Apoptotic/cell cycle arrest potential of dusty miller methanol extract against Paca-2 pancreatic cancer cells via upregulating Bax/Bcl2: A HPLC-ESI-MS/MS, GNPS-based molecular networking and network pharmacology studies.

Speciation analysis reveals intracellular distribution of arsenicals among subcellular organelles of arsenic-methylated human cells exposed to arsenite.

Impact of P-glycoprotein substrates on transendothelial transport of amyloid-β peptide in an in vitro model.

Introduction: Orthodontic tooth movement is governed by coordinated bone resorption and formation mediated primarily through the Receptor Activator of Nuclear Factor-κB Ligand – Osteoprotegerin signaling axis. Although mechanical force initiates this process, the biological rate of remodeling remains a limiting factor. CRISPR-based transcriptional activation presents a novel strategy to amplify force-induced molecular responses. Material and Methods: Human periodontal ligament stem cells were exposed to simulated compressive orthodontic force (2 g/cm²) and subjected to CRISPR-dCas9-VPR–mediated activation of the TNFSF11 (RANKL) promoter. Samples were divided into control, force-only, scramble control, and CRISPR-RANKL groups. Cell viability was assessed using CCK-8 assay, while gene and protein expression of RANKL, OPG, and RUNX2 were evaluated using RT-qPCR and ELISA. Results: Cell viability exceeded 90% across all groups, indicating no cytotoxic effects. CRISPR-mediated activation significantly enhanced RANKL expression under compressive force, producing a marked increase in the RANKL/OPG ratio compared with force alone (p < 0.001). RUNX2 expression was reduced under compression, consistent with osteoclastic dominance, and was unaffected by CRISPR modulation. Conclusion: CRISPR-dCas9-VPR–based activation of RANKL synergistically augments mechanical force–induced osteoclastic signaling in periodontal ligament stem cells. This proof-of-concept study highlights the potential of epigenetic modulation as a precision approach for biologically accelerating orthodontic tooth movement.

Introduction: Conventional vital pulp therapy relies primarily on calcium silicate-based cements that induce reparative dentin formation without restoring the native neurovascular architecture of the pulp tissue. Advances in tissue engineering using stem cells and biodegradable scaffolds offer the potential for true pulp regeneration rather than mere preservation. Material and Methods: Human dental pulp stem cells were isolated from healthy third molars and encapsulated within gelatin methacryloyl hydrogel constructs. In this randomized laboratory trial, 60 standardized human tooth slices were allocated into three groups: negative control (empty), positive control (Biodentine™), and test group (human dental pulp stem cells-gelatin methacryloyl construct). Cell viability, proliferation, odontogenic differentiation, and angiogenic potential were assessed using Live/Dead staining, Cell counting Kit-8 colorimetric assay, and quantitative reverse transcription polymerase chain reaction analysis of dentin sialophosphoprotein, dentin matrix acidic phosphoprotein 1, and vascular endothelial growth factor expression. Results: The quantitative reverse transcription polymerase chain reaction constructs demonstrated high cytocompatibility, with cell viability exceeding 94% at day 7. Proliferation was significantly greater in the test group compared with Biodentine at day 7 (p < 0.01). Odontogenic marker expression was comparable between the test and Biodentine groups, while vascular endothelial growth factor expression was markedly higher in the test group group, indicating superior angiogenic potential. Conclusion: Stem cell–laden gelatin methacryloyl constructs exhibit enhanced regenerative properties compared with conventional bioceramic materials in an ex vivo tooth slice model. These findings support the translational potential of hydrogel-based regenerative strategies as next-generation approaches for vital pulp therapy.

Prokaryotic and eukaryotic L-lactate dehydrogenases catalyze both the reduction of pyruvate to L-lactate and the reverse reaction generating the α-ketoacid. Remarkably, the energetic metabolism of human malignant cells is sustained by lactate dehydrogenase A (hLDH-A), the catalytic action of which is committed to its homotetrameric form (denoted hLDH-5), and is coupled to glycolysis. Therefore, hLDH-A represents a substantial druggable target, demanding the search for effective inhibitors of this enzyme. Here we report on the inhibition of hLDH-A exerted by dicarboxylates, whose performance is strictly dependent on their carbon chain length. In particular, the best performers were tetradecanedioic acid, hexadecanedioic acid, and crocetin (a polyunsaturated dicarboxylate), whose addition to assay mixtures strongly inhibited the activity of hLDH-A. Moreover, the inhibition of hLDH-A by hexadecanedioic acid was more effective against the monomeric enzyme than towards its tetrameric counterpart, suggesting that this dicarboxylate interferes with the assembly of hLDH-5. Furthermore, docking simulations support that long-, but not short-chain dicarboxylates, effectively bind to a specific site of monomeric hLDH-A, plausibly preventing its assembly into catalytically-competent hLDH-5. Overall, our observations indicate long-chain dicarboxylates as efficient inhibitors of hLDH-A, prompting to test their action in cellulo.

Activation of WNT signaling in human pluripotent stem cells efficiently drives lateral mesoderm specification and subsequent cardiomyocyte differentiation. Stabilization of the WNT effector β-catenin induces mesodermal genes such as TBXT (Brachyury) and triggers an epithelial-mesenchymal transition (EMT). Although mechanical forces are essential for embryonic development, the role of actomyosin contractility during human mesoderm specification remains unclear. We show that increasing contractility through constitutively active Rho kinase or myosin light-chain kinase unexpectedly blocks β-catenin-dependent mesoderm induction and prevents EMT. In contrast, pharmacological or genetic suppression of contractility enhances Brachyury expression and advances EMT onset by 24 h. While β-catenin signaling alone promotes colony-level contractility, we find that contractility must be reduced prior to WNT activation to promote mesoderm specification, indicating a sensitization effect at the pluripotent state. Mechanistically, reduced tension decreases junctional β-catenin and increases nuclear active β-catenin, identifying actomyosin contractility as a key regulator of lineage commitment following WNT pathway activation.

The conserved Ypt/Rab GTPases regulate all steps of the intracellular transport pathways. In yeast and human cells, Ypt1/Rab1 regulate early steps of secretion and autophagy, whereas Ypt31/Rab11 regulate a late step in secretion, and the TRAPP complexes act as their activators. The Ypt and transport step specificity of TRAPP complexes is currently controversial. Here, we use in vivo analyses of mutations in subunits of yeast TRAPP complexes to determine these specificities. First, deletion of the TRAPPIII-specific subunit Trs85 does not affect transport through the Golgi nor the localization of TRAPPI or Ypt1 to the Golgi. Second, conditional depletion of essential subunits of TRAPPI and TRAPPII shows that they are required for early and late secretion steps, respectively. Thus, TRAPPI and TRAPPII activate Ypt1 and Ypt31 in early- and late-Golgi, respectively, ascribing TRAPPIII in Ypt1 activation only in autophagy. This distinct activator assignment provides a mechanism for the dual function of Ypt1/Rab1 in secretion and autophagy, where Rab1 has been implicated in disease.

Distinct cytotoxicity of water disinfection byproducts in human uroepithelium and Chinese hamster ovary cells.

Facilitating siRNA delivery into mammalian cells via the LDL receptor.

Exposure of human corneal epithelial cells to microplastic particles induces a phase-specific cytokine response.

Insights into the cytotoxicity of Ganoderma adspersum mycelium extracts against human renal cancer cell lines

Arsenic exposure induces stemness in human normal breast epithelial cells via the E2F2/FZD10 axis.

Anaplastic thyroid cancer (ATC) is a highly malignant tumor with poor prognosis and limited therapeutic options, creating an urgent need for novel treatments. This study aimed to investigate the inhibitory effect of Vemurafenib (Ve) combined with Panobinostat (Pa) on human ATC cells (FRO and ARO) and its underlying mechanism. Four groups were established: Control, Ve, Pa, and Ve+Pa. Cell proliferation and drug synergy were analyzed using CCK-8 assay, colony formation assay, and CompuSyn software. Cell migration, invasion, apoptosis, and glucose consumption were detected by Transwell assay, wound healing assay, apoptosis assay, and glucose consumption assay, respectively. Molecular markers were examined via RT-qPCR, Western blotting, and immunofluorescence. CompuSyn analysis verified the synergistic effect of Ve+Pa in FRO and ARO cells. Compared with the other three groups, the Ve+Pa group showed significantly suppressed cell proliferation, migration, invasion, and glucose consumption, as well as enhanced apoptosis. Moreover, the mRNA and protein expression of the sodium iodide symporter (NIS) and iodine metabolism-related molecules was upregulated, whereas glucose transporter 1 (GLUT1) expression was downregulated. Ve combined with Pa exerts a synergistic inhibitory effect on the growth and metastasis of FRO and ARO cells, while promoting apoptosis and cellular redifferentiation. This combination may provide a potential therapeutic strategy for ATC.

Free Hemoglobin Increases Endothelial Adhesiveness to Red Blood Cells via a Toll-like Receptor 4-Dependent Mechanism.

