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traction force microscopy

Trung Dung Nguyen, YuanTong Gu
It is well-known that cell adhesion is important in many biological processes such as cell migration and proliferation. A better understanding of the cell adhesion process will shed insight into these cellular biological responses as well as cell adhesion-related diseases treatment. However, there is little research which has attempted to investigate the process of cell adhesion and its mechanism. Thus, this paper aims to study the time-dependent adhesion properties of single living chondrocytes using an advanced coupled experimental-numerical approach...
November 28, 2016: Scientific Reports
Rebecca L Lamason, Effie Bastounis, Natasha M Kafai, Ricardo Serrano, Juan C Del Álamo, Julie A Theriot, Matthew D Welch
Spotted fever group (SFG) rickettsiae are human pathogens that infect cells in the vasculature. They disseminate through host tissues by a process of cell-to-cell spread that involves protrusion formation, engulfment, and vacuolar escape. Other bacterial pathogens rely on actin-based motility to provide a physical force for spread. Here, we show that SFG species Rickettsia parkeri typically lack actin tails during spread and instead manipulate host intercellular tension and mechanotransduction to promote spread...
October 20, 2016: Cell
M Moazzem Hossain, Guangyi Zhao, Moon-Sook Woo, James H-C Wang, Jian-Ping Jin
Cell traction force (CTF) plays a critical role in controlling cell shape, enabling cell motility, and maintaining cellular homeostasis in many biological processes such as angiogenesis, development, wound healing, and cancer metastasis. Calponin is an actin filament-associated cytoskeletal protein in smooth muscles and multiple types of non-muscle cells. An established biochemical function of calponin is the inhibition of myosin ATPase in smooth muscle cells. Vertebrates have three calponin isoforms. Among them, calponin 2 is expressed in epithelial cells, endothelial cells, macrophages, myoblasts and fibroblasts, and plays a role in regulating cytoskeleton activities such as cell adhesion, migration and cytokinesis...
October 13, 2016: Biochemistry
Richard De Mets, Katharina Hennig, Lionel Bureau, Martial Balland
Mechanical interactions between cells and their microenvironment are crucial for fundamental biological processes ranging from migration to differentiation. This has led, over the last decades, to the development of new ways to culture cells. Living cells are now grown not only on glass coverslips, where they completely lose the mechanical and geometrical constraints coming from their microenvironment, but also on soft patterned substrates that mimic the rigidity and spatial information of their in vivo niches...
October 18, 2016: Biomaterials Science
Clément Roux, Alain Duperray, Valérie M Laurent, Richard Michel, Valentina Peschetola, Claude Verdier, Jocelyn Étienne
When crawling on a flat substrate, living cells exert forces on it via adhesive contacts, enabling them to build up tension within their cytoskeleton and to change shape. The measurement of these forces has been made possible by traction force microscopy (TFM), a technique which has allowed us to obtain time-resolved traction force maps during cell migration. This cell 'footprint' is, however, not sufficient to understand the details of the mechanics of migration, that is how cytoskeletal elements (respectively, adhesion complexes) are put under tension and reinforce or deform (respectively, mature and/or unbind) as a result...
October 6, 2016: Interface Focus
Martin Bergert, Tobias Lendenmann, Manuel Zündel, Alexander E Ehret, Daniele Panozzo, Patrizia Richner, David K Kim, Stephan J P Kress, David J Norris, Olga Sorkine-Hornung, Edoardo Mazza, Dimos Poulikakos, Aldo Ferrari
The mechanical wiring between cells and their surroundings is fundamental to the regulation of complex biological processes during tissue development, repair or pathology. Traction force microscopy (TFM) enables determination of the actuating forces. Despite progress, important limitations with intrusion effects in low resolution 2D pillar-based methods or disruptive intermediate steps of cell removal and substrate relaxation in high-resolution continuum TFM methods need to be overcome. Here we introduce a novel method allowing a one-shot (live) acquisition of continuous in- and out-of-plane traction fields with high sensitivity...
September 29, 2016: Nature Communications
Michiel Fokkelman, Hayri E Balcıoğlu, Janna E Klip, Kuan Yan, Fons J Verbeek, Erik H J Danen, Bob van de Water
Cancer cells migrate from the primary tumour into surrounding tissue in order to form metastasis. Cell migration is a highly complex process, which requires continuous remodelling and re-organization of the cytoskeleton and cell-matrix adhesions. Here, we aimed to identify genes controlling aspects of tumour cell migration, including the dynamic organization of cell-matrix adhesions and cellular traction forces. In a siRNA screen targeting most cell adhesion-related genes we identified 200+ genes that regulate size and/or dynamics of cell-matrix adhesions in MCF7 breast cancer cells...
