Matrix metalloproteinases and their role in periodontal diseases

The matrix metalloproteinases (MMPs) are a family of structurally and functionally related endoproteinases that are collectively capable of degrading most of the components of the extracellular matrix (ECM) 1, 2. They are calcium-dependent, zinc-containing endopeptidases, which are involved in tissue remodeling and degradation of the ECM, including collagens, elastins, gelatin, matrix glycoproteins, and proteoglycans. MMPs are excreted by a variety of connective tissue and pro-inflammatory cells including fibroblasts, osteoblasts, endothelial cells, macrophages, neutrophils, and lymphocytes. These enzymes are expressed as zymogens, which are subsequently processed by other proteolytic enzymes (such as serine proteases, furin, plasmin, and others) to generate the active forms. Under normal physiological conditions, the proteolytic activity of the MMPs is controlled primarily at any of the following three known stages: transcription, activation of the zymogens, and inhibition of the active forms by various tissue inhibitors of MMPs (TIMPs). These enzymes share common functional domains and activation mechanisms as they depend on Ca2+ and Zn2+ ions and are active at neutral pH.

Formation and activation of MMPs

The MMPs are secreted as latent enzymes and require activation. They are produced as zymogens, with a signal sequence and propeptide segment that must be removed during activation. Their proteolysis is tightly regulated to prevent tissue damage.


MMPs share a common structure that comprises of four main domains: the propeptide, catalytic, haemopexin-like, and transmembrane domains 3 (Figure 10.1). The propeptide domain contains a cysteine residue that binds zinc in the active site to form the cysteine switch. The binding of cysteine in the catalytic domain blocks the……………


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These domains are,

  • A short signal sequence is followed in order by a propeptide which endows the virgin enzyme with catalytic latency
  • A catalytic domain which contains the active site and the catalytic machinery
  • A proline-rich hinge region and
  • A pexin-like COOH-terminal domain which plays a role in determining substrate specificity.

Structure of MMPs

Structure of MMP's

TMD=Transmembrane domain


HP= Haemopexin

Classification of MMPs

MMPs are able to degrade practically all ECM components and can be classified according to their substrate specificity as collagenases, gelatinases, stromelysins and matrilysins 6 (Figure 10.2). Collagenases degrade triple helical fibrillar collagens, which are the main components of bone and cartilage. Gelatinases degrade molecules in the basal lamina around capillaries, facilitate angiogenesis and neurogenesis, and contribute to instigating cell death. Stromelysins [MMP-3, MMP-10, MMP-11, and MMP-7 (also known as matrilysins)] are small proteases that degrade components of the ECM, although not the triple-helical fibrillar collagens.  Another type of MMPs are called as the MT-MMPs (Membrane Type Matrix Metalloproteinases) that are localized to the cell surface. Four of the MT-MMPs contain hydrophobic transmembrane domains (MMP-14, MMP-15, MMP-16, MMP-24), followed by a cytoplasmic domain. Two other members of the MT-MMP subfamily, MMP-17 (MT4-MMP) and MMP-25 (MT6-MMP) are anchored to the plasma membrane via a glycosyl-phosphatidyl inositol (GPI) anchor.

Classification of MMP’s

Classification of MMPs

Biological action of MMPs

The substrate specificity of the MMPs is not yet fully characterized. Known substrates include most of the ECM components (fibronectin, vitronectin, laminin, entactin, tenascin, aggrecan, myelin basic protein, etc.). The collagens (Types I, II, III, IV, V, VI, VII, VIII, IX, X, XIV) have all been shown to be the substrates for different MMPs, with greatly different efficacies. In addition to connective tissue and ECM components, proteinase inhibitors such as α1-proteinase inhibitor, antithrombin-III and α2-macroglobulin are selectively cleaved by MMPs (Table 10.1).

The Matrix Metalloproteinase family



Collagen Substrates

Additional Substrates 



Collagenase-1, Fibroblast Collagenase, Interstitial Collagenase, Tissue Collagenase.


Aggrecan, Gelatin, MMP-2,MMP-9


72 kDa Gelatinase, 72 kDa Gelatinase/Type IV Collagenase, Gelatinase  A.


Aggrecan, Elastin, Fibronectin, Gelatin,Laminin,MMP-9,MMP-13.


Procollagenase, SL-1, Stromelysin-1, Transin-1.


Aggrecan, Elastin, Fibronectin, Gelatin, Laminin, MMP-7, MMP-8, MMP-13.


