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Purpose of Study: Thrombosis-related complications are among the leading causes for morbidity and mortality in patients who depend on artificial organs. For the prediction of platelet behavior both the flow conditions inside the device and the thrombogenic properties of the blood-contacting surfaces must be considered. Platelet reactions under the influence of well-defined shear rates are experimentally evaluated and numerically simulated. The approach is intended for the analysis of VAD and oxygenator design.
Methods Used: A mathematical model of platelet activation, adhesion and aggregation has been implemented into a finite element CFD (Computational Fluid Dynamics) code. The approach is based on the advective and diffusive transport equations for resting and activated platelets and platelet released agonists. Experiments with citrate-anticoagulated freshly-drawn whole blood are performed in a perfusion flow chamber as well as in a system of rotating cylinders for Couette and Taylor-vortex flow. Different biomaterials are used. The activation, adhesion and aggregation are quantified using scanning electron microscopy and flow cytometry.
Summary of Results: Regions and flow conditions with a high potential for thrombus growth could be identified. The experiments clearly show the influence of the blood contacting material and governing shear rates. Numerical analysis can explain observed adhesion patterns and the degree of thrombus formation
Behbahani, Mehdi ; Mai, A. ; Waluga, C. ; Bergmann, B. ; Tran, L. ; Vonderstein, K. ; Behr, Marek ; Mottaghy, K.
Aims: Thrombotic complications due to activation of platelets and plasmatic clotting factors belong still to the most investigated topics in the field of study of patho-physiological mechanisms. Mathematical modeling of thrombotic reactions is established and validated in test cases. Aim of this study is to experimentally evaluate and computationally simulate platelets under the influence of well-defined shear flow conditions. Platelet behaviour and reactions are experimentally reproduced, measured and used for validation of the numerical simulation. Methods: A mathematical model of platelet activation, adhesion and aggregation has been implemented into a finite element CFD (Computational Fluid Dynamics) code. The approach is based on the advective and diffusive transport equations for resting platelets, activated platelets and platelet released agonists. Adhesion rates for the reactive surfaces depend on the hemocompatibility properties of the surface and the local shear rate. Experiments with citrate-anticoagulated freshly-drawn whole blood are performed in a perfusion flow chamber as well as in a system of rotating cylinders for Couette and Taylor-vortex flow. Different biomaterials are used. The activation, drop of platelet concentration, adhesion and aggregation are quantified using scanning electron microscopy (SEM) and flow cytometry. Results: Regions and flow conditions with a high potential for thrombus growth could be identified. The experiments clearly show the influence of the blood contacting material and flow properties. By means of SEM diverse platelet adhesion patterns are observed. Numerical analysis can explain the patterns and the degree of thrombus formation. Conclusion: The numerical method shows good agreement with experimental data indicating a possible prediction of initiation of activation and detection of the local adhesion areas in connection with the role of Von-Willebrand-Factor.
Behbahani, Mehdi ; Mai, A. ; Bergmann, B. ; Waluga, C. ; Behr, Marek ; Tran, L. ; Vonderstein, K. ; Mottaghy, K.
Behbahani, Mehdi ; Behr, Marek ; Hormes, M. ; Steinseifer, U. ; Arora, D. ; Coronado, O. ; Pasquali, M.
