Analyzing Interactions Between Cells And Extracellular Matrix By Atomic Force Microscopy

Interactions of cells with the extracellular matrix (ECM) have important roles in various physiological and pathological processes, including tissue morphogenesis during embryonic development, wound healing and tumor invasion. Although most of the proteins involved in cell-ECM interactions have been identified, the underlying mechanisms and involved signaling pathways are incompletely understood. Here, atomic force microscope-based imaging and single-cell force measurements were used to characterize the interactions of different cell types with ECM proteins. The interplay between cells and ECM is complex. However, two interaction types, protein-protein and protein-carbohydrate, predominate. Integrins, adhesion receptors for ECM, mediate the former, galectins, a family of animal lectins, the latter. In the second chapter of this thesis, the contributions of both receptor families to the interactions of epithelial MDCK cells with ECM proteins are presented. It was found that galectins-3 and 9 are highly expressed in MDCK cells and required for optimal long-term adhesion (90 minutes) to ECM proteins collagen-I and laminin-111. Interestingly, early adhesion (< 2 minutes) to laminin-111, was integrin-independent and instead mediated by carbohydrate interactions and galectins. In contrast, early adhesion to collagen-I was exclusively mediated by integrins. Moreover, cells frequently entered an enhanced adhesion state, marked by a significant increase in the force required for cell detachment. Although adhesion was mediated by integrins, adhesion enhancement was especially observed in cells depleted for galectin-3. It was proposed that galectin-3 influences integrin-mediated adhesion complex formation by altering receptor clustering. To control their attachment to ECM proteins, cells regulate integrin receptors. One regulatory process is integrin crosstalk, where the binding of one type of integrin influences the activity of another type. In the third chapter, the implementation of a single-cell force spectroscopy assay to identify such crosstalks and gain insight into their mechanisms is described. In this assay the interactions of integrin receptors being specifically attached to one ligand are characterized in dependence of another ligand-bond receptor pair. With this assay a crosstalk between collagen-binding integrin α1β1 and fibronectin-binding integrin α5β1 was identified in HeLa cells. This crosstalk was directional from integrin α1β1 to integrin α5β1 and appeared to regulate integrin α5β1 by inducing its endocytosis. In the fourth and final chapter, mechanisms of matrix-induced cell alignment were studied by imaging cells on two-dimensional matrices assembled of highly aligned collagen fibrils. Integrin α2β1 was identified as the predominant receptor mediating cell polarization. Time-lapse AFM demonstrated that during alignment cells deform the matrix by reorienting individual collagen fibrils. Cells deformed the collagen matrix asymmetrically, revealing an anisotropy in matrix rigidity. When matrix rigidity was rendered uniform by chemical cross-linking or when the matrix was formed from collagen fibrils of reduced tensile strength, cell polarization did not occur. This suggested that both the high tensile strength and pliability of collagen fibrils contribute to the anisotropic rigidity of the matrix and lead to directional cellular traction and cell polarization. During alignment, cellular protrusions contacted the collagen matrix from below and above. This complex entanglement of cellular protrusions and collagen fibrils may further promote cell alignment by maximizing cellular traction. The work presented here adds to the understanding of cell-ECM interactions. Atomic force microscopy imaging allowed characterizing the behavior of cells on nanopatterned collagen matrices whereas single-cell force spectroscopy revealed insights into the regulation of cell adhesion by galectins. Furthermore, methodological advances in the single-cell force spectroscopy assay allowed the intracellular regulation of receptor molecules to be studied. The work demonstrates that atomic force microscopy is a versatile tool to study cell-ECM interactions

Design and Optimization of Compressor Airfoils by Using Class Function / Shape Function Methodology

International audienceThis paper describes a new design method for high Reynolds number subsonic compressor blade sections for industrial gas turbines and compressors. The focus is on the middle and end stages, where the Reynolds numbers are about 2 to 6 × 10 6 and the Mach numbers between 0.4 and 0.8. The new design method combines i) a parametric geometry definition method, ii) a fast blade-to-blade flow solver, and iii) an optimization tool with a suitable objective function. The development of a new blade section is based on a conventional NACA-65 design, subsequently modified to an optimized CSM profile, where CSM means Class Function / Shape Function Methodology. The new profile shapes are obtained by superimposing a camber line and a thickness distribution. Both the camber line and the thickness distribution are prescribed as analytical functions to cut down the CPU-time for geometry set up and to guarantee smooth geometries. Numerical calculations are performed by applying the two-dimensional blade-to-blade solver MISES. The optimization method used in this paper is the single-objective genetic algorithm (SOGA) from the DAKOTA library. The objective function consists of 5 components and takes into account the whole loss polar. The corresponding computing time is relatively short-that is 1 to 2 days. At high Reynolds number, the new profiles show decreased design point losses and increased operating limits compared to corresponding results using conventional NACA-65 profiles. In addition, the present results show close agreement with those produced by so-called high performance profiles of the relevant literature

Modelling of A Boundary Layer Ingesting Propulsor

Boundary layer ingestion is a promising method to decrease the propulsive power consumption of an aircraft, and therefore the fuel consumption. This leads to a reduced environmental impact and an improved cost-efficiency. To get a better understanding of this method and to estimate its benefits, the modelling of a propulsor located at the upper rear centerbody of a blended wing body aircraft is presented in this paper. A parallel compressor model approach is used to analyse the impact of the ingested low velocity fluid which leads to a non-uniform inflow. The required boundary layer data are generated with an analysis tool for 2D subsonic airfoils. Some parameter variations are conducted with the developed programme to study their impact on the power saving potential. In addition, a simple estimation for the benefit of embedded aeroengines is given. Despite the drawback from fan efficiency due to the inflow distortion, the results show a significant decrease in required propulsive power for boundary layer ingestion in combination with integrated engines