Matrilysin, Matrin, PUMP-1 Protease, Uterine Metalloproteinase.


Aggrecan, Elastin, Fibronectin, Gelatin, Laminin, MMP-1, MMP-2, MMP-9.



Collagenase–2, Neutrophil Collagenase, PMNL Collagenase.


Aggrecan, Elastin, Fibronectin, Gelatin, Laminin.



92 kDa Gelatinase, 92 kDa Gelatinase/Type IVCollagenase, Gelatinase B.


Aggrecan, Elastin, Fibronectin, Gelatin.



SL-2, Stromelysin-2, Transin-2.


Aggrecan, Elastin, Fibronectin, Gelatin, Laminin, MMP-1, MMP-8.


SL-3, Stromelysin-3, ST-3


Aggrecan, Fibronectin, Laminin


HME, Macrophage Metalloelastase MME


Elastin, Fibronectin, Gelatin, Laminin




Aggrecan, Gelatin


Xenopus Collagenase-4, xCol4






Fibronectin, Aggrecan, COMP, Laminin, Gelatin.




Aggrecan, Amelogenin, COMP


Matrilysin-2, Endometase


Gelatin, Fibronectin


Membrane-Type Metalloproteinase-14


Aggrecan, Elastin, Fibronectin, Gelatin,Laminin,MMP-2,MMP-13


Membrane-Type Metalloproteinase-15


Fibronectin, Gelatin, Laminin, MMP-2


Membrane-Type Metalloproteinase-16




Membrane-Type Metalloproteinase-17


Fibrin, Gelatin


MMP-25, Leukolysin


Gelatin, Fibronectin, Laminin-I

Up regulators of MMP production

  • IL-1β
  • Tumor necrosis factor- α
  • Epidermal growth factor
  • Platelet derived growth factor (PDGF)
  • Transforming growth factor – α (TGF-α)

Down regulators of MMP expression

  • Interferon-γ
  • TNF-β
  • Glucocorticoids

Description about individual MMPs 7:


MMP-1, also known as interstitial collagenase, can cleave collagen Types I, II, III, VII, VIII, and X helices to yield characteristic one-quarter-three-quarter products. MMP-1 is secreted as a latent pro-enzyme (57 kDa) that is subsequently processed to a MMP-1 41/42 kDa active forms. MMP-1 can be activated in vivo by proteinases (e.g., plasmin, trypsin). MMP-3 is also an important activator of MMP-1 in vivo. Although MMP sequences are conserved across species and there is >50% homology among MMP-1, MMP-8, and MMP-13, they can be distinguished from other MMPs by their domain structure and substrate specificity.


MMP-2, also known as the 72 kDa gelatinase/ Type IV collagenase and gelatinase A, cleaves a number of substrates including gelatins, Type I, III, IV, V, VII, and X collagen, fibronectin, elastin, and proteoglycan. It is synthesized as a 72 kDa pro-enzyme that is proteolytically processed to the 66 kDa active form.


MMP-3, also known as stromelysin-1 and transin-1, cleaves a number of substrates including, cartilage proteoglycan, Type II, III, IV, V, IX, X and XI collagen, as well as fibronectin and laminin. It is a major activator of MMP-1. MMP-3 is secreted as 57 and 59 kDa pro-enzymes that can be activated to a 45 kDa active MMP-3.


MMP-7, also known as matrilysin and PUMP-1 cleaves a number of substrates including Type IV and X collagen, elastin, fibronectin, gelatin, laminin, and proteoglycans. It is smallest of all the MMPs, consisting of only a propeptide domain and a catalytic domain. It lacks the hemopexin-like domain common to other members of the MMPs. It is secreted as a 28 kDa pro-enzyme that can be activated by MMP-3 to form a 19 kDa active MMP-7. The active MMP-7 can activate pro-MMP-1 and pro-MMP-9.


MMP-8, also known as collagenase-2 and neutrophil collagenase, preferentially cleaves Type I over Type II and III collagen. It is closely related to the fibroblast collagenases (47% homology). MMP-8 is secreted as an 85 kDa pro-enzyme that can be activated by trypsin, but not by cell bound plasmin, in a two-step process. Both steps are inhibited by TIMP-1 and TIMP-2, which bind to the catalytic domain of the enzyme. The active enzyme is generally unstable at 37°C, breaking down into two fragments of 40 kDa and ~27 kDa. The 40 kDa fragment has been identified as the biologically active catalytic domain.


