NP-treated Cells Research Articles

Mycofabrication of bimetal oxide nanoparticles (CuO/TiO2) using the endophytic fungus Trichoderma virens: Material properties and microbiocidal effects against bacterial pathogens

Bimetallic oxide nanoparticles (NPs) have gained the interest of researchers owing to the synergistic mechanism of action by the elements present. Mycosynthesis of nanostructures is a promising, facile, and an environmentally friendly approach. In the present study, CuO/TiO2 NPs were produced using the cell-free filtrate of the endophytic fungus Trichoderma virens. The reaction of the fungal filtrate with the two metal salts, CuSO4.5H2O and TiO2, resulted in the mycosynthesis of the CuO/TiO2 NPs. The prepared bimetal oxide NPs were characterized using XRD, FTIR spectroscopy, EDX, DLS, zeta potential analysis, XPS, Raman scattering, FESEM, and HRTEM. The mycosynthesized CuO/TiO2 NPs exhibited diffraction peaks characteristic of the monoclinic CuO and tetragonal rutile TiO2 crystal planes. Homogeneity and stability were observed as a result of the biological moieties present in the fungal filtrate, which capped the CuO/TiO2 NPs. The prepared CuO/TiO2 NPs were distributed as flake/plate-like and quasi-spherical/nanorod CuO and TiO2 NPs, respectively. A broad-spectrum antibacterial effect was recorded, with zones of inhibition as follows: foodborne pathogens Escherichia coli (14.34 mm), Salmonella enterica serovar Typhimurium (11.32 mm), and Staphylococcus aureus (9.63 mm); and phytopathogenic bacteria involving Clavibacter michiganensis subsp. michiganensis (Cmm, 12.16 mm), C. michiganensis subsp. capsici (Cmc, 11.26 mm), wild type and streptomycin-resistant Pectobacterium carotovorum subsp. carotovorum (Pcc, 11.06 and 12.37 mm, respectively), and wild type and streptomycin-resistant Xanthomonas citri pv citri (Xcc, 12.48 and 12.69 mm, respectively). The minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of the CuO/TiO2 NPs were determined using the broth microdilution assay. NP-treated bacterial cells at sub-MICs had rough surfaces with membrane defects and exhibited shrinkage, reduction of cell edges, cracks, and fractures on their surfaces. This study provides insights into the potential of the mycosynthesized CuO/TiO2 NPs as alternative nano-antimicrobial agents, which could be employed for future biomedical, agricultural, and pharmaceutical applications.

Brucine Entrapped Titanium Oxide Nanoparticle for Anticancer Treatment: An In Vitro Study.

Backgroundand Objective. The public's health has been seriously threatened by cervical cancer during recent times. In terms of newly diagnosed cases worldwide, it ranks as the ninth most prevalent malignancy. Multiple investigations have proven that nanoparticles can effectively combat cancer due to their small dimensions and extensive surface area. In the meantime, bioactive compounds which are biocompatible are being loaded onto nanoparticles to promote cancer therapy. The current study investigates the anticancerous potential of Brucine-entrapped titanium oxide nanoparticles (TiO2 NPs) in cervical cancer cell line (HeLa). Materials and Methods. The physiochemical, structural, and morphological aspects of Brucine-entrapped TiO2 NPs were evaluated by UV-visible spectrophotometer, Fourier transform-infrared spectroscopy (FT-IR), dynamic light scattering (DLS), scanning electron microscopy (SEM), and energy dispersive X-ray (EDAX). The cytotoxic effect against the HeLa cell line was assessed using a tetrazolium-based colorimetric assay (MTT), a trypan blue exclusion (TBE) assay, phase contrast microscopic analysis, and a fluorescence assay including ROS and DAPI staining. Furthermore, estimation of antioxidant markers includes catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD). Results. The UV spectrum at 266 nm revealed the formation of TiO NPs. The FT-IR peaks confirmed the effective entrapment of brucine with TiO2 NPs. The average size (100.0 nm) of Brucine-entrapped TiO2 NPs was revealed in DLS analysis. The micrograph of the SEM revealed the formation of ellipsoidal to tetragonal-shaped NPs. The Ti, O, and C signals were observed in EDAX. In MTT assay, Brucine-entrapped TiO2 NPs showed inhibition of cell proliferation in a dose-wise manner and IC50 was noticed at the concentration of 30 µg/mL. The percentage of viable cells gradually reduced in the trypan blue exclusion assay. The phase contrast microscopic analysis of Brucine-entrapped TiO2 NP-treated cells showed cell shrinkage, cell wall deterioration, and cell blebbing. The intracellular ROS level was increased in a dose-wise manner when compared to control cells in ROS staining. The condensed nuclei and apoptotic cells were increased in treated cells, as noted in DAPI staining. In treated cells, the antioxidant markers such as CAT, GSH, and SOD levels were substantially lower compared to the control cells. Conclusion. The synthesized Brucine entrapped TiO2 NPs exhibited remarkable anticancer activity against the HeLa cell line.

