The extracellular matrix (ECM) is a complex, non-cellular network of secreted macromolecules that surrounds and supports cells within all tissues and organs, providing essential structural scaffolding and biochemical signaling cues.[1] Composed primarily of proteins, glycans, proteoglycans, and other components, the ECM forms an intricate, tissue-specific meshwork that maintains organ integrity and modulates cellular processes.[2][3][4]The composition of the ECM varies by tissue type but generally includes fibrous proteins such as collagens and elastins for tensile strength and elasticity, as well as hydrated proteoglycans and glycosaminoglycans that create a gel-like environment for nutrient diffusion and hydration.[2][4] In specialized structures like the basement membrane, the ECM integrates laminins, type IV collagen, and nidogens to anchor epithelial and endothelial cells to underlying connective tissue.[5] These components are dynamically assembled and remodeled through cellular secretion and enzymatic activity, ensuring adaptability to physiological demands.[2][6]Beyond mechanical support, the ECM plays critical regulatory roles in cell adhesion, migration, proliferation, differentiation, and survival by interacting with cell surface receptors like integrins, thereby transmitting biomechanical and biochemical signals that influence tissue development, homeostasis, and repair.[6][7] It also contributes to mechanotransduction, where physical properties such as stiffness and topography guide cellular responses in processes ranging from embryogenesis to wound healing.[2][4]Dysregulation of ECM composition or remodeling is implicated in numerous diseases, including fibrosis, cancer progression, and cardiovascular disorders, where altered stiffness or excessive deposition promotes pathological cell behaviors such as invasion and inflammation.[8][9] In cancer, for instance, the ECM facilitates tumor growth and metastasis by providing a supportive niche and signaling pathways that enhance malignancy.[9] Understanding ECM dynamics thus holds significant potential for therapeutic interventions in regenerative medicine and disease treatment.[10]
Composition
Structural Proteins
The extracellular matrix (ECM) relies on structural proteins to provide tensile strength and organizational framework to tissues. Collagen, the most abundant protein in the animal kingdom, accounts for approximately one-third of the total protein content in the human body and is primarily synthesized by fibroblasts, which are specialized connective tissue cells responsible for producing and maintaining the ECM.[11][12]Collagens comprise a family of at least 28 types in vertebrates, formed by the assembly of 46 distinct chains, with fibrillar collagens such as types I, II, and III being the predominant forms that self-assemble into long, rope-like fibrils to confer mechanical stability.[3] Network-forming collagens, including types IV and VI, create sheet-like or beaded filament structures that support basal laminae and interstitial matrices, respectively.[3] At the molecular level, collagen molecules adopt a characteristic triple helix structure, where three polypeptide chains rich in glycine, proline, and hydroxyproline residues wind into a rigid rod approximately 300 nm long and 1.5 nm in diameter.[13]Collagen biosynthesis begins intracellularly in fibroblasts, where procollagen chains undergo posttranslational modifications such as hydroxylation of proline and lysine residues, followed by glycosylation in the endoplasmic reticulum and Golgi apparatus, before secretion as a procollagen trimer into the extracellular space.[12] Extracellularly, propeptides are cleaved to initiate spontaneous self-assembly into fibrils, which are then stabilized by enzymatic cross-linking mediated by lysyl oxidase, an enzyme that oxidizes lysine residues to form allysine, enabling covalent bonds that enhance fibril tensile strength.[14] These fibrils typically exhibit diameters ranging from 50 to 200 nm, varying by tissue type to optimize mechanical properties.[15]Elastin serves as the key structural protein imparting elasticity to the ECM, forming resilient fibers essential for tissues that undergo repeated deformation. Elastin fibers consist primarily of the hydrophobic protein tropoelastin, which polymerizes around a scaffold of fibrillin-rich microfibrils to create a cross-linked network capable of extending up to 150% of its length and recoiling with near-perfect reversibility.[16] The elastic recoil mechanism arises from the entropic elasticity of elastin's disordered, hydrophobic chains, which minimize conformational entropy when stretched and rapidly recover upon release, enabling efficient energy storage and dissipation in dynamic environments.[16]Tropoelastin monomers, synthesized by fibroblasts and smooth muscle cells, are secreted and self-aggregate on the cell surface before deposition onto fibrillin microfibrils, where lysyl oxidase catalyzes oxidative deamination of lysine residues to form unique tetrafunctional cross-links such as desmosine and isodesmosine.[17][18] These cross-links stabilize the amorphous elastin core, preventing degradation and ensuring long-term elasticity, with desmosine concentrations particularly high in load-bearing tissues.[19]In stretchable tissues like arteries and lungs, elastin fibers enable cyclic expansion and contraction; for instance, in arterial walls, they allow vessels to withstand pulsatile blood flow while maintaining recoil to propel blood forward, comprising up to 50% of the dry weight in elastic lamellae.[16]
Glycoproteins and Proteoglycans
Glycoproteins and proteoglycans are key non-collagenous components of the extracellular matrix (ECM) that mediate dynamic interactions between cells and the structural scaffold, primarily through their carbohydrate moieties and protein domains. These molecules facilitate cell adhesion, modulate growth factor binding, and contribute to tissue organization by integrating with collagen fibrils for enhanced mechanical stability. Unlike the rigid structural proteins, glycoproteins and proteoglycans enable adaptive responses due to their modular architectures and glycosylated extensions.Fibronectin, a prominent ECM glycoprotein, exhibits a modular structure composed of repeating type I, type II, and type III domains that enable diverse binding interactions. The type I and III domains primarily mediate self-assembly into fibrils, while type II domains facilitate collagen binding, and specific type III domains (e.g., III_{12-14}) interact with heparin and integrins. Fibronectin exists in multiple isoforms generated by alternative splicing, notably plasma fibronectin, which lacks the extra type III domain B (EDA) and is soluble in circulation, versus cellular fibronectin, which includes EDA and supports fibrillogenesis during tissue remodeling. These isoforms bind integrins such as α5β1 for cell adhesion, collagen types I and IV for matrix assembly, and heparin for regulating bioavailability of signaling molecules.