Design Decisions for a Powertrain Combination of Electric Motor and Propeller for an Electric Aircraft

For the sizing of an electric motor, the diameter and length of the installation space are important parameters that significantly influence and limit the characteristics of the electric motor. From an aerodynamic point of view, it is not technically advantageous to choose a propeller hub diameter that is larger than mechanically necessary. The hub diameter defines the installation space ans thus the outer diameter for the electric motor driving the propeller. The research question that arises is this: Is there an optimum diameter for the hub and thus the electric motor and how is it limited? The present work should help to establish a better understanding of the system combination of electric motor and propeller, especially the choice of the diameter and further geometrical parameters of the electric motor. A parameter study investigating aeromechanical and electromagnetic aspects has been performed and the results are combined to achieve the best solution in terms of efficiency and power density. The investigations show that an increase in the diameter causes little disadvantage from an aerodynamic point of view, but the improvements to the electric motor are significant only up to the point where the diameter-to-length ratio becomes too high and end-region effects become more significant. The installation space for the electric motor could be determined using a hub-to-tip ratio of 0.16 to 0.20, which is applicable to any propeller. In addition, it could be shown that a co-optimisation of the speed throughout a mission profile with regard to the efficiency is affected by the propeller alone

The Multipole Resonance Probe: Simultaneous Determination of Electron Density and Electron Temperature Using Spectral Kinetic Simulation

The investigation of the spectral kinetic model of the Multipole Resonance Probe (MRP) is presented and discussed in this paper. The MRP is a radio-frequency driven probe of the particular spherical design, which is suitable for the supervision and control of low-temperature plasma. The importance of the kinetic effects was introduced in the previous study of the spectral kinetic model of the idealized MRP. Such effects particularly dominate the energy loss in a low-pressure regime. Unfortunately, they are absent in the Drude model. With the help of the spectral kinetic scheme, those energy losses can be predicted, and it enables us to obtain the electron temperature from the FWHM in the simulated resonance curve. Simultaneously, the electron density can be derived from the simulated resonance frequency. Good agreements in the comparison between the simulation and the measurement demonstrate the suitability of the presented model

Aerodynamic and aeroelastic effects of design-based geometry variations on a low-pressure compressor

In modern aircraft engines, the low-pressure compressor (LPC) is subjected to a flow characterized by strong wakes and secondary flows from the upstream fan. This concerns ultra-high bypass ratio (UHBR) turbofan engines, in particular. This paper presents the aerodynamic and aeroelastic sensitivities of parametric variations on the LPC, driven by the design considerations in the upstream fan. The goal of this investigation was to determine the influence of design-based geometry parameter variations on the LPC performance under realistic inlet flow distributions and the presence of an s-duct. Aerodynamic simulations are conducted at the design and off-design operating points with the fan outflow as the inlet boundary conditions. Based on the aerodynamic results, time-linearized unsteady simulations are conducted to evaluate the vibration amplitude at the resonance operating points. First, the bypass ratio is varied by reducing the channel height of the LPC. The LPC efficiency decreases by up to 1.7% due to the increase in blockage of the core flow. The forced response amplitude of the rotor decreases with increasing bypass ratio due to increased aerodynamic damping. Secondly, the fan cavity leakage flow is considered as it directly affects the near hub fan flow and thus the inflow of the LPC. This results in an increased total-pressure loss for the s-duct due to mixing losses. The additional mixing redistributes the flow at the s-duct exit leading to a total-pressure loss reduction of 4.3% in the first rotor at design point. This effect is altered at off-design conditions. The vibration amplitude at low speed resonance points is increased by 19% for the first torsion and 26% for second bending. Thirdly, sweep and lean are applied to the inlet guide vane (IGV) upstream of the LPC. Despite the s-duct and the variable inlet guide vane (VIGV) affecting the flow, the three-dimensional blade design achieves aerodynamic and aeroelastic improvements of rotor 1 at off-design. The total-pressure loss reduces by up to 18% and the resonance amplitude more than 10%. Only negligible improvements for rotor 1 are present at the design point. In a fourth step, the influence of axial gap size between the stator and the rotor rows in the LPC is examined in the range of small variations which shows no distinct aerodynamic and aeroelastic sensitivities. This finding not only supports previous studies, but it also suggests a correlation between mode shapes and locally increased excitaion with increasing axial gap size. As a result, potential design improvements in future fan-compressor design are suggested. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) license (https://creativecommons.org/licenses/by-nc-nd/4.0/)

Transient Dynamic System Behavior of Pressure Actuated Cellular Structures in a Morphing Wing

High aspect ratio aircraft have a significantly reduced induced drag, but have only limited installation space for control surfaces near the wingtip. This paper describes a multidisciplinary design methodology for a morphing aileron that is based on pressure-actuated cellular structures (PACS). The focus of this work is on the transient dynamic system behavior of the multi-functional aileron. Decisive design aspects are the actuation speed, the resistance against external loads, and constraints preparing for a future wind tunnel test. The structural stiffness under varying aerodynamic loads is examined while using a reduced-order truss model and a high-fidelity finite element analysis. The simulations of the internal flow investigate the transient pressurization process that limits the dynamic actuator response. The authors present a reduced-order model based on the Pseudo Bond Graph methodology enabling time-efficient flow simulation and compare the results to computational fluid dynamic simulations. The findings of this work demonstrate high structural resistance against external forces and the feasibility of high actuation speeds over the entire operating envelope. Future research will incorporate the fluid–structure interaction and the assessment of load alleviation capability