Search Results (94)

Background/Objectives: Retinoic acid has been shown to inhibit melanoma progression; however, its underlying mechanisms remain unclear. In this study, we investigated the role of the retinoic acid-inducible gene TIG3 in regulating melanoma cell growth, as well as elucidating its involvement in apoptosis. Methods: The expression of TIG3 in melanoma tissues was analyzed using a cDNA microarray. Cell viability and cell death were measured using the WST-1 and LDH assay kits, respectively. The gene expression changes that were induced by TIG3 were identified through RNA sequencing, while apoptosis-related pathways were examined using a human apoptosis protein array. The protein expression levels were further validated using Western blot analysis. Results: TIG3 expression was significantly downregulated in melanoma tissues. The overexpression of TIG3 in melanoma cells led to reduced cell viability and increased cell death. TIG3 suppressed the expression of several apoptosis-regulating proteins, including PON2, Fas, cIAP-1, Claspin, Clusterin, HTRA2, and Livin, while promoting the expression of cleaved Caspase-3. Supplementation with cIAP-1, HTRA2, or Livin partially reversed TIG3-induced Caspase-3 expression and cell death. Conclusions: Our findings suggest that TIG3 may contribute to the anti-melanoma effects of retinoic acid, with IAP family proteins playing a key role in the TIG3-mediated regulation of melanoma cell survival. Full article

Background: Glioblastoma (GBM) resists current therapies due to its rapid proliferation, diffuse invasion, and heterogeneous cell populations. We previously showed that a single cold atmospheric plasma discharge tube (DT) reduces GBM viability via broad-spectrum electromagnetic (EM) emissions. Here, we tested whether two DTs arranged in a helmet configuration could generate overlapping EM fields to amplify the anti-tumor effects without thermal injury. Methods: The physical outputs of the single- and dual-DT setups were characterized by infrared thermography, broadband EM field probes, and oscilloscope analysis. Human U87-MG cells were exposed under the single or dual configurations. The viability was quantified with WST-8 assays mapped across 96-well plates; the intracellular reactive oxygen species (ROS), membrane integrity, apoptosis, and mitochondrial potential were assessed by multiparametric flow cytometry. Our additivity models compared the predicted versus observed dual-DT cytotoxicity. Results: The dual-DT operation produced constructive EM interference, elevating electric and magnetic field amplitudes over a broader area than either tube alone, while temperatures remained <39 °C. The single-DT exposure lowered the cell viability by ~40%; the dual-DT treatment reduced the viability by ~60%, exceeding the additive predictions. The regions of greatest cytotoxicity co-localized with the zones of highest EM field overlap. The dual-DT exposure doubled the intracellular ROS compared with single-DT and Annexin V positivity, confirming oxidative stress-driven cell death. The out-of-phase operation of the discharge tubes enabled the localized control of the treatment regions, which can guide future treatment planning. Conclusions: Two synchronously operated plasma discharge tubes synergistically enhanced GBM cell killing through non-thermal mechanisms that coupled intensified overlapping EM fields with elevated oxidative stress. This positions modular multi-DT arrays as a potential non-invasive adjunct or alternative to existing electric-field-based therapies for glioblastoma. Full article

