Imaging mass spectrometry differentiates the effects of doxorubicin formulations on non-targeted tissues.

Administration of cytotoxic agents like doxorubicin (DOX) is restrained by the effects on different non-targeted/non-cancerous tissues, which instigates the development of nano-enabled drug delivery systems, among others. In this study, imaging mass spectrometry (IMS) was selected to examine the effects of DOX nanoformulations on non-targeted tissues. Chemical alterations induced by liposomal (LPS) and poly (lactic-co-glycolic acid) (PLG) nanoformulations were assessed against the ones induced by the conventional (CNV) formulation. Kidney cryosections of the treated and control Wistar rats were used as a model of the non-targeted tissue and analyzed by MALDI TOF IMS in the 200-1000 Da m/z range. Principal component analysis (PCA) and Volcano plots of the average mass spectra demonstrated a large overlap between treatments. However, the Venn diagram of significant m/z values revealed a nanoformulation-specific fingerprint consisting of 59 m/z values, which set them apart from the CNV formulation characterized by the fingerprint of 22 significant m/z values. Fingerprint m/z values that were putatively annotated by metabolome database search were linked to apoptosis, cell migration and proliferation. In CNV and PLG cases, false discovery rate adjusted ANOVA showed no differences in the spatial distribution of fingerprint m/z values between the histological substructures like glomeruli and convoluted tubules indicating their tissue-nonselective effect. LPS caused the least significant changes in m/z values and some of the LPS-specific fingerprint m/z values were primarily distributed in the glomeruli. The IMS based procedure successfully differentiated the effects of DOX formulations on the model non-targeted tissue, thus indicating the importance of IMS in effective drug development.

Utilization of Olive Pomace in Green Synthesis of Selenium Nanoparticles: Physico-Chemical Characterization, Bioaccessibility and Biocompatibility.

Olive pomace extract (OPE) was investigated as a potential surface modifier for the development of the green synthesis process of selenium nanoparticles (SeNPs). In order to evaluate them as potential nutraceuticals, the obtained nanosystems were characterized in terms of size distribution, shape, zeta potential, stability in different media, gastrointestinal bioaccessibility and biocompatibility. Systems with a unimodal size distribution of spherical particles were obtained, with average diameters ranging from 53.3 nm to 181.7 nm, depending on the type of coating agent used and the presence of OPE in the reaction mixture. The nanosystems were significantly affected by the gastrointestinal conditions. Bioaccessibility ranged from 33.57% to 56.93% and it was significantly increased by functionalization of with OPE. Biocompatibility was investigated in the HepG2 and Caco2 cell models, proving that they had significantly lower toxicity in comparison to sodium selenite. Significant differences were observed in cellular responses depending on the type of cells used, indicating differences in the mechanisms of toxicity induced by SeNPs. The obtained results provide new insight into the possibilities for the utilization of valuable food-waste extracts in the sustainable development of nanonutraceuticals.

Functionalization of selenium nanoparticles with olive polyphenols - impact on toxicity and antioxidative activity.

Selenium nanoparticles (SeNPs) represent novel selenium (Se) formulation characterized by improved biocompatibility and a wider therapeutic range in comparison to inorganic Se. The aim of this work was to investigate the possibilities of functionalization of SeNPs with olive pomace extract (OPE), rich in health-promoting polyphenols, and to obtain innovative forms of nutraceuticals. Cytotoxic and antioxidative activities of four types of SeNPs (polyvinylpyrrolidone stabilized (PVP SeNPs), polysorbate stabilized (PS SeNPs), polyvinylpyrrolidone stabilized and functionalized using OPE (f PVP SeNPs) and polysorbate stabilized and functionalized using OPE (f PS SeNPs) were investigated. SeNPs showed lower toxicity on human hepatocellular carcinoma (HepG2) and human colorectal adenocarcinoma (Caco2) cells compared to selenite. Functionalization with polyphenols significantly improved their direct antiradical (f PVP SeNPs: 24.4 ± 1.84 and f PS SeNPs: 30.9 ± 2.47 mg TE/mmol Se) and reducing properties (f PVP SeNPs: 50 ± 3.16 and f PS SeNPs: 53.6 ± 3.22 mg GAE/mmol) compared to non-functionalized SeNPs. The significant impact of tested SeNPs on intracellular antioxidative mechanisms has been observed and it was dependent on both cell type and physico-chemical properties of SeNPs, indicating the complexity of involved mechanisms.

