sign in sign up
<<
Home , Collagen, Elastin, Extracellular matrix, Tissue remodeling, Type I collagen

Science at a Glance 4195

The extracellular at a glance Introduction The (ECM) is the non- Christian Frantz1, Kathleen M. Stewart1 and Valerie M. Weaver1,2,* cellular component present within all tissues and 1Department of Surgery and Center for Bioengineering and Regeneration, University of California San organs, and provides not only essential physical Francisco, San Francisco, CA 94143, USA scaffolding for the cellular constituents but also 2 Department of , Department of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad initiates crucial biochemical and biomechanical Center of Regeneration Medicine and Research at UCSF, UCSF Helen Diller Comprehensive Center, University of California San Francisco, San Francisco, CA 94143, USA cues that are required for tissue , *Author for correspondence ([email protected]) differentiation and homeostasis. The importance

Journal of Cell Science 123, 4195-4200 of the ECM is vividly illustrated by the wide © 2010. Published by The Company of Biologists Ltd range of syndromes, which can be anything doi:10.1242/jcs.023820 from minor to severe, that arise from genetic abnormalities in ECM (Jarvelainen et al., 2009). Although, fundamentally, the ECM is composed of water, proteins and The Extracellular Matrix at a Glance , each tissue has an ECM with Christian Frantz, Kathleen M. Stewart and Valerie M. Weaver a unique composition and topology that is

1 ECM generated during tissue development through Examples of Fibrous proteins a dynamic and reciprocal, biochemical hydrogel Modular PGs SLRPs Cell-surface PGs Glypican and biophysical dialogue between the Fibrous Core Glycosamino- various cellular components (e.g. epithelial, glycan chains , , endothelial elements) 2 ECM and function and the evolving cellular and protein Normal Aged Wounded or fibrotic Tumor

Loss of cell− Loss of apical−basal polarity Loss of apical–basal polarity microenvironment. Indeed, the physical, topological, and biochemical composition of the Thinner BM BM ECM is not only tissue-specific, but is also

Metastatic markedly heterogeneous. Cell adhesion to the cell migration

EMT Epithelial cell Inflammatory cell Senescent fibroblast (growth arrest, ECM is mediated by ECM receptors, such as (apical−basal polarity) (infiltration from resistant to apoptotic signals) and lymph vessels) Fibrin blood clot Fibroblast (secreting ECM Growth factors MMP precursors and reorganizing , discoidin domain receptors and the ECM) Non-enzymatic crosslinking (differentiated fibroblast) Proteoglycans (PGs) Adipocyte (each and glycosaminoglycan Activated infiltrated Transformed Differentiated surrounded by a secreted chains cell (peritumoral fibroblasts, reactive (Harburger and Calderwood, 2009; inflammatory cell thick ) fibroblasts, cancer-associated Col I Epithelial-to-mesenchymal fibroblasts and ) EMT Myoepithelial cell transition Humphries et al., 2006; Leitinger and (in contact with BM) Fibronectin -crosslinking Hohenester, 2007; Xian et al., 2010). Adhesion Molecular composition mediates cytoskeletal coupling to the ECM and Col I Fibronectin Degenerated elastin network ↑ Fibronectin ↑ Col types I, III ↑ Fibronectin ↑ Col types I, III and IV ↑ Fibronectin ↑ Elastin is involved in cell migration through the ECM

PGs (examples) (Schmidt and Friedl, 2010). Moreover, the ↑ HA ↑ Fibrin ↑ PGs (decorin, , lumican, …) ↓ Total PGs ↑ Decorin

Journal of Cell Science ECM is a highly dynamic structure that is HA Decorin Aggrecan Perlecan constantly being remodeled, either

Biological and mechanical properties enzymatically or non-enzymatically, and its

Compliant meshwork, β TGF-β and VEGF ↑ EGF-like GFs Unbound TGF , VGEF, G resists tensile and G G G promote vessel G CH G recruits inflammatory cells molecular components are subjected to a myriad compressive stress ↓ Tensile strength growth and CHCH PAI G ↑ TGF-β, PDGF, bFGF, G GFs: EGF, TGF-β, bFGF G ↑ β G ↑ VEGF TGF- , VEGF, PDGF, ROSS ROS G EGF, bFGF, HGF G of post-translational modifications. Through G CH CH ↓ Mechanical stability, G G PAI G G tensile strength, elasticity ROS ECM stiffening, G CH reduced elasticity G ↑ G Stiffness these physical and biochemical characteristics G G G CCH G G G G CH the ECM generates the biochemical and G Glycosaminoglycan- ROS G G bound G PAII CH G (GH) G G CH G mechanical properties of each , such as its G G G G G Unbound GH G CH ROSS G ROS Glycosaminoglycan G MMP activity G Newly secreted GH ROS G tensile and compressive strength and elasticity, G chain 500 Pa 3000 Pa and also mediates protection by a buffering Increased stiffness

Extrinsic and intrinsic forces action that maintains extracellular homeostasis

Tensional ECM resistance Reciprocal ECM resistance Reciprocal ECM resistance homeostasis Migrating and and water retention. In addition, the ECM dividing cells ECM-contracting Stiff crosslinked PG-modified collagen collagen and myofibroblast elastin fibers directs essential morphological organization Stiff crosslinked collagen fibers stress Solid stress Proliferating Weak basement transformed and physiological function by binding growth membrane Stiff crosslinked collagen fibers factors (GFs) and interacting with cell-surface Exerted ECM-contracting force myofibroblast receptors to elicit and

3 Natural and synthetic engineered ECMs regulate . The biochemical Natural Modified or synthetic Soft rBM gel (e.g. ™) (Kleinman et al., Col I (Friess, 1998) Functionalized HA (Serban and Prestwich, 2008) PEG hydrogels and biomechanical, protective and 1986; Kleinman and Martin, 2005) (Lutolf and Hubbell, 2005) Self-assembling (Hauser and Zhang, 2010) R R Conjugated Multi-arm macromers conjugated R RGD peptides, G R with RGD, LDV, YIGSR Self-assembled -amphiphiles. RRRR R organizational properties of the ECM in a given R Col I, laminin, G Functionalized with RGF, RGD, LDV, fibronectin G R YIGSR. diameter~50 nm R G ligands R R G R R G Adhesion receptor Collagen concentration G ligands tissue can vary tremendously from one tissue to G R SIS scaffold (Badylak, 2007) Fibrin gel (fibrinogen + ) Key (Blombäck and Bark, 2004) Electrospun (, lactic acid and glycolic acid Functionalized polyacrylamide gel (Pelham and Wang, 1997) another (e.g. versus versus ) and R Remodeling sites ) (Zhang et al., 2009; McCullen et al., 2009) R G Col I, laminin, GFs Crosslinked with ECM R R G Embedded growth factors G R and fibronectin R molecules and peptides R R R Fiber diameter 50 nm−300 μm. even within one tissue (e.g. renal cortex versus R R G Mixed with Col I, fibrin, HA