Matrix metalloproteinase-9 (MMP-9), also known as gelatinase B and 92 kDa gelatinase/Type IV collagenase, exhibits a broad range of substrate specificity for native collagens including Types IV, V, VII, and X as well as gelatin, proteoglycans, and elastin. It is secreted as a 92 kDa pro-enzyme and can be activated by cathepsin G and MMP-3. MMP-9 can be inhibited by TIMP-1 which binds exclusively to pro-MMP-9.


Matrix metalloproteinase-10, also known as stromelysin-2 and transin-2, is a member of the stromelysin subfamily with similar substrate specificities as MMP-3. It is secreted as a 56 kDa pro-enzyme which is activated by proteolytic cleavage to active 47 kDa form. The active MMP-10 can activate pro-MMP-1, pro-MMP-7, pro-MMP-8, and pro-MMP-9. MMP-10 is expressed in a number of tumors, including oral squamous carcinoma, and head and neck squamous cell carcinomas.


MMP-11 or Stromelysin-3 is also known as STMY3, SL-3 and ST-3. Unlike other MMPs, human MMP-11 does not cleave ECM proteins, such as collagen and elastin. MMP-11 has been shown to cleave a variety of non-matrix substrates, including alpha-1-protease inhibitor.


MMP-12, also known as macrophage metalloelastase, is expressed as a 53 kDa pro-enzyme that is processed to a mature ~22 kDA enzyme. MMP-12 is produced initially as an inactive proform that requires proteolytic activation. Main substrates of this MMP are elastin, fibronectin, gelatin and laminin.


Matrix metalloproteinase-13 (MMP-13), also known as collagenase-3, is secreted as a 60 kDa pro-enzyme that is proteolytically processed to the 48 kDa active MMP-13 form. It shows specificity toward the interstitial collagens Type- I, II, III, IV and gelatin. It exhibits greater specificity towards collagen II and cleaves collagen I with comparable efficiency to MMP-1 and MMP-8. MMP-13 can be inhibited by TIMP-1, TIMP-2, and TIMP-3.

MMP-18, MMP-19, MMP-20 and MMP-23

MMP-18, also known as Xenopus collagenase-4 or xCol4, is an interstitial collagenase and is known as a Xenopus enzyme. Sequence comparison of MMP-18 with other MMPs suggests that MMP-18 is not a homolog of any known collagenase. It is highly expressed in amphibian metamorphosis, suggesting a developmental role in the larval tissue degeneration and adult organogenesis.

MMP-19 (RASI) was initially named as MMP-181. This MMP has a wide variety of substrate range including aggrecan, fibronectin, type I gelatin, and basement membrane components such as laminin, nidogen, and type IV collagen.

MMP-20 is also known as enamelysin. By virtue of its broad substrate specificity, MMP-20 has a wide range of targets including amelogenin, aggrecan, dentin, and type V collagen. It must be noted that MMP-20 is expressed almost exclusively by developing teeth, thus playing an important role in tooth development.

MMP-23 is a recently described MMP having gelatinase activity. Its secretion and activation are regulated by a single cleavage at the furin site.

Membrane-Type Matrix Metalloproteinases (MT-MMPs):

MT-MMPs are a unique subclass of MMPs, four of which have transmembrane and cytosolic domains at the C-terminus. The other two MT-MMPs do not have a cytosolic domain and are thought to be GPI (glycosylphosphatidylinositol) anchored to the cell surface. MT-MMPs contain a unique insertion of 11 amino acids between the propeptide and catalytic domains that is cleaved by furin-like enzymes to yield activated metalloproteinases. Activated MT1-, MT2-, and MT3-MMP cleave proMMP-2 and proMMP-13 to their active forms, and exhibit the ability to directly cleave ECM proteins such as collagen Type I and fibronectin.

Inhibition of MMP activity

MMP activity can be controlled at various levels: transcription (by cytokines), proteolytic activation of the zymogen form (via plasmin-dependent or MMP-dependent pathway), and inhibition of the active enzyme 8.

Regulation at transcriptional level:

This task is performed by cytokines. Cytokines are the chemical messengers that affect the surrounding or distant cells by up-regulating or down-regulating the protein synthesis by these cells. As already discussed, the genes for MMP synthesis are activated when…………………………


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Inhibition at the level of proteolytic activation of the zymogen form:

Regulation at the level of proteolytic activation of the zymogen form to an active enzyme makes comparatively a smaller component of the regulatory mechanism of MMPs. The major regulatory mechanisms include regulation of MMP formation at the transcriptional level and the inhibition of the active enzyme.

