scholarly journals A cytoskeletal clutch mediates cellular force transmission in a soft, three-dimensional extracellular matrix

2017 ◽  
Vol 28 (14) ◽  
pp. 1959-1974 ◽  
Author(s):  
Leanna M. Owen ◽  
Arjun S. Adhikari ◽  
Mohak Patel ◽  
Peter Grimmer ◽  
Natascha Leijnse ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Actin Cytoskeleton ◽  
Dynamic Equilibrium ◽  
Three Dimensional ◽  
Time Lapse ◽  
Dermal Fibroblasts ◽  
Force Transmission ◽  
Ecm Remodeling

The ability of cells to impart forces and deformations on their surroundings underlies cell migration and extracellular matrix (ECM) remodeling and is thus an essential aspect of complex, metazoan life. Previous work has resulted in a refined understanding, commonly termed the molecular clutch model, of how cells adhering to flat surfaces such as a microscope coverslip transmit cytoskeletally generated forces to their surroundings. Comparatively less is known about how cells adhere to and exert forces in soft, three-dimensional (3D), and structurally heterogeneous ECM environments such as occur in vivo. We used time-lapse 3D imaging and quantitative image analysis to determine how the actin cytoskeleton is mechanically coupled to the surrounding matrix for primary dermal fibroblasts embedded in a 3D fibrin matrix. Under these circumstances, the cytoskeletal architecture is dominated by contractile actin bundles attached at their ends to large, stable, integrin-based adhesions. Time-lapse imaging reveals that α-actinin-1 puncta within actomyosin bundles move more quickly than the paxillin-rich adhesion plaques, which in turn move more quickly than the local matrix, an observation reminiscent of the molecular clutch model. However, closer examination did not reveal a continuous rearward flow of the actin cytoskeleton over slower moving adhesions. Instead, we found that a subset of stress fibers continuously elongated at their attachment points to integrin adhesions, providing stable, yet structurally dynamic coupling to the ECM. Analytical modeling and numerical simulation provide a plausible physical explanation for this result and support a picture in which cells respond to the effective stiffness of local matrix attachment points. The resulting dynamic equilibrium can explain how cells maintain stable, contractile connections to discrete points within ECM during cell migration, and provides a plausible means by which fibroblasts contract provisional matrices during wound healing.

scholarly journals Mechanobiology of Epithelia From the Perspective of Extracellular Matrix Heterogeneity

2020 ◽  
Vol 8 ◽  
Author(s):  
Aleksandra N. Kozyrina ◽  
Teodora Piskova ◽  
Jacopo Di Russo
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Cell Sheets ◽  
Ecm Remodeling ◽  
Cellular Behavior ◽  
New Concepts ◽  
Matrix Heterogeneity ◽  
The Impact

Understanding the complexity of the extracellular matrix (ECM) and its variability is a necessary step on the way to engineering functional (bio)materials that serve their respective purposes while relying on cell adhesion. Upon adhesion, cells receive messages which contain both biochemical and mechanical information. The main focus of mechanobiology lies in investigating the role of this mechanical coordination in regulating cellular behavior. In recent years, this focus has been additionally shifted toward cell collectives and the understanding of their behavior as a whole mechanical continuum. Collective cell phenomena very much apply to epithelia which are either simple cell-sheets or more complex three-dimensional structures. Researchers have been mostly using the organization of monolayers to observe their collective behavior in well-defined experimental setups in vitro. Nevertheless, recent studies have also reported the impact of ECM remodeling on epithelial morphogenesis in vivo. These new concepts, combined with the knowledge of ECM biochemical complexity are of key importance for engineering new interactive materials to support both epithelial remodeling and homeostasis. In this review, we summarize the structure and heterogeneity of the ECM before discussing its impact on the epithelial mechanobiology.

Extracellular matrix (ECM) microstructural composition regulates local cell-ECM biomechanics and fundamental fibroblast behavior: a multidimensional perspective

2005 ◽  
Vol 98 (5) ◽  
pp. 1909-1921 ◽  
Author(s):  
A. M. Pizzo ◽  
K. Kokini ◽  
L. C. Vaughn ◽  
B. Z. Waisner ◽  
S. L. Voytik-Harbin
Keyword(s):  
Extracellular Matrix ◽  
Cell Fate ◽  
Three Dimensional ◽  
Local Strain ◽  
Β1 Integrin ◽  
Ecm Remodeling ◽  
Surface Areas ◽  
Local Cell ◽  
And Function