Thrombin mg/ml (See accompanying article for full citations.) renal medulla), as well as from one physiological state to another (normal versus Abbreviations: bFGF, basic ; BM, basement membrane; CH, chemokines; Col, collagen; EGF, endothelial growth factor; HGH, human growth hormone; HA, ; MMP, ; LDV, Leu-Asp-Val; PAI, plasminogen activator inhibitor; PDGF, -derived growth factor; PEG, polyethyleneglycol; rBM, reconstituted basement membrane; RGD, Arg-Gly-Asp; ROS, reactive oxygen species; SIS, small intestinal ; SRLPs, small - cancerous). In this Cell Science at a Glance rich proteoglycans; TGF-β, transforming growth factor β; VEGF, vascular endothelial growth factor; © Journal of Cell Science 2010 (123, pp. 4195–4200) YIGSR, article, we briefly describe the main molecular (See poster insert) components of the ECM and then compare and 4196 Journal of Cell Science 123 (24)

contrast the ECM within a normal simple epithelial tissue with that found within a Box 1. Structure and function of proteoglycans pathologically modified tissue, as exemplified Proteoglycans (PGs) are composed of glycosaminoglycan (GAG) chains covalently linked in aged tissue, wounded or fibrotic tissue and to a specific protein core (with the exception of hyaluronic acid) (Iozzo and Murdoch, 1996; tumors. We particularly focus on the Schaefer and Schaefer, 2010). PGs have been classified according to their core proteins, localization and GAG composition. The three main families are: small leucine-rich composition and architecture of the ECM and proteoglycans (SLRPs), modular proteoglycans and cell-surface proteoglycans (Schaefer interactions with its cellular constituents, and Schaefer, 2010). The GAG chains on the protein core are unbranched and describe in detail common post- chains composed of repeating units [sulfated N-aceltylglucosamine or translational modifications that evoke defined N-acetylgalactosamine, D-glucuronic or L-iduronic acid and galactose (–4 N- topological and viscoelasticity changes in the acetylglucosamine-1,3-galactose-1)] that can be divided further into sulfated (chondroitin tissue. We thereafter discuss the functional sulfate, and ) and non-sulfated (hyaluronic acid) GAGs consequences of ECM remodeling on cellular (Schaefer and Schaefer, 2010). These molecules are extremely hydrophilic and, behaviors including altered GF sensitivity accordingly, adopt highly extended conformations that are essential for hydrogel formation elicited by changes in ECM tension. Owing to and that enable matrices that are formed by these molecules to withstand high space limitations and because the basement compressive forces. Many genetic diseases have been linked to in PG (Jarvelainen et al., 2009; Schaefer and Iozzo, 2008). SLRPs have been involved in multiple membrane (BM) is a unique ECM that has been signaling pathways including binding to and activation of epidermal growth factor receptor reviewed in detail elsewhere (LeBleu et al., (EGFR), -like growth factor 1 receptor (IGFIR) and low-density lipoprotein-receptor- 2007), we focus here on the interstitial stroma of related protein 1 (LRP1), regulation of inflammatory response reaction, binding to and simple glandular epithelial tissues. We complete activation of TGF (Goldoni and Iozzo, 2008; Schaefer and Iozzo, 2008; Schaefer our review with a brief discussion of the and Schaefer, 2010). Modular PGs can modulate cell adhesion, migration and proliferation application of natural and synthetic ECMs that (Schaefer and Schaefer, 2010). Basement membrane modular PGs (perlecan, and can be used to either recapitulate the interstitial collagen type XVIII) have a dual function as pro- and anti-angiogenic factors (Iozzo et al., ECM in culture to study tissue behaviors or to 2009). Cell-surface PGs (syndecans and glypicans) can act as co-receptor facilitating deconstruct and analyze how specific ECM ligand encounters with signaling receptors (Schaefer and Schaefer, 2010). parameters (stiffness, fiber orientation, ligand presentation, dimensionality) provoke specific cellular behaviors. The bulk of interstitial collagen is transcribed pleiotrophic changes in cellular behavior and and secreted by fibroblasts that either reside in implicate FN as an extracellular mechano- Bits and pieces – molecular the stroma or are recruited to it from neighboring regulator (Smith et al., 2007). Indeed, ‘tensed’ composition of the ECM tissues (De Wever et al., 2008). By exerting FN modulates the catch bond ‘force-activation’ The ECM is composed of two main classes of tension on the matrix, fibroblasts are able to and adhesion assembly of 51-integrin macromolecules: proteoglycans (PGs) and organize collagen into sheets and cables through exposure of its synergy site (Friedland fibrous proteins (see Boxes 1 and 2) (Jarvelainen and, thus, can dramatically influence the et al., 2009). FN is also important for cell et al., 2009; Schaefer and Schaefer, 2010). The alignment of collagen fibers. Although within migration during development and has been

