Molecular-scale Topographic Cues Induce the Orientation and Directional Movement of Fibroblasts on Two-dimensional Collagen Surfaces

Poole, Kate; Khairy, Khaled; Friedrichs, Jens; Franz, Clemens M.; Cisneros, David A.; Howard, Jonathon; Müeller, Daniel J.

doi:10.1016/j.jmb.2005.03.064

Cited by 118 publications

(109 citation statements)

References 24 publications

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“…The matrix fibrils displayed the 67 nm D-periodicity frequently observed for collagen type I fibrils in vivo. These ordered collagen matrices are thin (ϳ3 nm), extremely flat (height variations Ͻ1 nm), and promote the adhesion and spreading of different cell types, including fibroblasts and epithelial and endothelial cells (Poole et al, 2005). These matrices are well suited for single-cell adhesion studies because their planarity and structural homogeneity guarantee that differences in adhesion are cell-specific and not caused by variations in the matrix composition or topography.…”

Section: ␣ 2 ␤ 1 -Mediated Cell Adhesion To Collagen Type I Matricesmentioning

confidence: 99%

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Taubenberger

Cisneros

Friedrichs

et al. 2007

MBoC

Self Cite

192

197

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We have characterized early steps of ␣ 2 ␤ 1 integrin-mediated cell adhesion to a collagen type I matrix by using single-cell force spectroscopy. In agreement with the role of ␣ 2 ␤ 1 as a collagen type I receptor, ␣ 2 ␤ 1 -expressing Chinese hamster ovary (CHO)-A2 cells spread rapidly on the matrix, whereas ␣ 2 ␤ 1 -negative CHO wild-type cells adhered poorly. Probing CHO-A2 cell detachment forces over a contact time range of 600 s revealed a nonlinear adhesion response. During the first 60 s, cell adhesion increased slowly, and forces associated with the smallest rupture events were consistent with the breakage of individual integrin-collagen bonds. Above 60 s, a fraction of cells rapidly switched into an activated adhesion state marked by up to 10-fold increased detachment forces. Elevated overall cell adhesion coincided with a rise of the smallest rupture forces above the value required to break a single-integrin-collagen bond, suggesting a change from single to cooperative receptor binding. Transition into the activated adhesion mode and the increase of the smallest rupture forces were both blocked by inhibitors of actomyosin contractility. We therefore propose a two-step mechanism for the establishment of ␣ 2 ␤ 1 -mediated adhesion as weak initial, single-integrin-mediated binding events are superseded by strong adhesive interactions involving receptor cooperativity and actomyosin contractility. INTRODUCTIONIntegrins are a family of ␣/␤-heterodimeric receptors involved in cell-cell adhesion and cell attachment to the extracellular matrix (Hynes, 1992). In adherent cells, integrins are often arranged into highly organized structures, such as focal complexes, focal adhesions, and fibrillar adhesions . In these mature adhesive contacts, integrins are linked to the actin cytoskeleton via their intracellular domains and a multitude of adapter proteins, such as vinculin, talin, and ␣-actinin (Zamir and Geiger, 2001). Whereas the molecular composition of integrin complexes and signaling pathways controlling their macroscopic assembly or disassembly have been analyzed in detail, less is known about the early molecular events leading to the formation of integrin-based adhesion.Generally, integrin-mediated adhesion is thought to be initiated on the cell membrane by the engagement of individual integrin dimers with their respective ligand (Lotz et al., 1989;Gallant and Garcia, 2006). The number of receptorligand pairs may then grow by increasing the cell-substrate contact area and by receptor diffusion within that zone. Subsequent adhesion strengthening occurs through integrin clustering and linkage to the cytoskeleton. In addition, the ligand affinity of the integrins may increase as a result of intracellular regulatory pathways (Hughes and Pfaff, 1998). At a later stage, the assembly of higher-order integrincytoskeletal complexes requires actomyosin-driven contractility under the control of the small guanosine triphosphatase (GTPase) RhoA (Chrzanowska-Wodnicka and Burridge, 1996) and its downstream target Rho-assoc...