Green synthesis of strontium-doped tin dioxide (SrSnO2) nanoparticles using the Mahonia bealei leaf extract and evaluation of their anticancer and antimicrobial activities

Abstract In this study, a simple green method was employed to produce strontium (Sr)-doped-tin-dioxide (SnO2) nanoparticles (SrSnO2 NPs) using the Mahonia bealei leaf extract. The synthesized NPs were characterized with XRD, FE-SEM, FTIR, and PL spectroscopy measurements. SrSnO2 NPs were analysed for antimicrobial and anticancer activities. The XRD analysis revealed that the synthesized samples exhibited a tetragonal rutile crystal structure type of tin oxide. The EDX spectrum conforms to the chemical composition and elemental mapping of SrSnO2 NP synthesis. At 632 cm−1, the O–Sn–O band was observed and chemical bonding was confirmed using an FTIR spectrum. The PL spectrum identified surface defects and oxygen vacancies. The SrSnO2 NPs were tested against both Gram-positive and Gram-negative human pathogens. The synthesized nanoparticles exhibited effective antibacterial properties. The anticancer effects of SrSnO2 nanoparticles were also assessed against MCF-7 cells, and growth was decreased with increasing concentrations of the nanoparticles. Dual staining revealed high apoptosis in SrSnO2 NP-treated MCF-7 cells, proving its apoptotic potential. To conclude, we synthesized and characterized potential SrSnO2 nanoparticles using a green approach from the Mahonia bealei leaf extract. Further, green SrSnO2 nanoparticles showed significant antibacterial and anticancer properties against breast cancer cells (MCF-7) through apoptosis, which suggests a healthcare application for these nanoparticles.

Pluronic-F-127-Passivated SnO2 Nanoparticles Derived by Using Polygonum cuspidatum Root Extract: Synthesis, Characterization, and Anticancer Properties.

Nanotechnology has emerged as the most popular research topic with revolutionary applications across all scientific disciplines. Tin oxide (SnO2) has been gaining considerable attention lately owing to its intriguing features, which can be enhanced by its synthesis in the nanoscale range. The establishment of a cost-efficient and ecologically friendly procedure for its production is the result of growing concerns about human well-being. The novelty and significance of this study lie in the fact that the synthesized SnO2 nanoparticles have been tailored to have specific properties, such as size and morphology. These properties are crucial for their applications. Moreover, this study provides insights into the synthesis process of SnO2 nanoparticles, which can be useful for developing efficient and cost-effective methods for large-scale production. In the current study, green Pluronic-coated SnO2 nanoparticles (NPs) utilizing the root extracts of Polygonum cuspidatum have been formulated and characterized by several methods such as UV-visible, Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray (EDAX), transmission electron microscope (TEM), field emission-scanning electron microscope (FE-SEM), X-ray diffraction (XRD), photoluminescence (PL), and dynamic light scattering (DLS) studies. The crystallite size of SnO2 NPs was estimated to be 45 nm, and a tetragonal rutile-type crystalline structure was observed. FESEM analysis validated the NPs' spherical structure. The cytotoxic potential of the NPs against HepG2 cells was assessed using the in vitro MTT assay. The apoptotic efficiency of the NPs was evaluated using a dual-staining approach. The NPs revealed substantial cytotoxic effects against HepG2 cells but failed to exhibit cytotoxicity in different liver cell lines. Furthermore, dual staining and flow cytometry studies revealed higher apoptosis in NP-treated HepG2 cells. Nanoparticle treatment also inhibited the cell cycle at G0/G1 stage. It increased oxidative stress and promoted apoptosis by encouraging pro-apoptotic protein expression in HepG2 cells. NP treatment effectively blocked the PI3K/Akt/mTOR axis in HepG2 cells. Thus, green Pluronic-F-127-coated SnO2 NPs exhibits enormous efficiency to be utilized as an talented anticancer agent.