Innovation in oral implantology is constantly on the move, with a constant search for new biomaterials to overcome many of the limitations of the biomaterials used in current implantable medical devices. This study explores the biocompatibility of an innovative 5% calcium-to-zirconia (Ca-SZ) coating deposited by PVD on TA6V substrates for use in oral implantology. In order to determine the contribution of the Ca-SZ coating, an in vitro biocompatibility study was carried out to assess the potential influence of the Ca-SZ coating (1) on the viability and proliferation of saos-2 and HaCaT cells over a short-term exposure period of 96 h, (2) on the synthesis of pro-inflammatory cytokines, and (3) on the synthesis of osteogenic differentiation markers over a long-term exposure period of 21 days, in comparison with reference biomaterials. The sampling consisted of n = 3 biological replicates, and a p-value of <0.05 was used as the threshold for statistical significance. Viability and proliferation kinetics to WST-1 and CyQUANT NF, respectively, showed improved viability/proliferation of Ca-SZ exposed to both cell lines independently. The TNF-alpha and IL-6 assays revealed reduced levels of cytokines compared with the reference biomaterials, including the control groups. In parallel, in Saos-2 cells exposed to Ca-SZ for 21 days under osteogenic conditions increased expression of osteogenic markers, such as the synthesis of soluble collagens, alkaline phosphatase (ALP), osteopontin, and osteocalcin, reflecting dynamic and facilitated osteoblastic differentiation, which was supported by the formation of hydroxyapatite (HA) crystals observed by SEM micrograph and confirmed by EDS mapping. In conclusion, Ca-SZ demonstrates an overall better biocompatibility compared with reference biomaterials, linked to a bioactive interaction of calcium, promoting cell proliferation and differentiation for optimal osteointegration, underlining its potential as a relevant innovation for next-generation implants. Full article

The use of different nanoparticles (NPs) is increasing in a wide variety of everyday products. Nevertheless, most studies concerning NP risk assessment have evaluated exposure scenarios involving a single kind of NP. A stepwise study distinguishing between the effects resulting from exposure to one kind of NP and those resulting from different co-exposure scenarios to ${\mathrm{Al}}_2{\mathrm{O}}_3$ and ${\mathrm{CeO}}_2$ NPs at concentrations below acute toxicity was conducted with different analytical techniques. As a starting point, WST-1 viability assays were performed to assess whether the chosen exposure concentrations resulted in any acute loss of viability, which would hamper further insight into the cellular response to NP exposure. Then, data on NP dissolution and uptake were obtained via single-particle inductively coupled plasma–mass spectrometry (spICP-MS) and microwave-assisted ICP-MS. Additionally, time-of-flight secondary ion mass spectrometry (ToF-SIMS) was performed to check for differences in the biological response to the exposure scenarios at the single-cell level. It was found that the proposed combined techniques provide insight into changes in biological responses as well as cellular metal contents among the exposure scenarios. In this work, a comprehensive tiered analytical strategy for evaluating the biological responses to challenging exposure scenarios is provided. The results highlight the necessity of selecting situations more closely resembling real life—including concentrations below acute toxicity and potential interactions due to multiple NPs—when estimating potential health risks. These findings thus provide a foundation and an incentive for further research into the complex processes leading to the observed effects. Full article

Background/Objectives: The persistent threat of emerging respiratory RNA viruses like SARS-CoV-2 and Influenza A virus (IAV) necessitates the continuous development of effective, safe, broadly acting, and generally accessible antiviral agents. Current treatments often face limitations such as early administration requirements, resistance development, and limited global access. Natural products, like European black elderberry (Sambucus nigra L.; S. nigra) fruit extract and quinine, have been used historically against viral infections. In this study, we investigated the antiviral efficacy of a standardized black elderberry fruit extract containing 3.2% anthocyanins (EC 3.2) and, as a second natural antiviral product, quinine, against IAV and SARS-CoV-2 in vitro. Methods: Madin–Darby Canine Kidney II (MDCKII) cells were infected with IAV PR-8, while human Calu-3 lung epithelial cells were infected with Wuhan-type SARS-CoV-2. Cells were treated with varying concentrations of EC 3.2 and quinine either as mono- or combinational therapy. Viral replication was assessed using quantitative RT-PCR, and cell viability was evaluated using WST-1 assays. Results: Our results demonstrate, for the first time, that both EC 3.2 and quinine individually inhibited IAV replication in a dose-dependent manner, with IC₅₀ values of approximately 1:400 for EC 3.2 and 250 nM for quinine. Most importantly, the combinational treatment exhibited a strong synergistic antiviral effect, as confirmed by the Bliss independence model (synergy scores of 14.7 for IAV, and 27.8 for SARS-CoV-2), without affecting cell viability. Conclusions: These findings suggest that the combined use of black elderberry extract and quinine might serve as an effective antiviral strategy against IAV and SARS-CoV-2, particularly since the synergistic effect allows for lower doses of each product while retaining therapeutic efficacy. In summary, this combinational in vitro approach, when expanded to other respiratory RNA viruses and confirmed in clinical studies, has the potential to open a promising avenue for pandemic preparedness. Full article