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Section: ␣ 2 ␤ 1 -Mediated Cell Adhesion To Collagen Type I Matricesmentioning

confidence: 99%

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Taubenberger

Cisneros

Friedrichs

et al. 2007

MBoC

Self Cite

192

197

View full text Add to dashboard Cite

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“…This level of organization is thought to be important because ECM molecules are multifaceted, containing multiple domains with repeated motifs whose presentation to the cell is both complex and can be changed by protein folding and unfolding (Poole et al, 2005;Martino et al, 2009). We also know that the ability of integrins to cluster, a necessary prerequisite for recruitment of proteins to the FA complex, is affected by spacing of the presented ligand and has downstream consequence for integrin-linked signalling cascades (Irvine et al, 2002;Selhuber-Unkel et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Lateral spacing of adhesion peptides influences human mesenchymal stem cell behaviour

Frith

Mills

Cooper-White

2012

Journal of Cell Science

142

157

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SummaryMesenchymal stem cells (MSCs) have attracted great interest in recent years for tissue engineering and regenerative medicine applications due to their ease of isolation and multipotent differentiation capacity. In the past, MSC research has focussed on the effects of soluble cues, such as growth factors and cytokines; however, there is now increasing interest in understanding how parameters such as substrate modulus, specific extracellular matrix (ECM) components and the ways in which these are presented to the cell can influence MSC properties. Here we use surfaces of self-assembled maleimide-functionalized polystyrene-blockpoly(ethylene oxide) copolymers (PS-PEO-Ma) to investigate how the spatial arrangement of cell adhesion ligands affects MSC behaviour. By changing the ratio of PS-PEO-Ma in mixtures of block copolymer and polystyrene homopolymer, we can create surfaces with lateral spacing of the PEO-Ma domains ranging from 34 to 62 nm. Through subsequent binding of cysteine-GRGDS peptides to the maleimide-terminated end of the PEO chains in each of these domains, we are able to present tailored surfaces of controlled lateral spacing of RGD (arginine-glycine-aspartic acid) peptides to MSCs. We demonstrate that adhesion of MSCs to the RGD-functionalized block-copolymer surfaces is through specific attachment to the presented RGD motif and that this is mediated by a5, aV, b1 and b3 integrins. We show that as the lateral spacing of the peptides is increased, the ability of the MSCs to spread is diminished and that the morphology changes from well-spread cells with normal fibroblastic morphology and defined stress-fibres, to less-spread cells with numerous cell protrusions and few stress fibres. In addition, the ability of MSCs to form mature focal adhesions is reduced on substrates with increased lateral spacing. Finally, we investigate differentiation and use qRT-PCR determination of gene expression levels and a quantitative alkaline phosphatase assay to show that MSC osteogenesis is reduced on surfaces with increased lateral spacing while adipogenic differentiation is increased. We show here, for the first time, that the lateral spacing of adhesion peptides affects human MSC (hMSC) properties and might therefore be a useful parameter with which to modify hMSC behaviour in future tissue engineering strategies.

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“…In addition, we succeeded in generating a fluorescently labeled matrix. This highly ordered collagen-I matrix simplifies the visualization of cellular responses to the substrate Jiang et al, 2004;Poole et al, 2005). For this, a combination of fluorescence and atomic force microscopy (AFM) was used (Fig.…”

Section: Introductionmentioning

confidence: 99%

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

Kirmse

Otto

Ludwig

2011

Journal of Cell Science

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SummaryIt is becoming increasingly evident that the micromechanics of cells and their environment determine cell fate and function as much as soluble molecular factors do. We hypothesized that extracellular matrix proteolysis by membrane type 1 matrix metalloproteinase (MT1-MMP) depends on adhesion, force generation and rigidity sensing of the cell. Melanoma cells (MV3 clone) stably transfected with MT1-MMP, or the empty vector as a control, served as the model system. 21 integrins (cell adhesion), actin and myosin II (force generation and rigidity sensing) were blocked by their corresponding inhibitors (21 integrin antibodies, Cytochalasin D, blebbistatin). A novel, anisotropic matrix array of parallel, fluorescently labeled collagen-I fibrils was used. Cleavage and bundling of the collagen-I fibrils, and spreading and durotaxis of the cells on this matrix array could be readily discerned and quantified by a combined set-up for fluorescence and atomic force microscopy. In short, expression of the protease resulted in the generation of structural matrix defects, clearly indicated by gaps in the collagen lattice and loose fiber bundles. This key feature of matrix remodeling depended essentially on the functionality of 21 integrin, the actin filament network and myosin II motor activity. Interference with any of these negatively impacted matrix cleavage and three-dimensional matrix entanglement of cells.

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Molecular-scale Topographic Cues Induce the Orientation and Directional Movement of Fibroblasts on Two-dimensional Collagen Surfaces

Cited by 118 publications

References 24 publications

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Lateral spacing of adhesion peptides influences human mesenchymal stem cell behaviour

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

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Molecular-scale Topographic Cues Induce the Orientation and Directional Movement of Fibroblasts on Two-dimensional Collagen Surfaces

Cited by 118 publications

References 24 publications

Revealing Early Steps of α2β1Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α2β1Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Lateral spacing of adhesion peptides influences human mesenchymal stem cell behaviour

Interdependency of cell adhesion, force generation and extracellular proteolysis in matrix remodeling

Contact Info

Product

Resources

About

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

Revealing Early Steps of α₂β₁Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy