Search Results (114)

Background/Objectives: Brain pericytes are contractile mural cells of the neurovascular unit whose responses to endothelin-1 (ET-1) are mediated primarily by endothelin type A (ETA) receptors. ET-1/ETA signaling has been implicated in pathological pericyte contraction in neurovascular disorders such as Alzheimer’s disease and in cerebrovascular dysfunction after subarachnoid hemorrhage, motivating pharmacological evaluation of selective ETA receptor antagonists at the pericyte level. Clazosentan is a selective ETA receptor antagonist used clinically for cerebral vasospasm; however, its pharmacodynamic effects on brain pericytes remain insufficiently characterized. Methods: Pericyte impedance-based contractile and recovery responses to ET-1 were evaluated by xCELLigence real-time cell index analysis. Pericyte viability, morphology, and ETA protein abundance were examined using Cell Counting Kit-8 assay, immunocytochemistry, and Western blotting. As a secondary barrier-related assessment, transendothelial electrical resistance (TEER) was measured in primary rat brain endothelial cell-based in vitro blood–brain barrier models. Results: ET-1 induced impedance-based contractile responses with a concentration-related trend, followed by recovery responses, and increased ETA protein abundance in a time- and concentration-related manner; clazosentan significantly attenuated ET-1-induced ETA upregulation. Repeated ET-1 exposure was associated with an enhanced subsequent ET-1-induced impedance-based contractile response and a more sustained response, suggesting contractile sensitization; both effects were significantly attenuated by clazosentan. Clazosentan did not overtly disrupt TEER-assessed barrier properties. Conclusions: These findings suggest that ET-1/ETA signaling may shift brain pericytes toward a sensitized contractile response state and that selective ETA blockade by clazosentan attenuates this process without overtly disrupting TEER-assessed barrier properties under the present in vitro conditions. These observations support further pharmacological characterization of clazosentan as a modulator of pericyte ET-1 responses. Full article

Salvia officinalis L. (S. officinalis) or common sage has long been valued in traditional medicine for its broad therapeutic properties. Our study explored the antioxidant potential, the cytotoxic activity and the effects on smooth muscle (SM) contractility of a traditionally prepared sage aqueous infusion. A chromatographic analysis revealed the major phenolic constituents of the infusion, rosmarinic acid being the most abundant compound (38.16 mg/mL), followed by rutin (3.58 mg/g), caffeic acid (3.00 mg/g) and hesperidin (1.84 mg/g). Three spectrophotometric methods were used to determine the antioxidant potential of the plant extract: a DPPH assay for radical scavenging activity (IC₅₀ 21.77 ± 0.98 µg/mL), a CUPRAC assay (2.32 ± 0.04 mmol Trolox equivalents/g) and a total reducing capacity assay (1.92 ± 0.01 mmol Trolox equivalents/g) for evaluating reducing properties. A cytotoxicity was assessed in primary SM cells (isolated from rat gastric tissue) using the xCELLigence real-time cell analysis system, where the infusion demonstrated dose-dependent cytotoxicity with an IC₅₀ of 0.34 ± 0.12 mg/mL. Concentrations of 0.37 mg/mL rapidly reduced viability, while lower concentrations showed mild or negligible effects. Functional experiments with isolated rat gastric SM cells revealed a dose-dependent enhancement of both tonic and phasic contractile activity. Our results showed that the serotonergic pathways were mainly implicated in mediating these effects, while the cholinergic pathways were not involved with them. Together, these findings validate the traditional use of S. officinalis and point to its role in modulating SM function and providing antioxidant benefits, warranting further investigation into its mechanisms and therapeutic potential. Full article

Silver (Ag) and gold (Au) nanoparticles (NPs) are widely used in biomedicine, electronics, and catalysis, but their potential toxicity raises occupational health concerns. This study assessed the cytotoxicity and cellular interactions of Ag and Au NPs in human bronchial epithelial cells (BEAS-2B) using a standardized OECD three-tiered approach, alongside characterization of lung-deposited surface area (LDSA) concentrations during NP synthesis, which remained within ranges typically reported in occupational environments. Transmission electron microscopy revealed that AgNPs formed irregular clusters (~8.7 nm primary size, >30 nm aggregates), whereas AuNPs remained spherical (~13.4 nm). Real-time cytotoxicity analysis (xCELLigence) showed acute toxicity of AgNPs at 5 μg/cm², while AuNPs exhibited no cytotoxic effects. Dark-field and 3D hyperspectral imaging demonstrated that some AgNPs were internalized by BEAS-2B cells, whereas AuNPs remained mostly on the cell surface, indicating that uptake alone does not determine cytotoxicity. The greater dissolution potential of AgNPs and possible release of Ag⁺ ions may contribute to the enhanced cytotoxic effects observed in comparison to AuNPs, as suggested in previous studies. Although oxidative stress, mitochondrial dysfunction, and related cellular mechanisms were not directly assessed in the present study, the findings demonstrate differential cellular responses following nanoparticle exposure under realistic occupational exposure conditions. These results contribute to understanding nanoparticle–cell interactions and support the need for further mechanistic investigations to inform safer nanomaterial use. Full article