[20][21]Laminin, another major ECM glycoprotein, forms heterotrimeric complexes with α, β, and γ chains, resulting in cross-shaped structures essential for basement membrane integrity. Over 15 isoforms exist, such as laminin-111 (α1β1γ1), which is prevalent in early embryonic basement membranes and promotes epithelial cell polarization through binding to integrins α3β1 and α6β1. The globular domains at the termini interact with nidogen (entactin) to link laminin networks to type IV collagen, while the central rod-like regions enable polymerization into sheets that provide a substrate for cell attachment and migration. Laminin isoforms like laminin-511 (α5β1γ1) predominate in mature basement membranes, supporting long-term tissue stability.[22][23]Proteoglycans consist of a core protein substituted with one or more glycosaminoglycan (GAG) chains, covalently attached via tetrasaccharide linkers to serine residues in Ser-Gly motifs, enabling them to bridge ECM components and cells. The core proteins determine specificity; for instance, aggrecan, a large chondroitin sulfate proteoglycan, features multiple GAG attachment sites and interacts with hyaluronan via its N-terminal globular domains to form massive aggregates that imbue cartilage with compressive resistance. Decorin, a small leucine-rich proteoglycan, binds collagen fibrils via its core protein to regulate fibril diameter and spacing. Cell-associated proteoglycans include the syndecan family, which are transmembrane with heparan sulfate and chondroitin sulfate chains that cluster at cell surfaces to transduce signals, and the glypican family, anchored by glycosylphosphatidylinositol (GPI) tails, which modulate growth factor gradients in the pericellular space. Specific examples include versican, a chondroitin sulfate proteoglycan abundant in cartilage where it influences matrix assembly and chondrocyte proliferation through isoform-specific GAG content, and perlecan, a heparan sulfate proteoglycan in basement membranes that stabilizes networks by binding laminin and nidogen while sequestering growth factors.[24][25][26]Biosynthesis of proteoglycans begins in the endoplasmic reticulum where the core protein is synthesized and folded, followed by O-linked glycosylation in the Golgi apparatus. Initiation occurs via xylosyltransferase adding xylose to serine, extended by galactosyltransferases and glucuronyltransferase to form the linker tetrasaccharide. Elongation of GAG chains then proceeds through alternating additions by glycosyltransferases, such as chondroitin polymerase (ChSy family) for chondroitin sulfate or heparan synthase (EXT1/EXT2 complex) for heparan sulfate, with sulfotransferases modifying the chains for functional diversity. These enzymatic steps ensure precise GAG length and composition tailored to tissue demands.[25][27]
Glycosaminoglycans and Other Polysaccharides
Glycosaminoglycans (GAGs) and other polysaccharides form a significant class of non-protein components in the extracellular matrix (ECM), primarily existing as unbound chains that contribute to tissue hydration, lubrication, and electrostatic interactions through their polyanionic properties.[28] These molecules, including both non-sulfated and sulfated variants, create a hydrated gel-like environment in the ECM by attracting and retaining water molecules via hydrogen bonding and ionic interactions.[29]Hyaluronic acid (HA), the principal non-sulfated GAG, is a linear polysaccharide composed of repeating disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine linked by alternating β-1,4 and β-1,3 glycosidic bonds.[30] With molecular weights reaching up to 10 million Da, HA exhibits exceptional viscoelastic properties essential for ECMlubrication and resilience.[31] It is synthesized at the plasma membrane by hyaluronan synthases (HAS1, HAS2, and HAS3), which extrude the growing polymer into the extracellular space without the need for core protein attachment.[32] HA's hydrophilic nature enables it to bind up to 1000 times its weight in water, facilitating ECM hydration and space-filling functions.[29]Sulfated GAGs, present as free chains in the ECM alongside their proteoglycan-bound forms, include heparan sulfate (HS), chondroitin sulfate (CS), and keratan sulfate (KS), each characterized by distinct sulfation patterns that enhance their polyanionic character. Heparan sulfate consists of repeating disaccharide units of uronic acid (either D-glucuronic or L-iduronic acid) and D-glucosamine, with sulfation occurring variably at the 2-O position of the uronic acid and N-, 3-O-, and 6-O positions of the glucosamine, creating domains of high and low sulfation.[33] Chondroitin sulfate features disaccharide repeats of D-glucuronic acid and N-acetyl-D-galactosamine, sulfated predominantly at the 4-O (CS-A isomer) or 6-O (CS-C isomer) positions of the galactosamine residue, with the ratio of these isomers varying by tissue to modulate charge distribution.[34] Keratan sulfate is composed of repeating galactose β-1,4-linked to N-acetyl-D-glucosamine units, with sulfation mainly at the 6-O position of glucosamine and occasionally at the 6-O position of galactose.[35]Heparin-like molecules, structurally similar to highly sulfated HS, occur as free polyanionic polysaccharides in certain ECM contexts, exhibiting the highest degree of sulfation among GAGs and thus the greatest negative charge density.[36] The polyanionic nature of these sulfated GAGs and polysaccharides arises from their carboxylate and sulfate groups, enabling them to bind substantial water volumes—comparable to HA—through osmotic swelling and ion entrapment.[29]Sulfation patterns in GAGs critically influence their charge density, which governs interactions with cations such as Na⁺ and Ca²⁺; higher sulfation increases anionic sites, promoting stronger binding primarily to carboxylate groups on uronic acids, while monovalent Na⁺ ions contribute to overall hydration shells.[37] For instance, in chondroitin sulfate, increased sulfation correlates with enhanced Ca²⁺ affinity, stabilizing compact conformations that affect ECM ion balance.[38] These charge-based interactions underscore the role of free GAG chains in modulating ECM electrostatic environments, distinct from their occasional attachment to proteoglycans for amplified functionality.[28]
Extracellular Vesicles and Additional Components
