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Fibrin binds to collagen and provides a bridge for αVβ3 integrin-dependent contraction of collagen gels.

Reyhani V, Seddigh P, Guss B, Gustafsson R, Rask L, Rubin K - Biochem. J. (2014)

Bottom Line: This allowed murine myoblast C2C12 cells to contract the collagenous composite gel via αVβ3 integrin.A specific competitive inhibitor blocking the Col-I-binding site for fibrinogen abolished the organization of fibrin into discernable fibrils, as well as the C2C12-mediated contraction of Col I gels.Our data show that fibrin can function as a linkage protein between Col I fibres and cells, and suggest that fibrin at inflammatory sites indirectly connects αVβ3 integrins to Col I fibres and thereby promotes cell-mediated contraction of collagenous tissue structures.

View Article: PubMed Central - PubMed

Affiliation: *Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, BMC Box 582, SE-751 23 Uppsala, Sweden.

ABSTRACT
The functional significance of fibrin deposits typically seen in inflammatory lesions, carcinomas and in healing wounds is not fully understood. In the present study, we demonstrate that fibrinogen/fibrin specifically bound to native Col I (collagen type I) and used the Col I fibre network as a base to provide a functional interface matrix that connects cells to the Col I fibres through αVβ3 integrins. This allowed murine myoblast C2C12 cells to contract the collagenous composite gel via αVβ3 integrin. We show that fibrinogen specifically bound to immobilized native Col I at the site known to bind matrix metalloproteinase-1, discoidin domain receptor-2 and fibronectin, and that binding had no effect on Col I fibrillation. A specific competitive inhibitor blocking the Col-I-binding site for fibrinogen abolished the organization of fibrin into discernable fibrils, as well as the C2C12-mediated contraction of Col I gels. Our data show that fibrin can function as a linkage protein between Col I fibres and cells, and suggest that fibrin at inflammatory sites indirectly connects αVβ3 integrins to Col I fibres and thereby promotes cell-mediated contraction of collagenous tissue structures.

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Immunogold staining of fibrinogen/fibrin in C2C12-mediated fibrin/collagen gel contraction(A–D) Fibrin was detected in close proximity to cell-membrane protrusions of C2C12 cells cultured in fibrin/Col I gels. The fibril structures around the cell surfaces were analysed using SEM, and back-scattered SEM was used to detect the gold particles (15 nm diameter). The gold particles are seen as white spots in (C) or black spots in the inverted image in (D). (E–H) Representative micrographs of C2C12-α2β1 cells in pure Col I gels that were treated and analysed identically with C2C12 cells in fibrin/Col I gels (A–D).
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Figure 3: Immunogold staining of fibrinogen/fibrin in C2C12-mediated fibrin/collagen gel contraction(A–D) Fibrin was detected in close proximity to cell-membrane protrusions of C2C12 cells cultured in fibrin/Col I gels. The fibril structures around the cell surfaces were analysed using SEM, and back-scattered SEM was used to detect the gold particles (15 nm diameter). The gold particles are seen as white spots in (C) or black spots in the inverted image in (D). (E–H) Representative micrographs of C2C12-α2β1 cells in pure Col I gels that were treated and analysed identically with C2C12 cells in fibrin/Col I gels (A–D).


Fibrin binds to collagen and provides a bridge for αVβ3 integrin-dependent contraction of collagen gels.

Reyhani V, Seddigh P, Guss B, Gustafsson R, Rask L, Rubin K - Biochem. J. (2014)

Immunogold staining of fibrinogen/fibrin in C2C12-mediated fibrin/collagen gel contraction(A–D) Fibrin was detected in close proximity to cell-membrane protrusions of C2C12 cells cultured in fibrin/Col I gels. The fibril structures around the cell surfaces were analysed using SEM, and back-scattered SEM was used to detect the gold particles (15 nm diameter). The gold particles are seen as white spots in (C) or black spots in the inverted image in (D). (E–H) Representative micrographs of C2C12-α2β1 cells in pure Col I gels that were treated and analysed identically with C2C12 cells in fibrin/Col I gels (A–D).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109839&req=5

Figure 3: Immunogold staining of fibrinogen/fibrin in C2C12-mediated fibrin/collagen gel contraction(A–D) Fibrin was detected in close proximity to cell-membrane protrusions of C2C12 cells cultured in fibrin/Col I gels. The fibril structures around the cell surfaces were analysed using SEM, and back-scattered SEM was used to detect the gold particles (15 nm diameter). The gold particles are seen as white spots in (C) or black spots in the inverted image in (D). (E–H) Representative micrographs of C2C12-α2β1 cells in pure Col I gels that were treated and analysed identically with C2C12 cells in fibrin/Col I gels (A–D).
Bottom Line: This allowed murine myoblast C2C12 cells to contract the collagenous composite gel via αVβ3 integrin.A specific competitive inhibitor blocking the Col-I-binding site for fibrinogen abolished the organization of fibrin into discernable fibrils, as well as the C2C12-mediated contraction of Col I gels.Our data show that fibrin can function as a linkage protein between Col I fibres and cells, and suggest that fibrin at inflammatory sites indirectly connects αVβ3 integrins to Col I fibres and thereby promotes cell-mediated contraction of collagenous tissue structures.

View Article: PubMed Central - PubMed

Affiliation: *Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, BMC Box 582, SE-751 23 Uppsala, Sweden.

ABSTRACT
The functional significance of fibrin deposits typically seen in inflammatory lesions, carcinomas and in healing wounds is not fully understood. In the present study, we demonstrate that fibrinogen/fibrin specifically bound to native Col I (collagen type I) and used the Col I fibre network as a base to provide a functional interface matrix that connects cells to the Col I fibres through αVβ3 integrins. This allowed murine myoblast C2C12 cells to contract the collagenous composite gel via αVβ3 integrin. We show that fibrinogen specifically bound to immobilized native Col I at the site known to bind matrix metalloproteinase-1, discoidin domain receptor-2 and fibronectin, and that binding had no effect on Col I fibrillation. A specific competitive inhibitor blocking the Col-I-binding site for fibrinogen abolished the organization of fibrin into discernable fibrils, as well as the C2C12-mediated contraction of Col I gels. Our data show that fibrin can function as a linkage protein between Col I fibres and cells, and suggest that fibrin at inflammatory sites indirectly connects αVβ3 integrins to Col I fibres and thereby promotes cell-mediated contraction of collagenous tissue structures.

Show MeSH
Related in: MedlinePlus