The extracellular matrix (ECM) provides the principal means by which mechanical information is communicated between tissue and cellular levels of function. These mechanical signals play a central role in controlling cell fate and establishing tissue structure and function. However, little is known regarding the mechanisms by which specific structural and mechanical properties of the ECM influence its interaction with cells, especially within a tissuelike context. This lack of knowledge precludes formulation of biomimetic microenvironments for effective tissue repair and replacement. The present study determined the role of collagen fibril density in regulating local cell-ECM biomechanics and fundamental fibroblast behavior. The model system consisted of fibroblasts seeded within collagen ECMs with controlled microstructure. Confocal microscopy was used to collect multidimensional images of both ECM microstructure and specific cellular characteristics. From these images temporal changes in three-dimensional cell morphology, time- and space-dependent changes in the three-dimensional local strain state of a cell and its ECM, and spatial distribution of β1-integrin were quantified. Results showed that fibroblasts grown within high-fibril-density ECMs had decreased length-to-height ratios, increased surface areas, and a greater number of projections. Furthermore, fibroblasts within low-fibril-density ECMs reorganized their ECM to a greater extent, and it appeared that β1-integrin localization was related to local strain and ECM remodeling events. Finally, fibroblast proliferation was enhanced in low-fibril-density ECMs. Collectively, these results are significant because they provide new insight into how specific physical properties of a cell’s ECM microenvironment contribute to tissue remodeling events in vivo and to the design and engineering of functional tissue replacements.

scholarly journals Nicotine Upregulates Metalloproteinase Function in Vascular Smooth Muscle Cells Inducing Cell Migration

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 376-376
Author(s):  
Ann Centner ◽  
Abigail Cullen ◽  
Gloria Salazar
Keyword(s):  
Extracellular Matrix ◽  
Animal Model ◽  
Cell Migration ◽  
Plaque Rupture ◽  
Tissue Inhibitors Of Metalloproteinases ◽  
Phenotypic Switching ◽  
Ecm Remodeling ◽  
Plaque Instability ◽  
Health Promoting

Abstract Objectives We hypothesize that nicotine plays a role in the progression of atherosclerosis by inducing VSMC phenotypic switching and cell migration from the media to an inner layer–the intima. This forms a new layer called the neointima, which becomes a plaque with a lipid-rich necrotic core and fibrous VSMC cap. The cap confers protection against plaque rupture and stroke or heart attack. However, VSMCs can contribute to cap thinning by secreting metalloproteinases (MMPs) which degrade the extracellular matrix (ECM) creating a plaque prone to rupture. Methods Our lab will test blackberry extract. Blackberries contain health-promoting phytochemicals called polyphenols and other polyphenol extracts have been shown to inhibit MMPs. Specifically, blackberry extract will be added to nicotine treated cells and to animal model diets. Results In VSMCs, we found that nicotine significantly increased the expression of several MMPs, while decreasing the expression of multiple tissue inhibitors of metalloproteinases (TIMPs). MMP activity measured by zymography was also upregulated by nicotine suggesting that nicotine modulates extracellular matrix (ECM) remodeling by increasing MMP expression and activity. Consistent with ECM remodeling, nicotine promoted cell migration, which was associated with increased reactive oxygen species (ROS) levels. Conclusions These data suggest nicotine promotes VSMC migration into the intima and contributes to plaque instability by fibrous cap weakening. However, future studies are needed to demonstrate nicotine changes plaque composition in vivo. To negate detrimental effects, our lab will test blackberry extract. Blackberries contain health-promoting phytochemicals called polyphenols and other polyphenol extracts have been shown to inhibit MMPs. Specifically, blackberry extract will be added to nicotine treated cells and to animal model diets. Funding Sources Florida Department of Health.

scholarly journals Capturing relevant extracellular matrices for investigating cell migration

F1000Research ◽  
2015 ◽  
Vol 4 ◽  
pp. 1408 ◽  
Author(s):  
Patricia Keely ◽  
Amrinder Nain
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Three Dimensional ◽  
3D Culture ◽  
Extracellular Matrices ◽  
Matrix Composition ◽  
Disease States ◽  
Stromal Microenvironment ◽  
Different Tissues

Much progress in understanding cell migration has been determined by using classic two-dimensional (2D) tissue culture platforms. However, increasingly, it is appreciated that certain properties of cell migration in vivo are not represented by strictly 2D assays. There is much interest in creating relevant three-dimensional (3D) culture environments and engineered platforms to better represent features of the extracellular matrix and stromal microenvironment that are not captured in 2D platforms. Important to this goal is a solid understanding of the features of the extracellular matrix—composition, stiffness, topography, and alignment—in different tissues and disease states and the development of means to capture these features

Induction of in Vivo Ectopic Hematopoiesis by a Three-Dimensional Structured Extracellular Matrix Derived from Decellularized Cancellous Bone

2019 ◽  
Vol 5 (11) ◽  
pp. 5669-5680 ◽  
Author(s):  
Naoko Nakamura ◽  
Tsuyoshi Kimura ◽  
Kwangwoo Nam ◽  
Toshiya Fujisato ◽  
Hiroo Iwata ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cancellous Bone ◽  
Three Dimensional

scholarly journals Artificial Tumor Microenvironments in Neuroblastoma

Cancers ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1629
Author(s):  
Colin H. Quinn ◽  
Andee M. Beierle ◽  
Elizabeth A. Beierle
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Therapeutic Interventions ◽  
3D Bioprinting ◽  
Culture Studies ◽  
In Vivo Models ◽  
Novel Treatments ◽  
Tumor Microenvironments ◽  
Novel Approach

In the quest to advance neuroblastoma therapeutics, there is a need to have a deeper understanding of the tumor microenvironment (TME). From extracellular matrix proteins to tumor associated macrophages, the TME is a robust and diverse network functioning in symbiosis with the solid tumor. Herein, we review the major components of the TME including the extracellular matrix, cytokines, immune cells, and vasculature that support a more aggressive neuroblastoma phenotype and encumber current therapeutic interventions. Contemporary treatments for neuroblastoma are the result of traditional two-dimensional culture studies and in vivo models that have been translated to clinical trials. These pre-clinical studies are costly, time consuming, and neglect the study of cofounding factors such as the contributions of the TME. Three-dimensional (3D) bioprinting has become a novel approach to studying adult cancers and is just now incorporating portions of the TME and advancing to study pediatric solid. We review the methods of 3D bioprinting, how researchers have included TME pieces into the prints, and highlight present studies using neuroblastoma. Ultimately, incorporating the elements of the TME that affect neuroblastoma responses to therapy will improve the development of innovative and novel treatments. The use of 3D bioprinting to achieve this aim will prove useful in developing optimal therapies for children with neuroblastoma.

Cell-extracellular matrix interactions under in vivo conditions during interstitial cell migration in Hydra vulgaris

Development ◽  
1994 ◽  
Vol 120 (2) ◽  
pp. 425-432 ◽  
Author(s):  
X. Zhang ◽  
M.P. Sarras
Keyword(s):  
Extracellular Matrix ◽  
Cell Migration ◽  
Synthetic Peptide ◽  
Interstitial Cell ◽  
Extracellular Matrix Components ◽  
Matrix Interactions ◽  
Apical Half ◽  
Matrix Components ◽  
Extracellular Matrix Interactions

Interstitial cell (I-cell) migration in hydra is essential for establishment of the regional cell differentiation pattern in the organism. All previous in vivo studies have indicated that cell migration in hydra is a result of cell-cell interactions and chemotaxic gradients. Recently, in vitro cell adhesion studies indicated that isolated nematocytes could bind to substrata coated with isolated hydra mesoglea, fibronectin and type IV collagen. Under these conditions, nematocytes could be observed to migrate on some of these extracellular matrix components. By modifying previously described hydra grafting techniques, two procedures were developed to test specifically the role of extracellular matrix components during in vivo I-cell migration in hydra. In one approach, the extracellular matrix structure of the apical half of the hydra graft was perturbed using beta-aminopropionitrile and beta-xyloside. In the second approach, grafts were treated with fibronectin, RGDS synthetic peptide and antibody to fibronectin after grafting was performed. In both cases, I-cell migration from the basal half to the apical half of the grafts was quantitatively analyzed. Statistical analysis indicated that beta-aminopropionitrile, fibronectin, RGDS synthetic peptide and antibody to fibronectin all were inhibitory to I-cell migration as compared to their respective controls. beta-xyloside treatment had no effect on interstitial cell migration. These results indicate the potential importance of cell-extracellular matrix interactions during in vivo I-cell migration in hydra.