Extracellular vesicles (EVs) represent a diverse class of membrane-bound nanoparticles secreted by cells into the extracellular matrix (ECM), contributing to its dynamic composition beyond traditional protein and polysaccharide elements.[39] These vesicles are categorized primarily into exosomes and microvesicles based on their biogenesis and size. Exosomes, ranging from 30 to 100 nm in diameter, originate from the endosomal pathway, where intraluminal vesicles form within multivesicular bodies that subsequently fuse with the plasma membrane for release.[39][40] Microvesicles, larger at 100 to 1000 nm, arise directly from outward budding of the plasma membrane.[39] Both types encapsulate a variety of cargo, including proteins, lipids, and microRNAs (miRNAs), which they transport to mediate intercellular communication within the ECM environment.[39][41]In specialized tissues like bone, mineral components integrate with the organic ECM to provide rigidity and strength. Hydroxyapatite, with the chemical formula Ca₁₀(PO₄)₆(OH)₂, constitutes the primary inorganic phase, formed through the regulated deposition of calcium phosphate crystals within the matrix.[42] This mineralization process is tightly controlled by non-collagenous proteins such as osteopontin and bone sialoprotein, which influence crystal nucleation, inhibit excessive growth, and ensure proper orientation of hydroxyapatite along collagen fibrils.[43][44]Additional non-structural elements in the ECM include multimeric adhesive proteins and matricellular proteins that fine-tune matrix organization without serving primary load-bearing roles. Thrombospondin, a large multimeric glycoprotein, promotes cell adhesion and modulates interactions between cells and ECM components like collagen and fibronectin.[45] Matricellular proteins, exemplified by SPARC (secreted protein acidic and rich in cysteine, also known as osteonectin), act as transient regulators of ECM assembly by binding to structural proteins and growth factors, thereby influencing matrix deposition and remodeling during tissue development and repair.[45][46]Recent research since 2020 has advanced understanding of EVs' contributions to ECM dynamics, particularly in remodeling processes. EVs facilitate matrix degradation and synthesis by delivering enzymes and signaling molecules that alter ECM composition in contexts like bonehomeostasis.[47] Isolation of these vesicles commonly employs ultracentrifugation, which separates them based on density gradients to yield pure populations for analysis.[48] Furthermore, EVs hold promise as biomarkers for ECM-associated pathologies, as their protein and RNA cargo reflects alterations in matrixintegrity and can be detected non-invasively in biofluids.[49]
Functions
Mechanical Support and Elasticity
The extracellular matrix (ECM) provides mechanical support to tissues through its structural components, enabling them to withstand physical stresses without permanent deformation. Collagen fibrils, organized in a hierarchical cable-like structure, primarily confer tensile strength to the ECM, acting as load-bearing elements that resist pulling forces. In tendons, these fibrils achieve Young's moduli ranging from 1 to 10 GPa, allowing the tissue to support high tensile loads during movement.[50][51]Elasticity in the ECM arises from elastin networks, which facilitate reversible deformation and rapid recoil, essential for dynamic tissues like arteries and lungs. Elastin fibers can undergo strains up to 150% without damage, driven by entropic coiling mechanisms where random chain conformations return to a high-entropy state upon unloading.[52][53] This property ensures tissues recover shape after stretching, preventing fatigue.Viscoelasticity of the ECM, combining viscous damping and elastic recovery, is contributed by hyaluronic acid (HA) and proteoglycans, which absorb energy and dissipate stress through fluid interactions and molecular friction. In HA-based hydrogels mimicking ECM, shear moduli typically range from 0.1 to 10 kPa, providing time-dependent responses that buffer sudden impacts.[54][55]Tissue-specific ECM properties vary widely; for instance, bone's mineralized collagen matrix imparts high rigidity with Young's moduli up to 10 GPa, while cartilage's proteoglycan-rich ECM enables compressibility with moduli around 100-150 kPa in superficial zones. Cross-linking density in collagen networks significantly influences overall stiffness, as increased enzymatic cross-links enhance resistance to deformation.[55][56] These mechanical behaviors often follow linear elasticity principles, described by Hooke's law:\sigma = E \varepsilonwhere \sigma is stress, E is the Young's modulus, and \varepsilon is strain, applicable to small deformations in fibrillar ECM components.[57]
Cell Adhesion and Migration
The extracellular matrix (ECM) facilitates cell adhesion primarily through integrin receptors, which are transmembrane αβ heterodimers that bind specific motifs in ECM proteins such as the arginine-glycine-aspartic acid (RGD) sequence found in fibronectin and certain laminin isoforms. These integrins, including α5β1 for fibronectin and α3β1 or α6β1 for laminin, undergo conformational changes upon ligand binding, enabling intracellular connections to the actin cytoskeleton.[58] This binding initiates the assembly of focal adhesions, dynamic multiprotein complexes where talin links the integrin β-subunit tail to actin filaments, recruiting vinculin to reinforce the linkage and stabilize adhesion under mechanical stress.[59] Focal adhesions thus serve as sites for force transmission, allowing cells to sense and respond to ECM topography and rigidity.Beyond focal adhesions, specialized adhesion complexes further mediate ECM interactions during attachment and motility. Hemidesmosomes, prominent in epithelial tissues, anchor basal keratinocytes to the basement membrane via α6β4 integrins binding laminin-332, coupled to intermediate filaments through plectin and BP230 proteins for robust, stable attachment resistant to shear forces.[60] In contrast, invadopodia are transient, actin-rich protrusions formed by invasive cells, such as metastatic cancer cells, that locally degrade ECM through recruitment of metalloproteases like MT1-MMP, enabling directional penetration and migration.[61] These structures highlight the ECM's role in balancing stable adhesion with dynamic remodeling.Cell migration on the ECM involves guided motility modes, including durotaxis, where cells preferentially move toward regions of increasing substrate stiffness via integrin clustering and focal adhesion reinforcement on stiffer matrices, as observed in fibroblasts navigating tissue gradients.