Statins are widely prescribed to those with atherosclerosis to lower cholesterol and reduce cardiovascular risk, but their influence on plaque calcification remains unclear. While clinical data associate statin use with increased calcium scores and reduced cardiovascular events, their effect on rupture-prone microcalcifications has not been clinically assessed due to imaging limitations. In vitro studies report conflicting outcomes on statin-induced calcification in various cell types. To address this gap, we investigated the impact of atorvastatin on calcification across multiple plaque-relevant cell types using both monolayer cultures and a tissue-engineered 3D plaque model. Human mesenchymal stromal cells (MSCs) were isolated from iliac crest bone chips and differentiated into smooth muscle (mSMCs) cells using TGF-β1, while human vena saphena cells (HVSCs) were obtained from bypass surgery material. Monolayer cultures of MSCs, mSMCs, and HVSCs were exposed to calcifying medium with or without atorvastatin (0.1-1 μM) for 2 weeks. For the 3D model, cells were embedded in fibrin gels, cultured to form a collagenous matrix for 2 weeks, then calcified for an additional 2 weeks with or without atorvastatin (1-10 μM). In both monolayer and the 3D model, atorvastatin inhibited calcification in MSCs, while it induced calcification in mSMCs. For HVSCs, atorvastatin reduced calcification in 2D monolayer cultures, while it had no visible effect in 3D. This study highlights the cell type-specific effects of atorvastatin on calcification, underscoring the need to consider cellular heterogeneity when evaluating its impact on plaque stability.

Human stem cells from apical papilla (hSCAPs) are a promising ectomesenchyme‑ derived cell source with neuronal differentiation potential. Under 3D-neurospheres induction, hSCAPs can be induced into neural stem cells (NSCs) and further committed toward neuronal lineages. This study aimed to demonstrate the differentiation of NSCs‑hSCAPs into neuronal‑like cells undergoing maturation through 3D-neurospheres formation and subsequent neurogenic induction. The characterized hSCAPs were induced into NSCs via 3D-neurospheres formation. The intraspheroidal cells were verified for early neural stemness properties and subsequently dissociated and cultured under neurogenic induction conditions for 7 days. Neuronal differentiation was evaluated by identification of Nissl substance, immunofluorescent analysis of neuronal‑associated proteins, quantitative mRNA expression, and depolarization‑evoked intracellular Ca2+ imaging. Following 3D neurosphere induction, hSCAPs formed clusters of intraspheroidal cells exhibiting typical NSCs characteristics, including high expression of Nestin and SOX2 and self‑reaggregation ability. After 7 days of neurogenic induction, the differentiated cells displayed distinct neuronal‑like morphologies, reduced expression of early neuronal markers (Nestin/NES and SOX2/SOX2), and increased expression of early neuronal differentiation-associated markers (Beta‑III tubulin/TUBB3) at both protein and mRNA levels. The synaptic vesicle‑associated gene (SV2A) was highly detected at the mRNA level. Furthermore, depolarization‑evoked Ca2+ responses after KCl stimulation were observed, indicating membrane excitability and voltage‑gated calcium channel in the differentiated cells. NSCs‑hSCAPs possess the capacity to generate neuronal‑like cells undergoing maturation via 3D-neurospheres formation and neurogenic induction.

Human adipose stem cell-derived extracellular vesicles (hASC-EVs) have gained attention as potential cell-free therapeutics in regenerative medicine due to their immunomodulatory properties and low immunogenicity. Despite this promise, their immunotoxicity profile remains insufficiently characterized, particularly across species and genetic backgrounds. This study systematically assessed immune responses to repeated high-dose intravenous administration of hASC-EVs in two murine strains-C57BL/6 (inbred) and ICR (outbred)-and in human peripheral blood mononuclear cells (hPBMCs) in vitro. Flow cytometry of murine blood and spleen samples revealed transient, strain-dependent shifts in immune cell populations, including neutrophils, monocytes, macrophages, B cells, and NK cells. Notably, C57BL/6 mice exhibited more pronounced fluctuations than ICR mice, reflecting the role of host genetics in EV-induced immunomodulation. In contrast, hPBMCs exposed to equivalent concentrations of hASC-EVs displayed no significant changes in cell viability, immune cell subset composition, or activation markers over a 24-hour period. While a mild, transient increase in CD86+ monocytes was observed at 6h, this effect normalized by 12h. These results suggest that hASC-EVs induce minimal and reversible immune responses in vivo and are immunologically inert in human immune cells under the tested conditions. The strain- and species-specific differences observed emphasize the limitations of rodent-only models for predicting human immunotoxicity and support the incorporation of human immune cell assays into preclinical safety assessments of EV-based therapeutics.

Genotoxicity assessment of nylon and polyethylene microparticles in Caco‑2 cells.

Neohesperidin promotes osteogenic differentiation of human periodontal ligament stem cells under inflammatory stress.