Journal of Cell Science main fibrous ECM proteins are collagens, a given tissue, collagen fibers are generally a implicated in cardiovascular disease and tumor elastins, and (see panel 1 heterogeneous mix of different types, one type (Rozario and DeSimone, 2010; of the poster) (Alberts et al., 2007). PGs fill the of collagen usually predominates. Tsang et al., 2010). Like FN, other ECM majority of the extracellular interstitial space Collagen associates with elastin, another proteins such as tenascin exert pleiotrophic within the tissue in the form of a hydrated gel major ECM fiber. Elastin fibers provide recoil to effects on cellular behavior, including the (Box 1) (for details, see Jarvelainen et al., 2009). tissues that undergo repeated stretch. promotion of fibroblast migration during PGs have a wide variety of functions that reflect Importantly, elastin stretch is crucially limited (Trebaul et al., 2007; Tucker and their unique buffering, hydration, binding and by tight association with collagen fibrils (Wise Chiquet-Ehrismann, 2009). Indeed, levels of force-resistance properties. For example, in the and Weiss, 2009). Secreted tropoelastin (the C and W are elevated in the stroma glomerular BM, perlecan has a role in precursor of elastin) molecules assemble into of some transformed tissues where they can glomerular filtration (Harvey and Miner, 2008; fibers and become highly crosslinked to one inhibit the interaction between syndecan4 and Morita et al., 2005). By constrast, in ductal another via their residues by members of FN to promote tumor growth and metastasis epithelial tissues, decorin, biglycan and lumican the (LOX) family, which (Tucker and Chiquet-Ehrismann, 2009). associate with collagen fibers to generate a include LOX and LOXL (Lucero and Kagan, molecular structure within the ECM that is 2006). Elastin fibers are covered by The definition of normal – the ECM essential for mechanical buffering and hydration , mainly , and tissue homeostasis and that, by binding GFs, provides an easy, which are also essential for the integrity of the Normal glandular epithelial tissues are enzymatically accessible repository for these elastin fiber (Wise and Weiss, 2009). composed of a simple layer of epithelial cells factors (Iozzo and Murdoch, 1996). A third fibrous protein, fibronectin (FN) is that adopt apical–basal polarity, where the basal Collagen is the most abundant fibrous protein intimately involved in directing the organization side contacts the BM and the apical side is within the interstitial ECM and constitutes up to of the interstitial ECM and, additionally, has a opposite the fluid-filled lumen. In some 30% of the total protein mass of a multicellular crucial role in mediating cell attachment and glandular epithelium there is a basal or . Collagens, which constitute the main function. FN can be stretched several times over myoepithelial cell layer that separates the structural element of the ECM, provide tensile its resting length by cellular traction forces luminal epithelium from the interstitial ECM strength, regulate cell adhesion, support (Smith et al., 2007). Such force-dependent (Barsky and Karlin, 2005). Epithelial tissue and migration, and direct tissue unfolding of FN exposes cryptic integrin- homeostasis depends on the maintenance of development (Rozario and DeSimone, 2010). binding sites within the molecule that result in tissue organization and a dynamic dialogue with Journal of Cell Science 123 (24) 4197

a surrounding stroma composed primarily of non-activated fibroblasts and , and a Box 2. Collagen and fibronectin synthesis steady-state population of transiting, non- To date, 28 types of collagen have been identified in (Gordon and Hahn, 2010). stimulated leukocytes (Ronnov-Jessen et al., The majority of collagen molecules form a triple-stranded helix that subsequently can 1996). Thus, non-activated tissue fibroblasts assemble into supramolecular complexes, such as fibrils and networks, depending on the type of collagen. Fibrous collagens form the backbone of the collagen bundles within secrete and organize type I and III collagens, the interstitial tissue stroma, whereas network collagens are incorporated into the basal elastin, fibronectin, tenascin and a repertoire of membrane (BM). Synthesis of collagen type I involves a number of enzymatic post- PGs (hyaluronic acid and decorin), which all translational modifications (Gordon and Hahn, 2010; Myllyharju and Kivirikko, 2004), mainly maintain the structural and functional integrity the of and lysine residues, of lysine and the of of the interstitial ECM. Most glandular N- and C-terminal propeptides. Following their cleavage, collagen fibrils are strengthened epithelial tissues including , saliva gland, by the covalent crosslinking between lysine residues of the constituent collagen molecules , and prostate are in a state of tensional by lysyl oxidases (LOX) (Myllyharju and Kivirikko, 2004; Robins, 2007). homeostasis so that their normal state is highly FN is secreted as a dimer joined by two C-terminal disulfide bonds and has several mechanically compliant (Paszek and Weaver, binding sites to other FN dimers, to collagen, to and also to cell-surface integrin 2004). The ECM in a compliant tissue is receptors (Pankov and Yamada, 2002). Cell-surface binding of the soluble FN dimer is essential for its assembly into longer fibrils. Moreover, cell contraction through the composed of a relaxed meshwork of type I and actomyosin and the resulting integrin clustering promotes FN–fibril assembly III collagens and elastin that, together with FN, by exposing cryptic binding sites, thus allowing them to bind one another (Leiss et al., form a relaxed network of fibers that are 2008; Mao and Schwarzbauer, 2005; Vakonakis and Campbell, 2007). surrounded by and embedded in a hydrogel of glycosaminoglycan-chain-containing PGs (Bosman and Stamenkovic, 2003). because of elevated MMP-mediated events that characterize a wound response is Consequently, the relaxed network of collagen degradation and reduced BM protein synthesis vascular damage and the formation of a fibrin and elastin fibers allow the healthy ECM to (Callaghan and Wilhelm, 2008). Moreover, the clot, which stimulates monocyte infiltration to resist a wide range of tensile stresses. A resident fibroblasts in aged tissues are growth- the damaged ECM. Upon binding to ECM- functionally competent normal tissue can also arrested and resistant to apoptotic cues, which is degradation products and cytokines, monocytes easily resist compressive stresses because of the indicative of (Campisi and d’Adda rapidly differentiate into (Clark, binding of the hydrated glycosaminoglycan di Fagagna, 2007). Indeed, senescent fibroblasts 2001). These activated macrophages, in turn, (GAG) network to the fibrous ECM molecules typically express elevated levels of FN, MMPs, secrete and release multiple GFs, MMPs (Scott, 2003). Thus, the tissue ECM is a highly GFs, interleukins and cytokines, as well as high and cytokines that promote and dynamic entity that continuously undergoes levels of the plasminogen activator inhibitor stimulate fibroblast migration and proliferation regulated remodeling, whose precise (PAI) (Coppe et al., 2010) and mitochondrial- (Schultz and Wysocki, 2009). Thereafter, orchestration is crucial to the maintenance of related reactive oxygen species (ROS) recruited fibroblasts begin to synthesize and normal function (Egeblad et al., 2010; Kass et (Untergasser et al., 2005) and, as a result, are deposit large quantities of ECM proteins, al., 2007). Tissue homeostasis is mediated by the frequently in a state of chronic inflammation. including collagen type I and III, FN and

