LOXL2 was successfully silenced in DPSCs and inhibited DPSC proliferation and migration. LOXL2 silencing in Hypo-Exos partially reduced promotion of HUVEC migration and tube formation and inhibited the expression of angiogenesis-associated genes. Thus, LOXL2 is one of various factors mediating the angiogenic effects of Hypo-Exos.
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Exossomos , Humanos , Exossomos/metabolismo , Proliferação de Células/genética , Neovascularização Fisiológica/genética , Células Endoteliais da Veia Umbilical Humana , Células-Tronco , Aminoácido Oxirredutases/genéticaRESUMO
To alleviate the greenhouse effect, increasing the carbon storage of forest and harvested wood products is an effective way. As one of the most important components of forest resources, bamboo plays an important role in carbon sequestration. China possesses the richest bamboo resources in the world. In this study, we developed a comprehensive carbon model including the supply chain from bamboo forest to final products to estimate the carbon pool for China's bamboo industry from 1993 to 2018. To examine the changes in carbon storage, we conducted a spatiotemporal analysis for 15 provinces employing a simple linear regression model. Additionally, a stochastic simulation was employed to test the parameter uncertainty in carbon estimation. Results show the carbon storage of China's bamboo industry has steadily improved, including both bamboo forest and products. In 2018, the carbon storage of bamboo vegetation, soil, and ground layer reached 448.30 TgC, 396.75 TgC, and 11.20 TgC, respectively. The carbon storage of bamboo products also increased gradually, with the storage and emission being 55.33 TgC and 1.70 TgC, respectively. The total carbon storage of China's bamboo industry in 2018 is 909.88 TgC. Moreover, the spatiotemporal analysis quantifies the annual change of the carbon storage for each province. It reveals the spatiotemporal change pattern of total bamboo carbon storage as similar to the bamboo forest. The provinces with more carbon storage tend to increase more rapidly per year. The bamboo industry is immensely significant for both the economy and the environment.
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Sequestro de Carbono , Carbono , Carbono/análise , China , Florestas , SoloRESUMO
Current therapies for chronic pain can have insufficient efficacy and lead to side effects, necessitating research of novel targets against pain. Although originally identified as an oncogene, Tropomyosin-related kinase A (TrkA) is linked to pain and elevated levels of NGF (the ligand for TrkA) are associated with chronic pain. Antibodies that block TrkA interaction with its ligand, NGF, are in clinical trials for pain relief. Here, we describe the identification of TrkA-specific inhibitors and the structural basis for their selectivity over other Trk family kinases. The X-ray structures reveal a binding site outside the kinase active site that uses residues from the kinase domain and the juxtamembrane region. Three modes of binding with the juxtamembrane region are characterized through a series of ligand-bound complexes. The structures indicate a critical pharmacophore on the compounds that leads to the distinct binding modes. The mode of interaction can allow TrkA selectivity over TrkB and TrkC or promiscuous, pan-Trk inhibition. This finding highlights the difficulty in characterizing the structure-activity relationship of a chemical series in the absence of structural information because of substantial differences in the interacting residues. These structures illustrate the flexibility of binding to sequences outside of-but adjacent to-the kinase domain of TrkA. This knowledge allows development of compounds with specificity for TrkA or the family of Trk proteins.
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Inibidores de Proteínas Quinases/química , Inibidores de Proteínas Quinases/farmacologia , Receptor trkA/antagonistas & inibidores , Receptor trkA/química , Sequência de Aminoácidos , Sítios de Ligação , Cristalografia por Raios X , Avaliação Pré-Clínica de Medicamentos , Humanos , Cinética , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/genética , Modelos Moleculares , Conformação Proteica , Inibidores de Proteínas Quinases/síntese química , Receptor trkA/genética , Receptor trkB/antagonistas & inibidores , Receptor trkB/química , Receptor trkB/genética , Receptor trkC/antagonistas & inibidores , Receptor trkC/química , Receptor trkC/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/efeitos dos fármacos , Proteínas Recombinantes/genética , Relação Estrutura-Atividade , Ressonância de Plasmônio de SuperfícieRESUMO
The reliability-based sensitivity analysis requires to recursively evaluate a multivariate structural model for many failure probability levels. This is in general a computationally intensive task due to irregular integrations used to define the structural failure probability. In this regard, the performance function is first approximated by using the multiplicative dimensional reduction method in this paper, and an approximation for the reliability-based sensitivity index is derived based on the principle of maximum entropy and the fractional moment. Three examples in the literature are presented to examine the performance of this entropy-based approach against the brute-force Monte-Carlo simulation method. Results have shown that the multiplicative dimensional reduction based entropy approach is rather efficient and able to provide reliability estimation results for the reliability-based sensitivity analysis of a multivariate structural model.