Background and Objectives: Dysfunction of the airway epithelial barrier is increasingly recognized as an early pathogenic mechanism in allergic respiratory diseases. Although individual aeroallergens such as ragweed (RW) pollen and house dust mite (HDM) are known to impair epithelial integrity, the effects of combined exposure, more reflective of real-world conditions, remain insufficiently characterized. This study aimed to evaluate the impact of single versus combined allergen exposure on airway epithelial barrier function using a multimodal experimental approach. Materials and Methods: Differentiated normal human bronchial epithelial (NHBE) cells were exposed to RW (100 µg/mL), HDM (100 µg/mL), or a combined extract (RW + HDM; total 100 µg/mL). Barrier function under air–liquid interface conditions was assessed by transepithelial electrical resistance (TEER), while real-time cellular responses were evaluated using xCELLigence impedance monitoring. Structural alterations were examined by occludin-based immunofluorescence imaging, and transcriptional changes associated with epithelial stress and inflammation were analyzed by RT-qPCR. Results: Allergen exposure induced time- and concentration-dependent impairment of epithelial barrier function. Combined exposure resulted in the most pronounced and sustained reduction in TEER and impedance measurements. These functional changes were accompanied by disruption of tight junction organization and coordinated transcriptional modulation of genes involved in inflammatory and stress responses. Conclusions: Combined exposure to RW and HDM extracts induced more severe and persistent epithelial barrier dysfunction than individual allergens. These findings support the role of the airway epithelium as a central regulator of allergic airway disease and highlight barrier disruption as an early pathogenic event. The multimodal framework applied in this study provides an integrated platform for investigating epithelial responses to complex environmental exposures. Full article

Endometriosis is characterized by enhanced cellular proliferation, migration, and resistance to apoptosis, contributing to lesion persistence and progression. Targeting cellular plasticity and mesenchymal-associated functions may therefore represent a promising therapeutic strategy. Here, we investigated the effects of Pelargonium graveolens essential oil (PGEO) on proliferation, apoptosis, migration, cytoskeletal organization, transcriptional regulation, and metabolic alterations in human endometriotic 12Z cells. PGEO treatment suppressed proliferative capacity in a concentration-dependent manner and significantly impaired cell migration, accompanied by reduced β-tubulin expression and decreased levels of mesenchymal-associated markers CD73 and CD105. Increased GRP78 expression together with ultrastructural alterations, including cytoplasmic vacuolization and mitochondrial and endoplasmic reticulum changes, indicated activation of cellular stress responses. Although transcriptional analysis revealed increased CCND1 and PIK3CA mRNA levels, these changes did not parallel the observed suppression of proliferation, suggesting compensatory regulatory responses. Untargeted metabolomic profiling revealed alterations in energy metabolism characterized by increased levels of glycolysis-related metabolites, reduced levels of several amino acids including glutamine and histidine, and changes in lipid-associated metabolites. Collectively, these findings demonstrate that PGEO suppresses proliferative and migratory behavior in endometriotic cells while modulating cytoskeletal, transcriptional, and metabolic pathways, highlighting its potential as a candidate for further investigation in endometriosis-targeted therapeutic strategies. Full article

Human breast cancer (HBC) is the most common and often lethal malignancy in women. Canine mammary tumors (CMTs) share significant molecular and clinical characteristics with HBC, which makes dogs a valuable spontaneous model for the study of HBC. HBC chemotherapy treatment relies mainly on carboplatin, which is effective but, in turn, highly toxic. Here we tested enrofloxacin, a Minichromosomal Maintenance Complex Component (MCM2) inhibitor, for its ability to increase tumor cell sensitivity to platinum-based drugs, thus suggesting a potential synergistic therapeutic strategy. CMT samples were used to establish primary cell cultures. Cells were treated with carboplatin, enrofloxacin, and their combination at different concentrations. Cytotoxic and antiproliferative effects were assessed using xCELLigence and MTT assays. Single-drug treatments exert limited effects on cell proliferation, while enrofloxacin significantly enhances carboplatin efficacy, leading to a complete growth arrest within 48 h. The MTT assay confirms a strong synergistic effect of the two drugs, whereas the Dose Reduction Index analysis indicates that carboplatin could be decreased without losing effectiveness. These findings suggest that combined therapy could represent a more effective and less toxic option for HBC and CMTs. This work also strengthens the possible use of the canine model for cancer studies within a One Health framework. Full article