[62] Haptotaxis directs migration along gradients of ECM adhesiveness, such as varying fibronectin densities, prompting cells to follow higher ligand concentrations through biased lamellipodial extension.[63] Matrix metalloproteinases (MMPs), secreted by migrating cells, degrade ECM barriers to create paths, with MMP-2 and MMP-9 cleaving collagen and gelatin to facilitate invasion while exposing cryptic binding sites that promote further adhesion.[64] These dynamics are exemplified in wound healing, where keratinocytes and fibroblasts migrate collectively across provisional fibronectin-rich matrices, remodeling ECM to close gaps, and in embryonic development, where neural crest cells traverse basement membranes via integrin-MMP interactions to reach target sites.[65][66]Quantitatively, atomic force microscopy (AFM) measurements reveal that individual integrin-ECM bonds withstand rupture forces of 20-40 piconewtons (pN), sufficient for cells to generate traction during migration without bond failure under physiological loads.[67] This adhesion strength scales with cluster size in focal adhesions, enabling cells to exert forces up to several nanonewtons for effective pathfinding.[68]
Biochemical Signaling
The extracellular matrix (ECM) plays a pivotal role in biochemical signaling by acting as a reservoir and presenter of growth factors, sequestering them to regulate their bioavailability and spatiotemporal presentation to cells. Heparan sulfate (HS) proteoglycans, key ECM components, bind and stabilize growth factors such as fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF), preventing their diffusion and degradation while facilitating high-affinity interactions with cell surface receptors upon localized release. These interactions are modulated by the sulfation patterns of HS chains, which confer specificity to ligand binding and signaling activation. Release of sequestered growth factors occurs primarily through enzymatic degradation of the ECM, involving proteases that cleave HS proteoglycans and liberate bioactive molecules to influence processes like angiogenesis and tissue remodeling.Matrix metalloproteinases (MMPs), a family of zinc-dependent endopeptidases exemplified by MMP-2 (gelatinase A) and MMP-9 (gelatinase B), drive much of this signaling by cleaving ECM components to release bound growth factors and generate bioactive fragments known as matrikines. These matrikines, derived from proteins like collagen and elastin, function as signaling peptides that bind specific receptors on cells, promoting responses such as inflammation, proliferation, and apoptosis independent of growth factor release. MMP activity is precisely regulated by endogenous inhibitors, including tissue inhibitors of metalloproteinases (TIMPs), which form complexes with active MMPs to prevent excessive ECM degradation and maintain signaling homeostasis. Dysregulation of MMPs and TIMPs has been implicated in pathological signaling, where unbalanced matrikine production contributes to disease progression.Beyond growth factor release, ECM proteolysis unmasks cryptic sites—sequestered peptide sequences within structural proteins like collagen—that become exposed and bioactive upon cleavage, thereby initiating novel signaling cascades. For example, MMP-mediated breakdown of collagen type IV reveals hidden motifs that interact with integrins and growth factor receptors, amplifying cellular responses to injury or stress. This exposure of cryptic sites transforms the ECM from a passive scaffold into a dynamic signaling platform, where fragmented peptides act as endogenous regulators of cell behavior.Specific examples illustrate the ECM's signaling versatility: transforming growth factor-β (TGF-β) is stored in a latent complex with its latency-associated peptide (LAP), which covalently binds to fibrillin microfibrils in the ECM, controlling TGF-β activation through mechanical or proteolytic cues that disrupt the LAP-fibrillin interaction. Similarly, HS proteoglycans modulate Wnt signaling by clustering Wnt ligands and co-receptors on the cell surface, enhancing gradient formation and pathway specificity during development and homeostasis; distinct HS sulfation patterns differentially regulate canonical versus non-canonical Wnt branches.
Biological Roles
In Development and Morphogenesis
During embryonic development, the extracellular matrix (ECM) undergoes dynamic temporal changes to support tissue patterning and organ formation. Initially, a provisional ECM rich in fibronectin and hyaluronic acid (HA) predominates, providing a flexible scaffold that facilitates cell migration and proliferation in the early embryo.[69] As development progresses, this provisional matrix is remodeled into a mature, collagen-rich structure that offers greater mechanical stability for tissue maturation.[70]Basement membranes, specialized ECM sheets, assemble through self-organization of laminins and type IV collagens, crosslinked by nidogens and proteoglycans, to separate epithelial layers from underlying mesenchyme and guide organogenesis.[71]In morphogenetic processes, ECM components direct branching morphogenesis in organs such as the lungs and kidneys. Laminin gradients within the basement membrane regulate ureteric bud branching in the kidney and epithelial tip outgrowth in the lung, influencing directional cell proliferation and invasion.[72] Similarly, neural crest cells migrate along fibronectin-rich ECM tracks, where dynamic assembly of fibrillar fibronectin by leader cells creates haptotactic cues that coordinate collective stream migration during craniofacial and peripheral nervous system development.[73]ECM remodeling is mediated by specific enzymes that enable tissue shaping. Members of the ADAMTS family, such as ADAMTS-12, cleave versican and other proteoglycans in the provisional matrix, promoting chondrocyte proliferation and cartilage matrix organization during skeletal development.[74] Hyaluronidases, including TMEM2, degrade HA to reduce matrix viscosity, facilitating mesenchymal cell expansion and tissue hydration in expanding embryonic structures like the heart and neural tube.[75]Genetic models highlight ECM's essential roles in development. Knockout of collagen XVIII in mice disrupts eye morphogenesis, leading to abnormal hyaloid vessel regression, retinal vascular outgrowth defects, and anterior segment anomalies due to impaired basement membrane integrity in ocular tissues.[76] In species comparisons, Drosophila's basement membrane analogs, composed of collagen IV and laminin, support epithelial morphogenesis and organ shaping, with hemocyte-secreted components reinforcing ECM assembly akin to vertebrate processes.[77]Key developmental events rely on ECM dynamics for patterning. During gastrulation, localized ECM remodeling, including fibronectin fibrillogenesis, coordinates mesendoderm invagination and convergent extension movements in vertebrates like zebrafish.[78] In somitogenesis, ECM components such as fibronectin and laminin contribute to somite boundary formation, ensuring periodic segmentation of the vertebral column.[79]