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CONTEXT: Shikonin (SHI), an active component extracted from Radix Arnebiae, has been reported to possess anti-inflammatory properties in various cells. However, its effect on lipopolysaccharide (LPS)-stimulated human periodontal ligament cells (hPDLCs) is unknown. OBJECTIVE: To investigate the effects of SHI on the expression of inflammatory related cytokines in LPS-stimulated hPDLCs. MATERIALS AND METHODS: The effects of SHI (0.125, 0.25, 0.5, 1, and 2 µg/mL) on hPDLCs proliferation for 1, 3 and 7 days were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of interleukin-1 (IL-1), IL-6, tumor necrosis factor-α (TNF-α), matrix metalloproteinase-2 (MMP-2), MMP-9 and cyclooxygenase-2 (COX-2) were detected in hPDLCs following SHI treatment (0.25 and 0.5 µg/mL) using Quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR). The signaling pathways triggered by SHI in hPDLC were evaluated using western blotting. RESULTS: LD50 of SHI is 1.7 µg/mL (day 1) and 1.1 µg/mL (day 3 and 7) in hPDLCs. No morphological changes were observed when hPDLCs were treated with LPS only (1 µg/mL) or LPS with SHI (0.25 and 0.5 µg/mL). Data from qRT-PCR suggests that SHI attenuates LPS-induced increases of IL-1, IL-6, TNF-α, MMP-2, MMP-9 and COX-2 in hPDLCs. Down-regulation of phosphorylated extracellular signal-regulated kinase (ERK) and nuclear factor-κB (NF-κB), and up-regulation of I-κB, were observed in LPS-stimulated hPDLCs after exposed to SHI at 0.25 or 0.5 µg/mL. DISCUSSION AND CONCLUSIONS: SHI possesses anti-inflammatory effects in LPS-stimulated hPDLCs via phospho-ERK and NF-κB/I-κB signaling pathways; this suggests that SHI may hold potential as an anti-inflammatory agent against periodontitis.
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Anti-Inflamatórios/farmacologia , Naftoquinonas/farmacologia , Ligamento Periodontal/efeitos dos fármacos , Periodontite/tratamento farmacológico , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Humanos , Lipopolissacarídeos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Metaloproteinases da Matriz/metabolismo , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Ligamento Periodontal/patologia , Periodontite/induzido quimicamente , Periodontite/metabolismo , Periodontite/patologia , FosforilaçãoRESUMO
High mobility group box 1 protein (HMGB1) is a chromatin protein which can be released extracellularly, eliciting a pro-inflammatory response and promoting tissue repair process. This study aimed to examine the expression and distribution of HMGB1 and its receptor RAGE in inflamed dental pulp tissues, and to assess its effects on proliferation, migration and cytoskeleton of cultured human dental pulp cells (DPCs). Our data demonstrated that cytoplasmic expression of HMGB1 was observed in inflamed pulp tissues, while HMGB1 expression was confined in the nuclei in healthy dental pulp. The mRNA expression of HMGB1 and RAGE were significantly increased in inflamed pulps. In in vitro cultured DPCs, expression of HMGB1 in both protein and mRNA level was up-regulated after treated with lipopolysaccharide (LPS). Exogenous HMGB1 enhanced DPCs migration in a dose-dependent manner and induced the reorganization of f-actin in DPCs. Our results suggests that HMGB1 are not only involved in the process of dental pulp inflammation, but also play an important role in the recruitment of dental pulp stem cells, promoting pulp repair and regeneration.