Background: Obesity is a recognized risk factor for developing breast cancer (BC), but factors involved remain unclear. We investigated if breast adipose tissue from healthy women, BRCA1/2 mutation carriers and BC patients, can stimulate BC cell line migration and activation. Methods: adipose tissue conditioned medium (ATCM), was prepared from breast adipose tissue from healthy subjects (naïve; group 1 (n = 20)), BRCA1/2 mutation carriers (group 2 (n = 22)) and BC patients (group 3 (n = 38)). ATCM effect on migration of BC cell lines MCF-7, SK-BR-3 and MDA-MB-231 was measured with xCELLigence (ACEA Biosciences, San Diego, CA, USA) cell migration assay. Activation of migration was determined by measuring filopodia activation. Migration and filopodia activation were related to body mass index (BMI) and BC subtypes. Luminex multiplex assay was performed to examine the secretory profile of adipose tissue. Results: ATCM from group 1 induced migration and filopodia activation in MCF-7 and MDA-MB-231, but not in SK-BR-3. ATCM from group 2 induced filopodia activation but no migration. ATCM from group 3 induced less migration in MCF-7 than ATCM from group 1. Higher BMI was associated with increased ATCM-induced activation in MCF-7 (group 1) and MDA-MB-231 (group 2). ATCM from group 1 and 2 showed a metabolic secretory profile, whereas group 3 showed higher pro-angiogenic and inflammatory cytokines. Conclusions: This study shows that breast adipose tissue from healthy women, BRCA1/2 mutation carriers and BC patients, can stimulate BC cell line migration and activation. This effect is related to BC subtype and BMI. These data improve insight in adipose tissue as factor in BC development. Full article

Background/Objectives: Toothpaste ingredients such as strontium chloride (SrCl₂) and potassium carbonate (K₂CO₃) are recognized for their desensitizing and remineralizing effects but may be absorbed through the oral mucosa. Their potential cytotoxic and cardiotoxic properties, however, remain inadequately characterized. Here, we investigated the effects of SrCl₂ and K₂CO₃ on mouse-induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes (iPSC-CMs). Methods: Cells were exposed to varying concentrations of each compound for up to 72 h. Real-time cell analysis (xCELLigence RTCA Cardio system) was used to assess proliferation, and flow cytometry was used to evaluate cell viability. Functional properties of iPSC-CMs were examined using multi-electrode array (MEA) recordings and xCELLigence-based impedance measurements. Cardiac marker expression was examined via immunofluorescence and quantitative RT-PCR. Results: Both SrCl₂ and K₂CO₃ affected iPSC proliferation and reduced viability in a dose- and time-dependent manner, accompanied by altered embryoid body (EB) morphology and increased cell death. In iPSC-CMs, both compounds downregulated key cardiac genes and disrupted spontaneous beating activity, with effects intensifying at higher concentrations. Conclusions: These results demonstrate that SrCl₂ and K₂CO₃ induced dose-dependent cytotoxic and arrhythmogenic effects on iPSCs and iPSC-CMs. At elevated concentrations, these compounds impair iPSC-CM function and may pose safety concerns upon chronic exposure. Further mechanistic and long-term in vivo studies are warranted to assess their potential cardiotoxic risk in consumer oral care products. Full article

Hyaluronic acid (HA) is a natural biopolymer widely used in wound dressings for its supportive role in the healing process. In this study, we investigated the occurrence of microorganisms in chronic wounds and evaluated the antimicrobial activity of newly synthesized HA–Starch-based materials enriched with acidifying agents. Microbial isolates obtained from chronic wounds were tested for susceptibility using the agar diffusion method. The prepared materials exhibited significant antimicrobial activity against both reference strains and multidrug-resistant clinical isolates. Further characterization by scanning electron microscopy and elemental analysis confirmed uniform microfiber morphology and the expected elemental composition of the fibers. Cytotoxicity assessments performed using the xCELLigence system demonstrated the potential safety of developed materials. Overall, the results indicate that HA–Starch-based materials containing acidifying compounds exhibit strong in vitro antimicrobial activity against chronic-wound isolates, supporting their potential for further evaluation in wound care applications. Full article

