RESUMEN
CD109 antigen on the endothelial cell surface plays an important role in vascular pathology. The aim of the work was to investigate the effect of the immobilization of CD109 antigen with specific antibodies on nanomechanical properties of human umbilical endothelial cells (HUVECs) using atomic force microscopy in quantitative nanomechanical property mapping mode (PeakForce QNM). Anti-CD109 antibodies induced significant stiffening of the cell surface Me(LQ; UQ): in 1.45(1.07;2.29) times with respect to control cells for fixed cells and in 4.9(3.6;5.9) times with respect to control cells for living cells, and changes in the spatial distribution of cell surface mechanical properties. The changes in the HUVEC's mechanical properties were accompanied by the activation of the TGF-/Smad2/3 signaling pathway and reorganization of the vimentin and actin cytoskeletal elements. Our findings show that blocking CD109 antigen using anti-CD109 antibodies leads in HUVECs to the processes similar to that occur after cell TGF-ß-signaling activation. Therefore, we suggest that CD109 antigen may be involved in regulating the mechanical behavior of endothelial cells.
Asunto(s)
Células Endoteliales , Transducción de Señal , Humanos , Actinas/metabolismo , Membrana Celular/metabolismo , Citoesqueleto/metabolismo , Células Endoteliales/metabolismo , Microscopía de Fuerza Atómica/métodos , Transducción de Señal/fisiología , Factores de Transcripción/metabolismoRESUMEN
Endothelial cells, due to heterogeneity in the cell structure, can potentially form an inhomogeneous on structural and mechanical properties of the inner layer of the capillaries. Using quantitative nanomechanical mapping mode of atomic force microscopy, the parameters of the structural, elastic, and adhesive properties of the cell surface for living and glutaraldehyde-fixed human umbilical vein endothelial cells were studied. A significant difference in the studied parameters for three cell surface zones (peripheral, perinuclear, and nuclear zones) was established. The perinuclear zone appeared to be the softest zone of the endothelial cell surface. The heterogeneity of the endothelial cell mechanical properties at the nanoscale level can be an important mechanism in regulating the endothelium functions in blood vessels.
Asunto(s)
Membrana Celular/fisiología , Núcleo Celular/fisiología , Citoesqueleto/fisiología , Células Endoteliales de la Vena Umbilical Humana/fisiología , Mecanotransducción Celular , Adhesividad , Células Cultivadas , Módulo de Elasticidad , Humanos , Microscopía de Fuerza Atómica , Nanotecnología , Fenotipo , Análisis de la Célula Individual , Estrés MecánicoRESUMEN
Hereditary spherocytosis (HS), an erythrocyte membranopathy, is a heterogeneous disease, even at the level of the erythrocyte population. The paper aims at studying the mechanical properties (the Young's modulus, median and RMS roughness of friction force maps; fractal dimension, lacunarity and spatial distribution parameters of lateral force maps) of the cell surface layer of the erythrocytes of two different morphologies (discocytes and spherocytes) in HS using atomic force microscopy. The results of spatial-spectral and fractal analysis showed that the mechanical property maps of the HS spherocyte surface were more structurally homogeneous compared to the maps of HS discocytes. HS spherocytes also had a reduced RMS roughness and lacunarity of the mechanical property maps. The Young's modulus and averaged friction forces over the microscale HS spherocyte surface regions were approximately 20% higher than that of HS discocytes. The revealed significant difference at the nano- and microscales in the structural and mechanical properties of main (discoidal and spheroidal) morphological types of HS erythrocytes can potentially cause blood flow disturbance in the vascular system in HS.
Asunto(s)
Eritrocitos/patología , Fenómenos Mecánicos , Microtecnología , Nanotecnología , Esferocitosis Hereditaria/sangre , Fenómenos Biomecánicos , Niño , Módulo de Elasticidad , Femenino , Fricción , Humanos , MasculinoRESUMEN
Fractal dimension (DF) is one of the important parameters in the description of object's properties in different fields including biology and medicine. The present paper is focused on the application of the fractal dimension (the box counting dimension) in the analysis of the properties of cell surface on the base of its images obtained by atomic force microscopy (AFM). Fractal dimension of digital 3D AFM images depends on interpoint distances determined by the scanning step in the XY-plane and Z-scale factor t. We have studied the dependence of DF of AFM images on the Z-scale factor (DF=φ(t)) with purpose to reveal the features of the dependence and its usefulness in the analysis of the maps of surface properties. Using the model digital surfaces such as the plane, sinusoidal surfaces and "hilly" surface, we revealed that the sizes and spatial frequency of surface structural elements determined the basic features of the dependence (the parameters of peaks on the curve DF=φ(t)) and the element of chance in the localization of the structural elements on the surface had no significant influence on the dependence. Our findings demonstrate that the dependence of the fractal dimension on the Z-scale factor characterizes the structure of the AFM images more comprehensively than the roughness Ra and fractal dimension DF evaluated at a certain t. The dependence DF=φ(t) can be considered as a novel characteristic of AFM images. On analyzing the AFM images (lateral force maps) of glutaraldehyde-fixed adhered human fibroblasts and A549 human lung epithelial cells we found the significant difference in the dependences DF=φ(t) for different cell types that could be related to the difference of structural and mechanical surface properties of the studied cells.