Journal of Cell Science coordinated secretion of fibroblast metallopro- Indeed, the combination of chronic hyaluronic acid. The elevated mechanical stress teinases (MMPs) (Mott and Werb, 2004); this is inflammation and elevated MMPs, PAI and associated with this profound ECM deposition counterbalanced by tissue inhibitors of metallo- ROS destroy the integrity of the elastin network can induce the transdifferentiation of fibroblasts proteinases (TIMPs) (Cruz-Munoz and Khokha, and modify the collagen fiber network, whereas and other tissue-resident cells – i.e. epithelial-to- 2008) and the controlled activity of other reduced levels of tissue-associated GAGs mesenchymal transition (EMT) of epithelial enzymes, such as LOX, and also transglutami- compromise the integrity of the BM (Callaghan cells – or of circulating bone marrow-derived nases that crosslink and, consequently, stiffen and Wilhelm, 2008; Calleja-Agius et al., 2007; mesenchymal stem cells into myofibroblasts the ECM (Lucero and Kagan, 2006). A plethora Nomura, 2006). Nevertheless, and somewhat (Schultz and Wysocki, 2009; Velnar et al., of GFs that are bound to the ECM direct these paradoxically, in an aging tissue, collagen fibers 2009). Myofibroblasts, which have a high processes (Friedl, 2010; Hynes, 2009; Macri et are frequently – inappropriately – crosslinked capacity to synthesize ECM components and are al., 2007; Murakami et al., 2008; Oehrl and through glycation, by byproducts of lipid highly contractile, can promote the formation of Panayotou, 2008). These ECM-bound GFs oxidation and through exposure to UV light large, rigid collagen bundles that, if crosslinked differentially modulate cell growth and (Robins, 2007). The combination of elevated and by LOX enzymes, mechanically strengthen and migration and, when released, comprise part of a inappropriate collagen crosslinking contributes stiffen the tissue (Szauter et al., 2005). This tightly controlled feedback circuit that is to tissue stiffening so that an aged tissue is wounded ‘stiffened’ microenvironment disrupts essential for normal tissue homeostasis (Hynes, mechanically weaker and less elastic but also the BM that surrounds the epithelium and 2009). more rigid than a young tissue (Calleja-Agius compromises tissue integrity with loss of et al., 2007; Robins, 2007). This aberrant apical–basal polarity and destabilized cell–cell Stiffening up – the ECM and tissue mechanical state can severely compromise ECM adhesions. The remodeled ECM also promotes aging organization, and modify epithelial organization the directional migration of cells within the As a tissue ages the levels of junctional proteins and function, potentially promoting age-related tissue towards the wound site (Schafer and such as cadherin, catenin or occludin decrease diseases such as cancer (Coppe et al., 2010; Werner, 2008). In some instances, the release of and this loss can compromise junctional Freund et al., 2010; Sprenger et al., 2008). transforming growth factor  (TGF-) by integrity as revealed by the appearance of gaps tension and MMPs induces EMT of the resident between the epithelial cells (Akintola et al., Tensional homeostasis and epithelium (Schultz and Wysocki, 2009; Wipff 2008; Bolognia, 1995). Old tissue is also Acute injury activates the fibrogenic machinery et al., 2007; Xu et al., 2009). In a healthy tissue, characterized by a thinning of the BM, probably and induces . One of the first once the wound has been repopulated, strict 4198 Journal of Cell Science 123 (24)

feedback mechanisms are initiated that ensure ECM-embedded GFs (Bosman and epithelial or endothelial behavior and to restoration of tissue homeostasis and resolution Stamenkovic, 2003; Kessenbrock et al., 2010). distinguish between the ‘normal’ and of fibrosis (Schultz and Wysocki, 2009; Velnar The release of GFs, including vascular ‘malignant’ behavior of some tissues, has a et al., 2009). Under extreme conditions, such as endothelial growth factor (VEGF), enhances complex and rudimentarily defined repeated injury or when normal feedback vascular permeability and promotes new vessel composition, and fails to reconstruct the mechanisms are compromised, continuous growth, which generates interstitial tissue physical state of the native interstitial ECM. ECM synthesis, deposition and remodeling . Thus, an amplifying circuitry between Fibrin has also been used as natural ensue and myofibroblasts remain, in which tumor-associated ECM stiffening, an ensuing biodegradable scaffold with reasonable success TIMP production prevails over MMP synthesis. reciprocal ECM resistance that is induced by in vascular , but lacks the These aberrant conditions promote chronic resident tumor cells, and myoepithelial and cell- mechanical strength and durability of native vascular remodeling and enhanced ECM generated contractility act as a vicious, positive- interstitial ECM (Blomback and Bark, 2004; crosslinking that eventually leads to aberrant feedback loop to potentiate tumor growth Shaikh et al., 2008). By contrast, fibrosis and an inability of the tissue to heal and survival. This induces angiogenesis and is reasonably useful and can be combined with properly. This aberrant wound healing scenario and, eventually, fosters metastasis rBM, purified laminin or FN to reconstitute is characterized by the altered mechanical (Butcher et al., 2009; Erler and Weaver, 2009; some of the biological aspects of normal and stability and reduced elasticity that is typical of Paszek and Weaver, 2004; Paszek et al., 2005). diseased interstitial ECM (Friess, 1998; scarred tissue (Kisseleva and Brenner, 2008). In Gudjonsson et al., 2002). Moreover, collagen extreme cases, a can also Where do we go from here? type I readily assembles into a mechanically promote a tumor phenotype (De Wever et al., Challenges encountered with natural tense network of fibrils that can be oriented, 2008). and synthetic ECMs functionally modified, and enzymatically or Considering the importance of the ECM to so chemically crosslinked and stiffened. Thus Tumors – a tough situation many fundamental cellular processes, a myriad collagen I gels are useful substrates to assess Cancer is the loss of tissue organization and of tissue-culture models have been developed to the role of collagen and FN stiffness, and aberrant behavior of the cellular components. study the interplay between its biochemical and organization on the pathogenesis of tumor Cell transformation results from genetic biophysical properties, and to understand the progression and invasion (Levental et al., 2009; mutations and epigenetic alterations. Yet, molecular origins of cellular behaviors Provenzano et al., 2009). Nevertheless, collagen tumors have also been likened to that regulated by ECM ligation. With respect to gels are quite heterogeneous, and modifying fail to heal (Schafer and Werner, 2008). Thus, assessing the fundamental of cell their architecture changes their organization, the tumor stroma exhibits some of the character- adhesion and its effects on cell behavior, the pore size and ligand concentration, thereby istics found in an unresolved wound (Bissell and majority of cancer researchers have relied on complicating the interpretation of data Radisky, 2001). For example, tumors are char- coating tissue culture dishes (whether or generated from experiments conducted by using acteristically stiffer than the surrounding normal glass) with purified preparations or mixtures of this natural scaffold (Johnson et al., 2007). To tissue. The stiffening of tumors is induced by ECM proteins in order to obtain 2D monolayers overcome this issue, tissue engineers and ECM deposition and remodeling by resident (Kuschel et al., 2006). To address the issue of biomaterial specialists have generated denuded