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Quimiotaxia , Polpa Dentária/metabolismo , Proteína HMGB1/metabolismo , Sequência de Bases , Proliferação de Células , Primers do DNA , Polpa Dentária/citologia , Humanos , Reação em Cadeia da PolimeraseRESUMO
Silicon nanowire field effect (SiNW-FET) biosensors have been successfully used in the detection of nucleic acids, proteins and other molecules owing to their advantages of ultra-high sensitivity, high specificity, and label-free and immediate response. However, the presence of the Debye shielding effect in semiconductor devices severely reduces their detection sensitivity. In this paper, a three-dimensional stacked silicon nanosheet FET (3D-SiNS-FET) biosensor was studied for the high-sensitivity detection of nucleic acids. Based on the mainstream Gate-All-Around (GAA) fenestration process, a three-dimensional stacked structure with an 8 nm cavity spacing was designed and prepared, allowing modification of probe molecules within the stacked cavities. Furthermore, the advantage of the three-dimensional space can realize the upper and lower complementary detection, which can overcome the Debye shielding effect and realize high-sensitivity Point of Care Testing (POCT) at high ionic strength. The experimental results show that the minimum detection limit for 12-base DNA (4 nM) at 1 × PBS is less than 10 zM, and at a high concentration of 1 µM DNA, the sensitivity of the 3D-SiNS-FET is approximately 10 times higher than that of the planar devices. This indicates that our device provides distinct advantages for detection, showing promise for future biosensor applications in clinical settings.
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Técnicas Biossensoriais , Nanofios , Ácidos Nucleicos , Silício/química , Transistores Eletrônicos , DNA , Técnicas Biossensoriais/métodos , Nanofios/químicaRESUMO
Acute kidney injury (AKI) is a frequently occurring severe disease with high mortality. Cystatin C (Cys-C), as a biomarker of early kidney failure, can be used to detect and prevent acute renal injury. In this paper, a biosensor based on a silicon nanowire field-effect transistor (SiNW FET) was studied for the quantitative detection of Cys-C. Based on the spacer image transfer (SIT) processes and channel doping optimization for higher sensitivity, a wafer-scale, highly controllable SiNW FET was designed and fabricated with a 13.5 nm SiNW. In order to improve the specificity, Cys-C antibodies were modified on the oxide layer of the SiNW surface by oxygen plasma treatment and silanization. Furthermore, a polydimethylsiloxane (PDMS) microchannel was involved in improving the effectiveness and stability of detection. The experimental results show that the SiNW FET sensors realize the lower limit of detection (LOD) of 0.25 ag/mL and have a good linear correlation in the range of Cys-C concentration from 1 ag/mL to 10 pg/mL, exhibiting its great potential in the future real-time application.
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Técnicas Biossensoriais , Nanofios , Insuficiência Renal , Humanos , Silício , Cistatina C , Transistores Eletrônicos , Biomarcadores , Técnicas Biossensoriais/métodosRESUMO
With the intensification of conflicts between different ecosystem services, how to achieve a win-win situation between socio-economic development and ecological protection is an important issue that needs to be addressed nowadays. In particular, how to better quantify and assess the intensity of ecosystem service trade-offs and their relative benefits, and to identify the influencing factors are issues that need to be studied in depth. Based on the INVEST model, this paper analyzed the evolution of spatial and temporal patterns of ecosystem services such as Carbon Storage (CS), Food Production (FP), Habitat Quality (HQ), and Water Yield (WY) in the Shandong Yellow River Basin (SYRB) in 2000, 2010 and 2020. Next, we quantitatively measured the trade-off intensity and revealed the key influencing factors of the trade-off intensity evolution using automatic linear models, root mean square deviation, and geographically weighted regressions. Subsequently, we further analyzed the impact of the correlation between environmental and socio-economic factors on the trade-off intensity of ecosystem services. The results indicated that the temporal and spatial changes of the four main ecosystem services in SYRB area were inconsistent. WY showed a fluctuating trend, with a large interannual gap. CS and FP are on the rise, while HQ is on the decline. Spatially, WY and HQ showed a decreasing distribution from the center to the periphery, while FP and CS showed a decreasing distribution from the southwest to the northeast. The location characteristics of SYRB's four ecosystem services and their trade-offs were obvious. FP had absolute location advantage in ecosystem service trade-offs. Most of the four ecosystem services showed significant trade-offs, and the trade-off intensity had significant spatial heterogeneity, but the trade-off between FP and CS was relatively weak. At the same time, there were also differences in the trends of trade-off intensities. Counties with low trade-off intensity were mostly located in mountainous areas; these areas are less disturbed by human activities, and most of them are areas without prominent services. Counties with high trade-off intensities were mostly concentrated in areas with relatively developed agriculture; these areas are more disturbed by human activities and are mostly prominent in FP. The trade-off intensity of ecosystem services in SYRB was affected by several factors together, and there were difference characteristics in the degree and direction of influence of each factor. Moreover, these influencing factors have gradually changed over 20 years. In terms of the spatial distribution at the county scale, the influence areas of the dominant drivers of different trade-off types varied greatly, among which the areas with NDVI, CON, and PRE as the dominant factors were the largest. In the future, in effectively balancing regional economic development and ecological environmental protection, quantifiable correspondence strategies should be developed from the administrative perspective of counties and regions based on comprehensive consideration of the locational advantages of each ecosystem service and changes in trade-offs.
