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Microrheology modeling with COMSOL Multiphysics package
Koláček, Jakub ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis focuses on modeling Brownian motion using the COMSOL Multiphysics package and its Particle Tracing module. The aim of the work is to design and create elementary models that will be able to suitably simulate the movement of microparticles in viscous and viscoelastic environments, which can later be used for modeling passive microrheology. Within this work, Matlab scripts were created for the calculation of MSD from the simulation results, validation of the viscous model was performed on experimental data and elementary models for the simulation of the viscoelastic environment were also designed. Two different approaches were chosen for the design of these models, namely the use of rigid obstacles under the assumption of a discrete environment and a mathematical model assuming continuous environment. Data from the viscous model showed good agreement with the experimental results. The results of viscoelastic simulations are presented, and further possible development of these models is discussed. The continuous mathematical model is considered closest to modeling viscoelastic behavior because of a characteristic curvature that was observed in the evaluation of MSD.
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Derivatization of Sodium Hyaluronate as a Possible Tool for Increasing of the Stability of Model Artificial Synovial Fluid
Hrochová, Eliška ; Mravec, Filip (referee) ; Kalina, Michal (advisor)
This master thesis deals with the optimization of the procedure of modification of hyaluronic acid structure for the use in the artificial synovial liquids. Based on the literature research, the amino acid alanine was used for the modification of carboxylic group in the glucuronic acid. The main subject of study is the improvement of the stability and mechanical properties of synovial liquid. DLS microrheology, macrorheology, thermogravimetric analysis (TGA), multi-angle light scattering with flow-field flow fractionation (AF4-MALS) and infrared spectroscopy (FTIR) were used for characterization. The theoretical part of this theses submits review of the musculoskeletal system, role of hyaluronic acid in metabolism and summary of synovial liquid. The experimental part focuses on the measurement of the stability and mechanical properties of three artificial samples (first with no modification, second with modified hyaluronic acid and third with modified hyaluronic acid and chondroitin sulphate). These samples were compared with real horse synovial fluid and artificial viscosupplement Orthovisc®.
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Modelling of particle thermal motion
Orság, Miroslav ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
The goal of this thesis was to get familiar with the basics of mathematical description of the thermal motion of particles in a given media, and with other possibilities of the software package COMSOL Multiphysics. A model for viscous and viscoelastic environments was created, a uniform and user friendly system for simulation and calculation of MSD and system for data conversion from FCS to MSD. Furthermore, the possibilities of the model for use in microrheology were assessed and another procedure in the implementation of the COMSOL program in the characterization of gels was proposed.
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Influence of particle size on microreology experiments using fluorescence correlation spectroscopy
Valovič, Stela ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
This diploma thesis deals with microrheology measured via the fluorescence correlation spectroscopy. As microrheological probes, fluorescently marked nanoparticles of 5 sizes in the range of 10-100 nm, were used. The particles had been immersed in a variety of concentrated glycerol solutions and agarose gels of different concentrations, and the FCS measurement revealed a diffusion coefficient of individual particles in each environment. Based on the coefficient, the viscosity of the glycerol needed to stop the particles could be determined. Particles of 10 nm size were not stopped even by the 100 wt% glycerol. In the case of the agarose gels, a combination of higher agarose concentration and larger particles resulted in an increase in the diffusion coefficient to an unlikely high value. This was caused probably by an agarose autofluorescence and the value indicates stopping of the particles in the given agarose gel. Later, the data acquired by the FCS measurement were converted to MSD curves using MATLAB software. The thesis discusses the influence of the experimental parameters on the shape of the MSD curve. The results showed that the number of particles and autocorrelation function have the most significant effect.
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Preparation and Characterization of Mechanical Properties of Artificial Synovial Liquids
Hrochová, Eliška ; Sedláček, Petr (referee) ; Kalina, Michal (advisor)
This bachelor thesis deals with the optimization of preparation of artificial synovial liquids. The main subjects of study are the mechanical properties of real and artificial synovial fluid samples. DLS microrheology, thermogravimetric analysis (TGA) and infrared spectroscopy (FTIR) were used for characterization. The theoretical part of this thesis is represented by a literature research of a methods of preparation of artificial synovia and summary of definitions of rheological terms. The experimental part focuses on the preparation and characterization of the artificial synovial liquids originating from the published patent US 8716204. In the framework of the bachelor thesis, this preparation was optimized in several partial steps (method of dispersion of components, choice of molecular weight of hyaluronic acid, nature and ionic strength of used dispersion medium). The prepared optimized sample of synovial fluid was in following experimental characterization steps compared in terms of material characteristics and mechanical properties with the real horse synovial fluid sample. By modifying the process based on the aforementioned patent, an artificial synovial fluid could be formed. That meets the viscoelastic nature of the real matrix and that is stable over time.