Journal of Cell Science fibroblasts, and by increased contractility of the ECM rigidity, functionalized polyacrylamide ECM scaffold from various tissues (Macchiarini transformed epithelium (Butcher et al., 2009; (PA) gels crosslinked with reconstituted et al., 2008). These scaffolds, combined with Levental et al., 2009). Moreover, chemokines basement membrane (rBM) – generated from colonies of seeded stem cells, can reconstitute and GFs (De Wever et al., 2008) induce Engelbreth-Holm-Swarm mouse carcinoma normal tissues with reasonable fidelity (Lutolf et inflammation and modify the repertoire of (commercially available as Matrigel™), al., 2009). ECMs have also been isolated and infiltrating T lymphocytes (Tan and Coussens, collagen type I, FN or ECM peptides – has extracted from various tissues, such as small 2007). Tissue inflammation potentiates stromal become the standard approach (Johnson et al., intestine, skin (from cadavers), pancreas and fibroblast activation and induces their trans- 2007; Pelham and Wang, 1997). Yet, none of breast (Rosso et al., 2005), and these ECMs have differentiation into myofibroblasts, thus these strategies faithfully recapitulates the been used to engineer skin grafts (Badylak, exacerbating and promoting tissue behavior of cells within tissues, which demand 2007), enhance wound healing and to study (De Wever et al., 2008; Desmouliere et al., not only a 3D format, but an ECM that can be tumor progression. One such example is given 2004). Myofibroblasts deposit copious readily remodeled. To address the aspect of 3D by porcine-derived small intestinal submucosa quantities of ECM proteins, secrete GFs and and ECM remodeling, researchers have used (SIS), which has proven clinical success for exert strong contraction forces on the ECM (De natural ECM and reconstituted ECM gels to treating patients with hernias (Franklin et al., Wever et al., 2008; Desmouliere et al., 2004). As recapitulate specific aspects of tissue-specific 2002) (reviewed in Badylak, 2007). Although a consequence, newly deposited and remodeled differentiation and architecture (see poster, these purified ECMs certainly have useful collagen and elastin fibers are reoriented and, panel 3). For instance, the rBM, which mimics applications, their use is limited in scope owing thereafter, crosslinked by LOX and transglutam- some of the biochemical and biophysical to the need for well-defined microenvironments inase, thus generating larger, more-rigid fibrils properties of endogenous epithelial basement in tissue regeneration and stem cell transplanta- that further stiffen the tissue ECM (Butcher et membranes, has been used frequently in 3D tion in which animal byproducts and al., 2009; Erler and Weaver, 2009; Levental organotypic culture assays, for xenograft contaminants are limited. Moreover, to et al., 2009; Lucero and Kagan, 2006; Payne et manipulations or tissue engineering, and to understand the molecular and biophysical al., 2007; Rodriguez et al., 2008). MMPs, which study tissue-specific morphogenesis (e.g. mechanisms by which the ECM elicits diverse are secreted and activated by tumor cells and branching, acini formation) and differentiation effects on and by myofibroblasts (De Wever et al., 2008; (Kleinman and Martin, 2005; Kleinman et al., morphogenesis it is crucial to use chemically Kessenbrock et al., 2010), also remodel the BM 1986). Unfortunately, BM preparations such as and physically defined, modular ECMs that can surrounding the tumor and release and activate Matrigel™, although useful for studying normal be reliably reproduced. In this respect, synthetic Journal of Cell Science 123 (24) 4199