Background: Chemotherapy remains one of the main approaches for treating malignant tumors, but repeated exposure to cytostatics leads to multidrug resistance (MDR), increasing tumor aggressiveness and reducing therapeutic efficacy. Identifying adjuvant agents that restore tumor sensitivity to drugs while minimizing toxicity is a cornerstone challenge today. This study aimed to investigate the potential of mesyl phosphoramidate antisense oligonucleotides (µ-ASOs) targeting miR-17, miR-21, and miR-155 as agents for enhancing the efficacy of cisplatin (Cis) and doxorubicin (Dox) in MDR-positive human epidermoid carcinoma KB-8-5 cells. Methods: Optimal regimens for the simultaneous application of µ-ASOs and Dox or Cis in KB-8-5 cells, including a concentration-dependent analysis and the type of compound interaction in combinations (synergy/additivity/antagonism), were studied using the MTT assay. Antiproliferative effects of the combinations were assessed using the real-time cell monitoring xCELLigence system. The potential molecular mechanism underlying KB-8-5 cell sensitization to cytostatics was investigated using RT-PCR and Western blot hybridization, supported by bioinformatic reconstruction of the gene network. Results: The most effective combinations including µ-ASOs targeting miR-21 and miR-17 together with Cis or Dox demonstrated additive to moderately synergistic effects on KB-8-5 cell viability (HSA synergy score = 4.8–8.7). The co-application of µ-ASOs allowed a 5- to 20-fold reduction in the dose of cytostatics, while maintaining a strong antiproliferative effect of 70–95%. Sensitization of KB-8-5 cells to Cis or Dox following µ-ASO treatment was mediated by a 1.5- to 3-fold decrease in the levels of the well-known MDR marker ABCB1 as well as the newly identified MDR-associated targets ZYX, TUBA4A, and SEH1L. Conclusions: miRNA-targeted mesyl phosphoramidate oligonucleotides are effective tools for overcoming resistance to the clinically approved chemotherapeutics cisplatin and doxorubicin. The relationship between miR-21, miR-17, and miR-155 and the novel MDR markers such as SEH1L, TUBA4A, and ZYX was revealed, thereby expanding the current understanding of the molecular mechanisms underlying tumor cell resistance to chemotherapy. Full article

The adrenocorticotropic hormone (ACTH) is a key regulator of adrenal cortex function, promoting glucocorticoid synthesis and modulating cell proliferation. However, the role of extracellular steroid availability in shaping ACTH responses is still not fully defined. In this study, the functional and transcriptomic effects of ACTH were investigated in primary rat adrenocortical cells cultured under standard conditions and under simulating serum starvation (charcoal-stripped serum). The cells were treated with ACTH (10 nM), and proliferation was monitored using xCELLigence RTCA, while corticosterone secretion was assessed via ELISA. The RNA extracted from these samples was then utilised for the purpose of microarray-based gene expression profiling. The present study revealed that charcoal-stripped serum markedly improved ACTH-induced corticosterone output, suggesting that the absence of endogenous steroids sensitises cells to ACTH stimulation possibly by removing negative feedback constraints. This enhanced steroidogenic response was accompanied by a significant suppression of proliferation, confirming that the stimulation of specialised functions (such as steroid secretion) reduces proliferative capacity of adrenocortical cells. Transcriptomic data revealed that the steroids stimulating effect on corticosterone output was mainly mediated via steroid biosynthetic and lipid metabolic processes while inhibitory effect on proliferation rate was mediated mainly by cell adhesion molecules. These results suggest that, in primary culture of rat adrenocortical cells, the stimulatory effect of ACTH on their specialised function (corticosteroid secretion) simultaneously reduces their basal function, which is their proliferation process. Changes in this type are also observed in cells cultured in steroid-depleted conditions. Full article

Background: Medullary thyroid carcinoma is a rare neuroendocrine tumor with limited therapeutic options, as current kinase inhibitors are often associated with significant toxicity and drug resistance. This study aimed to explore novel treatment strategies by testing targeted agents alone and in combination. Methods: Human medullary thyroid carcinoma TT cells with RET mutations were treated with Sorafenib, Lapatinib, and Bevacizumab. Cell proliferation was monitored in real time using the xCELLigence system, and apoptosis was assessed by flow cytometry. Results: Sorafenib and Lapatinib each showed strong, dose-dependent cytotoxic effects, with Lapatinib demonstrating the greatest potency. Bevacizumab alone exhibited minimal cytotoxic activity, but when combined with Sorafenib or Lapatinib it significantly enhanced their effects, even at concentrations that were only partially effective individually. The Lapatinib–Bevacizumab combination produced the most potent inhibition of cell viability, comparable to high-dose monotherapy. Conclusions: These findings suggest that combining kinase inhibitors with Bevacizumab may enhance antitumor activity, allow the use of lower drug doses, and overcome resistance, representing a promising therapeutic strategy for medullary thyroid carcinoma that warrants further investigation in clinical settings. Full article