Phyto-Assisted Synthesis of Nanoselenium-Surface Modification and Stabilization by Polyphenols and Pectins Derived from Agricultural Wastes.

Raw and purified mandarin peel-derived pectins were characterized and combined with olive pomace extract (OPE) in the green synthesis of selenium nanoparticles (SeNPs). SeNPs were characterized in terms of size distribution and zeta potential, and their stability was monitored during 30 days of storage. HepG2 and Caco-2 cell models were used for the assessment of biocompatibility, while antioxidant activity was investigated by the combination of chemical and cellular-based assays. SeNP average diameters ranged from 171.3 nm up to 216.9 nm; smaller SeNPs were obtained by the utilization of purified pectins, and functionalization with OPE slightly increased the average. At concentrations of 15 mg/L SeNPs were found to be biocompatible, and their toxicity was significantly lower in comparison to inorganic selenium forms. Functionalization of SeNPs with OPE increased their antioxidant activity in chemical models. The effect was not clear in cell-based models, even though all investigated SeNPs improved cell viability and protected intracellular reduced GSH under induced oxidative stress conditions in both investigated cell lines. Exposure of cell lines to SeNPs did not prevent ROS formation after exposure to prooxidant, probably due to low transepithelial permeability. Future studies should focus on further improving the bioavailability/permeability of SeNPs and enhancing the utilization of easily available secondary raw materials in the process of phyto-mediated SeNP synthesis.

Interaction of Differently Coated Silver Nanoparticles With Skin and Oral Mucosal Cells.

Silver nanoparticles (AgNP) can be found in different consumer products and various medical devices due to their excellent biocidal properties. Despite extensive scientific literature reporting biological effects of AgNP, there is still a lack of scientific evidence on how different surface functionalization affects AgNP interaction with the human skin and the oral epithelium. This study aimed to investigate biological consequences following the treatment of HaCaT and TR146 cells with AgNP stabilized with negatively charged sodium bis(2-ethylhexyl)-sulfosuccinate (AOT), neutral polyvinylpyrrolidone (PVP), and positively charged poly-l-lysine (PLL). All AgNP were characterized by means of size, shape and surface charge. Interactions with biological barriers were investigated in vitro by determining cell viability, particle uptake, oxidative stress response and DNA damages following AgNP treatment. Results showed a significant difference in cytotoxicity depending on the surface coating used for AgNP stabilization. All three types of AgNP induced apoptosis, oxidative stress response and DNA damages in cells, but AOT- and PVP-coated AgNP exhibited lower toxicity than positively charged PLL-AgNP. Considering the number of data gaps related to the safe use of nanomaterials in biomedicine, this study highlights the importance of particle surface functionalization that should be considered during design and development of future AgNP-based medical products.

Response of platelets to silver nanoparticles designed with different surface functionalization.

Despite increasing use of silver nanoparticles (AgNPs) in different medicinal products, knowledge about their effects on hemostasis and platelets functionality is still scarce. Published scientific reports provide neither data on oxidative stress response of platelets to AgNPs nor information about the effects of AgNPs physicochemical properties on functionality and activation of platelets. This study aimed to explore the role of AgNPs surface functionalization on cell viability, particle uptake, oxidative stress response, and activation of platelets. Small sized, spherical AgNPs were surface functionalized by negatively charged sodium bis(2-ethylhexyl) sulphosuccinate (AOT), neutral polymer polyvinylpyrrolidone (PVP), positively charged polymer poly-l-lysine (PLL) and bovine serum albumin (BSA). Platelet viability, activation and particle uptake were evaluated by flow cytometry. Oxidative stress response was evaluated by measuring the levels of intracellular glutathione (GSH), peroxy and superoxide radicals using assays based on fluorescence dies. Cytotoxicity and uptake of AgNPs to platelets were found to be dose-dependent in a following order PLL-AgNP >> > BSA-AgNP > AOT-AgNP > PVP-AgNP. Particle internalization was further confirmed by transmission electron microscopy. Treatment of platelets with AgNPs induced superoxide radical formation, depletion of GSH and hyperpolarization of the mitochondrial membrane. Small, but statistically significant increase of P-selectin expression in cells treated with all AgNPs compared to non-treated controls evidenced AgNPs-induced activation of platelets. Increased PAC-1 expression was found only in platelets treated with PLL-AgNPs. Obtained results demonstrate that different surface decoration of AgNPs determines their biological effects on platelets highlighting the importance of careful design of AgNPs-based medicinal products regarding their biocompatibility and functionality.

