Moderate Static Magnetic Field (6 mT)-Induced Lipid Rafts Rearrangement Increases Silver NPs Uptake in Human Lymphocytes

One of the most relevant drawbacks in medicine is the ability of drugs and/or imaging agents to reach cells. Nanotechnology opened new horizons in drug delivery, and silver nanoparticles (AgNPs) represent a promising delivery vehicle for their adjustable size and shape, high-density surface ligand attachment, etc. AgNPs cellular uptake involves different endocytosis mechanisms, including lipid raft-mediated endocytosis. Since static magnetic fields (SMFs) exposure induces plasma membrane perturbation, including the rearrangement of lipid rafts, we investigated whether SMF could increase the amount of AgNPs able to pass the peripheral blood lymphocytes (PBLs) plasma membrane. To this purpose, the effect of 6-mT SMF exposure on the redistribution of two main lipid raft components (i.e., disialoganglioside GD3, cholesterol) and on AgNPs uptake efficiency was investigated. Results showed that 6 mT SMF: (i) induces a time-dependent GD3 and cholesterol redistribution in plasma membrane lipid rafts and modulates gene expression of ATP-binding cassette transporter A1 (ABCA1), (ii) increases reactive oxygen species (ROS) production and lipid peroxidation, (iii) does not induce cell death and (iv) induces lipid rafts rearrangement, that, in turn, favors the uptake of AgNPs. Thus, it derives that SMF exposure could be exploited to enhance the internalization of NPs-loaded therapeutic or diagnostic molecules

Microscopies at the nanoscale for nano-scale drug delivery systems

One of the frontier of nanoscience is undoubtedly represented by the use of nanotechnologies in the pharmaceutical research. During the last decades a big family of nanostructures that have a surface-acting action, such as NanoParticles (NPs), lipid nanocarriers and many more, have been developed to be used as Drug Delivery Systems (DDSs). However, these nanocarriers opened also new frontiers in nanometrology, requiring an accurate morphological characterization, near atomic resolution, before they are really available to clinicians to ascertain their elemental composition, to exclude the presence of contaminants introduced during the synthesis procedure and to ensure biocompatibility. Classical Transmission (TEM) and Scanning Electron Microscopy (SEM) techniques frequently have to be adapted for an accurate analysis of formulation morphology, especially in case of hydrated colloidal systems. Specific techniques such as environmental scanning microscopy and/or cryo preparation are required for their investigation. Analytical Electron Microscopy (AEM) techniques such as Electron Energy-Loss Spectroscopy (EELS) or Energy-Dispersive X-ray Spectroscopy (EDXS) are additional assets to determine the elemental composition of the systems. Here we will discuss the importance of Electron Microscopy (EM) as a reliable tool in the pharmaceutical research of the 21st century, focalizing our attention on advantages and limitations of different kind of NPs (in particular silver and carbon NPs, cubosomes) and vesicles (liposomes and niosomes)

the dialogue between died and viable cells in vitro and in vivo bystander effects and 1h nmr based metabolic profiling of soluble factors

AbstractThe bystander effect (BE) is an important biological phenomenon that induces damages in distant and not directly affected by a chemical/physical stress cells. This effect, well known in ionizing radiation treatment, relies on reactive signals released by exposed cells and transmitted via cell–cell interaction or culture medium. In this study, cycloheximide (CHX)-induced apoptotic U937 cells and untreated THP-1 cells were chosen to investigate the chemical-induced BE. The effects of apoptotic U937 cells culture medium, Conditioned Medium (CM), on THP-1 cells were evaluated by morphological and immunohistochemical analysis performed by light microscopy; 1D 1H and 2D J-resolved (JRES) NMR metabolomic analysis has been used to characterize the molecules involved in the BE. In summary, this study indicates that: CM of CHX-treated U937 cells induces a time-dependent induction of toxicity, probably apoptotic cell death, and macrophagic differentiation in THP-1 cells; CM contains different metabolites respect fresh culture medium; CM recruits in vivo activated fibroblasts, endothelial cells, macrophages and mononuclear inflammatory cells in rat calf muscles. These data suggest that CHX exposed cells could cause BE through the release, during the apoptotic process, of soluble factors into the medium that could be exploited in anticancer protocols

