Joanna Gromadzka-Ostrowska

Time‐dependent biodistribution and excretion of silver nanoparticles in male Wistar rats

Silver nanoparticles (AgNPs) are the most commonly used nanoparticles owing to their antimicrobial properties. The motivation of the present study was (1) to analyze the effect of silver particle size on rat tissue distribution at different time points, (2) to determine the accumulation of AgNPs in potential rat target organs, (3) to analyze the intracellular distribution of AgNPs and (4) to examine the excretion of AgNPs by urine and feces. AgNPs were characterized by dynamic light scattering (DLS), zeta potential measurements, BET surface area measurements, transmission and scanning electron microscopy. AgNPs (20 and 200 nm) were administered intravenously (i.v.) to male Wistar rats at a dose of 5 mg kg–1 of body weight. Biological material was sampled 24 h, 7 and 28 days after injection. Using inductively coupled plasma‐mass spectrometry (ICP‐MS) and transmission electron microscopy (TEM) it was observed that AgNPs translocated from the blood to the main organs and the concentration of silver in tissues was significantly higher in rats treated with 20 nm AgNPs as compared with 200 nm AgNPs. The highest concentration of silver was found in the liver after 24 h. After 7 days, a high level of silver was observed in the lungs and spleen. The silver concentration in the kidneys and brain increased during the experiment and reached the highest concentration after 28 days. Moreover, the highest concentration of AgNPs was observed in the urine 1 day after the injection, maintained high for 14 days and then decreased. The fecal level of silver in rats was the highest within 2 days after AgNPs administration and then decreased. Copyright

Silver nanoparticles effects on epididymal sperm in rats.

The motivation of our study was to examine the acute effects of intravenously administered a single bolus dose of silver nanoparticles (AgNPs) on rat spermatogenesis and seminiferous tubules morphology. In the treated rats compared to the vehicle treated control animals, the experiments revealed a size-dependent (20nm and 200nm), dose-dependent (5 and 10mg/kg body mass) and time-dependent (24h, 7 and 28days) decrease the epididymal sperm count measured by histological methods. In parallel AgNPs injection increased the level of DNA damage in germ cells, as measured by alkaline comet assay. Histological examination of the testes showed change in the testes seminiferous tubule morphometry in 200nm Ag NPs treated rats. No change of body weight, adipose tissue distribution and the frequency of abnormal spermatozoa was observed. Twenty nanometers AgNP appeared to be more toxic than 200nm ones.

Composition differences between organic and conventional meat: a systematic literature review and meta-analysis

Demand for organic meat is partially driven by consumer perceptions that organic foods are more nutritious than non-organic foods. However, there have been no systematic reviews comparing specifically the nutrient content of organic and conventionally produced meat. In this study, we report results of a meta-analysis based on sixty-seven published studies comparing the composition of organic and non-organic meat products. For many nutritionally relevant compounds (e.g. minerals, antioxidants and most individual fatty acids (FA)), the evidence base was too weak for meaningful meta-analyses. However, significant differences in FA profiles were detected when data from all livestock species were pooled. Concentrations of SFA and MUFA were similar or slightly lower, respectively, in organic compared with conventional meat. Larger differences were detected for total PUFA and n-3 PUFA, which were an estimated 23 (95 % CI 11, 35) % and 47 (95 % CI 10, 84) % higher in organic meat, respectively. However, for these and many other composition parameters, for which meta-analyses found significant differences, heterogeneity was high, and this could be explained by differences between animal species/meat types. Evidence from controlled experimental studies indicates that the high grazing/forage-based diets prescribed under organic farming standards may be the main reason for differences in FA profiles. Further studies are required to enable meta-analyses for a wider range of parameters (e.g. antioxidant, vitamin and mineral concentrations) and to improve both precision and consistency of results for FA profiles for all species. Potential impacts of composition differences on human health are discussed.

Chapter Five - Toxicity of Silver Nanomaterials in Higher Eukaryotes

Abstract The rapid expansion of nanotechnology promises to have significant benefits to society, yet there is increasing concern that exposure to nanoparticles (particles typically in vitro and in vivo uptake, biodistribution, and toxicity of AgNPs. Emphasis is placed on the systematization of data over animal and cell models, organs examined, doses applied, the type of particle administration, and the time of examination.

