Endothelial cells form the lining of the walls of blood vessels and are continuously subjected to mechanical stimuli from the blood flow. Microtubule-organizing center (MTOC),<br>also known as centrosome is a structure found in eukaryotic cells close to the nucleus. MTOC relocates relative to the nucleus when endothelial cells are exposed to shear stress which determines their polarization, thus it plays a critical role in cell migration and wound healing. The nuclear lamina, a mesh-like network that lies underneath the nuclear membrane, is composed of lamins, type V intermediate filament proteins. Mutations in LMNA gene that encodes A-type lamins cause the production of a mutant form of lamin A called progerin and leads to a rare premature aging disease known as Hutchinson-Gilford Progeria Syndrome<br><div>(HGPS). The goal of this study is to investigate how fluid flow affects the cytoskeleton of endothelial cells.</div><div><br></div>This thesis consists of two main sections; computational mechanical modeling and laboratory experimental work. The mechanical model was implemented using Ansys Workbench software as a tensegrity-based cellular model in order to simulate the state of an endothelial cell under the effects of induced shear stress from the blood fluid flow. This tensegrity-based cellular model - composed of a plasma membrane, cytoplasm, nucleus, microtubules, and<br><div>actin filaments - aims to understand the effects of the fluid flow on the mechanics of the cytoskeleton. In addition, the laboratory experiments conducted in this study examined the MTOC-nuclear orientation of endothelial cells under shear stress with the presence of wound healing. Wild-type lamin A and progerin-expressing BAECs were studied under static and sheared conditions.</div><div><br></div><div> Moreover, a custom MATLAB code was utilized to measure the MTOC-nuclear orientation</div>angle and classification. Results demonstrate that shear stress leads to different responses of the MTOC orientation between the wild-type and progerin-expressing cells around the vertical wound edge. Future directions for this study involve additional experimental work together with the improved simulation results to confirm the MTOC orientation<br>relative to the nucleus under shear stress.

This diploma thesis focuses on computational modeling of the cell mechanical tests. The goal of this thesis is to build a cell model and to simulate compression test on this model. If necessary, the model should be adjusted so the model reflects real cell behavior. It was created the cell model reflecting cytoplasm, nucleus, membrane and cell cytoskeleton. Cytoskeleton was modeled as tensegrity structure. After this, the pressure test was simulated on this model. The behavior of the cell model and real cell was compared using the stress force. The stress force - cell deformation curve was markedly different for the cell model and the real cell. For this reason, the cytoplasm material model was adjusted. The difference between the curves was acceptable after this modification. It was found during computations that the cytoskeleton model influence on the cell load is minimal. These results does not reflects real cell behavior, which means that the model is considered inadequate for performing stress load simulation.

The master’s thesis deals with computational modelling of mechanical testing of isolated cells, particularly of single-axle tensile test. The aim is to imitate the real deformed shape known from experiments. At first, the structure of each cell component is described and analyzed according to their significance for mechanical behavior. The outline of basic mechanical tests used for cell testing is discussed next. A structural computational model comprising all components significant for mechanical purposes is created for the modelling. Those components are nucleus, cytoplasm, cell membrane and cytoskeleton. Due to the problems with convergence the model was divided into two parts. The first one treats separately the shape of cytoskeleton and the second one treats the shape of communicating components (nucleus, cytoplasm and cell membrane). Both of those partial models succeed in reaching the deformations according to the experiments.

The master’s thesis deals with the computational modelling of the mechanical testing of isolated smooth muscle cells. The main aims are to create computational model of a cell, to simulate single-axis tensile test and to modify the model so that the model reflects real mechanical response. The model of the cell includes cytoplasm, nucleus, cell membrane and cytoskeleton which is modelled as a tensegrite structure. On this model the tensile test was simulated in case of the cell with cytoskeleton and the cell with distributed the cytoskeleton. Force-elongation curves, which were obtained from this simulation, were compared with experimental data which were taken from literature. Tensile properties were measured on freshly isolated cells from rat thoracic aorta, cultured cells, and cells treated with cytochalasin D to disrupt their actin filaments. It was found that the cytoskeleton influence on the cell load in computational model was smaller than in the real cell. Therefore the model was modified by changing material propreties and geometry so that the model of the cell corresponded with the different types of experimentally measured cells.