A new method for determining the viscoelastic properties of materials was introduced and investigated. Results of three groups of samples obtained using one particle microrheology method, DLS microrheology method and classical rheology method were compared to be sure of correctness of measurements. As a model system were chosen mixtures of glycerol of different viscosities. In case of samples containing glycerol, results were also compared with tabulated values. Hyaluronan of various molecular weights was used as a biopolymer and polystyrene particles were used for microrheology. It was confirmed, that viscosity values of biopolymer samples obtaining by each method are comparable.

The main aim of the doctoral thesis was study of passive microrheological techniques as advanced methods for characterisation of viscoelastic properties of soft material. These techniques are able to provide certain advantaged or additional information on classical rheology. Several techniques from the wide range of passive microrheological group such as one particle video – microrhelogy with IDL data procesing, microrheology based on light scattering – DLS or FCS microrheology have been applied in practice during my PhD studies. New Matlab script has been also invented to simply obtain information about viscosity of low volume samples. Aplicability of techniques were studied on several various bicolloids such as solutions or gels of hyaluronic acid, on study of properties at the interfaces or on gelation proces studies of agarose. Obtained data were compared classical rheology results. Suitability of each technique for investigated systems or appropriate reccomendations for further measurements were didcussed based on obtained data. Main advantages and limites of passive microrheology techniques were also described in comparison wih classical rheology method.

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.

Diploma thesis was aimed to deal with evaluation of microrheology method in the research of biopolymer-protein. Used biopolymer was sodium hyaluronate and proteins were trypsin and chymotrypsin. For measuring of microrheology were used particles with different radius (0,5 m and 1 m). It was found, that both substances have viscous charakter. Passive microrheology is suitable for measuring the viscoelastic properties of biopolymers.

This master thesis is focused on the evaluation of the influence of particle surface properties on the results of microrheological measurements with biopolymer solutions. Hyaluronan has been chosen as negatively charged polymer, chitosan as positively charged polymer and glycerol and its solutions of various concentrations were used as homogenous model systems. Dynamic light scattering and single particle tracking microrheology were chosen from passive microrheological techniques. Particles with various surface modifications (neutral, positive surface charge and negative surface charge) were used for the experiments. The results of microrheological techniques were then compared with classic rheology and moreover the glycerol results were compared with tabulated values.

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.

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®.

The behavior of the system consisted by mixture of two different molecular weight hyaluronates and surfactant was investigated. Mixtures were 17 kDa hyaluronate with 1,46 MDa, 73 kDa with 1,46 MDa, 300 kDa with 1,46 MDa, 806 kDa with 1,46 MDa and 1800 kDa with 1,46 MDa. These compounds were always mixed in the weight ratios 70:30, 50:50 and 30:70. As the surfactant cetrimonium bromide and TWEEN 20 were used. Interactions were studied in aqueous solution with different ionic strength. Sudan red was used as hydrophobic dye. In all experimental series with cetrimonium bromide was observed phenomenon of discontinuous separated phases, described as pearls. Samples containing pearls were tested on stability, were dried and rehydrated back, as were also heated in solution. Subsequently, the particle size was measured in the remaining sample after pearls were filtrated. Mixtures of hyaluronate were characterized by measuring the viscosity using rheology microrheology. It was found that these compounds are heterogeneous and each sample point is not the same viscosity.