S9.4b Trimetallic Cu-Zn-Fe nanoparticles induced apoptosis and cell cycle arrest in multidrug-resistant Candida auris S9.4 Free oral presentations (late breaking), September 23, 2022, 4:45 PM - 6:15 PM

Background Candida species are opportunistic fungus that can cause serious infections, particularly in immunocompromised population. The number of fungal infections has increased steadily with Candida species being responsible for ˃ 70% of these instances, particularly in hospitalized patients with significant underlying conditions. Pharmacological resistance in Candida species and the advent of Candida auris have elevated candidiasis to a major public health concern. Candida auris is an emerging multidrug-resistant fungus that can cause catastrophic bloodstream infections and high fatality rates, particularly in hospitalized patients with major medical issues. Antifungal study of trimetallic nanoparticles (NPs) of various types have been studied as a therapy option for efficient and safe control of candidiasis. These NPs were highlighted for being environmentally friendly and sustainable synthetic preparative possibilities.ObjectiveThis work aimed to synthesize and characterize novel Cu-Zn-Fe trimetallic NPs and determine their in vitro antifungal activity and mechanism of antifungal action against Candida auris isolates.MethodsThe synthesis and characterization of Cu-Zn-Fe trimetallic NPs was done by standard methods. The antifungal capability of these NPs were determined by calculating minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) following CLSI recommended guidelines. Susceptibility on planktonic cells and biofilms was further confirmed by MuseTM cell count and viability assay and scanning electron microscopy (SEM) respectively. For insight antifungal mechanisms, apoptosis and cell cycle arrest were studied by exploring different apoptotic markers and MuseTM cell analyzer.ResultsCharacterizations by Fourier-transform infrared spectroscopy (FTIR), diffuse reflectance UV-visible spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) determine the successful biosynthesis of Cu-Zn-Fe trimetallic NPs. Susceptibility assay confirmed the fungicidal activity of Cu-Zn-Fe NPs with MIC and MFC values of 12.5 and 25 μg/ml respectively. These results were further confirmed by viability assay reporting the cell viability of 45.5%, 13.5%, and 1.8% when C. auris cells were treated with 1/2 MIC, MIC, and 2MIC respectively. Cell cycle analysis revealed that 91.2% of healthy developing untreated control cells were in G0/G1 phase, whereas 5.2% and 3.7% of cells were in the S phase and G2/M phase, respectively. In contrast, NP-treated cells were observed to be arrested in S phase with 49.3% cells at 2MIC. To study the physiology of cell death caused by NPs, we investigated mitochondrial membrane potential (∆ψm), with live cells having stable (∆ψm) whereas treated cells showed loss of (∆ψm). Another important parameter of apoptosis in yeast cells is the release of cytochrome C from the mitochondria to the cytosol and NP-treated cells resulting in decreased mitochondrial cytochrome C and elevated cytosolic cytochrome C levels. Both results confirmed the potential test NPs in causing apoptotic cell death in C. auris.ConclusionThe trimetallic (Cu-Zn-Fe) nanoparticles displayed strong antifungal activity against C. auris, with a potential to arrest the cell cycle at S-phase, which could be linked to the DNA damage. Important yeast apoptotic markers suggested that the test NPs have a potential to cause apotosis in C. auris. All these findings suggest the potential of these trimetallic NPs to be taken to the next level of research in the development of novel antifungal medications.

Zinc oxide nanoparticles inhibit osteosarcoma metastasis by downregulating β-catenin via HIF-1α/BNIP3/LC3B-mediated mitophagy pathway.

Osteosarcoma (OS) therapy faces many challenges, especially the poor survival rate once metastasis occurs. Therefore, it is crucial to explore new OS treatment strategies that can efficiently inhibit OS metastasis. Bioactive nanoparticles such as zinc oxide nanoparticles (ZnO NPs) can efficiently inhibit OS growth, however, the effect and mechanisms of them on tumor metastasis are still not clear. In this study, we firstly prepared well-dispersed ZnO NPs and proved that ZnO NPs can inhibit OS metastasis-related malignant behaviors including migration, invasion, and epithelial-mesenchymal transition (EMT). RNA-Seqs found that differentially expressed genes (DEGs) in ZnO NP-treated OS cells were enriched in wingless/integrated (Wnt) and hypoxia-inducible factor-1 (HIF-1) signaling pathway. We further proved that Zn2+ released from ZnO NPs induced downregulation of β-catenin expression via HIF-1α/BNIP3/LC3B-mediated mitophagy pathway. ZnO NPs combined with ICG-001, a β-catenin inhibitor, showed a synergistic inhibitory effect on OS lung metastasis and a longer survival time. In addition, tissue microarray (TMA) of OS patients also detected much higher β-catenin expression which indicated the role of β-catenin in OS development. In summary, our current study not only proved that ZnO NPs can inhibit OS metastasis by degrading β-catenin in HIF-1α/BNIP3/LC3B-mediated mitophagy pathway, but also provided a far-reaching potential of ZnO NPs in clinical OS treatment with metastasis.

