The aim of this study was to evaluate the pro-osteogenic properties of sol-gel-derived silica/zirconium dioxide coatings functionalized with 1 mM of metformin. The matrices were applied on 316L stainless steel using dip-coating technique. First of all, physicochemical properties of biomaterials were evaluated. Surface morphology and topography was determined using energy-dispersive X-ray spectroscopy and atomic force microscopy. The chemical composition was evaluated using Fourier transform infrared spectroscopy. Further, wettability and surface free energy were characterized. Cytocompatibility of biomaterials was tested in vitro using model of human multipotent mesenchymal stromal cells isolated from adipose tissue. The influence of biomaterials on cells morphology and proliferation was determined. Osteogenic effect of obtained biomaterials was evaluated in terms of their influence on secretory activity of human multipotent mesenchymal stromal cells isolated from adipose tissue and matrix mineralization. Analysis was performed in relation to the control cultures i.e. maintained on pure SS316L substrate and SS316L covered with silica/zirconium dioxide. Obtained results indicate that silica/zirconium dioxide_metformin coatings ameliorated metabolic and proliferative activity of human multipotent mesenchymal stromal cells isolated from adipose tissue, as well as promoted their proper growth and adhesion. The human multipotent mesenchymal stromal cells isolated from adipose tissue cultured on biomaterials were characterized by typical fibroblast-like morphology. The addition of metformin to the silica/zirconium dioxide coatings improved functional differentiation of human multipotent mesenchymal stromal cells isolated from adipose tissue. Osteogenic cultures on silica/zirconium dioxide_metformin were characterized by formation of well-developed osteonodules rich in calcium and phosphorous. Moreover, human multipotent mesenchymal stromal cells isolated from adipose tissue cultured on silica/zirconium dioxide_metformin synthesized increased amount of alkaline phosphatase, bone morphogenetic protein 2 and osteopontin, both on messenger RNA and protein level. Obtained biomaterials modulate cellular plasticity of human multipotent mesenchymal stromal cells isolated from adipose tissue promoting their osteogenic differentiation, thus may find application in broadly defined tissue engineering.
Spirulina platensis (SP) is a blue-green microalga that has recently raised attention not only as a nutritional component, but also as a source of bioactivities that have therapeutic effects and may find application in medicine, including cancer treatment. In the present study we determined the cytotoxic effect of S. platensis filtrates (SPF) on human colon cancer cell line Caco-2. Three concentrations of SPF were tested—1.25%, 2.5%, and 5% (v/v). We have found that the highest concentration of SPF exerts the strongest anti-proliferative and pro-apoptotic effect on Caco-2 cultures. The SPF negatively affected the morphology of Caco-2 causing colony shrinking and significant inhibition of metabolic and proliferative activity of cells. The wound-healing assay showed that the SPF impaired migratory capabilities of Caco-2. This observation was consistent with lowered mRNA levels for metalloproteinases. Furthermore, SPF decreased the transcript level of pro-survival genes (cyclin D1, surviving, and c-Myc) and reduced the autocrine secretion of Wnt-10b. The cytotoxic effect of SPF involved the modulation of the Bax and Bcl-2 ratio and a decrease of mitochondrial activity, and was related with increased levels of intracellular reactive oxygen species (ROS) and nitric oxide (NO). Moreover, the SPF also caused an increased number of cells in the apoptotic sub-G0 phase and up-regulated expression of mir-145, simultaneously decreasing expression of mir-17 and 146. Obtained results indicate that SPF can be considered as an agent with anti-cancer properties that may be used for colon cancer prevention and treatment.
The aim of this work was to assess the effect of different stabilizing agents, for example, polyethylenimine (PEI) and polyvinylpyrrolidone (PVP), on gold nanoparticles (AuNPs) and their influence on equine platelet activation and release of particular growth factors. The gold nanoparticles were produced by chemical reduction of chloroauric acid. UV-Vis spectroscopy confirmed the presence of gold nanoparticles in investigated solutions. The AuNPs were incubated with whole blood at various concentrations. The morphology of platelets in PRP prepared from the blood incubated with AuNPs was characterized by scanning transmission electron microscopy, whereas the concentrations of growth factors and cytokines were evaluated by ELISA assays. The most promising results were obtained with equine platelets incubated with 5% AuNPs stabilized by PEI, which lead to secretion of bone morphogenetic protein 2 (BMP-2), vascular endothelial growth factor (VEGF), and fibroblast growth factor 1 (FGF-1) and simultaneously cause decrease in concentration of interleukin-1 alpha (IL-1α). The qRT-PCR confirmed ELISA test results. The incubation with 5% AuNPs stabilized by PEI leads to upregulation of BMP-2 and VEGF transcripts of mRNA level and to downregulating expression of interleukin-6 (IL-6). Obtained data shed a promising light on gold nanoparticle application for future regenerative medicine application.
The aim of studies was to evaluate the microstructure and elemental compositions of alpaca hair. The hair from eight alpacas, which came from different part of Poland, were analyzed by scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). The quantitive analysis was performed for nutritionally important elements (calcium and phosphorous), elements, which are part of building blocks of hair structure (silicon and sulfur) and trace elements (copper, zinc, selenium). Moreover, the elemental mapping was carried out. The SEM images showed that microstructure of hair is irregular and rough. The average diameter of hair is 32,27 ± 4,06 µm. The obtained results showed content of elements in hair and also their distribution in the structure of hair. This findings can be a good benchmark for further studies. .
The rapid aging of the population results in increased number of metabolic and degenerative disorders, especially in the elderly.Thus, a novel approach in the fields of orthopedic and reconstructive surgery for bone regeneration is strongly desirable. A new perspective in the therapy of bone fractures is tissue engineering which combines living cells with biomaterials to develop modern substitutes that can restore tissue functions. Metallic biomaterials, including stainless steel and pure titanium, have been extensively used for the fabrication of surgical implants over decades. Chemical modification of material surface for example incorporation of chemotactic factors may significantly improve the therapeutic effect. In this paper we describe titanium substrate modifications with ZrO2/SiO2 coating functionalized with resveratrol using a sol – gel, dip-coating technique. Moreover, we established the effects of fabricated scaffolds on adipose stem cells isolated from elderly patients. Using fluorescence imaging, polymerase chain reaction (PCR)and cytotoxicity tests, we established that 0.5 Res_ZrO2/SiO2 significantly reduced apoptosis and accumulation of oxidative stress factors in adipose derived stem cells (ASC). Thus exploitation of fabricated biomaterials in regenerative medicine as a strategy for rejuvenate ASC from elderly patients in vivo, seems fully justified.
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