Quantitative characterization of endothelial cell morphologies depending on shear stress in different blood vessels of domestic pigs using a focused ion beam and high resolution scanning electron microscopy (FIB-SEM)

Pham, Tam T.; Maenz, Stefan; Lüdecke, Claudia; Schmerbauch, Christoph; Settmacher, Utz; Bossert, Jörg; Zanow, Jürgen

doi:10.1016/j.tice.2014.12.005

Cited by 7 publications

(3 citation statements)

References 36 publications

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“…Focused ion beam scanning electron microscopy (FIB‐SEM) is now the election technique for three‐dimensional (3D) visualization of biological structures (cells) at nanoscale resolution . FIB‐SEM tomography allows 3D imaging at the subcellular level and is considered a breakthrough for ultrastructural volume reconstruction.…”

Section: Introductionmentioning

confidence: 99%

FIB‐SEM tomography of human skin telocytes and their extracellular vesicles

Crețoiu

Gherghiceanu

Hummel

et al. 2015

J Cellular Molecular Medi

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We have shown in 2012 the existence of telocytes (TCs) in human dermis. TCs were described by transmission electron microscopy (TEM) as interstitial cells located in non-epithelial spaces (stroma) of many organs (see www.telocytes.com). TCs have very long prolongations (tens to hundreds micrometers) named Telopodes (Tps). These Tps have a special conformation with dilated portions named podoms (containing mitochondria, endoplasmic reticulum and caveolae) and very thin segments (below resolving power of light microscopy), called podomers. To show the real 3D architecture of TC network, we used the most advanced available electron microscope technology: focused ion beam scanning electron microscopy (FIB-SEM) tomography. Generally, 3D reconstruction of dermal TCs by FIB-SEM tomography revealed the existence of Tps with various conformations: (i) long, flattened irregular veils (ribbon-like segments) with knobs, corresponding to podoms, and (ii) tubular structures (podomers) with uneven calibre because of irregular dilations (knobs) – the podoms. FIB-SEM tomography also showed numerous extracellular vesicles (diameter 438.6 ± 149.1 nm, n = 30) released by a human dermal TC. Our data might be useful for understanding the role(s) of TCs in intercellular signalling and communication, as well as for comprehension of pathologies like scleroderma, multiple sclerosis, psoriasis, etc.

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Section: Introductionmentioning

confidence: 99%

FIB‐SEM tomography of human skin telocytes and their extracellular vesicles

Crețoiu

Gherghiceanu

Hummel

et al. 2015

J Cellular Molecular Medi

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“…Doxycycline treatment also specifically inhibited thrombin-induced tumor cell migration in control cells to a greater extent than PAR1 low-expressing cells or PAR1-negative cells (Figure 2G and 2H and Supplementary Figure 2). Cells treated with doxycycline undergo significant changes in morphology, including pseudopod disappearance and cell shrinking, as detected by SEM (Figure 2I) [16]. In invasion assays, compared with the control group, the invasion ability increased significantly after the addition of thrombin.…”

Section: Resultsmentioning

confidence: 99%

Doxycycline directly targets PAR1 to suppress tumor progression

et al. 2017

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Doxycycline have been reported to exert anti-cancer activity and have been assessed as anti-cancer agents in clinical trials. However, the direct targets of doxycycline in cancer cells remain unclear. In this study, we used a chemical proteomics approach to identify the Protease-activated receptor 1 (PAR1) as a specific target of inhibition of doxycycline. Binding assays and single-molecule imaging assays were performed to confirm the inhibition of doxycycline to PAR1. The effect of doxycycline on multi-omics and cell functions were assessed based on a PAR1/thrombin model. Molecular docking and molecular dynamic simulations revealed that doxycycline interacts with key amino acids in PAR1. Mutation of PAR1 further confirmed the computation-based results. Moreover, doxycycline provides highly selective inhibition of PAR1 signaling in tumors in vitro and in vivo. Using pathological clinical samples co-stained for doxycycline and PAR1, it was found that doxycycline fluorescence intensity and PAR1 expression shown a clear positive correlation. Thus, doxycycline may be a useful targeted anti-cancer drug that should be further investigated in clinical trials.

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“…These results are consistent with the finding 38 that shear stress decreases with a decrease in cell height for the same applied shear rate. [49][50][51][52] The maximum shear stress values are, however, lower than those obtained in computational studies. 36,52 In those studies, atomic force microscopy (AFM) or fluorescence imaging was used to measure cell topography and, in turn, generate computational grids for the endothelial cells.…”

Section: B Shear Stress and Pressure Variationsmentioning

confidence: 99%

In vitro measurements of hemodynamic forces and their effects on endothelial cell mechanics at the sub-cellular level

et al. 2018

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This paper presents micro-particle tracking velocimetry measurements over cultured bovine aortic endothelial cell monolayers in microchannels. The objective was to quantify fluid forces and cell morphology at the sub-cellular scale for monolayers subjected to steady shear rates of 5, 10, and 20 dyn/cm 2. The ultimate goal of this study was to develop an experimental methodology for in vitro detailed study of physiologically realistic healthy and diseased conditions. Cell topography, shear stress, and pressure distributions were calculated from sets of velocity fields made in planes parallel to the microchannel wall. For each experiment, measurements were made in 3 h intervals for 18 h. It was found that there is a three-dimensional change in cell morphology as a result of applied shear stress. That is, cells flatten and become more wedge shaped in the stream direction while conserving volume by spreading laterally, i.e., in the cross-stream direction. These changes in cell morphology are directly related to local variations in fluid loading, i.e., shear stress and pressure. This paper describes the first flow measurements over a confluent layer of endothelial cells that are spatially resolved at the sub-cellular scale with a simultaneous temporal resolution to quantify the response of cells to fluid loading.

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Quantitative characterization of endothelial cell morphologies depending on shear stress in different blood vessels of domestic pigs using a focused ion beam and high resolution scanning electron microscopy (FIB-SEM)

Cited by 7 publications

References 36 publications

FIB‐SEM tomography of human skin telocytes and their extracellular vesicles

FIB‐SEM tomography of human skin telocytes and their extracellular vesicles

Doxycycline directly targets PAR1 to suppress tumor progression

In vitro measurements of hemodynamic forces and their effects on endothelial cell mechanics at the sub-cellular level

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