Influence of type I collagen surface density on fibroblast spreading, motility, and contractility.

We examine the relationships of three variables (projected area, migration speed, and traction force) at various type I collagen surface densities in a population of fibroblasts. We observe that cell area is initially an increasing function of ligand density, but that above a certain transition level, increases in surface collagen cause cell area to decline. The threshold collagen density that separates these two qualitatively different regimes, approximately 160 molecules/ microm(2), is approximately equal to the cell surface density of integrin molecules. These results suggest a model in which collagen density induces a qualitative transition in the fundamental way that fibroblasts interact with the substrate. At low density, the availability of collagen binding sites is limiting and the cells simply try to flatten as much as possible by pulling on the few available sites as hard as they can. The force per bond under these conditions approaches 100 pN, approximately equal to the force required for rupture of integrin-peptide bonds. In contrast, at high collagen density adhesion, traction force and motility are limited by the availability of free integrins on the cell surface since so many of these receptors are bound to the surface ligand and the force per bond is very low.

[1]  S. Breit,et al.  Microplate reader-based quantitation of collagens. , 1992, Analytical biochemistry.

[2]  S. Carter,et al.  Haptotaxis and the Mechanism of Cell Motility , 1967, Nature.

[3]  K. Jacobson,et al.  Imaging the traction stresses exerted by locomoting cells with the elastic substratum method. , 1996, Biophysical journal.

[4]  D A Lauffenburger,et al.  Mathematical model for the effects of adhesion and mechanics on cell migration speed. , 1991, Biophysical journal.

[5]  R. Marchant,et al.  Force measurements on the molecular interactions between ligand (RGD) and human platelet αIIbβ3 receptor system , 2001 .

[6]  Joyce Y. Wong,et al.  Directed Movement of Vascular Smooth Muscle Cells on Gradient-Compliant Hydrogels† , 2003 .

[7]  B. Geiger,et al.  Interaction of fibronectin-coated beads with attached and spread fibroblasts. Binding, phagocytosis, and cytoskeletal reorganization. , 1986, Experimental cell research.

[8]  Micah Dembo,et al.  Focal adhesion kinase is involved in mechanosensing during fibroblast migration , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A. Huttenlocher,et al.  Integrin-mediated adhesion regulates cell polarity and membrane protrusion through the Rho family of GTPases. , 2001, Molecular biology of the cell.

[10]  M. Dembo,et al.  Traction force microscopy of migrating normal and H-ras transformed 3T3 fibroblasts. , 2001, Biophysical journal.

[11]  M A Horton,et al.  Single integrin molecule adhesion forces in intact cells measured by atomic force microscopy. , 1999, Biochemical and biophysical research communications.

[12]  M. Dembo,et al.  Stresses at the cell-to-substrate interface during locomotion of fibroblasts. , 1999, Biophysical journal.

[13]  A. Kleinzeller,et al.  Current Topics in Membranes and Transport , 1970 .

[14]  Y. Wang,et al.  Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. , 1998, Methods in enzymology.

[15]  M. Dembo,et al.  Distinct roles of frontal and rear cell-substrate adhesions in fibroblast migration. , 2001, Molecular biology of the cell.

[16]  Sean P. Palecek,et al.  Erratum: Integrin–ligand binding properties govern cell migration speed through cell–substratum adhesiveness , 1997, Nature.

[17]  K. Beningo,et al.  Nascent Focal Adhesions Are Responsible for the Generation of Strong Propulsive Forces in Migrating Fibroblasts , 2001, The Journal of cell biology.

[18]  A. Harris,et al.  Behavior of cultured cells on substrata of variable adhesiveness. , 1973, Experimental cell research.

[19]  Douglas A. Lauffenburger,et al.  Measurement of individual cell migration parameters for human tissue cells , 1992 .

[20]  R. Tranquillo,et al.  A stochastic model for adhesion-mediated cell random motility and haptotaxis , 1993, Journal of mathematical biology.

[21]  S. Dzik,et al.  The immunological synapse: A molecular machine controlling T cell activation , 2000 .

[22]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[23]  L. Addadi,et al.  Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates , 2001, Nature Cell Biology.

[24]  Dunn Ga,et al.  Characterising a kinesis response: time averaged measures of cell speed and directional persistence. , 1983 .

[25]  K. Yamada,et al.  The interaction of plasma fibronectin with fibroblastic cells in suspension. , 1985, The Journal of biological chemistry.

[26]  D E Ingber,et al.  Cytoskeletal filament assembly and the control of cell spreading and function by extracellular matrix. , 1995, Journal of cell science.

[27]  Sean P. Palecek,et al.  Integrin-ligand binding properties govern cell migration speed through cell-substratum adhesiveness , 1997, Nature.

[28]  D A Lauffenburger,et al.  Maximal migration of human smooth muscle cells on fibronectin and type IV collagen occurs at an intermediate attachment strength , 1993, The Journal of cell biology.

[29]  Micah Dembo,et al.  Measurements of cell-generated deformations on flexible substrata using correlation-based optical flow. , 2003, Methods in enzymology.

[30]  K. Beningo,et al.  Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions. , 2002, Methods in cell biology.

[31]  D E Ingber,et al.  Integrin binding and cell spreading on extracellular matrix act at different points in the cell cycle to promote hepatocyte growth. , 1994, Molecular biology of the cell.

[32]  Y. Wang,et al.  Cell locomotion and focal adhesions are regulated by substrate flexibility. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Shawn M. Sweeney,et al.  Mapping the Ligand-binding Sites and Disease-associated Mutations on the Most Abundant Protein in the Human, Type I Collagen* , 2002, The Journal of Biological Chemistry.

[34]  R Langer,et al.  Switching from differentiation to growth in hepatocytes: Control by extracellular matrix , 1992, Journal of cellular physiology.

[35]  S. Timoshenko,et al.  Theory of elasticity , 1975 .

[36]  Zhibing Hu,et al.  New method for measuring Poisson's ratio in polymer gels , 1993 .

[37]  W. H. Reid,et al.  The Theory of Elasticity , 1960 .

[38]  G. Dunn,et al.  Characterising a kinesis response: time averaged measures of cell speed and directional persistence. , 1983, Agents and actions. Supplements.

[39]  M. Dembo,et al.  The Thermodynamics of Cell Adhesion , 1987 .

[40]  M. Dembo,et al.  Cell movement is guided by the rigidity of the substrate. , 2000, Biophysical journal.