Genotoxicity and gene expression modulation of silver and titanium dioxide nanoparticles in mice.

Abstract Recently, we showed that silver nanoparticles (AgNPs) caused apoptosis, necrosis and DNA strand breaks in different cell models in vitro. These findings warranted analyses of their relevance in vivo. We investigated the genotoxic potential and gene expression profiles of silver particles of nano- (Ag20, 20 nm) and submicron- (Ag200, 200 nm) size and titanium dioxide nanoparticles (TiO2-NPs, 21 nm) in selected tissues from exposed male mice including the gonades. A single dose of 5 mg/kg bw nanoparticles was administered intravenously to male mice derived from C57BL6 (WT) and 8-oxoguanine DNA glycosylase knock-out (Ogg1−/− KO). Testis, lung and liver were harvested one and seven days post-exposure and analyzed for DNA strand breaks and oxidized purines employing the Comet assay with Formamidopyrimidine DNA glycosylase (Fpg) treatment, and sperm DNA fragmentation by the sperm chromatin structure assay (SCSA). Based on an initial screening of a panel of 21 genes, seven genes were selected and their expression levels were analyzed in all lung and testis tissues sampled from all animals (n = 6 mice/treatment group) using qPCR. AgNPs, in particular Ag200, caused significantly increased levels of DNA strand breaks and alkali labile sites in lung, seven days post-exposure. Fpg-sensitive lesions were significantly induced in both testis and lung. The transcript level of some key genes; Atm, Rad51, Sod1, Fos and Mmp3, were significantly induced compared to controls, particularly in lung samples from Ag200-exposed KO mice. We conclude that the Ag200 causes genotoxicity and distinct gene expression patterns in selected DNA damage response and repair related genes.

Effect of crop protection and fertilization regimes used in organic and conventional production systems on feed composition and physiological parameters in rats.

Very little is known about the effects of an organic or conventional diet on animal physiology and health. Here, we report the effect of contrasting crop protection (with or without chemosynthetic pesticides) and fertilization (manure or mineral fertilizers) regimes on feed composition and growth and the physiological parameters of rats. The use of manure instead of mineral fertilizers in feed production resulted in lower concentrations of protein (18.8 vs 20.6%) and cadmium (3.33 vs 4.92 μg/100 g) but higher concentrations of polyphenols (1.46 vs 0.89 g/100 g) in feeds and higher body protein (22.0 vs 21.5%), body ash (3.59 vs 3.51%), white blood cell count (10.86 vs 8.19 × 10³/mm³), plasma glucose (7.23 vs 6.22 mmol/L), leptin (3.56 vs 2.78 ng/mL), insulin-like growth factor 1 (1.87 vs 1.28 μg/mL), corticosterone (247 vs 209 ng/mL), and spontaneous lymphocyte proliferation (11.14 vs 5.03 × 10³ cpm) but lower plasma testosterone (1.07 vs 1.97 ng/mL) and mitogen stimulated proliferation of lymphocytes (182 vs 278 × 10³ cpm) in rats. There were no main effects of crop protection, but a range of significant interactions between fertilization and crop protection occurred.

Effects of high-fat diets on body composition, hypothalamus NPY, and plasma leptin and corticosterone levels in rats

We investigated the adipogenic effects of high-fat diets with different fat sources, as well as their influence on hypothalamus NPY content (NPY) and plasma corticosterone (Cs) and leptin (Lep) concentrations. In a 6-wk experiment performed in 28 male Wistar rats, high-fat diets (approx 60% of energy as fat) containing sunflower oil (S), rapeseed oil (R), palm oil (P), or lard (L) as the fat source were applied. Carcass composition was analyzed by standard methods; NPY, Cs, and Lep were determined by RIA method. Gas chromatography was applied to measure fatty acid composition of dietary fats. S group had the highest body fat content and Lep and the lowest Cs and NPY, while L rats had high Lep and the highest Cs and NPY. We conclude that the fatty acid composition of dietary fat is of high importance where effects of high fat diets on adiposity, as well as on plasma levels of both hormones and hypothalamus NPY content are concerned.

Hepato- and gastro- protective activity of purified oat 1-3, 1-4-β-d-glucans of different molecular weight.

Protective and antioxidant properties of highly purified oat β-glucans of high and low molecular weight in liver and stomach were evaluated. The novelty in approach was to determine whether dietary β-glucans affect the parameters of oxidative stress directly in the stomach and indirectly in the liver, especially in inflammation states. Physicochemical properties e.g. viscoelastic was found as strictly dependent from molecular weight of oat β-glucans hence its metabolic activity could also show dependence. Three groups of rats were fed control diet and diet supplemented with low and high molecular weights oat β-glucans. Animals were divided into controls and individuals with experimentally induced intestinal inflammation. Most active in increasing of total antioxidant status was low molecular weight β-glucan. High molecular weight β-glucan supplementation inhibits lipid oxidation the most in LPS treated animals. The results obtained from experiment encourage for dietary intervention with oat β-glucans for stomach and liver protection during existing enteritis.

Progressive effects of silver nanoparticles on hormonal regulation of reproduction in male rats.

The growing use of silver nanoparticles (AgNPs) in various applications, including consumer, agriculture and medicine products, has raised many concerns about the potential risks of nanoparticles (NPs) to human health and the environment. An increasing body of evidence suggests that AgNPs may have adverse effects of humans, thus the aim of this study was to investigate the effects of AgNPs on the male reproductive system. Silver particles (20nm AgNPs (groups Ag I and Ag II) and 200nm Ag sub-micron particles (SPs) (group Ag III)) were administered intravenously to male Wistar rats at a dose of 5 (groups Ag I and Ag III) or 10 (group Ag II) mg/kg of body weight. The biological material was sampled 24h, 7days and 28days after injection. The obtained results revealed that the AgNPs had altered the luteinising hormone concentration in the plasma and the sex hormone concentration in the plasma and testes. Plasma and intratesticular levels of testosterone and dihydrotestosterone were significantly decreased both 7 and 28days after treatment. No change in the prolactin and sex hormone-binding globulin concentration was observed. Exposure of the animals to AgNPs resulted in a considerable decrease in 5α-reductase type 1 and the aromatase protein level in the testis. Additionally, expression analysis of genes involved in steroidogenesis and the steroids metabolism revealed significant down-regulation of Star, Cyp11a1, Hsd3b1, Hsd17b3 and Srd5a1 mRNAs in AgNPs/AgSPs-exposed animals. The present study demonstrates the potential adverse effect on the hormonal regulation of the male reproductive function following AgNP/AgSP administration, in particular alterations of the sex steroid balance and expression of genes involved in steroidogenesis and the steroids metabolism.

Modifications of Western-type diet regarding protein, fat and sucrose levels as modulators of steroid metabolism and activity in liver

The aim of the study was to evaluate whether the modification of the Western-type diet (high-fat, high-sucrose diet rich in saturated fatty acids) considering macronutrients content would influence hepatic metabolism and activity of steroids. For 3 weeks Wistar rat were fed the Western-type diet (21% fat, 35% sucrose, 19% protein, lard) and its modifications regarding dietary protein (10 and 19%), fat (5 and 21%) and sucrose (0 and 35%) levels. The steroid 5α-reductase type 1 (Srd5a1) and androgen receptor (Ar) gene expression as well as testosterone (T) conversion towards 5α-reduced derivatives in liver were positively correlated with body weight gain. The Western-type diets with decreased protein content regardless of the sucrose level exerted the most negative effect on the antioxidant system decreasing catalase (Cat), sodium dismutase (Sod1) and glutathione peroxidase (Gpx1) gene expression as well as Cat and Gpx activity and total antioxidant status, simultaneously intensifying lipid peroxidation. The impaired antioxidant system was accompanied by decreased level of hepatic T metabolism towards estrogens: 17β-estradiol (E2) and estriol, and increased estrogen receptor type 1 (Esr1) gene expression. Liver Esr1 mRNA level was differently correlated with T (positively) and E2 (negatively) plasma levels. Whereas the fat reduction in Western-type diet restored the plasma proportion between T and E2. In conclusion it could be stated that Western-type diet modification relating to protein, sucrose and fat content can influence hepatic steroid metabolism and activity; however the estrogens and androgens metabolism in liver would be connected with impairment of liver function or catabolic activity, respectively.