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Conservação dos Recursos Naturais , Ecossistema , Humanos , Conservação dos Recursos Naturais/métodos , Rios , Agricultura , ChinaRESUMO
Accumulated hard tissue debris (AHTD) is an inevitable by-product during endodontic treatment and is difficult to remove completely using traditional syringe and needle irrigation (SNI). Adjunctive irrigation is proposed to assist the clean-up of AHTD. This systematic review and meta-analysis aimed to evaluate the AHTD removal efficacy of different root canal irrigation devices using micro-computed tomography (Micro-CT). A literature search was carried out within the main scientific databases until 20 June 2022. All results were screened with detailed eligibility criteria. Eleven studies were included for analysis. SNI, passive ultrasonic irrigation (PUI), negative pressure systems, sonically activated irrigation (SAI), mechanical-activated system and laser-activated irrigation (LAI) were assessed. PUI is superior to SNI for debris removal and LAI has better AHTD removal performance than PUI. The negative pressure system and mechanical-activated system were proved to be less effective. Registration: PROSPERO (CRD42021273892).
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In situ X-ray computed tomography (X-ray CT) is used to investigate the effects of characteristic microstructural features on the pitting initiation and propagation in austenitic stainless steel specimens prepared with laser powder bed fusion (LPBF) additive manufacturing. In situ X-ray CT in probing the mechanism and kinetics of localized corrosion is demonstrated by immersing two LPBF specimens with different porosities in an aggressive ferric chloride solution for the evaluation of corrosion. X-ray CT images are acquired from the specimens after every 8 hours of immersion over an extended period of time (216 hours). Corrosion pit growth is then quantitatively analyzed with a data-constrained modeling method. The pitting growth mechanism of LPBF stainless steel is found to be different from that of conventional stainless steels. More specifically, the mechanism of corrosion pit initiation is closely correlated with the original lack of fusion porosity (LOF) distribution on the surface of the specimens and preferential pit propagation through the LOF pores inside the specimens. Pit growth kinetics are derived from pit volume changes determined through 3D data analysis. The pit growth kinetics in LPBF specimens are found to vary in the initial pit formation, competitive pit propagation, and the dominant pit growth stages.
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Erythropoietin (EPO) is a 34-kDa glycoprotein that possesses the potential for angiogenesis, as well as anti-inflammatory and anti-apoptotic properties. The present study aimed to examine the effect of EPO on the angiogenesis of dental pulp cells (DPCs) and to explore the underlying mechanisms of these effects. It was demonstrated that EPO not only promoted DPCs proliferation but also induced angiogenesis of DPCs in a paracrine fashion. EPO enhanced the angiogenic capacity by stimulating DPCs to secrete a series of angiogenic cytokines. ELISA confirmed that high concentrations of EPO increased the production of MMP-3 and angiopoietin-1 but decreased the secretion of IL-6. Furthermore, EPO activated the ERK1/2 and p38 signaling pathways in DPCs, while inhibition of these pathways diminished the angiogenesis capacity of DPCs. The present study suggested that EPO may have an important role in the repair and regeneration of dental pulp.
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Rational modification of a previously identified spirohydantoin lead structure has identified a series of potent spiroazaoxindole CGRP receptor antagonists. The azaoxindole was found to be a general replacement for the hydantoin that consistently improved in vitro potency. The combination of the indanylspiroazaoxindole and optimized benzimidazolinones led to highly potent antagonists (e.g., 25, CGRP K(i)=40pM). The closely related compound 27 demonstrated good oral bioavailability in dog and rhesus.