In Tissue Homeostasis and Repair
The extracellular matrix (ECM) maintains tissuehomeostasis through a balanced process of continuous synthesis and degradation, ensuring structural integrity and functional adaptability in mature tissues. In steady-state conditions, fibroblasts remain largely quiescent, producing ECM components at a rate that matches their enzymatic breakdown by matrix metalloproteinases (MMPs) and other proteases. For instance, type I collagen, a major ECM protein in skin, exhibits a half-life of approximately 15 years, reflecting the slow turnover necessary for long-term tissue stability. This equilibrium prevents excessive accumulation or loss of matrix, supporting organ function without overt remodeling. Disruptions in this balance, such as altered protease activity, can lead to pathological changes, underscoring the ECM's role in physiological maintenance.[80]In wound repair, the ECM dynamically participates across overlapping phases to restore tissue integrity. During the inflammatory phase, recruited immune cells release MMPs that degrade damaged ECM, clearing debris and facilitating leukocyte infiltration to control infection. In the subsequent proliferative phase, fibroblasts deposit a provisional matrix rich in fibronectin, fibrin, and hyaluronan, which provides a scaffold for granulation tissue formation and angiogenesis. The remodeling phase, lasting weeks to months, involves collagen cross-linking and realignment by lysyl oxidase, culminating in scar formation that replaces the provisional matrix with a more organized but often less elastic structure. These ECM-mediated events parallel developmental processes in their orchestration of cellular responses, though adapted for adult recovery.[81][82]The ECM also sustains stem cell niches critical for tissue renewal. In the hematopoietic stem cell (HSC) niche within bone marrow, osteopontin—a glycoprotein ECM component secreted by osteoblasts—acts as a negative regulator, limiting HSC proliferation and maintaining quiescence to preserve the stem cell pool. Similarly, in intestinal crypts, gradients of laminin isoforms in the basement membrane ECM guide stem cell positioning and differentiation, with higher concentrations at the crypt base promoting self-renewal while lower levels toward the villus apex favor maturation. These niche-specific ECM cues ensure controlled regeneration without exhaustion of progenitor pools.[83][84]Aging impairs ECM homeostasis through progressive stiffening and loss of elasticity, primarily driven by advanced glycation end-products (AGEs). AGEs form via non-enzymatic glycation of long-lived proteins like collagen, creating irreversible cross-links that increase matrix rigidity and disrupt fibril assembly. In skeletal muscle, for example, age-associated ECM stiffening correlates directly with elevated AGE adducts and collagen content, compromising tissue compliance and contributing to reduced regenerative capacity. These changes accumulate over decades, altering mechanotransduction and exacerbating functional decline in multiple organs.[85][86]Recent advances post-2020 have leveraged ECM components for enhanced tissue repair strategies. ECM-derived hydrogels, incorporating decellularized matrix from dermal or adipose sources, serve as bioactive wound dressings that promote moist healing, reduce inflammation, and accelerate re-epithelialization in chronic wounds. For instance, collagen-based hydrogels functionalized with antimicrobial peptides have demonstrated improved outcomes in diabetic ulcers by mimicking native matrix signaling. Complementing this, decellularized ECM scaffolds in tissue engineering preserve native architecture and bioactive cues, enabling vascularized constructs for skin and cardiac regeneration with demonstrated biocompatibility and reduced immunogenicity compared to synthetic alternatives. As of 2025, ongoing clinical trials are expanding dECM applications to myocardial infarction and cartilage regeneration.[87][88][89]
Mechanical Sensing and Response
Cells sense the mechanical properties of the extracellular matrix (ECM) through mechanotransduction, a process where physical forces from the ECM are converted into biochemical signals that influence cellular behavior and gene expression. This sensing primarily occurs via integrin-mediated adhesions, which connect the ECM to the actin cytoskeleton, allowing cells to probe matrix stiffness and topography. Upon engagement with ECM ligands, integrins cluster to form focal adhesions, where mechanical forces activate downstream signaling pathways.[90]A key mechanotransduction pathway involves focal adhesion kinase (FAK), which is activated by tensile forces transmitted through integrins and talin-vinculin linkages at focal adhesions. Force application unfolds FAK's autoinhibitory domains, enabling autophosphorylation at tyrosine 397 and recruitment of Src kinase, which amplifies signaling to regulate cell migration and survival. Parallel to FAK, the Hippo pathway effectors YAP and TAZ are mechanosensitive; on stiff matrices, nuclear translocation of YAP/TAZ is promoted via integrin-FAK-RhoA-actin signaling, driving transcription of genes involved in proliferation and differentiation. In contrast, soft matrices retain YAP/TAZ in the cytoplasm through Hippo kinase activation, favoring other fates.[91][92][93]Matrix stiffness profoundly affects gene expression and cell fate. For instance, mesenchymal stem cells (MSCs) on stiff matrices (25–40 kPa, mimicking bone) upregulate RUNX2, a master osteogenic transcription factor, via YAP/TAZ and ERK signaling, promoting osteogenesis. Conversely, soft matrices (0.1–1 kPa, akin to brain tissue) suppress RUNX2 and activate neurogenic markers like β-III tubulin through mechanosensitive ion channels and reduced YAP activity, directing neurogenesis. In myogenesis, aligned ECM topography enhances myoblast fusion into multinucleated myotubes by orienting cytoskeletal tension and cadherin signaling, as seen in fibronectin-coated substrates. Adipogenesis is favored on compliant substrates (1–10 kPa), where low stiffness limits YAP nuclear entry, allowing PPARγ upregulation and lipid accumulation in preadipocytes.