Nonuniform strain of human soleus aponeurosis-tendon complex during submaximal voluntary contractions in vivo

2003 ◽  
Vol 95 (2) ◽  
pp. 829-837 ◽  
Author(s):  
Taija Finni ◽  
John A. Hodgson ◽  
Alex M. Lai ◽  
V. Reggie Edgerton ◽  
Shantanu Sinha
Keyword(s):  
Achilles Tendon ◽  
Maximal Voluntary Contraction ◽  
Three Dimensional ◽  
Motor Units ◽  
Magnetic Resonance Images ◽  
Voluntary Contraction ◽  
Force Transmission ◽  
Voluntary Contractions ◽  
Nonuniform Strain

The distribution of strain along the soleus aponeurosis tendon was examined during voluntary contractions in vivo. Eight subjects performed cyclic isometric contractions (20 and 40% of maximal voluntary contraction). Displacement and strain in the apparent Achilles tendon and in the aponeurosis were calculated from cine phase-contrast magnetic resonance images acquired with a field of view of 32 cm. The apparent Achilles tendon lengthened 2.8 and 4.7% in 20 and 40% maximal voluntary contraction, respectively. The midregion of the aponeurosis, below the gastrocnemius insertion, lengthened 1.2 and 2.2%, but the distal aponeurosis shortened 2.1 and 2.5%, respectively. There was considerable variation in the three-dimensional anatomy of the aponeurosis and muscle-tendon junction. We suggest that the nonuniformity in aponeurosis strain within an individual was due to the presence of active and passive motor units along the length of the muscle, causing variable force along the measurement site. Force transmission along intrasoleus connective tissue may also be a significant source of nonuniform strain in the aponeurosis.

Interaction between vascular endothelial cells and vascular intimal spindle-shaped cells in vitro

1983 ◽  
Vol 60 (1) ◽  
pp. 89-102
Author(s):  
D de Bono ◽  
C. Green
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Vascular Endothelial Cells ◽  
Three Dimensional ◽  
Vascular Endothelial ◽  
Pure Cultures ◽  
Species Specific ◽  
Endothelial Growth Factor

The interactions between human or bovine vascular endothelial cells and fibroblast-like vascular intimal spindle-shaped cells have been studied in vitro, using species-specific antibodies to identify the different components in mixed cultures. Pure cultures of endothelial cells grow as uniform, nonoverlapping monolayers, but this growth pattern is lost after the addition of spindle cells, probably because the extracellular matrix secreted by the latter causes the endothelial cells to modify the way they are attached to the substrate. The result is a network of tubular aggregates of endothelial cells in a three-dimensional ‘polylayer’ of spindle-shaped cells. On the other hand, endothelial cells added to growth-inhibited cultures of spindle-shaped cells will grow in sheets over the surface of the culture. Human endothelial cells grown in contact with spindle-shaped cells have a reduced requirement for a brain-derived endothelial growth factor. The interactions of endothelial cells and other connective tissue cells in vitro may be relevant to the mechanisms of endothelial growth and blood vessel formation in vivo, and emphasize the potential importance of extracellular matrix in controlling endothelial cell behaviour.

Local, Three-Dimensional Strain Measurements Within Largely Deformed Extracellular Matrix Constructs

2004 ◽  
Vol 126 (6) ◽  
pp. 699-708 ◽  
Author(s):  
Blayne A. Roeder ◽  
Klod Kokini ◽  
J. Paul Robinson ◽  
Sherry L. Voytik-Harbin
Keyword(s):  
Extracellular Matrix ◽  
Local Level ◽  
Three Dimensional ◽  
Collagen Matrices ◽  
Mechanical Loads ◽  
Strain Measurements ◽  
3D Collagen ◽  
Vitro Model

The ability to create extracellular matrix (ECM) constructs that are mechanically and biochemically similar to those found in vivo and to understand how their properties affect cellular responses will drive the next generation of tissue engineering strategies. To date, many mechanisms by which cells biochemically communicate with the ECM are known. However, the mechanisms by which mechanical information is transmitted between cells and their ECM remain to be elucidated. “Self-assembled” collagen matrices provide an in vitro-model system to study the mechanical behavior of ECM. To begin to understand how the ECM and the cells interact mechanically, the three-dimensional (3D) mechanical properties of the ECM must be quantified at the micro-(local) level in addition to information measured at the macro-(global) level. Here we describe an incremental digital volume correlation (IDVC) algorithm to quantify large (>0.05) 3D mechanical strains in the microstructure of 3D collagen matrices in response to applied mechanical loads. Strain measurements from the IDVC algorithm rely on 3D confocal images acquired from collagen matrices under applied mechanical loads. The accuracy and the precision of the IDVC algorithm was verified by comparing both image volumes collected in succession when no deformation was applied to the ECM (zero strain) and image volumes to which simulated deformations were applied in both 1D and 3D (simulated strains). Results indicate that the IDVC algorithm can accurately and precisely determine the 3D strain state inside largely deformed collagen ECMs. Finally, the usefulness of the algorithm was demonstrated by measuring the microlevel 3D strain response of a collagen ECM loaded in tension.

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