Design, Synthesis and Characterization of a PEGylated Stanozolol for Potential Therapeutic Applications

Stanozolol (STZ) is a drug used to treat serious disorders like aplastic anemia and hereditary angioedema. It is also indicated as an adjunct therapy for the treatment of vascular disorders and growth failures. Encouraging results obtained using animal models demonstrated that STZ increases bone formation and mineralization, thus improving both density and biomechanical properties. Like natural androgens, such as TST and 5α-dihydrotestosterone (5α-DHT), STZ binds androgen receptor (AR) to activate AR-mediated signalling. Despite its therapeutic effects, this synthetic anabolic-androgenic steroid (AAS), or 5α-DHT derivative, due to its high lipophilicity, is poor soluble in water. Thus, to increase the water solubility and stability of STZ, as well as its bioavailability and efficacy, an innovative PEGylated STZ (STZ conjugated with (MeO-PEG-NH2)10kDa, (MeO-PEG-NH)10kDa-STZ) was synthesized. As confirmed by chromatography (RP-HPLC) and spectrometry (ATR-FTIR, 1H-NMR, elemental CHNS(O) analysis, MALDI-TOF/TOF) analyses, a very pure, stable and soluble compound was obtained. Acetylcholinesterase (AChE) competitive ELISA kit demonstrated that the resulting PEGylated STZ competes against biological TST, especially at lower concentrations. Cytotoxicity of increasing concentrations (1, 10, 25 or 50 µM) of STZ and/or (MeO-PEG-NH)10kDa-STZ was also evaluated for up 80 h by performing the MTT assay on human osteosarcoma Saos-2 cells, which express AR and are responsive to STZ. PEGylation mitigated cytotoxicity of STZ, by increasing the cell viability values, especially at higher drug concentrations. Furthermore, these results suggest that (MeO-PEG-NH)10kDa-STZ is a promising and reliable drug to be used in clinical conditions in which TST is required.Peer reviewe

Nutraceutical Vegetable Oil Nanoformulations for Prevention and Management of Diseases

The scientific community is becoming increasingly interested in identifying, characterizing, and delivering nutraceuticals, which constitutes a multi-billion-dollar business. These bioactive agents are claimed to exhibit several health benefits, including the prevention and treatment of diseases such as arthritis, cancer, osteoporosis, cataracts, Alzheimer’s, and Huntington’s diseases, heart, brain and metabolic disorders, etc. Nutraceuticals are typically consumed as part of a regular human diet and are usually present within foods, comprising vegetable oil, although at low levels and variable composition. Thus, it is difficult to control the type, amount and frequency of their ingestion by individuals. Nanoformulations about vegetable oil-based bioactive compounds with nutraceutical properties are useful for overcoming these issues, while improving the uptake, absorption, and bioavailability in the body. The purpose of this current study is to review papers on such nanoformulations, particularly those relevant for health benefits and the prevention and management of diseases, as well as bioactives extracted from vegetable oils enhancing the drug effectiveness, retrieved through bibliographic databases by setting a timespan from January 2000 to April 2020 (about 1758 records)

Nanonutraceuticals Delivery

Technological innovation, environmental sustainability, health, and wellness are the trajectories explored by current research to identify new strategies for a general improvement of human quality of life [...

Environmental Factors Affecting Phagocytosisof Dying Cells: Smoking and StaticMagnetic Fields