Background/Objectives: The Centaurea genus is characterized by many species, a broad biological diversity, and a rich secondary metabolite content. These species exhibit various biological activities, including antioxidant, anti-inflammatory, antimicrobial, antiproliferative, and wound-healing properties. However, there are limited anticancer research studies available on the species. This study aims to investigate the potential cytotoxic effects of dichloromethane (CRD) and methanol (CRM) extracts obtained from the root of the endemic Centaurea lycaonica to clarify the mechanism of apoptosis by the intrinsic pathway on the human endometrial cancer cell line RL95-2 based on phytochemical analysis. Methods: The cytotoxicity studies were performed using a Real-Time Cell Analyzer (xCELLigence) and the MTT assay. The activities of caspase 3, caspase 9, Bax, and Bcl-2 were evaluated to investigate the molecular mechanism of apoptosis. LC-HRMS determined the phytochemical content of extracts. Results: CRD and CRM had a concentration-dependent effect in increasing caspase 3 and 9 activities and Bax/Bcl-2 ratios compared to the control with low IC₅₀ values. Conclusions: Apoptosis induction was more pronounced with CRM, which was enriched in hesperidin; this association warrants targeted validation with purified standards. Full article

Primary human endothelial cells represent an essential tool to model endothelial dysfunction and to screen interventions for its treatment. Here, we developed a protocol for the synchronous isolation of primary human saphenous vein endothelial cells (HSaVEC), human internal thoracic artery endothelial cells (HITAEC), and human microvascular endothelial cells (HMVEC) from SV and ITA utilized as conduits during coronary artery bypass graft surgery and from subcutaneous adipose tissue excised while providing an access to the heart. Treatment by collagenase type IV and magnetic separation with anti-CD31-antibody-coated beads ensured relatively high efficiency of the isolation (≈60% for HSaVEC, ≈50% for HITAEC, and ≈20% for HMVEC) and high purity (≥99%) of isolated ECs within ≈2 weeks (HSaVEC), ≈2–3 weeks (HITAEC), and ≈3–4 weeks (HMVEC). A colorimetric assay of cell viability and proliferation, as well as real-time bioimpedance monitoring using the xCELLigence instrument, demonstrated high proliferative activity in HSaVEC, HITAEC, and HMVEC, whilst the in vitro tube formation assay indicated their angiogenic potential. The isolation of HSaVEC, HITAEC, and HMVEC from patients undergoing coronary artery bypass graft surgery is a promising option to investigate endothelial heterogeneity, to interrogate endothelial responses to various stresses, and to pinpoint the optimal approaches for restoring endothelial homeostasis, thereby reproducing them within the bedside-to-bench-to-bedside concept. Full article

Tamoxifen is a well-established selective estrogen receptor modulator (SERM) widely used in breast cancer treatment, yet its efficacy varies across tumor types. To enhance its antitumor potential, we previously synthesized and investigated novel ferrocene-linked (T5, T15) derivatives. This publication is a close continuation of this work, introducing a new indene-based (T6) derivative. Objectives: The main aim of this study was to further broaden our knowledge of the mechanism behind the increased antitumor effect of the ferrocene-linked drugs (T5 and T15) and compare it with a new, indene-based tamoxifen derivative, T6. The indene moiety was selected as a rigid, hydrophobic aromatic unit to probe pharmacological effects independent of ferrocene’s redox activity. Methods: The compounds were tested on MCF7, MDA-MB231 and PANC1 cells. Cell viability was assessed with the AlamarBlue assay and the xCELLigence SP system. Reactive oxygen species (ROS) production was measured with the ROS Glo assay. Flow cytometry and RT-qPCR experiments were conducted to assess apoptosis and ROS regulation as well. Results: The modified compounds demonstrated an increased cell-viability-decreasing effect in breast (MCF7, MDA-MB-231) and pancreatic (PANC1) cancer cell lines, influencing both estrogen-receptor-dependent and -independent pathways. T6 led to G2/M phase arrest in PANC1 cells. Beyond cell cycle disruption, these derivatives significantly elevated ROS levels, contributing to apoptosis. Conclusions: Our findings suggest that these structural modifications retain tamoxifen’s pharmacophore properties while expanding its mechanism of action, particularly through universal interactions independent of the ER status of tumor cells. The enhanced antitumor effects highlight the potential of these derivatives as promising candidates for improved cancer therapies. Full article