Silver nanoparticles (AgNP) can migrate to tissues and cells of the body, as well as to agglomerate, which reduces the effectiveness of their use for the antimicrobial protection of the skin. Graphene oxide (GO), with a super-thin flake structure, can be a carrier of AgNP that stabilizes their movement without inhibiting their antibacterial properties. Considering that the human skin is often the first contact with antimicrobial agent, the aim of the study was to assess whether the application of the complex of AgNP and GO is biocompatible with the skin model in in vitro studies. The conducted tests were performed in accordance with the criteria set in OECD TG439. AgNP-GO complex did not influence the genotoxicity and metabolism of the tissue. Furthermore, the complex reduced the pro-inflammatory properties of AgNP by reducing expression of IP-10 (interferon gamma-induced protein 10), IL-3 (interleukin 3), and IL-4 (interleukin 4) as well as MIP1β (macrophage inflammatory protein 1β) expressed in the GO group. Moreover, it showed a positive effect on the micro- and ultra-structure of the skin model. In conclusion, the synergistic effect of AgNP and GO as a complex can activate the process of epidermis renewal, which makes it suitable for use as a material for skin contact.

Finding an effective muscle regeneration technique is a priority for regenerative medicine. It is known that the key factors determining tissue formation include cells, capable of proliferating and/or differentiating, a niche (surface) allowing their colonization and growth factors. The interaction between these factors, especially between the surface of the artificial niche and growth factors, is not entirely clear. Moreover, it seems that the use of a complex of complementary growth factors instead of a few strictly defined ones could increase the effectiveness of tissue maturation, including muscle tissue. In this study, we evaluated whether graphene oxide (GO) nanofilm, chicken embryo muscle extract (CEME), and GO combined with CEME would affect the differentiation and functional maturation of muscle precursor cells, as well as the ability to spontaneously contract a pseudo-tissue muscle. CEME was extracted on day 18 of embryogenesis. Muscle cells obtained from an 8-day-old chicken embryo limb bud were treated with GO and CEME. Cell morphology and differentiation were observed using different microscopy methods. Cytotoxicity and viability of cells were measured by lactate dehydrogenase and Vybrant Cell Proliferation assays. Gene expression of myogenic regulatory genes was measured by Real-Time PCR. Our results demonstrate that CEME, independent of the culture surface, was the main factor influencing the intense differentiation of muscle progenitor cells. The present results, for the first time, clearly demonstrated that the cultured tissue-like structure was capable of inducing contractions without externally applied impulses. It has been indicated that a small amount of CEME in media (about 1%) allows the culture of pseudo-tissue muscle capable of spontaneous contraction. The study showed that the graphene oxide may be used as a niche for differentiating muscle cells, but the decisive influence on the maturation of muscle tissue, especially muscle contractions, depends on the complexity of the applied growth factors.

Background: Bacterial skin infections, including Staphylococcus aureus, are a powerful and still not fully resolved problem. The aim of this research was to determine the possibility of using a complex of graphene oxide (GO) encrusted with silver nanoparticles as an effective antibacterial agent against S. aureus and to assess its pro-inflammatory properties. Methods: The tests were carried out in vitro on EpiDerm™ Skin, an artificial skin model (MatTek in vitro Life Science Laboratories, Slovak Republic), and the fibroblast cell line (HFF-2 from ATCC, USA). Both models were infected with S. aureus bacteria (ATCC 25923) and then treated with antibiotics or our experimental factors: silver nanoparticles (AgNPs, Nano-koloid, Poland), graphene oxide (GO, NanoPoz, Poland), and complex AgNP-GO (hydrocolloid created by self-assembly). Results: The antibacterial effectiveness of the AgNP-GO complex was equivalent to that of the antibiotic. In addition, an increase in the level of pro-inflammatory cytokines was observed under the influence of antibiotic administration, in contrast to the effect of AgNP-GO, which showed very limited pro-inflammatory activity. Conclusion: Hydrocolloid of the AgNP-GO complex, administered in the form of a liquid dressing, may act as an antibacterial agent and also reduce inflammation induced by S. aureus infection.