[94][95][96][97]Cells exhibit durotaxis, a directed migration toward stiffer ECM regions, driven by differential adhesion strengthening and actomyosin contractility on stiffness gradients, which is crucial for wound healing and cancer invasion. ECM thixotropy, the reversible fluidization under shear stress, facilitates cell migration by temporarily reducing matrix viscosity, allowing proteolysis and cytoskeletal pushing without permanent remodeling. These behaviors are studied using polyacrylamide (PAAm) hydrogels, tunable from 0.1 kPa (soft) to 40 kPa (stiff) by varying acrylamide and bis-acrylamide concentrations, coated with ECM proteins like collagen or fibronectin to mimic physiological cues. Atomic force microscopy (AFM) enables nanoscale stiffness mapping of native ECM, revealing heterogeneous mechanics that correlate with cellular responses in tissues.[98][99][100][101]In focal adhesions, force balance is often modeled using Hooke's law, where the applied force F equals the spring constant k times displacement d:F = k \cdot dThis equation describes how adhesions act as viscoelastic springs, with k reflecting ECM stiffness and linkage compliance, balancing cellular contractility against matrix resistance to propagate signals.[102]
Clinical Significance
Role in Diseases
Alterations in the extracellular matrix (ECM) play a central role in various pathologies, where dysregulated remodeling leads to tissue dysfunction and disease progression. In fibrotic conditions, excessive deposition of ECM components, particularly collagen, disrupts normal tissue architecture and impairs organ function. For instance, in liver cirrhosis and idiopathic pulmonary fibrosis (IPF), fibroblasts differentiate into myofibroblasts under the influence of transforming growth factor-β (TGF-β), driving persistent collagen synthesis and matrix stiffening.[103][104] This TGF-β-mediated activation sustains myofibroblast persistence, creating a feed-forward loop that exacerbates fibrosis and hinders resolution.[105]In cancer, ECM remodeling contributes to tumor progression through desmoplasia, where stromal fibroblasts deposit dense collagen networks that stiffen the surrounding matrix and facilitate invasion. In breast cancer, accumulation of hyaluronic acid (HA) in the tumor microenvironment enhances cell motility and promotes metastatic spread by altering matrix compliance.[106] Furthermore, ECM stiffening supports the formation of pre-metastatic niches, where primary tumor-derived factors prime distant sites for colonization via extracellular vesicle-mediated remodeling.[107][108]Genetic disorders often arise from direct defects in ECM structural proteins, leading to connective tissue fragility. Ehlers-Danlos syndrome (EDS) is characterized by mutations in collagen genes, such as COL5A1 and COL5A2, resulting in abnormal fibril assembly and reduced matrix integrity that manifests as skin hyperextensibility and joint hypermobility.[109]Marfan syndrome stems from mutations in the FBN1 gene encoding fibrillin-1, a key ECM glycoprotein, which disrupts microfibril formation and elastic fiber stability, predisposing individuals to aortic aneurysms.[110]Osteogenesis imperfecta involves mutations in type I collagen genes (COL1A1 or COL1A2), causing brittle bones due to defective ECM mineralization and reduced collagen secretion.[111]Inflammatory diseases like rheumatoid arthritis (RA) feature ECM degradation driven by matrix metalloproteinases (MMPs), which are upregulated in synovial fibroblasts and macrophages. MMP-1, MMP-3, and MMP-9 target collagens and proteoglycans, leading to cartilage erosion and joint destruction in RA.[112]Recent insights post-2020 highlight ECM involvement in emerging conditions. In severe COVID-19, viral infection triggers excessive ECM deposition in the lungs, with elevated collagen and fibronectin contributing to post-acute fibrosis through myofibroblast activation and TGF-β signaling.[113] In inflammatory bowel disease (IBD), gut commensal bacteria, such as those enriched in dysbiotic microbiomes, degrade ECM components like collagen IV via microbial proteases, exacerbating mucosal barrier breakdown and chronic inflammation.[114]
Diagnostic and Therapeutic Applications
The extracellular matrix (ECM) serves as a valuable source of biomarkers for diagnosing fibrotic and degenerative diseases, enabling non-invasive monitoring of tissue remodeling. Circulating collagen fragments, such as PRO-C3—a neo-epitope marker of type III collagen synthesis—have been established as predictors of fibrosis progression in conditions like liver fibrosis, correlating with disease severity and clinical outcomes in multiple cohorts.[115] Similarly, elevated levels of hyaluronic acid (HA) in synovial fluid indicate ECM disruption in osteoarthritis, reflecting increased turnover and inflammation that correlates with joint degeneration and pain severity.[116]Advanced imaging techniques exploit ECM components for precise visualization of tissue architecture and pathology. Second harmonic generation (SHG) microscopy provides label-free imaging of collagen fibers within the ECM, revealing fibrillar organization changes in tumors and fibrotic tissues, which aids in assessing disease progression without exogenous dyes.[117]Magnetic resonance imaging (MRI), particularly T2 and T1ρ mapping, quantifies ECM alterations in cartilage, such as glycosaminoglycan loss and collagen matrix degradation, offering a non-invasive tool for early osteoarthritis detection and monitoring therapeutic responses.[118]Therapeutic strategies targeting the ECM focus on modulating its remodeling to halt pathological progression. Matrix metalloproteinase (MMP) inhibitors like doxycycline have shown promise in preclinical studies by reducing ECM degradation in connective tissue disorders, such as hypermobile Ehlers-Danlos syndrome, where they restore collagen organization and mitigate fibroblast dysfunction.[119] Anti-fibrotic agents, including pirfenidone, inhibit ECM deposition in pulmonary fibrosis models by suppressing collagen synthesis and fibril formation, leading to reduced lung stiffness and improved function in preclinical and clinical settings.[120] ECM-mimicking scaffolds composed of collagen and HA composites support tissue repair by providing biomechanical cues and promoting cell adhesion, as demonstrated in dermal and cartilage regeneration applications.[121]In regenerative medicine, decellularized ECM (dECM) derived from native tissues serves as a bioactive scaffold for organoid culture, preserving matrix proteins to enhance stem celldifferentiation and organ-specific functionality in models of liver and intestinal regeneration.[122] Three-dimensional (3D) bioprinting using gelatin methacryloyl (GelMA) hydrogels incorporates ECM motifs to fabricate vascularized constructs, improving cell viability and matrix deposition for applications in skin and bone repair.[123]Emerging post-2020 approaches leverage nanotechnology and gene editing for precise ECM modulation. Nanomedicine platforms, such as enzyme-loaded nanoparticles, enable targeted ECM degradation in tumor microenvironments, enhancing drug penetration and immune cell infiltration in solid cancers like pancreatic ductal adenocarcinoma.[124]CRISPR/Cas9-based editing of genes affecting ECM composition in stem cells alters matrix properties in engineered tissues, offering potential for personalized regenerative therapies.[125]