2016: Scientific Reports
Alexandra S Piotrowski-Daspit, Celeste M Nelson
The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, which is regulated by a variety of soluble and physical signals in the microenvironment. Described here is a method created to study the process of branching morphogenesis by forming engineered three-dimensional (3D) epithelial tissues of defined shape and size that are completely embedded within an extracellular matrix (ECM). This method enables the formation of arrays of identical tissues and enables the control of a variety of environmental factors, including tissue geometry, spacing, and ECM composition...
2016: Journal of Visualized Experiments: JoVE
Erik T Valent, Geerten P van Nieuw Amerongen, Victor W M van Hinsbergh, Peter L Hordijk
In many pathological conditions the endothelium becomes activated and dysfunctional, resulting in hyperpermeability and plasma leakage. No specific therapies are available yet to control endothelial barrier function, which is regulated by inter-endothelial junctions and the generation of acto-myosin-based contractile forces in the context of cell-cell and cell-matrix interactions. However, the spatiotemporal distribution and stimulus-induced reorganization of these integral forces remain largely unknown. Traction force microscopy of human endothelial monolayers was used to visualize contractile forces in resting cells and during thrombin-induced hyperpermeability...
September 10, 2016: Experimental Cell Research
Mikheil Azatov, Silvia M Goicoechea, Carol A Otey, Arpita Upadhyaya
Cells organize actin filaments into higher-order structures by regulating the composition, distribution and concentration of actin crosslinkers. Palladin is an actin crosslinker found in the lamellar actin network and stress fibers, which are critical for mechanosensing of the environment. Palladin also serves as a molecular scaffold for α-actinin, another key actin crosslinker. By virtue of its close interactions with actomyosin structures in the cell, palladin may play an important role in cell mechanics...
2016: Scientific Reports
Jihan Kim, Christopher A R Jones, Nicholas Scott Groves, Bo Sun
Cells in three-dimensional (3D) environments exhibit very different biochemical and biophysical phenotypes compared to the behavior of cells in two-dimensional (2D) environments. As an important biomechanical measurement, 2D traction force microscopy can not be directly extended into 3D cases. In order to quantitatively characterize the contraction field, we have developed 3D reflectance traction microscopy which combines confocal reflection imaging and partial volume correlation postprocessing. We have measured the deformation field of collagen gel under controlled mechanical stress...
2016: PloS One
Andreas P Kourouklis, Kerim B Kaylan, Gregory H Underhill
Recent approaches have utilized microfabricated platforms to examine combinations of microenvironmental signals that regulate stem and progenitor cell differentiation. However, the majority of these efforts have focused on the biochemical properties of extracellular matrix (ECM) or soluble factors without simultaneously exploring the biomechanical effects of cell-substrate interactions. To address this need, we combined a high-throughput approach for the analysis of combinatorial ECM cues with substrates of modular stiffness and traction force microscopy...
August 2016: Biomaterials
Philip A Bondzie, Hui A Chen, Mei Zhen Cao, Julie A Tomolonis, Fangfang He, Martin R Pollak, Joel M Henderson
Several glomerular pathologies resulting from podocyte injury are linked to genetic variation involving the MYH9 gene, which encodes the heavy chain of non-muscle myosin-IIA (NM-IIA). However, the functional role of NM-IIA has not been studied extensively in podocytes. We hypothesized that NM-IIA is critical for maintenance of podocyte structure and mechanical function. To test this hypothesis, we studied murine podocytes in vitro subjected to blebbistatin inhibition of NM-II activity, or RNA interference-mediated, isoform-specific ablation of Myh9 gene and protein (NM-IIA) or its paralog Myh10 gene and protein (NM-IIB)...
August 2016: Cytoskeleton
Sjoerd van Helvert, Peter Friedl
The multistep process of cell migration requires cells to dynamically couple to extracellular interfaces and generate traction force or friction for displacement of the cell body. When deformed, biopolymer networks, including fibrillar collagen and fibrin, undergo a nonlinear elasticity change that is termed strain stiffening and is commonly measured by bulk rheology. It remains poorly characterized, however, whether forces generated by moving cells suffice to induce strain stiffening. To detect strain stiffening at the leading edge of normal and tumor cells moving across fibrillar type I collagen, we combined AFM nanoindentation and differential field probing with confocal reflection microscopy...