Analysis of Nanotoxicity with Integrated Omics and Mechanobiology.

Nanoparticles (NPs) in biomedical applications have benefits owing to their small size. However, their intricate and sensitive nature makes an evaluation of the adverse effects of NPs on health necessary and challenging. Since there are limitations to conventional toxicological methods and omics analyses provide a more comprehensive molecular profiling of multifactorial biological systems, omics approaches are necessary to evaluate nanotoxicity. Compared to a single omics layer, integrated omics across multiple omics layers provides more sensitive and comprehensive details on NP-induced toxicity based on network integration analysis. As multi-omics data are heterogeneous and massive, computational methods such as machine learning (ML) have been applied for investigating correlation among each omics. This integration of omics and ML approaches will be helpful for analyzing nanotoxicity. To that end, mechanobiology has been applied for evaluating the biophysical changes in NPs by measuring the traction force and rigidity sensing in NP-treated cells using a sub-elastomeric pillar. Therefore, integrated omics approaches are suitable for elucidating mechanobiological effects exerted by NPs. These technologies will be valuable for expanding the safety evaluations of NPs. Here, we review the integration of omics, ML, and mechanobiology for evaluating nanotoxicity.

Ultrasonic Atomizer-Driven Development of Biocompatible and Biodegradable Poly(d,l-lactide-co-glycolide) Nanocarrier-Encapsulated Suberoylanilide Hydroxamic Acid to Combat Brain Cancer.

The path to the discovery of anticancer drugs and investigating their potential activity has remained a quest for several decades. Suberoylanilide hydroxamic acid (SAHA), also known as "Vorinostat", is a well-known histone deacetylase inhibitor (HDACi) and has the potential to act as a therapeutic agent against tumorigenesis. Herein, we have fabricated SAHA incorporated into biocompatible and biodegradable poly(d,l-lactide-co-glycolide) PLGA nanoparticles (NPs) using a facile method of ultrasonic atomization and evaluated their anticancer property. We have explored their characteristics using dynamic light scattering (DLS), scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HR-TEM), encapsulation efficiency, and in vitro drug release and have investigated their efficacy on U87 glioblastoma (GBM) cells. SAHA-PLGA NPs synthesized were of average mean size of 80 ± 23 and 105 ± 6.0 nm observed through cryo-field-emission gun SEM and HR-TEM with a polydispersity index of 0.068 and a ζ-potential value of -13.26 mV. The encapsulation efficiency was 53%, with a sustained in vitro release up to 48 h. The in vitro assessment of SAHA-PLGA NPs for their anticancer activity on U87 GBM cells showed cellular cytotoxicity with an IC50 of 19.91 μM. SAHA-PLGA NP-treated cells also showed suppression in migration with 8.77 μM concentration, and cell growth inhibition was observed in the wound scratch assay for up to 24 h. The cellular uptake studies have been utilized by time-dependent experiments, revealing their cellular internalization. Taking this into account, our present experimental findings indicate that SAHA-PLGA NPs could play a significant role in enhancing the effectiveness and bioavailability and reducing adverse effects of cancer chemotherapy. It also highlights the inherent potential of these biocompatible entities for chemotherapeutic applications in biomedical and pharmaceutics.

Membrane Disruption Potential of Doped Cuprous Oxide Nanoparticles Against blaNDM-1 and mcr-1 Positive Colistin Resistant E. coli

Bacterial resistance against the last-resort antibiotic “colistin” is a growing problem worldwide. Novel strategies in the field of nanotechnology for the development of suitable antibacterial drug with minimal toxicities are need of the time. Here we synthesized, Cu2O nanoparticles (Cu2O-NPs) doped with 5% Zn metal ions by adopting solution-phase method using hydrazine reduction in aqueous medium. Antibacterial assays showed a promising activity against colistin-resistant Escherichia coli isolates. Leakage assay, propidium iodide uptake assay, EtBr uptake assay, and LIVE/DEAD® BacLight™ assay showed membrane damaging activity of Cu2O-NPs. Imaging by scanning electron microscopy of NP-treated bacterial cells confirmed membrane damaging potential of used Cu2O-NPs. Toxicity assessment by cell viability assay showed no cytotoxic activity to the HEK cell line. In conclusion, our results suggest that Cu2O-NPs doped with 5% Zn may exert their antimicrobial action by disruption of the bacterial cell membrane and probably generating reactive oxygen species (ROS). We suggest that Cu2O-NPs doped with 5% Zn can be used as effective antibacterial agent.