ECM in Diverse Organisms
In Animals
In animals, the extracellular matrix (ECM) exhibits remarkable diversity across tissues, tailored to specific mechanical and functional demands. In bone, the ECM is predominantly composed of mineralized collagen fibrils, primarily type I collagen, which provide rigidity and tensile strength through hydroxyapatite crystal deposition, enabling load-bearing functions. Cartilage ECM, in contrast, is rich in aggrecan proteoglycans and type II collagen, forming a hydrated gel that resists compressive forces while allowing flexibility in joints. Vascular tissues feature elastin lamellae within the ECM of arterial walls, facilitating elastic recoil and maintaining blood flow dynamics under pulsatile pressure.Evolutionarily, the ECM in animals shows deep conservation, with basement membranes—thin sheets of type IV collagen, laminins, and nidogens—present from sponges (Porifera) to mammals, underscoring their role in epithelial organization since the emergence of metazoa. Invertebrate ECMs are largely collagenous, as seen in the fibrous networks of cnidarians and echinoderms, though arthropods incorporate chitin as a key polysaccharide in their exoskeletal matrices for structural reinforcement. This collagen-centric framework predates more complex vertebrate innovations, highlighting progressive elaboration in multicellular animals.Comparatively, the ECM in fish scales consists of layered collagen and hydroxyapatite, which dictate skin innervation patterns and vascular distribution, differing from the mammalian dermis where type I collagen and elastin fibers support dermal-epidermal interactions and wound healing. In regeneration, the salamander limb ECM undergoes dynamic turnover, with matrix metalloproteinases degrading pre-existing structures to form a provisional matrix that guides blastema formation and tissue repatterning, enabling scar-free regrowth absent in most mammals.Pre-metazoan origins of animal-like ECM are evident in Filozoa, the clade including choanoflagellates, where ECM-like structures such as siliceous loricae and adhesive glycoproteins facilitate colonial aggregation and substrate adhesion, suggesting proto-ECM components that preceded full metazoan multicellularity.
In Plants and Fungi
In plants, the extracellular matrix is manifested primarily through the cell wall, a rigid structure that provides mechanical support and withstands turgor pressure to prevent cell rupture while enabling controlled expansion. Primary cell walls, formed during active growth, consist of cellulose microfibrils (15–40%) embedded in a matrix of hemicelluloses such as xyloglucans (20–30%) and pectic polysaccharides (30–50%), along with lesser amounts of arabinoxylans and structural proteins.[126] Secondary cell walls, deposited in mature cells for enhanced rigidity, incorporate lignin alongside cellulose and hemicelluloses, contributing to vascular tissue strength and overall plant architecture.[127] Hydroxyproline-rich glycoproteins like extensins function as structural analogs to animal ECM proteins, cross-linking to reinforce wall integrity and extensibility during development.[128] The apoplastic fluid in intercellular spaces contains secreted proteins, including cell wall-modifying enzymes, oxidoreductases, and stress-related factors, facilitating signaling and nutrient exchange akin to ECM functions.[129]In fungi, the cell wall acts as the primary extracellular matrix, comprising a chitin-β-glucan scaffold embedded with mannoproteins that form a protective outer layer. Chitin microfibrils and branched β-1,3-glucans constitute the inner rigid framework, cross-linked via β-1,6-glucans, while mannoproteins—glycoproteins with up to 50% carbohydrate content—provide adhesion and porosity regulation.[130][131] During hyphal growth, fungi secrete ECM-like matrices rich in polysaccharides and proteins, supporting biofilm formation and environmental adaptation.[132] These matrices enable hyphal adhesion to substrates and host tissues, contrasting with the more dynamic, collagen-dominated animal ECM by emphasizing rigidity for osmotic stability and protection.[133]Plant and fungal matrices share functional parallels with animal ECM in structural support and signaling but differ in composition and interaction mechanisms; for example, plants lack integrins and instead use wall-associated kinases (WAKs)—receptor-like kinases spanning the plasma membrane—to detect wall perturbations and trigger responses like cell expansion or defense.[134] Recent post-2020 studies reveal dynamic roles in stress adaptation: in plants, apoplastic remodeling under drought involves peroxidase-mediated ROS modulation in the cell wall to maintain integrity and enhance tolerance, as seen in maize where ZmPrx25 regulates extracellular oxidative balance.[135] In fungi, hyphal ECM secretions in biofilms bolster pathogenesis by encapsulating virulence factors and evading host immunity, with Paracoccidioides species demonstrating polysaccharide-rich matrices that promote tissue invasion.[136]
History
Early Discoveries
In the late 18th and early 19th centuries, the foundations of understanding the extracellular matrix were laid through histological descriptions of connective tissues. French anatomist Marie François Xavier Bichat, often regarded as a pioneer of tissue pathology, systematically classified human body structures into 21 distinct tissue types in his 1801 treatiseAnatomie générale appliquée à la physiologie à la médecine, including "cellular tissue" which encompassed what is now identified as connective tissue providing structural support to organs. Bichat's work, conducted without microscopy, emphasized the integration of these tissues in organ formation and pathology, shifting focus from organs to their compositional elements.Building on this, German pathologist Rudolf Virchow advanced the concept in his seminal 1858 lectures compiled as Cellular Pathology, where he described the "ground substance" as an intercellular matrix originating from cells but distinct from them, serving as the medium in which cells are embedded. Virchow viewed this amorphous ground substance as a product of cellular activity, integral to tissue integrity and pathological changes, thereby integrating it into his cellular theory of disease. This recognition marked a pivotal shift, portraying the extracellular material not merely as passive filler but as a dynamic component influenced by cellular processes.