Fibroblast contraction occurs on release of tension in attached collagen lattices: Dependency on an organized actin cytoskeleton and serum

Tomasek, James J.; Haaksma, Carol J.; Eddy, Robert J.; Vaughan, Melville B.

doi:10.1002/ar.1092320305

Cited by 228 publications

(215 citation statements)

References 33 publications

Supporting

Mentioning

201

Contrasting

Unclassified

Order By: Relevance

“…The contractility of whole cell populations was quantified with the use of stress-released collagen lattices (Mochitate et al, 1991;Tomasek et al, 1992;Grinnell et al, 1999b). Our results indicate a correlation between the level of ␣-SMA expression and fibroblast contraction.…”

Section: Introductionmentioning

confidence: 80%

Alpha-Smooth Muscle Actin Expression Upregulates Fibroblast Contractile Activity

Hinz

Celetta

Tomasek³

et al. 2001

MBoC

Self Cite

1,096

947

View full text Add to dashboard Cite

To evaluate whether ␣-smooth muscle actin (␣-SMA) plays a role in fibroblast contractility, we first compared the contractile activity of rat subcutaneous fibroblasts (SCFs), expressing low levels of ␣-SMA, with that of lung fibroblasts (LFs), expressing high levels of ␣-SMA, with the use of silicone substrates of different stiffness degrees. On medium stiffness substrates the percentage of cells producing wrinkles was similar to that of ␣-SMA-positive cells in each fibroblast population. On high stiffness substrates, wrinkle production was limited to a subpopulation of LFs very positive for ␣-SMA. In a second approach, we measured the isotonic contraction of SCF-and LF-populated attached collagen lattices. SCFs exhibited 41% diameter reduction compared with 63% by LFs. TGF␤1 increased ␣-SMA expression and lattice contraction by SCFs to the levels of LFs; TGF␤-antagonizing agents reduced ␣-SMA expression and lattice contraction by LFs to the level of SCFs. Finally, 3T3 fibroblasts transiently or permanently transfected with ␣-SMA cDNA exhibited a significantly higher lattice contraction compared with wild-type 3T3 fibroblasts or to fibroblasts transfected with ␣-cardiac and ␤-or ␥-cytoplasmic actin. This took place in the absence of any change in smooth muscle or nonmuscle myosin heavy-chain expression. Our results indicate that an increased ␣-SMA expression is sufficient to enhance fibroblast contractile activity. INTRODUCTIONEarly during healing of an open wound, resident dermal fibroblasts proliferate from the wound margin and migrate into the provisional matrix composed of a fibrin clot. About 1 week after wounding, the provisional matrix is replaced by neo-formed connective tissue, known as granulation tissue, essentially composed of small vessels, extracellular matrix, and fibroblastic cells that become activated and modulate into myofibroblasts. The main feature of myofibroblasts is represented by an important contractile apparatus similar to that of smooth muscle (Gabbiani et al., 1971), and in particular by the neo-expression of ␣-smooth muscle actin (␣-SMA), the actin isoform typical of vascular smooth muscle cells (Skalli et al., 1986). Myofibroblasts are recognized to play a central role in closing the wound tissue, through their capacity to produce a strong contractile force (for review see Grinnell, 1994;Rønnov-Jessen et al., 1996;Powell et al., 1999;Serini and Gabbiani, 1999), possibly generated within stress fibers, similar to those present in cultured fibroblasts (Skalli et al., 1986;Serini and Gabbiani, 1999). Because wound contraction takes place when de novo expressed ␣-SMA is incorporated in stress fibers (Darby et al., 1990), it has been suggested that this actin isoform plays an important role in granulation tissue contraction (for review see Serini and Gabbiani, 1999). Although stress fibers have been proposed to function as contractile organelles (Burridge, 1981;Katoh et al., 1998) and their presence has been correlated with the production of isometric tension (Harris et al., 1981), at p...

show abstract

Section: Introductionmentioning

confidence: 80%

Alpha-Smooth Muscle Actin Expression Upregulates Fibroblast Contractile Activity

Hinz

Celetta

Tomasek³

et al. 2001

MBoC

Self Cite

1,096

947

View full text Add to dashboard Cite

show abstract

“…8 The lattices were prepared in high attachment 6-well plates (Corning Life Sciences). For each well, 0.4 mL of fibroblast suspension containing 2.5 ϫ 10 5 cells were admixed with 0.4 mL of rat tail type I collagen solution (3.85 mg/mL; BD Biosciences) and placed into a plastic culture dish.…”

Section: Attached-released Collagen Contraction Assaymentioning

confidence: 99%

“…The attached-released assay is preferentially used to observe tension development. 8 The rate of contraction immediately after release (Ͻ10 minutes) indicates the relative tension development, whereas cell movements and matrix organization can be observed during longer periods after lattice release.…”