(2007). Biomolecular hydrogels formed and degraded via matrices have been developed that feature functional assessment in physiological culture site-specific enzymatic reactions. Biomacromolecules 8, defined and tunable compositions, organization, assays and animal models. Although only time 3000-3007. mechanics and ECM remodeling capabilities. can tell whether this new generation of Erler, J. T. and Weaver, V. M. (2009). Three-dimensional context regulation of metastasis. Clin. Exp. Metastasis 26, Indeed, in response to this need there has been will indeed prove useful, it is an 35-49. literally an explosion of publications describing appealing time to be an ECM biologist and our Franklin, M. E., Jr, Gonzalez, J. J., Jr, Michaelson, R. the generation and application of synthetic next challenge will be to embrace this P., Glass, J. L. and Chock, D. A. (2002). Preliminary experience with new bioactive prosthetic material for repair ECMs for tissue regeneration, and the reader is smorgasbord of enticing new tools – which of hernias in infected fields. Hernia 6, 171-174. referred to some excellent reviews on these hopefully will at last allow us to decipher the Freund, A., Orjalo, A. V., Desprez, P. Y. and Campisi, J. topics (Ayres et al., 2009; Dutta and Dutta, 2009; language of the matrix. (2010). Inflammatory networks during : causes and consequences. Trends Mol. Med. 16, 238-246. Lutolf and Hubbell, 2005; McCullen et al., This work was supported by the NIH grants Friedl, A. (2010). Proteoglycans: master modulators of 2009; Rosso et al., 2005; Zisch et al., 2003). One U54CA143836 and CA138818-01A1 and the DOD paracrine fibroblast-carcinoma cell interactions. Semin. Cell example is given by (PEG) grant W81XWH-05-1-0330 to V.M.W. Deposited in Dev. Biol. 21, 66-71. PMC for release after 12 months. Friedland, J. C., Lee, M. H. and Boettiger, D. (2009). hydrogels – frequently used biologically Mechanically activated integrin switch controls alpha5beta1 compatible synthetic matrices that support cell function. Science 323, 642-644. adhesion, viability and growth (Lutolf and References Friess, W. (1998). Collagen-biomaterial for drug delivery. Eur. J. Pharm. Biopharm. 45, 113-136. Hubbell, 2005). Although these matrices can be Akintola, A. D., Crislip, Z. L., Catania, J. M., Chen, G., Zimmer, W. E., Burghardt, R. C. and Parrish, A. R. Goldoni, S. and Iozzo, R. V. (2008). Tumor covalently modified with ECM ligands and (2008). Promoter methylation is associated with the age- microenvironment: modulation by decorin and related -degradable peptides and GFs dependent loss of N-cadherin in the kidney. Am. J. molecules harboring leucine-rich tandem motifs. Int. J. Physiol. Renal Physiol. 294, F170-F176. Cancer 123, 2473-2479. (Ehrbar et al., 2007; Zisch et al., 2003), they do Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K. Gordon, M. K. and Hahn, R. A. (2010). Collagens. Cell not mimic the organizational features of native and Walter, P. (2007). Molecular Biology of the Cell. Tissue Res. 339, 247-257. collagen gels and all too often their pore sizes London: Garland Science. Gudjonsson, T., Ronnov-Jessen, L., Villadsen, R., Rank, Ayres, C. E., Jha, B. S., Sell, S. A., Bowlin, G. L. and F., Bissell, M. J. and Petersen, O. W. (2002). Normal and strongly impede cell migration. By contrast, Simpson, D. G. (2009). Nanotechnology in the design of tumor-derived myoepithelial cells differ in their ability to peptide-based hydrogels, such as peptide- scaffolds: innovations in structure and function. interact with luminal breast epithelial cells for polarity and basement membrane deposition. J. Cell Sci. 115, 39-50. amphiphiles, assemble into secondary Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 2, 20-34. Badylak, S. F. (2007). The extracellular matrix as a biologic Harburger, D. S. and Calderwood, D. A. (2009). Integrin that recapitulate the collagen , and scaffold material. Biomaterials 28, 3587-3593. signalling at a glance. J. Cell Sci. 122, 159-163. readily support stem cell growth and viability, Barsky, S. H. and Karlin, N. J. (2005). Myoepithelial Harvey, S. J. and Miner, J. H. (2008). Revisiting the glomerular charge barrier in the molecular era. Curr. Opin. and direct multicellular morphogenesis cells: autocrine and paracrine suppressors of breast cancer progression. J. Biol. Neoplasia 10, 249- Nephrol. Hypertens. 17, 393-398. (Hauser and Zhang, 2010; Sieminski et al., 260. Hauser, C. A. and Zhang, S. (2010). Designer self- 2008; Smith and Ma, 2004; Ulijn and Bissell, M. J. and Radisky, D. (2001). Putting tumours in assembling peptide biological materials. Chem. context. Nat. Rev. Cancer 1, 46-54. Soc. Rev. 39, 2780-2790. Smith, 2008). These peptides-amphiphiles are Blombäck, B. and Bark, N. (2004). Fibrinopeptides and Humphries, J. D., Byron, A. and Humphries, M. J. amenable to modification by covalent binding of fibrin gel structure. Biophys. Chem. 112, 147-151. (2006). Integrin ligands at a glance. J. Cell Sci. 119, 3901- native proteins and MMP-degradable ECM Bolognia, J. L. (1995). Aging skin. Am. J. Med. 98, 99S- 3903. 103S. Hynes, R. O. (2009). The extracellular matrix: not just peptides. Alternatively, poly(lactic-co-glycolic Bosman, F. T. and Stamenkovic, I. (2003). Functional pretty fibrils. Science 326, 1216-1219. acid) (PLGA), a copolymer of glycolic acid and structure and composition of the extracellular matrix. J. Iozzo, R. V. and Murdoch, A. D. (1996). Proteoglycans of Pathol. 200, 423-428. the extracellular environment: clues from the gene and