Nineteen potential mimics of 8,9-epoxyeicosatrienoic acid (8,9-EET), a natural bioactive oxylipin, were synthesized and evaluated for their ability to protect renal mesangial cells against sorafenib-induced cell death in a water-soluble tetrazolium (WST-8) assay. All compounds were also evaluated as inhibitors of soluble epoxide hydrolase. As expected of a potent pan-kinase inhibitor the drug sorafenib caused a significant decrease in cell viability in HRMCs. Several analogs containing amide and oxamide groups in place of the epoxide showed efficacy in reducing sorafenib induced human renal mesangial cell (HRMC) death. Oxamide containing analogs proved particularly effective, with the most promising analog increasing cell viability five-fold over control at 1 µM. These analogs, containing an oxamide group as a bioisostere for the epoxide in 8,9-EET, did not display significant inhibitory activity towards soluble epoxide hydrolase. This preliminary structure–activity relationship analysis reveals the oxamide group as a promising bioisostere for the epoxide in the 8,9-position of the fatty acid chain, producing protective effects against sorafenib-induced cell death in HRMCs. Collectively, these findings demonstrate the potential for using epoxide mimics and particularly oxamides as 8,9-EET analogs as bioisosteres of the corresponding epoxide in a therapeutic strategy against sorafenib-induced glomerular nephrotoxicity. Full article

(1) Autologous chondrocyte implantation (ACI) is a prominent method for treating cartilage damage, but periosteal patches can cause chondrocyte leakage. This study evaluates the potential of a decellularized membrane derived from the cell-produced extracellular matrix of 1-day-old porcine cartilage (pcECM-DM) to act as a substitute for periosteal patches. (2) The interaction between young rabbit chondrocyte cells and pcECM-DM was assessed through cytotoxicity, differentiation, cell viability, cell migration, and adhesive ability. Rabbit chondrocyte cells, cultivated until passage two, were seeded onto a 6 mm diameter membrane. Assessments included DAPI-PKH26 staining, histological staining, live/dead assay, WST-1 assay, and proteomics analysis. (3) Results: DAPI-PKH26 staining showed successful adhesion and the uniform distribution of cells on the membrane. Safranin-O and H&E staining confirmed that the membrane supports chondrocyte adhesion and extracellular matrix production with high cell density and typical chondrocyte morphology. The live/dead assay demonstrated a high proportion of viable cells at 24 and 48 h, with increased cell proliferation over time. The WST-1 assay showed a significant increase in OD450 values, confirming cell proliferation and biocompatibility. Proteomic analysis revealed the significant enrichment of genes associated with extracellular matrix organization, cell adhesion, and cartilage development. (4) Conclusions: This novel biomaterial holds the potential to enhance cartilage regeneration and offer a viable alternative to periosteal patches. Full article