Potential antibiofilm activity of farnesol-loaded poly(DL-lactide-co-glycolide) (PLGA) nanoparticles against Candida albicans

Candida species are ubiquitous fungal pathogens and are the most common causes of mucosal and invasive fungal infections in humans. Especially Candida albicans commonly resides as a commensal in the mucosal tissues of approximately half of the human population. When the balance of the normal flora is disrupted or the immune defenses are compromised, Candida species can become pathogenic, often causing recurrent disease in susceptible individuals.The treatments available for Candida infection are commonly drug-based and can involve topical and systemic antifungal agents. However, the use of standard antifungal therapies can be limited because of toxicity, low efficacy rates, and drug resistance. Candida species ability to produce drug-resistant biofilm is an important factor in human infections, because microorganisms within biofilm benefit from various advantages over their planktonic counterparts including protection from antimicrobials and chemicals. These limitations emphasize the need to develop new and more effective antifungal agents. Natural products are attractive alternatives for this purpose due to their broad spectrum of biological activities. Farnesol is produced by many microorganisms and found in some essential oils. It has also a great attention as a quorum-sensing molecule and virulence factor. It has also antimicrobial potential due to its inhibitory effects on various bacteria and fungi. However, as it is a hydrophobic component, its solubility and biofilm inhibiting properties are limited.To overcome these shortcomings, nanoparticle-based drug delivery systems have been successfully used. For this purpose, especially using biodegradable polymeric nanoparticles has gained increasing attention owing to their biocompatibility and minimal toxicity. Poly (DL-lactide-co-glycolide) (PLGA) is the most widely used polymer in this area. In this study, farnesol is loaded to PLGA nanoparticles (F-PLGA NPs) by emulsion evaporation method and characterized by DLS, TEM, and FT-IR analyses. Our TEM findings indicate that the sizes of F-PLGA NPs are approximately 140 nm. The effects of F-PLGA NPs on planktonic cells and biofilm formation of C. albicans were compared with effects of farnesol alone. Farnesol inhibits the growth at a range of 53% at a concentration of 2.5 μL compared to the control group. This rate is 45% for F-PLGA NPs at the same concentration. However, although farnesol amount in F-PLGA is approximately 22.5% of the total volume, the observed effect is significant. In TEM examinations, planktonic Candida cells treated with farnesol showed relatively regular ultrastructural morphology. Few membrane and wall damage and electron density in the cytoplasm were determined. In F-PLGA NP-treated cells, increased irregular cell morphology, membrane and wall damages, and large vacuoles are observed. Our SEM and XTT data suggest that F-PLGA NPs can reduce the biofilm formation at lower concentrations than farnesol alone 57%, and our results showed that F-PLGA NPs are effective and biocompatible alternatives for inhibiting growth and biofilm formation of C. albicans, but detailed studies are needed.

Nanoparticles Affect the Expression Stability of Housekeeping Genes in Plant Cells.

PurposeWe report on the expression stability of several housekeeping/reference genes that can be used in the normalization of target gene expression in quantitative real-time PCR (qRT-PCR) analysis of plant cells challenged with metal nanoparticles (NPs).Materials and MethodsUniform cell suspension cultures of Hypericum perforatum were treated with 25 mg/l silver and gold NPs (14–15 nm in diameter). Cells were collected after 0.5, 4.0, and 12 h. The total RNA isolated from the cells was analyzed for the stability of ACT2, ACT3, ACT7, EF1-α, GAPDH, H2A, TUB-α, TUB-β, and 18S rRNA genes using qRT-PCR. The cycle threshold (Ct) values of the genes were analyzed using the geNorm, NormFinder, BestKeeper, and RefFinder statistical algorithms to rank gene stability. The stability of the top-ranked genes was validated by normalizing the expression of HYP1.ResultsThe expression of the tested housekeeping genes varied with treatment duration and NP types. EF1-α in gold NP treatment and TUB-α and EF1-α in silver NP treatment ranked among the top three positions. However, none of the genes retained their top ranking with time and across NP types.ConclusionEF1-α can be used as a reference for treatment involving both silver and gold NPs in H. perforatum cells. TUB-α can be used only for silver NP-treated cells. The expression instability of most of the housekeeping genes highlights the importance of systematic standardization of reference genes for NP treatment conditions to draw proper conclusions on the target gene expression.

