Carbon nanoparticles downregulate expression of basic fibroblast growth factor in the heart during embryogenesis

Mateusz Wierzbicki,1 Ewa Sawosz,1 Marta Grodzik,1 Anna Hotowy,1 Marta Prasek,1 Slawomir Jaworski,1 Filip Sawosz,2 André Chwalibog2 1Division of Nanobiotechnology, Warsaw University of Life Sciences, Warsaw, Poland; 2Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Copenhagen, Denmark Abstract: Carbon nanoparticles, with their high biocompatibility and low toxicity, have recently been considered for biomedical applications, including antiangiogenic therapy. Critical to normal development and tumor formation, angiogenesis is the process of forming capillary blood vessels from preexisting vessels. In the present study, we evaluated the effects of diamond and graphite nanoparticles on the development of chicken embryos, as well as vascularization of the chorioallantoic membrane and heart at the morphological and molecular level. Nanoparticles did not affect either body/heart weight or serum indices of the embryos’ health. However, vascularization of the heart and the density of branched vessels were significantly reduced after treatment with diamond nanoparticles and, to a lesser extent, graphite nanoparticles. Application of nanoparticles significantly downregulated gene and protein expression of the proangiogenic basic fibroblast growth factor, indicating that both diamond and graphite nanoparticles inhibit angiogenesis. Keywords: diamond, graphite, nanoparticles, vasculogenesis, bFGF, VEG

Nanoparticles of carbon allotropes inhibit glioblastoma multiforme angiogenesis in ovo

Marta Grodzik1, Ewa Sawosz1, Mateusz Wierzbicki1, Piotr Orlowski1, Anna Hotowy2, Tomasz Niemiec1, Maciej Szmidt3, Katarzyna Mitura4, André Chwalibog21Division of Biotechnology and Biochemistry of Nutrition, Warsaw University of Life Sciences, Warsaw, Poland; 2Department of Basic Animal and Veterinary Science, University of Copenhagen, Copenhagen, Denmark; 3Division of Histology and Embryology, Warsaw University of Life Sciences, Warsaw, Poland; 4Department of Biomedical Engineering, Koszalin University of Technology, Koszalin, PolandAbstract: The objective of the study was to determine the effect of carbon nanoparticles produced by different methods on the growth of brain tumor and the development of blood vessels. Glioblastoma multiforme cells were cultured on the chorioallantoic membrane of chicken embryo and after 7 days of incubation, were treated with carbon nanoparticles administered in ovo to the tumor. Both types of nanoparticles significantly decreased tumor mass and volume, and vessel area. Quantitative real-time polymerase chain reaction analysis showed downregulated fibroblast growth factor-2 and vascular endothelial growth factor expression at the messenger ribonucleic acid level. The present results demonstrate antiangiogenic activity of carbon nanoparticles, making them potential factors for anticancer therapy.Keywords: cancer, nanoparticle, embryo, angiogenesis, FGF-2, VEG

Toxicity studies of six types of carbon nanoparticles in a chicken-embryo model

Natalia Kurantowicz,1 Ewa Sawosz,1 Gabriela Halik,1 Barbara Strojny,1 Anna Hotowy,1 Marta Grodzik,1 Radosław Piast,2 Wanvimol Pasanphan,3 André Chwalibog4 1Department of Animal Nutrition and Biotechnology, Warsaw University of Life Sciences, 2Faculty of Chemistry, Warsaw University, Warsaw, Poland; 3Department of Materials Science, Faculty of Science, Kasetsart University, Bangkok, Thailand; 4Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Copenhagen, Denmark Abstract: In the present study, the toxicity of six different types of carbon nanoparticles (CNPs) was investigated using a chicken-embryo model. Fertilized chicken eggs were divided into the following treatment groups: placebo, diamond NPs, graphite NPs, pristine graphene, small graphene oxide, large graphene oxide, and reduced graphene oxide. Experimental solutions at a concentration of 500 µg/mL were administrated into the egg albumin. Gross pathology and the rate of survival were examined after 5, 10, 15, and 20 days of incubation. After 20 days of incubation, blood samples were collected and the weight of the body and organs measured. The relative ratio of embryo survival decreased after treatment all treatments except diamond NPs. There was no correlation between the rate of survival and the ζ-potential or the surface charge of the CNPs in solution. Body and organ weight, red blood-cell morphology, blood serum biochemical parameters, and oxidative damage in the liver did not differ among the groups. These results indicate that CNPs can remain in blood circulation without any major side effects, suggesting their potential applicability as vehicles for drug delivery or active compounds per se. However, there is a need for further investigation of their properties, which vary depending on production methods and surface functionalization. Keywords: nanoparticles, diamond, graphite, graphene, toxicity, red blood cells, oxidative stress, surface charg

Effect of heparan sulfate and gold nanoparticles on muscle development during embryogenesis