Cancer remains one of the most formidable diseases globally and continues to be a leading cause of mortality. While chemotherapeutic agents are crucial in cancer treatment, they often come with severe side effects. Furthermore, the development of acquired drug resistance poses a significant challenge in the ongoing battle against cancer. Combining these chemotherapeutic agents with plant-derived phenolic compounds offers a promising approach, potentially reducing side effects and counteracting drug resistance. Phytochemicals, the bioactive compounds found in plants, exhibit a range of health-promoting properties, including anticarcinogenic, antimutagenic, antiproliferative, antioxidant, antimicrobial, neuroprotective, and cardioprotective effects. Their ability to enhance treatment, coupled with their non-toxic, multi-targeted nature and synergistic potential when used alongside conventional drugs, underscores the growing importance of natural therapeutics. In this study, we investigated the anticancer effects of olaparib (OL), a small-molecule PARP inhibitor that has shown promising results in both preclinical and clinical trials, and gallic acid (GA), a phenolic compound, in olaparib-resistant human osteosarcoma U2OS cells (U2OS-PIR). Both parental U2OS and U2OS-PIR cell lines were treated with increasing concentrations of olaparib and gallic acid, and their cytotoxic effects were assessed using the WST-1 cell viability assay. The synergistic potential of OL and GA, based on their determined IC₅₀ values, was further explored in combination treatment. A colony survival assay revealed the combination’s ability to significantly reduce the colony-forming capacity of cancer cells. Additionally, the apoptotic effects of OL and GA, both individually and in combination, were examined in U2OS-PIR cells using acridine orange/ethidium bromide dual staining. The anti-angiogenic properties were assessed through a VEGF ELISA, while the expression of proteins involved in DNA damage and apoptotic signaling pathways was analyzed via Western blot. The results of this study demonstrate that gallic acid effectively suppresses cell viability and colony formation, particularly when used in combination therapy to combat OL resistance. Additionally, GA inhibits angiogenesis and induces DNA damage and apoptosis by modulating key apoptosis-related proteins, including cPARP, Bcl-2, and Bax. These findings highlight gallic acid as a potential compound for enhancing therapeutic efficacy in overcoming acquired drug resistance. Full article

Background: Sodium butyrate (NaBu), a short-chain fatty acid, modulates global gene expression through histone deacetylase (HDAC) inhibition, suppressing proliferation and inducing apoptosis in various cancers. Rutin (RUT), a polyphenolic flavonoid found in many plants, exhibits notable anticancer properties. Combining chemotherapeutic agents with natural polyphenols represents a promising strategy for cancer therapy. This study aims to evaluate, for the first time, the potential effects of NaBu and RUT combination therapy on metastatic castration-resistant prostate cancer (mCRPC) cells. Methods: PC-3 cells were treated with varying concentrations of NaBu, RUT, and their combinations. Cell viability was assessed using the WST-1 assay. Based on combination index values, selected treatments were further analyzed for apoptosis (Annexin V assay), intracellular reactive oxygen species (ROS) production, mRNA expression levels, and changes in cell and nuclear morphology. Results: The combined treatment of NaBu and RUT significantly reduced cell viability compared to individual treatments. Enhanced apoptotic induction and elevated ROS levels were observed in combination-treated cells, alongside notable changes in cellular and nuclear morphology and mRNA expression levels. Conclusions: NaBu and RUT combination therapy exhibits a synergistic anticancer effect in mCRPC cells by inhibiting cell viability, inducing apoptosis, and increasing ROS production. These findings suggest a promising therapeutic approach that warrants further investigation to elucidate the underlying molecular mechanisms and assess its potential in preclinical and clinical settings. Full article

Background: This study aims to evaluate the antibacterial effect of Lespedeza cuneata extract on Porphyromonas gingivalis (P. gingivalis), a principal bacterium in periodontal disease, and its impact on human gingival fibroblasts (HGFs). Methods: Dried Lespedeza cuneata was extracted using 70% ethanol, concentrated, and freeze-dried to obtain the Lespedeza cuneata extract in powder form. The antibacterial effect, indicated by the inhibition of P. gingivalis growth, was assessed by administering concentrations of 1, 3, 5, 10, 20, 30, and 40 mg/mL. After 24 h of anaerobic incubation, colony-forming units per milliliter (CFU/mL) were measured. Cytotoxicity on HGF cells was evaluated after treatment with WST-1 solution followed by incubation at 37 °C, 5% CO₂ for 2 h. Cell morphology and proliferation were assessed using the Sulforhodamine B (SRB) assay. Results: The antibacterial effect of Lespedeza cuneata extract was concentration-dependent, with 99.98% inhibition observed at 5 mg/mL, 99.99% at 10 mg/mL, and no detectable CFUs were observed at 40 mg/mL under the tested conditions. Evaluating the change in growth rate of HGF cells showed a decrease in cell viability as the concentration increased, and the application of Lespedeza cuneata extract at 10 mg/mL was found to be a safe and effective concentration with a half-maximal inhibitory concentration (IC50). Conclusion: Based on the antibacterial effect, cytotoxicity, and safety profile of Lespedeza cuneata extract, it holds potential as a natural extract material for the prevention, improvement, or treatment of periodontal disease. Additionally, validation of the practical approach will be necessary via a clinical applicability evaluation. Full article