Chloroquine as a promising adjuvant therapy for type 1 Diabetes Mellitus

Chloroquine (CQ) and hydroxychloroquine, are promising anti-inflammatory drugs for the treatment of Diabetes mellitus (DM) to prevent associated complications. Therefore, this study evaluated the anti-inflammatory effects of CQ-free and CQ-incorporated polylactic acid nanoparticles (NPs) in the peripheral blood mononuclear cells (PBMCs) of patients with type 1 Diabetes mellitus (T1DM). In total, 25 normoglycemic individuals and 25 patients with T1DM aged 10–16 years were selected and glycemic controls evaluated. After cell viability assessed by MTT assay, T1DM PBMCs were subjected to a CQ concentration of 10 µM in three different conditions: not treated (NT), treated with CQ, and treated with CQ NPs. The cells were incubated for 48 h, and the mRNA expressions of cytokines IL1B, IFNG, TNFA, IL12, and IL10 were determined by relative quantification through real-time PCR at 24 h intervals. IL1B expression decreased in CQ and CQ NP-treated cells after 48 h (p < 0.001) and 24 h (p < 0.05) of treatment, respectively. IFNG and IL12 expressions significantly decreased (p < 0.001) in cells treated with CQ and CQ NPs at 24 and 48 h compared to NT. TNFA and IL10 expressions significantly decreased after 48 h (p < 0.001) and 24 h (p < 0.002), respectively, by both CQ and CQ NPs treatment. Despite being a preliminary in vitro study, CQ has anti-inflammatory activity in the primary cells of T1DM patients and could represent an alternative and adjuvant anti-inflammatory therapy to prevent diabetes complications.

GSH-ZnS Nanoparticles Exhibit High-Efficiency and Broad-Spectrum Antiviral Activities via Multistep Inhibition Mechanisms.

Despite the good biocompatibility and antibacterial activity of zinc sulfide nanoparticles (ZnS NPs), whether they possess antiviral activity is still unclear. Here, GSH-modified ZnS NPs (GSH-ZnS NPs) were synthesized and their significant antiviral activity was demonstrated using the Arteriviridae family RNA virus, porcine reproductive and respiratory syndrome virus (PRRSV), as a model. Mechanistically, GSH-ZnS NPs were shown to reduce PRRSV-induced ROS production to prevent PRRSV multiplication, with no activating effect on the interferon (IFN) signal pathway, the first defense line against virus infection. Furthermore, isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomic analysis of GSH-ZnS NP-treated cells revealed the involvement of numerous crucial proteins in virus proliferation, with vitronectin (VTN) being confirmed as an efficient PRRSV antagonist here. Furthermore, GSH-ZnS NPs were found to have potent antiviral effects on the Herpesviridae family DNA virus, pseudorabies virus (PRV), the Coronaviridae family positive-sense RNA virus, porcine epidemic diarrhea virus (PEDV), and the Rhabdoviridae family negative-stranded RNA virus, vesicular stomatitis virus (VSV), indicating their broad-spectrum antiviral activity against viruses from different families with various genome types. Overall, GSH-ZnS NP is a prospective candidate for the development of antiviral nanomaterials and may serve as a model for investigation of potential host restriction factors in combination with proteomics.

Doxorubicin Delivered Using Nanoparticles Camouflaged with Mesenchymal Stem Cell Membranes to Treat Colon Cancer.

PurposeThe primary goal of the present study was to design doxorubicin (DOX)-loaded superparamagnetic iron oxide (SPIO) nanoparticles (NPs) coated with mesenchymal stem cell (MSC) membranes and explore their effect on colon cancer in vitro and in vivo.MethodsDOX-SPIO NPs were coated with MSC membranes using an extruder, and the morphological characteristics of MSC membrane-camouflaged nanodrug (DOX-SPIO@MSCs) evaluated by transmission electron microscopy (TEM) and NP-tracking analysis. Drug loading and pH response were assessed by UV spectrophotometry. Intracellular colocalization was analyzed using NP-treated MC38 cells stained with 3,3′-dioctadecyloxacarbocyanine perchlorate and Hoechst 33342. Cellular uptake was analyzed using an inverted fluorescence microscope and flow cytometry and cytotoxicity evaluated by cell counting kit-8 assay. Biological compatibility was assessed by hemolysis analysis, immunoactivation test and leukocyte uptake experiments. Furthermore, intravenous injection of chemotherapy drugs into MC38 tumor-bearing C57BL/6 mice was used to study anti-tumor effects.ResultsTypical core-shell NP structures were observed by TEM. Particle size remained stable in fetal bovine serum and phosphate-buffered saline (PBS). Compared with DOX-SPIO, DOX-SPIO@MSCs improved cellular uptake efficiency, enhanced anti-tumor effects, and reduced the immune system response. Animal experiments demonstrated that DOX-SPIO@MSCs enhanced tumor treatment efficacy while reducing systemic side effects.ConclusionOur experimental results demonstrate that DOX-SPIO@MSCs are a promising targeted nanocarrier for application in treatment of colon cancer.