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Modelling of particle movement in liquid and hydrogel media
Orság, Miroslav ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
Main goal of this thesis was to became familiar with general mathematical description of particle motion in fluid and hydrogel media in COMSOL software. Simulate a basic Brownian motion of particles with various diameter in media with different density. According to software options, simulate motion in polymer structure or in viscoelastic media. Consider an options of the model for use in microrheology and suggest next process in this field.
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The Utilization of Light Scattering Techniques for the Study on Hydrogel Gelation
Candráková, Simona ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
This bachelor thesis focuses on the study of hydrogel gelation using light scattering methods. For these purposes two different biopolymers (agarose and sodium alginate), with different sol to gelation phase transition, were selected. In the case of agarose, the gelation is caused by temperature change. On the other side, the gelation of sodium alginate is initiated by addition of polyvalent cations. In the experimental part of the work, agarose gelation was studied by DLS, temperature gradients of agarose solutions (40 – 30 °C) were measured. During the measurement the particle size distributions in the solutions having different concentrations were monitored as well as the temperature influence on the diffusion coefficient. The DLS method provided the values of the gelling temperatures of the solutions at a concentration of 0,5 wt.% and 1,0 wt.%. Furthermore, the DLS microrheology method was used, where temperature dependences of viscoelastic characteristics of agarose solutions (40 – 30 °C) were also measured, from which the gelling temperatures of agarose solutions of 0,1 wt.% and 0,5 wt.% were evaluated. The classical rheology was also used in the work to compare the viscoelastic behaviour of the samples and to determine the gelation point of the agarose solutions. By this method were determined gelling temperatures for all studied concentration of agarose (0,1 wt.%, 0,5 wt.% and 1,0 wt.%). The DLS method was also used to monitor the interactions of sodium alginate with the addition of Ca2+ ions, these interactions were then also evaluated and discussed in the experimental part.
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Time-Resolved Fluorescence in Research of Hyaluronan-Colloidal Systems Interactions
Mondek, Jakub ; Táborský, Petr (referee) ; Peter, Kapusta (referee) ; Pekař, Miloslav (advisor)
The aim of the doctoral thesis was to study advanced fluorescence techniques and its use in colloids or hyaluronan-surfactant systems and hydrogels based on hyaluronan, respectively. Steady-state and time-resolved fluorescence were used to study excited state proton transfer fluroescen probes in hyaluronan-surfactant systems to asses the influence of hyaluronan hydration to its interactions with oppositely charged surfactants. Firstly, different excited state proton transfer fluorescence probes were discussed to choose the most suitable candidate for next research. The influence of hyaluronan on inner environment of micells was determined based on the sensitivity of excited state proton transfer of chosen fluorescence probe 1-naphtol and, based on above mentioned experiments, the structure of hyaluronan hydration shell was discussed. The next part of doctoral thesis was focused on fluorescence lifetime correlation spectroscopy and on the development of method of nanorheology. Measured correlation functions were transformed to mean square displacement with developed MATLAB script. Firstly, the fluorescence method was compared with well described methods such as videomicrorheology and dynamic light scattering to asses the reliability of fluorescence correlation spectroscopy in microrheology. Secondly, nanorheology method was developed and its use in passive nanorheology of hyaluronan hydrogels was discussed. Based on mentioned experiments, the fluorescence correlation spectroscopy microrheology and nanorheology methods were optimized to use the methods in hydrogel research.
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Optimalization of techniques of microrheology for characterization of biopolymer hydrogels
Dušenková, Alica ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The main aim of the bachelor thesis is optimalization of microrheological techniques for characterization of biopolymer hydrogels. Hydrogels, based on thermoreversible biopolymer agarose, were selected for these experiments. The influence of incorporated poly(sodium 4-styrenesulfonate) on structure and viscoelastic properties of agarose hydrogels was investigated through diffusion coefficient and MSD curves. Microrheological properties were studied by fluorescence correlation spectroscopy, by using fluorescent beads, which were embedded in hydrogel systems.
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Advanced microrheological techniques in the research of hydrogels
Kábrtová, Petra ; Smilek, Jiří (referee) ; Mravec, Filip (advisor)
This diploma thesis deals with the use of fluorescence correlation spectroscopy technique for microrheological characterization of hydrogel in a system of hyaluronate-cetyltrimethylammonium bromide. Fluorescently labelled particles were used for microrheological FCS analysis. To optimize the method the most appropriate size of particles was chosen on the basis of Newtonian glycerol solutions analysis. Among other things, the discussion was focused on the influence of refractive index change of analysed solutions on analysis results. After hyaluronate solutions analysis it was possible to assess the biopolymer concentration and molecular weight impact on the FCS microrheology results, which could then be compared with analysis results of model hydrogels of hyaluronate and CTAB. Finally, usability and limitations of FCS microrheology have been discussed.
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