Laser-induced photothermal therapy using gold nanoparticles (AuNPs) has emerged as a promising approach to cancer therapy. However, optimizing various laser parameters is critical for enhancing the photothermal conversion efficacy of plasmonic nanomaterials. In this regard, the present study investigates the photothermal effects of dodecanethiol-stabilized hydrophobic ultrasmall spherical AuNPs (TEM size 2.2 ± 1.1 nm), induced by a 343 nm wavelength ultrafast femtosecond-pulse laser with a low intensity (0.1 W/cm²) for 5 and 10 min, on the cell morphology and viability of human hepatocellular carcinoma (HepG2) cells treated in vitro. The optical microscopy images showed considerable alteration in the overall morphology of the cells treated with AuNPs and irradiated with laser light. Infrared thermometer measurements showed that the temperature of the cell medium treated with AuNPs and exposed to the laser increased steadily from 22 °C to 46 °C and 48.5 °C after 5 and 10 min, respectively. The WST-1 assay results showed a significant reduction in cell viability, demonstrating a synergistic therapeutic effect of the femtosecond laser and AuNPs on HepG2 cells. The obtained results pave the way to design a less expensive, effective, and minimally invasive photothermal approach to treat cancers with reduced side effects. Full article

Graphene oxide-mediated photothermal therapy using femtosecond lasers has recently shown promise in treating hepatocellular carcinoma. However, significant work remains to optimize irradiation parameters for specific nanoparticle types and cancer cells to improve nanomaterial-mediated photothermal anticancer therapy. This study investigated the photothermal potential of nGO and nGO-PEG nanoparticles (NPs) combined with femtosecond laser irradiation at 515 nm and 1030 nm wavelengths, with varying power (0.1 and 0.2 W/cm²) and duration (5 and 10 min), to optimize photothermal therapy for hepatocellular carcinoma. Conversion efficiency of NPs, morphology and viability of HepG2 and normal MDCK cells after treatments were evaluated using an electronic thermometer, phase-contrast microscopy, and WST-1 assay. The results revealed that nGO-PEG NPs exhibited better photothermal efficiency than nGO, with 515 nm of irradiation inducing a temperature increase up to 19.1 °C compared to 4.7 °C with 1030 nm of light. Laser exposure to 515 nm significantly reduced HepG2 cell viability, with the most intense conditions (10 min at 0.2 W/cm²) causing a decrease of up to 58.2% with nGO and 43.51% with nGO-PEG. Normal MDCK cells showed minimal impact or a slight viability increase, especially with nGO-PEG. Combined treatment with laser irradiation and NPs induced significant morphological changes in HepG2 cells, including cell detachment and apoptotic-like characteristics, particularly with 1030 nm of irradiation. MDCK cells exhibited minimal morphological changes, with some recovery observed under lower energy conditions. These findings suggest that low-energy lasers and engineered nanomaterials could provide a minimally invasive approach to photothermal cancer therapy with reduced side effects. Full article