[137]Entering the early 20th century, improved optical techniques enabled more precise visualization of matrix components. In the 1920s, polarization microscopy revealed the fibrillar structure of collagen, demonstrating its birefringent properties due to aligned molecular chains, which distinguished it from other tissue elements.[138] This method, applied to connective tissues, highlighted collagen's organized, rope-like fibrils as key to tensile strength. Concurrently, elastin was isolated in a relatively pure form in 1925 through hydrolysis studies, confirming its distinct chemical composition and elastic properties separate from collagen.[139]Key contributions in the 1930s further characterized matrix elements. Pioneering cell biologist Albert Claude, through early tissue fractionation techniques at the Rockefeller Institute, developed methods for separating cellular components, laying groundwork for later understanding of matrix-cell interactions via differential centrifugation.[140] Simultaneously, the discovery of glycosaminoglycans (GAGs), then termed mucopolysaccharides, occurred in the 1930s when Karl Meyer and colleagues identified sulfated polysaccharides like chondroitin sulfate in cartilage and other matrices, revealing their acidic, hydrated nature essential for tissue resilience.[141]The advent of electron microscopy in the 1950s provided ultrastructural insights, particularly into basement membranes. Researchers such as David C. Pease and Baker in 1950, followed by Krakower and Greenspon in 1951, used transmission electron microscopy to visualize the glomerular basement membrane as a distinct, electron-dense layer approximately 300-500 nm thick, separating epithelial and endothelial cells in renal tissues.[142] These observations confirmed basement membranes as specialized matrix sheets composed of intertwined filaments, foundational to epithelial barriers. By the 1960s, the first purification of proteoglycans—complexes of GAGs bound to core proteins—was achieved, including isolations from bovine nasal cartilage, elucidating their macromolecular structure and role in matrix hydration.[143]
Modern Advances
The 1970s marked a pivotal shift in extracellular matrix (ECM) research with the identification and sequencing of key glycoproteins, including the isolation of laminin as a major basement membrane component in 1979. Fibronectin, first isolated from fibroblasts in the early 1970s, was sequenced through cDNA cloning efforts that revealed its modular structure comprising repeating amino acid domains essential for cell adhesion and migration.[144] By the late 1970s, partial amino acid sequencing confirmed fibronectin's role in linking cells to the ECM, laying groundwork for understanding tissue organization.[145]The 1980s brought molecular breakthroughs, including the discovery of integrins as transmembrane receptors mediating cell-ECM interactions. Integrins were first identified in the mid-1980s through studies on platelet aggregation and leukocyte adhesion, with the fibronectin receptor (α5β1 integrin) cloned and shown to bind specific ECM motifs like the Arg-Gly-Asp sequence.[145] Concurrently, matrix metalloproteinases (MMPs) were cloned, revealing their zinc-dependent proteolytic activity against ECM components such as collagen and laminin; the first human MMP (MMP-1, interstitial collagenase) cDNA was isolated in 1986, highlighting MMPs' role in ECM remodeling during development and pathology.[146]In the 1990s and 2000s, genetic models illuminated ECM functions in stem cell biology and tissue development. Knockout mice targeting ECM genes, such as the 1995 generation of laminin β2-deficient mutants, demonstrated severe glomerular defects and nephrotic syndrome, underscoring laminin's structural and signaling roles in basement membranes.[147] These models, alongside studies showing ECM stiffness directing mesenchymal stem cell differentiation into lineages like osteoblasts on rigid substrates versus adipocytes on soft ones, established the ECM as a niche regulator of stem cell fate and self-renewal.[148]The completion of the Human Genome Project in 2003 accelerated ECM research by enabling comprehensive annotation of matrisome genes—over 1,000 protein-coding genes encoding ECM and ECM-associated proteins—facilitating genotype-phenotype mapping in diseases.[149] This genomic resource supported large-scale analyses linking ECM variants to conditions like fibrosis and cancer, transforming ECM from a structural scaffold to a dynamically regulated network.The 2010s saw the rise of omics technologies in ECM profiling, with proteomics approaches defining the matrisome across tissues. Mass spectrometry-based methods, refined since 2010, identified over 200 core matrisome proteins in human organs, revealing tissue-specific compositions and post-translational modifications that influence bioavailability and function.[150] Mechanobiology advanced concurrently, with YAP/TAZ transcription factors emerging as key sensors of ECM mechanics; studies from 2011 onward showed that stiff ECM promotes YAP/TAZ nuclear translocation via integrin-actin linkages, driving proliferation and fibrosis.[151]Post-2020 developments integrated advanced tools for dissecting ECM dynamics. Single-molecule force spectroscopy using atomic force microscopy has quantified ECM protein unfolding forces, such as collagen's tensile strength exceeding 100 pN, providing atomic-scale insights into mechanical stability during tissue stress.[152]Artificial intelligence models now simulate ECM remodeling, with machine learning algorithms predicting stiffness-induced signaling cascades and matrisome alterations in aging tissues based on multi-omics data.[153] In longevityresearch, ECM dysregulation has been linked to cellular senescence, where fragmented collagens accumulate and impair rejuvenation; interventions modulating ECM composition show potential to extend lifespan in model organisms.[154]Yoshinori Ohsumi's 2016 Nobel Prize in Physiology or Medicine for elucidating autophagy mechanisms has implications for ECM turnover, as autophagy contributes to degrading misfolded ECM proteins and regulating lysosomal pathways in fibroblasts to help prevent fibrotic accumulation.[155] These advances underscore the ECM's integration with cellular homeostasis, paving the way for targeted therapies in regenerative medicine.