Cell surface modifications are fundamental for the correct and fast removal of apoptotic cells. These changes include the appearance of tethering molecules on the surface of apoptotic cells, the externalization of PS, oxidation of phospholipids and qualitative and quantitative changes in surface sugars and ICAM-3. Phagocytes, both professional and non-professional, use specific receptors that bind to the apoptotic cells either directly or through bridging molecules. In non-pathological conditions, apoptotic cells are normally cleared via an anti-inflammatory pathway. In contrast, the uptake and removal of necrotic cells normally involves inflammation and an immune response. Besides the “eat me” signals on the dying cells, phagocytes can also recognize “leave-me-alone” signals on healthy cells. The correct repertoire of molecules exposed on the cell surfaces prevents the engulfment of living undamaged cells. Thus, any factors influencing cell surface molecule expression on both phagocytes and/or apoptotic cells can in turn affect recognition of living and/or apoptotic cells. One such factor is cigarette smoke, which contain highly reactive carbonyls, which can modify proteins that directly or indirectly affect cellular functions. Moreover cigarette smoke is a major etiological factor in the development of COPD, in which apoptosis and defective PACs play a fundamental role. Another environmental factor that may interfere with the normal correct exposure of molecules on cell surfaces is exposure to (S)MFs. Despite the multiplicity of experimental conditions (i.e. in vitro or in vivo models, intensity and type of field, time of exposure, metabolic state of the cells, etc), converging data indicate that the primary site of action of (S)MFs and (E)MFs is the plasma membrane: i.e. they affect the electrochemical balance of the membrane, the distribution of membrane proteins and membrane receptors, cell-cell and cell-matrix junctions, sugar residue

Environmental Factors Affecting Phagocytosis of Dying Cells: Smoking and Static Magnetic Fields

Cell surface modifications are fundamental for the correct and fast removal of apoptotic cells. These changes include the appearance of tethering molecules on the surface of apoptotic cells, the externalization of PS, oxidation of phospholipids and qualitative and quantitative changes in surface sugars and ICAM-3. Phagocytes, both professional and non-professional, use specific receptors that bind to the apoptotic cells either directly or through bridging molecules. In non-pathological conditions, apoptotic cells are normally cleared via an anti-inflammatory pathway. In contrast, the uptake and removal of necrotic cells normally involves inflammation and an immune response. Besides the “eat me” signals on the dying cells, phagocytes can also recognize “leave-me-alone” signals on healthy cells. The correct repertoire of molecules exposed on the cell surfaces prevents the engulfment of living undamaged cells. Thus, any factors influencing cell surface molecule expression on both phagocytes and/or apoptotic cells can in turn affect recognition of living and/or apoptotic cells. One such factor is cigarette smoke, which contain highly reactive carbonyls, which can modify proteins that directly or indirectly affect cellular functions. Moreover cigarette smoke is a major etiological factor in the development of COPD, in which apoptosis and defective PACs play a fundamental role. Another environmental factor that may interfere with the normal correct exposure of molecules on cell surfaces is exposure to (S)MFs. Despite the multiplicity of experimental conditions (i.e. in vitro or in vivo models, intensity and type of field, time of exposure, metabolic state of the cells, etc), converging data indicate that the primary site of action of (S)MFs and (E)MFs is the plasma membrane: i.e. they affect the electrochemical balance of the membrane, the distribution of membrane proteins and membrane receptors, cell-cell and cell-matrix junctions, sugar residue

Comparative Analysis of Biological Effects Induced on Different Cell Types by Magnetic Fields with Magnetic Flux Densities in the Range of 1–60 mT and Frequencies up to 50 Hz

Moderate static magnetic fields (SMFs) are generated from sources such as new-generation electric trams and trains, electric arc welding, and magnetic resonance imaging (MRI) devices, as well as during the industrial production of aluminium, while extremely low frequency pulsed magnetic fields (ELF-PMFs) are produced by house power installations, household appliances, and high voltages transmission lines. Moderate SMFs and ELF-PMFs with magnetic flux densities (B) in the range of 1–60 mT and frequencies (f) up to 50 Hz are common MF exposure sources for the population. Even though humans are continually exposed to these MFs, to date no definitive endpoint has been drawn about their safety. In this review, the state of knowledge about the biological effects induced by these MFs on different cell types will be addressed. In our own observation, the putative modulation of Ca2+/H+ and Na+/H+ plasma membrane antiporters of human peripheral blood lymphocytes (PBLs) was found to occur after a 24 h exposure to a 6 mT SMF, and the bystander effect observed on U937 cells cultivated for up to 6 h in the conditioned medium harvested from human PBLs previously exposed for 24 h to the same MF (secondary necrosis induction) will be also herein discussed