Background: Essential oils exhibit several biological activities such as antimicrobial, antioxidant, proliferative, and anti-inflammatory. This study was aimed at investigating the antimicrobial effects and cytotoxic activities of niaouli, palmarosa, and clove essential oils. Methods: Content analyses of these essential oils were carried out by gas chromatography–mass spectrometry. The antibacterial activity was screened against methicillin-resistant S. aureus ATCC 43300, P. aeruginosa ATCC 27853, P. aeruginosa PAO1, S. aureus ATCC 25923, and 44 isolates (22 P. aeruginosa isolates, 4 S. aureus isolates, and 18 Staphylococcus spp. isolates) obtained from dogs with previous wound infections who were included in the current study. The antimicrobial effects of essential oils were investigated using disk diffusion and minimum inhibition/bactericidal concentration methods. Additionally, the antibiofilm, protease, elastase, and gelatinase activities of the essential oils were evaluated. Different concentrations of each essential oil ranging from 10 to 1000 µg/mL were also analyzed in terms of cell viability by WST-8 assay in primary canine fibroblast cells. Results: The fibroblast cell viabilities of palmarosa, niaouli, and clove oils at a 1000 µg/mL concentration were 75.4%, 96.39%, and 75.34%, respectively. All the EOs were found to have bactericidal effects with MBCs/MICs of 0.015 to 0.5 µL/mL against P. aeruginosa, Staphylococcus isolates (p < 0.001). Palmarosa was found to have the largest inhibition zone diameter (20.5 ± 6.6, 16.4 ± 2.3) compared to other essential oils in the disk diffusion test against Staphylococcus spp. and P. aeruginosa (p < 0.001). But none of the EOs reduced protease, elastase, and gelatinase activities, which are some of the virulence properties of the tested bacteria. Conclusions: These results showed that palmarosa, niaouli, and clove essential oils act as potential antibacterial agents for dogs against P. aeruginosa, methicillin-resistant S. aureus, and Staphylococcus spp., without damaging the skin. Full article

Background: Electromagnetic field therapy is gaining attention for its potential in treating bone disorders, with Extracorporeal Magnetotransduction Therapy (EMTT) emerging as an innovative approach. EMTT offers a higher oscillation frequency and magnetic field strength compared to traditional Pulsed Electromagnetic Field (PEMF) therapy, showing promise in enhancing fracture healing and non-union recovery. However, the mechanisms underlying these effects remain unclear. Results: This study demonstrates that EMTT significantly enhances osteoblast bone formation at multiple levels, from gene expression to extracellular matrix mineralization. Key osteoblastogenesis regulators, including SP7 and RUNX2, and bone-related genes such as COL1A1, ALPL, and BGLAP, were upregulated, with expression levels surpassing those of the control group by over sevenfold (p < 0.001). Enhanced collagen synthesis and mineralization were confirmed by von Kossa and Alizarin Red staining, indicating increased calcium and phosphate deposition. Additionally, calcium imaging revealed heightened calcium influx, suggesting a cellular mechanism for EMTT’s osteogenic effects. Importantly, EMTT did not compromise cell viability, as confirmed by live/dead staining and WST-1 assays. Conclusion: This study is the first to show that EMTT can enhance all phases of osteoblastogenesis and improve the production of critical mineralization components, offering potential clinical applications in accelerating fracture healing, treating osteonecrosis, and enhancing implant osseointegration. Full article

In this study, we assessed the impact of hepatocyte growth factor (HGF) on corneal endothelial cells (CECs), finding that HGF concentrations of 100–250 ng/mL significantly increased CEC proliferation by 30%, migration by 32% and improved survival under oxidative stress by 28% compared to untreated controls (p < 0.05). The primary objective was to identify non-fibrotic pharmacological strategies to enhance corneal endothelial regeneration, addressing a critical need in conditions like Fuchs’ endothelial dystrophy (FED), where donor tissue is scarce. To confirm the endothelial nature of the cultured CECs, Na⁺/K⁺-ATPase immunohistochemistry was performed. Proliferation rates were determined through BrdU incorporation assays, while cell migration was assessed via scratch assays. Cell viability was evaluated under normal and oxidative stress conditions using WST-1 assays. To ensure that HGF treatment did not trigger epithelial-mesenchymal transition, which could lead to undesirable fibrotic changes, α-SMA staining was conducted. These comprehensive methodologies provided robust data on the effects of HGF, confirming its potential as a therapeutic agent for corneal endothelial repair without inducing harmful EMT, as indicated by the absence of α-SMA expression. These findings suggest that HGF holds therapeutic promise for enhancing corneal endothelial repair, warranting further investigation in in vivo models to confirm its clinical applicability. Full article