Pancreatic cancer, due to its asymptomatic development and drug-resistance, is difficult to cure. As many metallic and carbon-based nanomaterials have shown anticancer properties, we decided to investigate their potential use as anticancer agents against human pancreatic adenocarcinoma. The objective of the study was to evaluate the toxic properties of the following nanomaterials: silver (Ag), gold (Au), platinum (Pt), graphene oxide (GO), diamond (ND), and fullerenol (C₆₀(OH)₄₀) against the cell lines BxPC-3, AsPC-1, HFFF-2, and HS-5. The potential cytotoxic properties were evaluated by the assessment of the cell morphology, cell viability, and cell membrane damage. The cancer cell responses to GO and ND were analysed by determination of changes in the levels of 40 different pro-inflammatory proteins. Our studies revealed that the highest cytotoxicity was obtained after the ND treatment. Moreover, BxPC-3 cells were more sensitive to ND than AsPC-1 cells due to the ND-induced ROS production. Furthermore, in both of the cancer cell lines, ND caused an increased level of IL-8 and a decreased level of TIMP-2, whereas GO caused only decreased levels of TIMP-2 and ICAM-1 proteins. This work provides important data on the toxicity of various nanoparticles against pancreatic adenocarcinoma cell lines.

The effects of CEME and it complex with GO injected in ovo on the growth and development of chicken embryo hindlimb muscle were investigated. First, the preliminary in vitro study on primary muscle precursor cell culture obtained from a nine-day-old chicken embryo was performed to assess toxicity (MTT assay) of CEME, GO (100 ppm) and it complex with different concentrations (1, 2, 5, and 10 wt.%). The effect on cell proliferation was investigated by BrdU assay. CEME at concentrations 1–5% increased cell proliferation, but not the complex with GO. In vitro cytotoxicity was highest in 10% and GO groups. Next, the main experiment with chicken embryos was performed with CEME, GO and it complex injected in ovo on day one of embryogenesis. On day 20 of embryogenesis survival, morphological development, histological structure of the muscle, and biochemical parameters of blood serum of the embryos were measured. No negative effect on mortality, body weight, or biochemistry of blood after use of CEME or GO-CEME complexes was observed. Interestingly, the slight toxicity of GO, observed in in vitro studies, was not observed in vivo. The use of CEME at the levels of 2% and 5% improved the structure of the lower limb muscle by increasing the number of cells, and the administration of 2% CEME increased the number of nuclei visible in the stained cross-section of the muscle. The complex GO-CEME did not further improve the muscle structure. The results indicate that CEME can be applied as an in ovo enhancer of muscle development in broilers.

The purpose of this study was to determine the nutritional suitability of WDGS in pigs’ feeding and production. Pigs were liquid fed and divided into 3 groups. Pigs in the control group were fed diets based on cereal grains, while the experimental groups were also given 10% or 15% WDGS, which partially replaced their cereal grains. During this study, the average daily gains (ADG), feed intake, chemical composition of meat, fatty acid profile of meat, and quality parameters of the carcass and meat were examined. The highest statistical weight gains were detected for the group WDGS 10% during the first stage of the fattening period. No statistical differences were detected for the final body weight, carcass traits, chemical composition of the meat or the composition of fatty acids such as SFAs, PUFAs, and MUFAs, with the exception of eicosenoic acid (C20:1n9). Pigs fed on 10% WDGS exhibited lower peroxidation of lipids (TBARS) than the control group or WDGS 15%. Similarly, water holding capacity (WHC) was the lowest for the group WDGS 10%. Of the meat coloration, redness (a*), yellowness (b*), and chroma (C*) were affected by the WDGS’ inclusion, where the highest values were observed for the group WDGS 10%. In conclusion, WDGS can be utilized in the liquid feeding of pigs for up to 15% of their DM.