Journal of Cell Science lactic acid (McCullen et al., 2009) that is Butcher, D. T., Alliston, T. and Weaver, V. M. (2009). A protein side offer novel perspectives in molecular diversity inherently biodegradable as it is hydrolyzed into tense situation: forcing tumour progression. Nat. Rev. and function. FASEB J. 10, 598-614. lactic acid and glycolic acid, has been developed Cancer 9, 108-122. Iozzo, R. V., Zoeller, J. J. and Nystrom, A. (2009). and can be readily conjugated to various ECM Callaghan, T. M. and Wilhelm, K. P. (2008). A review of Basement membrane proteoglycans: modulators Par and an examination of clinical methods in the Excellence of cancer growth and angiogenesis. Mol. Cells ligands and peptides, or coated with collagen or assessment of ageing skin. Part 2, Clinical perspectives and 27, 503-513. chitosan to support cell adhesion, viability and clinical methods in the evaluation of ageing skin. Int. J. Jarvelainen, H., Sainio, A., Koulu, M., Wight, T. N. and Penttinen, R. (2009). Extracellular matrix molecules: growth. Indeed, one of the most exciting recent Cosmet. Sci. 30, 323-332. Calleja-Agius, J., Muscat-Baron, Y. and Brincat, M. P. potential targets in pharmacotherapy. Pharmacol. Rev. 61, advances in the field has been the development (2007). Skin ageing. Menopause Int. 13, 60-64. 198-223. of modular biocompatible ECMs, which contain Campisi, J. and d’Adda di Fagagna, F. (2007). Cellular Johnson, K. R., Leight, J. L. and Weaver, V. M. (2007). senescence: when bad things happen to good cells. Nat. Rev. Demystifying the effects of a three-dimensional ligand-binding cassettes and have tunable Mol. Cell Biol. 8, 729-740. microenvironment in tissue morphogenesis. Methods Cell stiffness features that permit a precise patterning Clark, R. A. (2001). Fibrin and wound healing. Ann. NY Biol. 83, 547-583. of cell adhesion in 2D and 3D formats (Serban Acad. Sci. 936, 355-367. Kass, L., Erler, J. T., Dembo, M. and Weaver, V. M. Coppe, J. P., Desprez, P. Y., Krtolica, A. and Campisi, J. (2007). Mammary epithelial cell: influence of extracellular and Prestwich, 2008). The realization that ECM (2010). The senescence-associated secretory phenotype: the matrix composition and organization during development organization is a crucial aspect of cellular dark side of tumor suppression. Annu. Rev. Pathol. 5, 99- and tumorigenesis. Int. J. Biochem. Cell Biol. 39, 1987- behavior has led to the development of new 118. 1994. Cruz-Munoz, W. and Khokha, R. (2008). The role of Kessenbrock, K., Plaks, V. and Werb, Z. (2010). Matrix methodologies and generated ECMs whose tissue inhibitors of metalloproteinases in tumorigenesis and metalloproteinases: regulators of the tumor fiber size, orientation, stiffness, ligand-binding metastasis. Crit. Rev. Clin. Lab. Sci. 45, 291-338. microenvironment. Cell 141, 52-67. function and remodeling potential can be strictly De Wever, O., Demetter, P., Mareel, M. and Bracke, M. Kim, D. H., Lipke, E. A., Kim, P., Cheong, R., (2008). Stromal myofibroblasts are drivers of invasive Thompson, S., Delannoy, M., Suh, K. Y., Tung, L. and controlled and monitored – including cancer growth. Int. J. Cancer 123, 2229-2238. Levchenko, A. (2010). Nanoscale cues regulate the electrospun silk, and lactic-acid Desmouliere, A., Guyot, C. and Gabbiani, G. (2004). The structure and function of macroscopic cardiac tissue stroma reaction myofibroblast: a key player in the control constructs. Proc. Natl. Acad. Sci. USA 107, 565-570. (PLLA) and PLGA scaffolds (Zhang et al., of tumor cell behavior. Int. J. Dev. Biol. 48, 509-517. Kisseleva, T. and Brenner, D. A. (2008). Mechanisms of 2009). Anisotropically nanofabricated Dutta, R. C. and Dutta, A. K. (2009). Cell-interactive 3D- fibrogenesis. Exp. Biol. Med. (Maywood) 233, 109-122. substrates formed from scalable biocompatible scaffold; advances and applications. Biotechnol. Adv. 27, Kleinman, H. K. and Martin, G. R. (2005). Matrigel: 334-339. basement membrane matrix with biological activity. Semin. PEG (Kim et al., 2010; Smith et al., 2009) are Egeblad, M., Rasch, M. G. and Weaver, V. M. (2010). Cancer Biol. 15, 378-386. exciting new developments in the biomaterials Dynamic interplay between the collagen scaffold and tumor Kleinman, H. K., McGarvey, M. L., Hassell, J. R., Star, field, whose only major impediment to their . Curr. Opin. Cell Biol. 22, 697-706. V. L., Cannon, F. B., Laurie, G. W. and Martin, G. R. Ehrbar, M., Rizzi, S. C., Schoenmakers, R. G., Miguel, (1986). Basement membrane complexes with biological biological application appears to be a lack of B. S., Hubbell, J. A., Weber, F. E. and Lutolf, M. P. activity. 25, 312-318. 4200 Journal of Cell Science 123 (24)