Synergetic effects of thymoquinone-loaded porous PVPylated Fe3O4 nanostructures for efficient pH-dependent drug release and anticancer potential against triple-negative cancer cells.

Porous iron oxide nanostructures have attracted increasing attention due to their potential biomedical applications as nanocarriers for cancer and many other therapies as well as minimal toxicity. Herbal anti-cancer agent thymoquinone loaded on Fe3O4 nanoparticles is envisaged to offer solution towards cancer treatment. The purpose of the present study was to investigate the efficacy of thymoquinone-loaded PVPylated Fe3O4 magnetic nanoparticles (TQ-PVP-Fe3O4 NPs) against triple-negative breast cancer (TNBC) cells. The porous PVPylated Fe3O4 NPs were prepared by a simple solvothermal process, whereas the thymoquinone drug was loaded via the nanoprecipitation method. Fourier transform infrared (FTIR) spectroscopic analysis confirmed the molecular drug loading, and surface morphological observation further confirmed this. The quantity of thymoquinone adsorbed onto the porous PVPylated Fe3O4 NPs was studied by thermogravimetric analysis (TGA). The positive surface charge of TQ-PVP-Fe3O4 NPs facilitates the interaction of the NPs with cancer (MDA-MB-231) cells to enhance the biological functions. In addition, the anticancer potential of NPs involving cytotoxicity, apoptosis induction, reactive oxygen species (ROS) generation, and changes in the mitochondrial membrane potential (ΔΨm) of TNBC cells was evaluated. TQ-PVP-Fe3O4 NP-treated cells effectively increased the ROS levels leading to cellular apoptosis. The study shows that the synthesized TQ-PVP-Fe3O4 NPs display pH-dependent drug release in the cellular environment to induce apoptosis-related cell death in TNBC cells. Hence, the prepared TQ-PVP-Fe3O4 NPs may be a suitable drug formulation for anticancer therapy.

Bioaccumulation, growth performance, and transcriptomic response of Dictyosphaerium sp. after exposure to nonylphenol

Algal cultures of Dictyosphaerium sp. were treated with a wide range (0–8 mg/L) of nonylphenol (NP) applications to investigate the species' potential to perform bioremediation of NP-contaminated wastewater and explore the mechanism of NP toxicity in algal cell. The algal species exhibited a high tolerance for NP and efficiently removed even high concentrations of NP. The accumulation of NP in algal cells tended to increase and the percentage of NP adhered to the cell walls tended to decrease as the NP concentration increased. The thylakoid density and the pyrenoid size also tended to decrease as the NP concentration rose. The key genes involved in the antioxidative pathways and photosynthetic pathways were down-regulated in the NP-treated algal cells. Algal ribosome genes were especially sensitive to NP. The results indicated that NP could induce oxidative stress in algal cells and that Dictyosphaerium sp. showed potential as a bioremediation agent.

Bisphenol-A and Nonylphenol Induce Apoptosis in Reproductive Tract Cancer Cell Lines by the Activation of ADAM17.

Endocrine-disruptor chemicals (EDCs), such as bisphenol A (BPA) and nonylphenol (NP), have been widely studied due to their negative effects on human and wildlife reproduction. Exposure to BPA or NP is related to cell death, hormonal deregulation, and cancer onset. Our previous studies showed that both compounds induce A Disintegrin And Metalloprotease 17 (ADAM17) activation. Here, we show that BPA and NP induce apoptosis in prostate and ovary cancer cell lines, in a process dependent on ADAM17 activation. ADAM17 knockdown completely prevented apoptosis as well as the shedding of ADAM17 substrates. Both compounds were found to induce an increase in intracellular calcium (Ca2+) only in Ca2+-containing medium, with the NP-treated cells response being more robust than those treated with BPA. Additionally, using a phosphorylated protein microarray, we found that both compounds stimulate common intracellular pathways related to cell growth, differentiation, survival, and apoptosis. These results suggest that BPA and NP could induce apoptosis through ADAM17 by activating different intracellular signaling pathways that may converge in different cellular responses, one of which is apoptosis. These results confirm the capacity of these compounds to induce cell apoptosis in cancer cell lines and uncover ADAM17 as a key regulator of this process in response to EDCs.