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Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina , Indóis/síntese química , Compostos de Espiro/síntese química , Administração Oral , Animais , Disponibilidade Biológica , Cães , Descoberta de Drogas , Humanos , Indóis/farmacologia , Macaca mulatta , Oxindóis , Compostos de Espiro/farmacologia , Relação Estrutura-AtividadeRESUMO
In the process of bone tissue engineering, the osteoimmunomodulatory property of biomaterials is very important for osteogenic differentiation of stem cells, which determines the outcome of bone regeneration. Magnesium (Mg) is a biodegradable, biocompatible metal that has osteoconductive properties and has been regarded as a promising bone biomaterial. However, the high degradation rate of Mg leads to excessive inflammation, thereby restricting its application in bone tissue engineering. Importantly, different coatings or magnesium alloys have been utilized to lower the rate of degradation. In fact, a prior study proved that ß-TCP coating of Mg scaffolds can modulate the osteoimmunomodulatory properties of Mg-based biomaterials and create a favorable immune microenvironment for osteogenesis. However, the osteoimmunomodulatory properties of Mg ions themselves have not been explored yet. In this study, the osteoimmunomodulatory properties of Mg ions with involvement of macrophages and bone marrow stem cells (BMSCs) were systematically investigated. Microscale Mg ions (100 mg/L) were found to possess osteoimmunomodulatory properties that favor bone formation. Specifically, microscale Mg ions induced M2 phenotype changes of macrophages and the release of anti-inflammatory cytokines by inhibiting the TLR-NF-κB signaling pathway. Microscale Mg ions also stimulated the expression of osteoinductive molecules in macrophages while Mg ions/macrophage-conditioned medium promoted osteogenesis of BMSCs through the BMP/SMAD signaling pathway. These findings indicate that manipulating Mg ion concentration can endow the Mg biomaterial with favorable osteoimmunomodulatory properties, thereby providing fundamental evidence for improving and modifying the effect of Mg-based bone biomaterials.
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Células da Medula Óssea/efeitos dos fármacos , Imunomodulação/efeitos dos fármacos , Magnésio/farmacologia , Osteogênese/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Animais , Regeneração Óssea , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Camundongos , Células RAW 264.7 , Transdução de Sinais , Alicerces TeciduaisRESUMO
The previously disclosed spirohydantoin-based CGRP receptor antagonists were optimized for potency through modification of the benzimidazolone substituents. Compounds were identified which had minimal shift in the cAMP functional assay containing 50% human serum. Blockade of CGRP-mediated vasodilation was observed with these compounds in a rhesus pharmacodynamic assay and the in vivo potency correlated with the in vitro activity in the serum-shifted functional assay.
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Benzimidazóis/síntese química , Benzimidazóis/farmacologia , Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina , Indanos/química , Compostos de Espiro/síntese química , Compostos de Espiro/farmacologia , Animais , Benzimidazóis/sangue , Benzimidazóis/química , Técnicas de Química Combinatória , Humanos , Macaca mulatta , Estrutura Molecular , Compostos de Espiro/química , Relação Estrutura-AtividadeRESUMO
Periodontitis is a progressive inflammatory disease initiated by bacterial biofilm adhering to the tooth surface. If left untreated, periodontitis may lead to tooth loss and destruction of the alveolar bone. Regaining the lost alveolar bone is a clinical challenge because of the limited differentiation ability of osteoblasts in inflammatory environments. We have previously shown the anti-inflammatory and antiosteoclastogenic activities of parthenolide (PTL) in human periodontal ligament-derived cells by inhibiting nuclear factor kappa B (NF-κB) signaling, indicating its potential for periodontitis treatment. In this study, we further examined whether PTL could stimulate differentiation of osteoblasts from human alveolar bone in inflammatory conditions and investigated the involvement of the Wnt/ß-catenin signaling pathway during this process. The results showed that PTL significantly stimulated alkaline phosphatase activity, mineralization nodule formation, and osteogenesis-related gene/protein expression of osteoblasts under the stimulation of tumor necrosis factor-α (TNF-α). In addition, PTL inhibited the NF-κB/p50 pathway and resisted the inhibition of Wnt/ß-catenin signaling induced by TNF-α. Our results indicate that the stimulatory effect of PTL on the differentiation of osteoblasts in inflammatory environments may involve the activation of the Wnt/ß-catenin signaling pathway, and PTL may be a promising component for bone regeneration in periodontitis treatment.