Section: Fussel-15 Is Expressed In Activated Fibroblasts During Woundmentioning

confidence: 99%

Fussel-15, a New Player in Wound Healing, Is Deregulated in Keloid and Localized Scleroderma

Arndt

Schmidt

Wacker

et al. 2011

The American Journal of Pathology

View full text Add to dashboard Cite

Dermal wound healing depends on highly complex interplay among various cytokines and cell types. Disruption of this process can result in impaired healing in the form of excessive scarring, as is the case in fibrotic diseases such as keloid and scleroderma. In the present study, we found Fussel-15, a new member of the Ski/Sno family of TGF-␤/BMP signaling repressors, to be expressed in early wound healing and constantly overexpressed in keloid-derived and scleroderma-derived fibroblasts. Comparing the results of three-dimensional free-floating and attached-released in vitro wound healing assays, we observed that Fussel-15 is expressed during the migratory phase in the free-floating assay, indicating that Fussel-15 might play a role during fibroblast migration. Fussel-15-transfected fibroblasts showed greater migration ability in a scratch wound healing assay, compared with control-transfected cells. This migratory phenotype due to Fussel-15 was confirmed by increased peripheral F-actin localization and modifications in size, amount, and distribution of focal adhesion complexes, which were observed using F-actin and focal adhesion kinase (FAK) immunofluorescence staining, respectively. The present results suggest that expression of Fussel-15 during wound healing might promote fibroblast migration. Permanent expression of Fussel-15 in keloid and skin sclerosis fibroblasts could be involved in the pathogenesis of these conditions, but the molecular mechanism underlying this up-regulation remains to be determined.

show abstract

“…In fibroblast seeded fibrin gels, active retraction accounted for ∼75% of the total retraction (Grouf et al 2007;Balestrini and Billiar 2009) and in collagen gels seeded with fibroblasts, this was ∼80% (Tomasek et al 1992). Based on this literature, the active retraction in the TE constructs was less than expected.…”

Section: Passive and Active Components In Tissue Retractionmentioning

confidence: 88%

“…Active and passive phenomena have been investigated thoroughly in short-term studies with fibroblast-populated collagen and fibrin gel systems Marquez et al 2009;John et al 2010;Youssef et al 2011;Tomasek et al 1992;Grouf et al 2007;Balestrini and Billiar 2009). In tissue engineered heart valves, the ECM is developed during 4 weeks of tissue culture and is more mature than the generally investigated collagen and fibrin gels.…”

Section: Introductionmentioning

confidence: 99%

Passive and active contributions to generated force and retraction in heart valve tissue engineering

Vlimmeren

Driessen-Mol

Oomens

et al. 2012

Biomech Model Mechanobiol

View full text Add to dashboard Cite

In tissue engineered heart valves, cell-mediated stress development during culture results in leaflet retraction at time of implantation. This tissue retraction is partly active due to traction forces exerted by the cells and partly passive due to release of residual stress in the extracellular matrix and the cells. Within this study, we unraveled the passive and active contributions of cells and matrix to generated force and retraction in engineered heart valve tissues. Tissue engineered rectangular strips, fabricated from PGA/P4HB scaffolds and seeded with human myofibroblasts, were cultured for 4 weeks, after which the cellular contribution was changed at different levels. Elimination of the active cellular traction forces was achieved with Cytochalasin D and inhibition of the Rho-associated kinase pathway. Both active and passive cellular contributions were eliminated by lysation and/or decellularization of the tissue. Maximum cell activity was reached by increasing the fetal bovine serum concentration to 50%. The generated force decreased ∼20% after elimination of the active cellular component, ∼25% when the passive cellular component was removed as well and remained unaffected by increased serum concentrations. Passive retraction accounted for ∼60% of total retraction, of which ∼15% was residual stress in the matrix and ∼45% was passive cell retraction. Cell traction forces accounted for the remainder ∼40% of the retraction. Full activation of the cells increased retraction by ∼45%. These results illustrate the importance of the cells in the process of tissue retraction, not only actively retracting the tissue, but also in a passive manner to a large extent.

show abstract

Fibroblast contraction occurs on release of tension in attached collagen lattices: Dependency on an organized actin cytoskeleton and serum

Cited by 228 publications

References 33 publications

Alpha-Smooth Muscle Actin Expression Upregulates Fibroblast Contractile Activity

Alpha-Smooth Muscle Actin Expression Upregulates Fibroblast Contractile Activity

Fussel-15, a New Player in Wound Healing, Is Deregulated in Keloid and Localized Scleroderma

Passive and active contributions to generated force and retraction in heart valve tissue engineering

Contact Info

Product

Resources

About