Marlena Zielinska1,2, Ewa Sawosz1, Marta Grodzik1, Mateusz Wierzbicki1, Maria Gromadka1, Anna Hotowy3, Filip Sawosz3, Andrzej Lozicki1, Andrè Chwalibog31Division of Biotechnology and Biochemistry of Nutrition, Warsaw University of Life sciences, Warsaw, 2The Kielanowski Institute of Animal Physiology and Nutrition, Jablonna, Poland; 3Department of Basic Animal and Veterinary sciences, University of copenhagen, Frederiksberg, DenmarkPurpose: It was hypothesized that heparan sulfate (HS) as an essential compound for myogenesis and nanoparticles of gold (nano-Au) as highly reactive compounds can affect muscle development as a consequence of molecular regulation of muscle cell formation, and that these effects may be enhanced by a complex of HS conjugated with nano-Au. The objective of the present study was to determine the effect of administration of nano-Au, HS, and a nano-Au+HS complex on the morphological and molecular characteristics of breast muscle during embryogenesis.Methods: Chicken embryos were used as in vivo model. Fertilized chicken eggs (n = 350) were randomly divided into the control group and the groups treated with nano-Au, HS, and nano-Au+HS. The experimental solutions were given in ovo on the first day of incubation and the embryos were evaluated on day 20 of incubation. The methods included biochemical indi- ces in blood, immunohistochemistry, microscopy (transmission electron microscopy, scanning electron microscopy, confocal), and gene expression at the messenger ribonucleic acid and protein levels.Results: The treatments did not adversely affect mortality, organ weight, and homeostasis of the embryos. HS stimulated the development and maturation of breast muscle by increasing the number of nuclei, satellite cells, and muscle fibers and affected the expression of basic fibroblast growth factor-2 and paired-box transcription factor-7. Furthermore, the nano-Au+HS complex contributed to the increased number of myocytes and nuclei in chicken embryo muscles.Conclusion: The results indicate that the administration of HS and nano-Au affects muscle development and that this effect is enhanced by conjugating HS with nano-Au.Keywords: gold nanoparticles, heparan sulfate, muscle development, embryogenesis, chicke

Relations between pig growth and regulatory mechanism of pancreas - facts and hypotheses

The aim of the mini review is to more precisely determine the relationship between exocrine pancreas function and the growth of young pigs. In order to do so, it is necessary to clarify the components, pathways and messengers involved in the CCK-dependent neuro-hormonal regulation of the exocrine pancreas in the growing pigs. These mechanisms are largely unknown, and the role of CCK has been questioned since CCK receptors on the enzyme producing acinar cells are lacking. Our hitherto obtained results have shown the importance of CCK for the pancreatic enzyme secretion and that in pigs CCK functions via CCK-B receptor-mediated entero(duodenal)-pancreatic reflex and via CCK-A receptor mechanism which localization is unknown. Preliminary studies have indicated this later mechanism as being related to the growth performance of pigs. The planned studies will comprise a pharmacologic characterization and an immunohistologic localization of the components and messengers involved in the CCK-dependent neuro-hormonal pancreatic regulation using an appropriately catheterized pig model. Since the applied aim of the project is to correlate the growth of the pigs to the effectiveness of the pancreatic secretion and to the development of its regulatory mechanisms, the experiments will be performed on pigs having different growth and performance. It is expected that the discussion will provide important applicable understanding of pancreatic function and its impact on pig growth and performance and also highlight the mechanism of the inter-organ regulatory pathways of the exocrine pancreatic function

Indukcja metaloproteinazy-3 macierzy pozakomórkowej za pomocą terapii fotodynamicznej z zastosowaniem liposomowych preparatów kwasu aminolewulinowego i jego estru metylowego

Photodynamic therapy (PDT) showed promising results in treatment of malignant and non-malignant disorders. The PDT requires for a therapeutic effect the combined action of photosensitizer and light. PDT causes direct cytotoxicity to malignant cells and may also have both direct and indirect effects upon various non-malignant components of tumor microenvironment. Unfortunately, some photosensitizers reveal low selectivity in pathologic tissues. Previous studies indicated that aminolevulinic acid (ALA) and its methyl ester (metvix) encapsulated in liposomes improved the quality and optimized results of PDT. Matrix metalloproteinases (MMPs) are enzymes implicated in various diseases by enabling the spread of disease. MMP-3 in turn, exerts the protective role in the development of tumors. We report the effect of liposomal formulation of ALA and metvix-based photodynamic therapy on induction of matrix metalloproteinase 3 in animal tumor model. Our results showed strong expression of MMP-3 in tumor-bearing rat tumor cells after PDT with liposomal formulations. The expression of MMP-3 in all tumor-bearing rats treated with PDT was stronger than those observed in animals free of tumors and especially in those treated with free precursor-PDT. The effect of liposomes was found to be important and these formulations elicited stronger intensity of immunohistochemical reactions than ALA- or metvix –PDT without liposomes.Terapia fotodynamiczna (photodynamic therapy – PDT) daje obiecujące wyniki w leczeniu złośliwych i niezłośliwych chorób. PDT wymaga połączonego działania fotouczulacza i światła dla uzyskania efektu terapeutycznego. PDT wywołuje efekt cytotoksyczny w patologicznych komórkach. Może także działać bezpośrednio i pośrednio na różne składniki mikrośrodowiska nowotworu. Niestety, niektóre fotouczulacze mają niską selektywność w tkankach chorobowych. Poprzednie badania wykazały, że kwas aminolewulinowy (ALA) i jego ester metylowy (metvix) umieszczone w liposomach poprawiały jakość i wyniki PDT. Metaloproteinazy macierzy (MMPs) są enzymami wprzęgniętymi w różne choroby umożliwiającymi ich szerzenie się. Z kolei MMP-3 odgrywa rolę ochronną w rozwoju nowotworów. W pracy przedstawiono wpływ działania preparatów liposomowych ALA i metviksu w warunkach PDT na indukcję MMP-3 w zwierzęcym modelu nowotworowym. Uzyskane wyniki wskazują na silną ekspresję MMP- 3 w komórkach nowotworu po PDT z użyciem preparatów liposomowych. Ekspresja MMP-3 u wszystkich zwierząt zaszczepionych nowotworem i traktowanych PDT była silniejsza niż u zwierząt wolnych od nowotworu i szczególnie od tych leczonych PDT z użyciem wolnych prekursorów. Wpływ liposomów okazał się być istotny i przyczyniał się do silniejszej reakcji immunohistochemicznej niż PDT bez tych nośników