Search results for: R. Matejka

Authors: Špela Hajduk, Sean P. Berglund, Matejka Podlogar, Goran Dražić, Fatwa F. Abdi, Zorica C. Orel, Menny Shalom

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Graphitic carbon nitride materials (g-CN) have emerged as an attractive photocatalyst and electrocatalyst for photo and electrochemical water splitting reaction, due to their environmental benignity nature and suitable band gap. Many approaches were introduced to enhance the photoactivity and electronic properties of g-CN and resulted in significant changes in the electronic and catalytic properties. Here we demonstrate the synthesis of thin and homogenous g-CN layer on highly ordered ZnO nanowire (NW) substrate by growing a seeding layer of small supramolecular assemblies on the nanowires. The new synthetic approach leads to the formation of thin g-CN layer (~3 nm) without blocking all structure. Two different deposition methods of carbon nitride were investigated and will be presented. The amount of loaded carbon nitride significantly influences the PEC activity of hybrid material and all the ZnO/g-CNx electrodes show great improvement in photoactivity. The chemical structure, morphology and optical properties of the deposited g-CN were fully characterized by various techniques as X-ray powder spectroscopy (XRD), scanning electron microscopy (SEM), focused ion beam scanning electron microscopy (FIB-SEM), high-resolution scanning microscopy (HR-TEM) and X-ray photoelectron spectroscopy (XPS).

Keywords: carbon nitride, photoanode, solar water splitting, zinc oxide

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Authors: Tomas Barnak, Libor Matejka

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The aim of this study is to create a standard application using multi-criteria evaluation in the field of forensic engineering. This situation can occur in the professional assessment in several cases such as when it is necessary to consider more criteria variant of the structural subsystems, more variants according to several criteria based on a court claim, which requires expert advice. A problematic situation arises when it is necessary to clearly determine the ranking of the options according to established criteria, and reduce subjective evaluation. For the procurement in the field of construction which is based on the prepared text of the law not only economic criteria but also technical, technological and environmental criteria will be determined. This fact substantially changes the style of evaluation of individual bids. For the above-mentioned needs of procurement, the unification of expert’s decisions and the use of multi-criteria assessment seem to be a reasonable option. In the case of experimental verification when using multi-criteria evaluation of alternatives construction subsystem the economic, technical, technological and environmental criteria will be compared. The core of the solution is to compare a selected number of set criteria, application methods and evaluation weighting based on the weighted values assigned to each of the criteria to use multi-criteria evaluation methods. The sequence of individual variations is determined by the evaluation of the importance of the values of corresponding criteria concerning expertise in the problematic of outside wall constructional subsystems.

Keywords: criteria, expertise, multi-criteria evaluation, outside wall subsystems

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Authors: I. Karagjozova, B. Dejanova, J. Pluncevic, S. Petrovska, V. Antevska, L. Todorovska

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Introduction: Medical checking assessment is important in sports medicine. To follow the health condition in subjects who perform sports, body composition parameters, such as intracellular water, extracellular water, protein and mineral content, muscle and fat mass might be useful. The aim of the study was to show available parameters and to compare them to manual assessment. Material and methods: A number of 20 subjects (14 male and 6 female) at age of 20±2 years were determined in the study, 5 performed recreational sports, while others were professional ones. The mean height was 175±7 cm, the mean weight was 72±9 cm, and the body mass index (BMI) was 23±2 kg/m2. The measured compartments were as following: intracellular water (IW), extracellular water (EW), protein component (PC), mineral component (MC), skeletal muscle mass (SMM) and body fat mass (BFM). Lean balance were examined for right and left arm (LA), trunk (T), right leg (RL) and left leg (LL). The comparison was made between the calculation derived by manual made measurements, using Matejka formula and parameters obtained by body composition analyzer (BCA) - Inbody 720 BCA Biospace. Used parameters for the comparison were muscle mass (SMM), body fat mass (BFM). Results: BCA obtained values were for: IW - 22.6±5L, EW - 13.5±2 L, PC - 9.8±0.9 kg, MC - 3.5±0.3, SMM - 27±3 kg, BFM - 13.8±4 kg. Lean balance showed following values for: RA - 2.45±0.2 kg, LA - 2.37±0.4, T - 20.9±5 kg, RL - 7.43±1 kg, and LL - 7.49 ±1.5 kg. SMM showed statistical difference between manual obtained value, 51±01% to BCA parameter 45.5±3% (p<0.001). Manual obtained values for BFM was lower (17±2%) than BCA obtained one, 19.5±5.9% (p<0.02). Discussion: The obtained results showed appropriate values for the examined age, regarding to all examined parameters which contribute to overview the body compartments, important for sport performing. Due to comparison between the manual and BCA assessment, we may conclude that manual measurements may differ from the certain ones, which is confirmed by statistical significance.

Keywords: athletes, body composition, bio electrical impedance, sports medicine

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Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

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The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

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Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

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Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

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Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova

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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.

Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF

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