014 The arylhydrocarbon receptor agonist 4-n-nonylphenol switches non-regulatory T cells into a regulatory phenotype

4-n-nonylphenol (NP), an arylhydrocarbon receptor (AhR)-agonist, was found to inhibit the induction of contact hypersensitivity (CHS) in mice via induction of regulatory T cells (Treg). Although dendritic cells were identified as one of the major targets for NP in its immunosuppressive features, it is unknown whether NP can directly affect T cells as well. To address this issue, non-regulatory T cells (CD4+CD25-) were isolated from trinitrochlorobenzene (TNCB)-sensitized mice and stimulated with NP or left untreated. After 48 hours cells were injected intravenously into naïve recipients which were sensitized after injection; Treg (CD4+CD25+) served as a control. Ear challenge with TNCB was significantly suppressed in mice injected with NP-treated cells comparable to the Treg control. In contrast, injection of untreated CD4+CD25- did not affect CHS in the recipients. Flow cytometry analysis revealed a strong induction of the Treg markers Foxp3 and Garp in response to NP, suggesting that NP shifts non-regulatory T cells into a regulatory phenotype. However, NP-Treg did not suppress the elicitation phase of CHS since the ear challenge response was not suppressed upon intravenous injection of NP-Treg into already sensitized mice. When injected subcutaneously into the ears of sensitized mice before challenge the ear swelling response was significantly reduced, indicating that NP-Treg upon intravenous injection may migrate into the lymph nodes but not into the skin. This may be due to the expression of specific tissue homing receptors. To prove whether NP induces Treg also in the human system CD4+CD25- cells were isolated from peripheral blood mononuclear cells, incubated with NP and subjected to an in vitro suppression assay. NP-Treg suppressed the proliferative capacity of the responder cells, whereas unstimulated CD4+CD25- did not. Together this implies that activation of the AhR switches non-regulatory T cells into a regulatory phenotype which is functionally suppressive.

The uptake, retention and clearance of drug-loaded dendrimer nanoparticles in astrocytes - electrophysiological quantification.

Nanoparticle-based drug delivery systems may impose risks to patients due to potential toxicity associated with a lack of clearance from cells or prolonged carrier-cell retention. This work evaluates vesicular cell uptake, retention and the possible transfer of endocytosed methylprednisolone-loaded carboxymethylchitosan/poly(amidoamine) dendrimer nanoparticles (NPs) into secretory vesicles of rat cultured astrocytes. The cells were incubated with NPs and unitary vesicle fusions/fissions with the plasma membrane were monitored employing high-resolution membrane capacitance measurements. In the NP-treated cells the frequency of unitary exocytotic events was significantly increased. The presence of NPs also induces an increase in the size of exocytotic vesicles interacting with the plasma membrane, which exhibit transient fusion with prolonged fusion pore dwell-time. Live-cell confocal imaging revealed that once NPs internalize into endocytotic compartments they remain in the cell for 7 days, although a significant proportion of these merge with secretory vesicles destined for exocytosis. Co-localization studies show the route of clearance of NPs from cells via the exocytotic pathway. These findings bring new insight into the understanding of the intracellular trafficking and biological interactions of drug-loaded dendrimer NPs targeting astrocytes.

Zinc oxide nanoparticles induce apoptosis and autophagy in human ovarian cancer cells.

BackgroundZinc oxide nanoparticles (ZnO NPs) are frequently used in industrial products such as paint, surface coating, and cosmetics, and recently, they have been explored in biologic and biomedical applications. Therefore, this study was undertaken to investigate the effect of ZnO NPs on cytotoxicity, apoptosis, and autophagy in human ovarian cancer cells (SKOV3).MethodsZnO NPs with a crystalline size of 20 nm were characterized with various analytical techniques, including ultraviolet-visible spectroscopy, X-ray diffraction, transmission electron microscopy, Fourier transform infrared spectroscopy, and atomic force microscopy. The cytotoxicity, apoptosis, and autophagy were examined using a series of cellular assays.ResultsExposure of cells to ZnO NPs resulted in a dose-dependent loss of cell viability, and the characteristic apoptotic features such as rounding and loss of adherence, enhanced reactive oxygen species generation, and loss of mitochondrial membrane potential were observed in the ZnO NP-treated cells. Furthermore, the cells treated with ZnO NPs showed significant double-strand DNA breaks, which are gained evidences from significant number of γ-H2AX and Rad51 expressed cells. ZnO NP-treated cells showed upregulation of p53 and LC3, indicating that ZnO NPs are able to upregulate apoptosis and autophagy. Finally, the Western blot analysis revealed upregulation of Bax, caspase-9, Rad51, γ-H2AX, p53, and LC3 and downregulation of Bcl-2.ConclusionThe study findings demonstrated that the ZnO NPs are able to induce significant cytotoxicity, apoptosis, and autophagy in human ovarian cells through reactive oxygen species generation and oxidative stress. Therefore, this study suggests that ZnO NPs are suitable and inherent anticancer agents due to their several favorable characteristic features including favorable band gap, electrostatic charge, surface chemistry, and potentiation of redox cycling cascades.