Mechano-pharmacological testing of L-Type Ca²⁺ channel modulators via human vascular celldrum model
(2020)
Bayer, Robin ; Temiz Artmann, Aysegül ; Digel, Ilya ; Falkenstein, Julia ; Artmann, Gerhard ; Creutz, Till ; Hescheler, Jürgen
Background/Aims: This study aimed to establish a precise and well-defined working model, assessing pharmaceutical effects on vascular smooth muscle cell monolayer in-vitro. It describes various analysis techniques to determine the most suitable to measure the biomechanical impact of vasoactive agents by using CellDrum technology. Methods: The so-called CellDrum technology was applied to analyse the biomechanical properties of confluent human aorta muscle cells (haSMC) in monolayer. The cell generated tensions deviations in the range of a few N/m² are evaluated by the CellDrum technology. This study focuses on the dilative and contractive effects of L-type Ca²⁺ channel agonists and antagonists, respectively. We analyzed the effects of Bay K8644, nifedipine and verapamil. Three different measurement modes were developed and applied to determine the most appropriate analysis technique for the study purpose. These three operation modes are called, particular time mode" (PTM), "long term mode" (LTM) and "real-time mode" (RTM). Results: It was possible to quantify the biomechanical response of haSMCs to the addition of vasoactive agents using CellDrum technology. Due to the supplementation of 100nM Bay K8644, the tension increased approximately 10.6% from initial tension maximum, whereas, the treatment with nifedipine and verapamil caused a significant decrease in cellular tension: 10nM nifedipine decreased the biomechanical stress around 6,5% and 50nM verapamil by 2,8%, compared to the initial tension maximum. Additionally, all tested measurement modes provide similar results while focusing on different analysis parameters. Conclusion: The CellDrum technology allows highly sensitive biomechanical stress measurements of cultured haSMC monolayers. The mechanical stress responses evoked by the application of vasoactive calcium channel modulators were quantified functionally (N/m²). All tested operation modes resulted in equal findings, whereas each mode features operation-related data analysis.
Hypertension describes the pathological increase of blood pressure, which is most commonly associated with the increase of vascular wall stiffness [1]. Referring to the “Deutsche Bluthochdruck Liga” this pathology shows a growing trend in our aging society. In order to find novel pharmacological and probably personalized treatments, we want to present a functional approach to study biomechanical properties of a human aortic vascular model.
In this method review we will give an overview of recent studies which were carried out with the CellDrum technology [2] and underline the added value to already existing standard procedures known from the field of physiology.
Herein described CellDrum technology is a system to measure functional mechanical properties of cell monolayers and thin tissue constructs in-vitro. Additionally, the CellDrum enables to elucidate the mechanical response of cells to pharmacological drugs, toxins and vasoactive agents. Due to its highly flexible polymer support, cells can also be mechanically stimulated by steady and cyclic biaxial stretching.
Baumann, Marcus ; Schwarz, Sonja ; Kotliar, Konstantin ; Eynatten, Maximilian von ; Trucksaess, Arno ; Burckhardt, Klaus ; Lutz, Jens ; Heemann, Uwe ; Lanzl, Ines
Background
Minor changes in protein structure induced by small organic and inorganic molecules can result in significant metabolic effects. The effects can be even more profound if the molecular players are chemically active and present in the cell in considerable amounts. The aim of our study was to investigate effects of a nitric oxide donor (spermine NONOate), ATP and sodium/potassium environment on the dynamics of thermal unfolding of human hemoglobin (Hb). The effect of these molecules was examined by means of circular dichroism spectrometry (CD) in the temperature range between 25°C and 70°C. The alpha-helical content of buffered hemoglobin samples (0.1 mg/ml) was estimated via ellipticity change measurements at a heating rate of 1°C/min.
Results
Major results were:
1) spermine NONOate persistently decreased the hemoglobin unfolding temperature T u irrespectively of the Na + /K + environment,
2) ATP instead increased the unfolding temperature by 3°C in both sodium-based and potassium-based buffers and
3) mutual effects of ATP and NO were strongly influenced by particular buffer ionic compositions. Moreover, the presence of potassium facilitated a partial unfolding of alpha-helical structures even at room temperature.
Conclusion
The obtained data might shed more light on molecular mechanisms and biophysics involved in the regulation of protein activity by small solutes in the cell.
As a deduction from these results, we can conclude that proteins mainly in vitro, denaturate totally at a temperature between 57°C -62°C, and they also affected by NO and different ions types. In which mainly, NO cause earlier protein denaturation, which means that, NO has a destabilizing effect on proteins, and also different ions will alter the protein denaturation in which, some ions will cause earlier protein denaturation while others not.