August 31, 2016: ACS Applied Materials & Interfaces
Franziska Vielmuth, Ricarda G Schumann, Volker Spindler, Armin Wolf, Renate Scheler, Wolfgang J Mayer, Paul B Henrich, Christos Haritoglou
PURPOSE: To assess the stiffness of the human internal limiting membrane (ILM) and evaluate potential changes of mechanical properties following intravitreal ocriplasmin injection for vitreomacular traction. METHODS: This is an interventional comparative case series of 12 surgically excised ILM specimens consecutively obtained from 9 eyes of 9 patients after unsuccessful pharmacologic vitreolysis with ocriplasmin. During the same time period, 16 specimens from 13 other eyes without ocriplasmin treatment were harvested during vitrectomy and served as controls...
2016: Ophthalmologica. Journal International D'ophtalmologie
Angela J Argyropoulos, Patrick Robichaud, Rebecca Mutesi Balimunkwe, Gary J Fisher, Craig Hammerberg, Yan Yan, Taihao Quan
Alterations of the collagen, the major structural protein in skin, contribute significantly to human skin connective tissue aging. As aged-appearing skin is more common in diabetes, here we investigated the molecular basis of aged-appearing skin in diabetes. Among all known human matrix metalloproteinases (MMPs), diabetic skin shows elevated levels of MMP-1 and MMP-2. Laser capture microdissection (LCM) coupled real-time PCR indicated that elevated MMPs in diabetic skin were primarily expressed in the dermis...
2016: PloS One
Vincent Nier, Shreyansh Jain, Chwee Teck Lim, Shuji Ishihara, Benoit Ladoux, Philippe Marcq
We combine traction force data with Bayesian inversion to obtain an absolute estimate of the internal stress field of a cell monolayer. The method, Bayesian inversion stress microscopy, is validated using numerical simulations performed in a wide range of conditions. It is robust to changes in each ingredient of the underlying statistical model. Importantly, its accuracy does not depend on the rheology of the tissue. We apply Bayesian inversion stress microscopy to experimental traction force data measured in a narrow ring of cohesive epithelial cells, and check that the inferred stress field coincides with that obtained by direct spatial integration of the traction force data in this quasi one-dimensional geometry...
April 12, 2016: Biophysical Journal
Mahmut Selman Sakar, Jeroen Eyckmans, Roel Pieters, Daniel Eberli, Bradley J Nelson, Christopher S Chen
Planar in vitro models have been invaluable tools to identify the mechanical basis of wound closure. Although these models may recapitulate closure dynamics of epithelial cell sheets, they fail to capture how a wounded fibrous tissue rebuilds its 3D architecture. Here we develop a 3D biomimetic model for soft tissue repair and demonstrate that fibroblasts ensconced in a collagen matrix rapidly close microsurgically induced defects within 24 h. Traction force microscopy and time-lapse imaging reveal that closure of gaps begins with contractility-mediated whole-tissue deformations...
2016: Nature Communications
Ismaeel Muhamed, Jun Wu, Poonam Sehgal, Xinyu Kong, Arash Tajik, Ning Wang, Deborah E Leckband
This report elucidates an E-cadherin-based force-transduction pathway that triggers changes in cell mechanics through a mechanism requiring epidermal growth factor receptor (EGFR), phosphoinositide 3-kinase (PI3K), and the downstream formation of new integrin adhesions. This mechanism operates in addition to local cytoskeletal remodeling triggered by conformational changes in the E-cadherin-associated protein α-catenin, at sites of mechanical perturbation. Studies using magnetic twisting cytometry (MTC), together with traction force microscopy (TFM) and confocal imaging identified force-activated E-cadherin-specific signals that integrate cadherin force transduction, integrin activation and cell contractility...
May 1, 2016: Journal of Cell Science
David A Stout, Eyal Bar-Kochba, Jonathan B Estrada, Jennet Toyjanova, Haneesh Kesari, Jonathan S Reichner, Christian Franck
Mechanobiology relates cellular processes to mechanical signals, such as determining the effect of variations in matrix stiffness with cell tractions. Cell traction recorded via traction force microscopy (TFM) commonly takes place on materials such as polyacrylamide- and polyethylene glycol-based gels. Such experiments remain limited in physiological relevance because cells natively migrate within complex tissue microenvironments that are spatially heterogeneous and hierarchical. Yet, TFM requires determination of the matrix constitutive law (stress-strain relationship), which is not always readily available...
March 15, 2016: Proceedings of the National Academy of Sciences of the United States of America
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