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Diferenciação Celular/efeitos dos fármacos , Inflamação/metabolismo , Osteoblastos/efeitos dos fármacos , Sesquiterpenos/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Humanos , Osteoblastos/citologia , Osteoblastos/metabolismoRESUMO
UNLABELLED: Osteoarthritis (OA) is a chronic, incurable and destructive joint disease that is characterized by chondrocyte hypertrophy and cartilage degradation. Angiogenesis, mediated by the action of vascular endothelial growth factor (VEGF), is known to be a contributing factor in the pathogenesis of OA. In this study, we use a lentivirus-based approach to investigate whether VEGF knockdown would be beneficial to chondrogenesis and could prevent or slow down OA progression. We first profiled cytokines in human OA cartilage using cytokine antibody arrays. This revealed that as many as 21 angiogenesis-related cytokines were significantly upregulated in severe OA cartilage compared to mild OA samples. Next, we infected chondrocytes with VEGF small hairpin RNA (shRNA) lentivirus (LV-VEGF shRNA) and treated these cells with tumour necrosis factor alpha (TNF-α) to induce hypertrophy. The results showed that inhibition of VEGF not only enhanced chondrogenic differentiation, but also protected chondrocytes from TNF-α-induced hypertrophy. We also found that knockdown of VEGF suppressed TNF-α-induced phosphorylation of ERK1/2 in chondrocytes. Furthermore, using a surgically induced OA rat model, we showed that VEGF inhibition delayed OA progression in animals given intra-articular injection of LV-VEGF shRNA. In conclusion, in this study, we have shown that VEGF knockdown can enhance chondrogenesis and prevent OA progression, thus providing evidence that inhibition of VEGF may be a potential therapeutic approach for OA patients. KEY MESSAGES: Numerous pro-angiogenic factors are upregulated in severe OA cartilage. Inhibition of VEGF by shRNA protects chondrocytes from TNF-α-induced hypertrophy. Knockdown of VEGF suppresses TNF-α-induced phosphorylation of ERK1/2 in chondrocytes. VEGF inhibition delays OA progression in rat model in vivo. Inhibition of VEGF may be a potential therapeutic approach for OA patients.
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Cartilagem Articular/metabolismo , Neovascularização Patológica/prevenção & controle , Osteoartrite do Joelho/terapia , RNA Interferente Pequeno/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Cartilagem Articular/patologia , Proliferação de Células , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Condrócitos/patologia , Feminino , Regulação da Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Lentivirus/genética , Lentivirus/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Osteoartrite do Joelho/genética , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Fosforilação/efeitos dos fármacos , Cultura Primária de Células , RNA Interferente Pequeno/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais , Fator de Necrose Tumoral alfa/farmacologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Hypertrophic scarring/hypertrophic scars (HS) is a highly prevalent condition following burns and trauma wounds. Numerous studies have demonstrated that transforming growth factor-ß1 (TGFß1) plays an essential role in the wound healing process by regulating cell differentiation, collagen production and extracellular matrix degradation. The increased expression of TGF-ß1 is believed to result in the formation of HS. Shikonin (SHI), an active component extracted from the Chinese herb, Radix Arnebiae, has previously been found to downregulate the expression of TGF-ß1 in keratinocyte/fibroblast co-culture conditioned medium. In view of this, in this study, we aimed to further investigate the effects of SHI on TGF-ß1-stimulated hypertrophic scar-derived human skin fibroblasts (HSFs) and examined the underlying mechanisms. Cell viability and proliferation were measured using alamarBlue and CyQUANT assays. The total amount of collagen and cell contraction were examined using Sirius red staining and the cell contraction assay kit. Gene expression and signalling pathway activation were detected using reverse transcription-quantitative polymerase chain reaction and western blot analysis. Our results revealed that SHI reduced TGF-ß1induced collagen production through the ERK/Smad signalling pathway and attenuated TGF-ß1induced cell contraction by downregulating α-smooth muscle actin (αSMA) expression in the HSFs. The data from this study provide evidence supporting the potential use of SHI as a novel treatment for HS.