Antifungal activities of silver and selenium nanoparticles stabilized with different surface coating agents.

BACKGROUND: Extensive and growing use of different chemical pesticides that affect both the environment and human health raises a need for new and more suitable methods to deal with plant pathogens. Nanotechnology has enabled the use of materials at the nanoscale with exceptional functionality in different economic domains including agricultural production. This study aimed to evaluate antifungal potential of selenium nanoparticles (SeNPs) and silver nanoparticles (AgNPs) stabilized with different surface coatings and characterized by different surface charge on plant pathogenic fungi Macrophomina phaseolina, Sclerotinia sclerotiorum and Diaporthe longicolla. RESULTS: AgNPs were coated with three different stabilizing agents: mono citrate (MC-AgNPs), cetyltrimethyl ammonium bromide (CTAB-AgNPs) and polyvinylpyrrolidon (PVP-AgNPs). SeNPs were coated with poly-l-lysine (PLL-SeNPs), polyacrylic acid (PAA-SeNPs), and polyvinylpyrrolidon (PVP-SeNPs). Seven different concentrations (0.1, 0.5, 1, 5, 10, 50 and 100 mg L-1 ) of nanoparticles were applied. All AgNPs and SeNPs significantly inhibited the growth of the tested fungi. Among the tested NPs, PVP-AgNPs showed the best inhibitory effect on the tested plant pathogenic fungi, especially against S. sclerotiorum. The similar inhibition of the sclerotia formation was observed for S. sclerotiorum treated with PLL-SeNPs. CONCLUSION: Obtained results provides new insights on fungicide effect of AgNPs and SeNPs stabilized with different coating agents on different plant pathogens. Further work should focus on detailed risk/benefit ratio assessment of using SeNPs or AgNPs in agriculture taking into account whole agroecosystem. © 2020 Society of Chemical Industry.

Impact of surface functionalization on the toxicity and antimicrobial effects of selenium nanoparticles considering different routes of entry.

Selenium nanoparticles (SeNPs) were first designed as nutritional supplements, but they are attractive also for use in diagnostic and therapeutic systems owing to their biocompatibility and protective effects. This study aimed to examine if different SeNPs stabilization strategies affect their (i) antimicrobial activity against bacteria Escherichia coli and Staphylococcus aureus and yeast Saccharomyces cerevisiae and (ii) toxicity to human cells of different biological barriers i.e., skin, oral and intestinal mucosa. For surface stabilization, polyvinylpyrrolidone (PVP), poly-L-lysine (PLL) and polyacrylic acid (PAA) were used rendering neutral, positively and negatively charged SeNPs, respectively. The SeNPs (primary size ~80 nm) showed toxic effects in human cells in vitro and in bacteria S. aureus, but not in E. coli and yeast S. cerevisiae. Toxicity of SeNPs (24 h IC50) ranged from 1.4 to >100 mg Se/L, depending on surface functionalization (PLL > PAA > PVP) and was not caused by ionic Se. At subtoxic concentrations, all SeNPs were taken up by all human cell types, induced oxidative stress response and demonstrated genotoxicity. As the safety profile of SeNPs was dependent not only on target cells (mammalian cells, bacteria, yeast), but also on surface functionalization, these aspects should be considered during development of novel SeNPs-based biomedical products.

Synergistic effects of parabens and plastic nanoparticles on proliferation of human breast cancer cells.

Many personal care products on the market contain endocrine disrupting chemicals, including parabens. Parabens are well known chemical additives used as preservatives. They have been found in mammary glands and breast cancer tissues. At the same time, the general public is increasingly exposed to plastic micro- and nanoparticles generated during plastic production and waste disposal. Exposure to chemical cocktails is a realistic scenario of high public health interest, in which many types of compounds such as these two may exhibit synergistic or additive adverse effects. This study evaluated the effects of plastic nanoparticles, parabens, and their mixture on the viability and proliferation of two human breast cancer cell lines: MDA-MB 231, which lacks oestrogen receptors, and MCF-7, which expresses these receptors. Parabens increased proliferation of oestrogen-sensitive breast cancer cells, and this effect became synergistic in the presence of plastic nanoparticles. The mechanism behind synergy may be related to the translocation and adsorption properties of nanoplastics, which served as a Trojan horse to expose cells to parabens more efficiently. These preliminary findings support growing evidence warning about the urgent problem of human exposure to combinations of plastic waste and contingent chemicals.