Kuschel, C., Steuer, H., Maurer, A. N., Kanzok, B., Paszek, M. J., Zahir, N., Johnson, K. R., Lakins, J. N., Smith, L. A. and Ma, P. X. (2004). Nano-fibrous scaffolds Stoop, R. and Angres, B. (2006). Cell adhesion profiling Rozenberg, G. I., Gefen, A., Reinhart-King, C. A., for tissue engineering. Colloids Surf. B Biointerfaces 39, using extracellular matrix protein microarrays. Margulies, S. S., Dembo, M., Boettiger, D. et al. (2005). 125-131. Biotechniques 40, 523-531. Tensional homeostasis and the malignant phenotype. Smith, M. L., Gourdon, D., Little, W. C., Kubow, K. E., LeBleu, V. S., Macdonald, B. and Kalluri, R. (2007). Cancer Cell 8, 241-254. Eguiluz, R. A., Luna-Morris, S. and Vogel, V. (2007). Structure and function of basement membranes. Exp. Biol. Payne, S. L., Hendrix, M. J. and Kirschmann, D. A. Force-induced unfolding of fibronectin in the extracellular Med. (Maywood) 232, 1121-1129. (2007). Paradoxical roles for lysyl oxidases in cancer-a matrix of living cells. PLoS Biol. 5, e268. Leiss, M., Beckmann, K., Giros, A., Costell, M. and prospect. J. Cell Biochem. 101, 1338-1354. Sprenger, C. C., Plymate, S. R. and Reed, M. J. (2008). Fassler, R. (2008). The role of integrin binding sites in Pelham, R. J., Jr and Wang, Y. (1997). Cell Extracellular influences on tumour angiogenesis in the aged fibronectin matrix assembly in vivo. Curr. Opin. Cell Biol. locomotion and focal adhesions are regulated by host. Br. J. Cancer 98, 250-255. 20, 502-507. substrate flexibility. Proc. Natl. Acad. Sci. USA 94, Szauter, K. M., Cao, T., Boyd, C. D. and Csiszar, K. Leitinger, B. and Hohenester, E. (2007). Mammalian 13661-13665. (2005). Lysyl oxidase in development, aging and collagen receptors. Matrix Biol. 26, 146-155. Provenzano, P. P., Eliceiri, K. W. and Keely, P. J. (2009). pathologies of the skin. Pathol. Biol. 53, 448-456. Levental, K. R., Yu, H., Kass, L., Lakins, J. N., Egeblad, Shining new light on 3D cell motility and the metastatic Tan, T. T. and Coussens, L. M. (2007). Humoral immunity, M., Erler, J. T., Fong, S. F., Csiszar, K., Giaccia, A., process. Trends Cell. Biol. 19, 638-648. inflammation and cancer. Curr. Opin. Immunol. 19, 209- Weninger, W. et al. (2009). Matrix crosslinking forces Robins, S. P. (2007). Biochemistry and functional 216. tumor progression by enhancing integrin signaling. Cell significance of collagen cross-linking. Biochem. Soc. Trans. Trebaul, A., Chan, E. K. and Midwood, K. S. (2007). 139, 891-906. 35, 849-852. Regulation of fibroblast migration by tenascin-C. Biochem. Lucero, H. A. and Kagan, H. M. (2006). Lysyl oxidase: Rodriguez, C., Rodriguez-Sinovas, A. and Martinez- Soc. Trans. 35, 695-697. an oxidative enzyme and effector of cell function. Cell. Mol. Gonzalez, J. (2008). Lysyl oxidase as a potential Tsang, K. Y., Cheung, M. C., Chan, D. and Cheah, K. S. Life Sci. 63, 2304-2316. therapeutic target. Drug News Perspect. 21, 218-224. (2010). The developmental roles of the extracellular matrix: Lutolf, M. P. and Hubbell, J. A. (2005). Synthetic Ronnov-Jessen, L., Petersen, O. W. and Bissell, M. J. beyond structure to regulation. Cell Tissue Res. 339, 93-110. biomaterials as instructive extracellular microenvironments (1996). Cellular changes involved in conversion of normal Tucker, R. P. and Chiquet-Ehrismann, R. (2009). The for morphogenesis in tissue engineering. Nat. Biotechnol. to malignant breast: importance of the stromal reaction. regulation of tenascin expression by tissue 23, 47-55. Physiol. Rev. 76, 69-125. microenvironments. Biochim. Biophys. Acta 1793, 888-892. Lutolf, M. P., Gilbert, P. M. and Blau, H. M. (2009). Rosso, F., Marino, G., Giordano, A., Barbarisi, M., Ulijn, R. V. and Smith, A. M. (2008). Designing peptide Designing materials to direct stem-cell fate. Nature 462, Parmeggiani, D. and Barbarisi, A. (2005). Smart based nanomaterials. Chem. Soc. Rev. 37, 664-675. 433-441. materials as scaffolds for tissue engineering. J. Cell Physiol. Untergasser, G., Madersbacher, S. and Berger, P. (2005). Macchiarini, P., Jungebluth, P., Go, T., Asnaghi, M. A., 203, 465-470. Benign prostatic hyperplasia: age-related tissue-remodeling. Rees, L. E., Cogan, T. A., Dodson, A., Martorell, J., Rozario, T. and DeSimone, D. W. (2010). The extracellular Exp. Gerontol. 40, 121-128. Bellini, S., Parnigotto, P. P. et al. (2008). Clinical matrix in development and morphogenesis: a dynamic view. Vakonakis, I. and Campbell, I. D. (2007). Extracellular transplantation of a tissue-engineered airway. Lancet 372, Dev. Biol. 341, 126-140. matrix: from atomic resolution to . Curr. Opin. 2023-2030. Schaefer, L. and Iozzo, R. V. (2008). Biological functions Cell Biol. 19, 578-583. Macri, L., Silverstein, D. and Clark, R. A. (2007). Growth of the small leucine-rich proteoglycans: from to Velnar, T., Bailey, T. and Smrkolj, V. (2009). The wound factor binding to the pericellular matrix and its importance signal transduction. J. Biol. Chem. 283, 21305-21309. healing process: an overview of the cellular and molecular in tissue engineering. Adv. Drug Deliv. Rev. 59, 1366-1381. Schaefer, L. and Schaefer, R. M. (2010). Proteoglycans: mechanisms. J. Int. Med. Res. 37, 1528-1542. Mao, Y. and Schwarzbauer, J. E. (2005). Fibronectin from structural compounds to signaling molecules. Cell Wipff, P. J., Rifkin, D. B., Meister, J. J. and Hinz, B. fibrillogenesis, a cell-mediated matrix assembly process. Tissue Res. 339, 237-246. (2007). Myofibroblast contraction activates latent TGF- Matrix Biol. 24, 389-399. Schafer, M. and Werner, S. (2008). Cancer as an beta1 from the extracellular matrix. J. Cell Biol. 179, 1311- McCullen, S. D., Ramaswamy, S., Clarke, L. I. and overhealing wound: an old hypothesis revisited. Nat. Rev. 1323. Gorga, R. E. (2009). Nanofibrous composites for tissue Mol. Cell Biol. 9, 628-638. Wise, S. G. and Weiss, A. S. (2009). Tropoelastin. Int. J. engineering applications. Wiley Interdiscip. Rev. Nanomed. Schmidt, S. and Friedl, P. (2010). Interstitial cell Biochem. Cell Biol. 41, 494-497. Nanobiotechnol. 1, 369-390. migration: integrin-dependent and alternative adhesion Xian, X., Gopal, S. and Couchman, J. R. (2010). Morita, H., Yoshimura, A., Inui, K., Ideura, T., mechanisms. Cell Tissue Res. 339, 83-92. Syndecans as receptors and organizers of the extracellular Watanabe, H., Wang, L., Soininen, R. and Tryggvason, Schultz, G. S. and Wysocki, A. (2009). Interactions matrix. Cell Tissue Res. 339, 31-46. K. (2005). Heparan sulfate of perlecan is involved in between extracellular matrix and growth factors in wound Xu, J., Lamouille, S. and Derynck, R. (2009). TGF-beta- glomerular filtration. J. Am. Soc. Nephrol. 16, 1703-1710. healing. Wound Repair Regen. 17, 153-162. induced epithelial to mesenchymal transition. Cell Res. 19, Mott, J. D. and Werb, Z. (2004). Regulation of matrix Scott, J. E. (2003). Elasticity in extracellular matrix ‘shape 156-172. Journal of Cell Science biology by matrix metalloproteinases. Curr. Opin. Cell Biol. modules’ of , , etc. A sliding - Zhang, X., Reagan, M. R. and Kaplan, D. L. (2009). 16, 558-564. filament model. J. Physiol. 553, 335-343. Electrospun silk biomaterial scaffolds for regenerative Murakami, M., Elfenbein, A. and Simons, M. (2008). Serban, M. A. and Prestwich, G. D. (2008). Modular medicine. Adv. Drug Deliv. Rev. 61, 988-1006. Non-canonical fibroblast growth factor signalling in extracellular matrices: solutions for the puzzle. Methods 45, Zisch, A. H., Lutolf, M. P. and Hubbell, J. A. (2003). angiogenesis. Cardiovasc. Res. 78, 223-231. 93-98. Biopolymeric delivery matrices for angiogenic growth Myllyharju, J. and Kivirikko, K. I. (2004). Collagens, Shaikh, F. M., Callanan, A., Kavanagh, E. G., Burke, P. factors. Cardiovasc. Pathol. 12, 295-310. modifying enzymes and their mutations in humans, flies and E., Grace, P. A. and McGloughlin, T. M. (2008). Fibrin: worms. Trends Genet. 20, 33-43. a natural biodegradable scaffold in vascular tissue Nomura, Y. (2006). Structural change in decorin with skin engineering. Cells Tissues Organs 188, 333-346. aging. Connect. Tissue Res. 47, 249-255. Sieminski, A. L., Semino, C. E., Gong, H. and Oehrl, W. and Panayotou, G. (2008). Modulation of Kamm, R. D. (2008). Primary sequence of ionic self- Cell Science at a Glance on the Web growth factor action by the extracellular matrix. Connect. assembling peptide gels affects endothelial cell adhesion Tissue Res. 49, 145-148. and morphogenesis. J. Biomed. Mater. Res. A 87, Electronic copies of the poster insert are Pankov, R. and Yamada, K. M. (2002). Fibronectin at a 494-504. available in the online version of this article glance. J. Cell Sci. 115, 3861-3863. Smith, I. O., Liu, X. H., Smith, L. A. and Ma, P. X. at jcs.biologists.org. The JPEG images can Paszek, M. J. and Weaver, V. M. (2004). The tension (2009). Nanostructured polymer scaffolds for tissue be downloaded for printing or used as mounts: mechanics meets morphogenesis and malignancy. engineering and . Wiley Interdiscip. slides. J. Mammary Gland Biol. Neoplasia 9, 325-342. Rev. Nanomed. Nanobiotechnol. 1, 226-236.