Inhibition of the active enzyme:


α-Macroglobulins are potent inhibitors of MMP activity. Active MMPs are captured by α-Macroglobulins, by a unique venus-fly-trap mechanism acti­vated by cleavage of a bond in the “bait region.” This cleavage leads to the hydrolysis of a labile internal thiol-ester bond and covalent cross-linking of a nascent glutamyl residue to lysyl side chain exposed on the surface of the attacking proteinase. The rapid capture rates, particularly with collagenases, suggest that α-Macroglobulins, particularly α2- M, play an important role in the regulation of MMP activity in human periodontal diseases 16.

Tissue inhibitors of metalloproteinases (TIMPs):

TIMPs are important regulators of MMP activity. The TIMP family consists of: TIMP-1, TIMP-2, TIMP-3, and TIMP-4. Binding of the TIMPs to their specific MMPs results in the inhibition of MMP activity. TIMPs share a high degree of homology to each other, however, they may be either secreted extracellularly in soluble form (TIMP-1, TIMP-2, and TIMP-4) or bind to extracellular matrix components (TIMP-3). TIMPs have been shown to bind to the pro-enzyme forms of MMP-2 and MMP-9 with a high degree of specificity. This interaction provides an extra level of regulation by preventing activation. TIMP-1 binds to the pro-MMP-9, whereas TIMP-2 binds to the pro-MMP-2.


It is a 184 amino acid glycoprotein of 28.5 kDa, more widely distributed than the other TIMPs and inhibits the activity of all the active MMPs. Its binding to MMP-9 and MMP-1 occurs via a reversible non-covalent binding to the catalytic domain of the MMP protein.


It is a 194 amino acid unglycosylated protein of 21.5 kDa, which has 41% and 44% sequence homology to TIMP-1 and TIMP-3, respectively. It inhibits the activity of all active MMPs and regulates the activation of pro-MMP-2 by binding to its C-terminal region.


It is a 27 kDa glycoprotein, expressed by a variety of cells. It has an affinity for the components of the ECM and as a consequence, it is largely sequestered there. It forms a non-covalent, stoichiometric complex with both latent and active MMPs. TIMP-3 prevents the activation of pro-MMP-2 by MT2-MMP.


The chromosomal location for the gene coding for TIMP-4 is 3p25. It functions in a tissue specific manner in extracellular matrix homeostasis.

Role of MMP’s in periodontal diseases

The microbiological etiology of periodontal diseases is well defined. Lipopolysaccharides (LPS) derived from the bacteria have the capacity to activate junctional epithelial cells to release potent cytokines, such as interleukin-1, interleukin-8, tumor necrosis factor-α, prostaglandins, and proteases. These chemoattractant signals lead to the transmigration of leukocytes and monocytes/ macrophages. Further, these inflammatory cells stimulate the resident ligament cells, such as fibroblasts, macrophages, osteoblasts, keratinocytes, and endothelial cells to produce cytokines and MMPs.

As already stated, under normal physiological conditions, the activities of MMPs are precisely regulated at the level of transcription, activation of the precursor zymogens, interaction with specific ECM components, and inhibition by endogenous inhibitors 17, 18. However, under pathological conditions they play an important role in connective tissue degradation. Birkedal-Hansen et al. (1993) 19 have provided the following evidence to prove the involvement of MMP’s in periodontal diseases,

  • Cells isolated from normal and inflamed gingiva are capable of expressing a wide variety of MMPs in culture,
  • Several MMPs can be detected from the gingiva in vivo and,
  • Matrix MMP-8 and matrix MMP-3 are readily detected, in gingival crevicular fluid (GCF) from gingivitis and periodontitis patients.

Furthermore, the authors proposed following mechanisms responsible for periodontal destruction based on MMPs and TIMPs interactions,

  • Imbalance between MMPs and TIMPs causes irreversible connective tissue breakdown.
  • MMP-8 (Neutrophil collagenase) exists in elevated amounts in the GCF collected from inflamed periodontal pockets and is converted to the active form by the plaque, host and microbial derived proteases.
  • Periodontal ligament collagen fibers attached to the root surface are destroyed by MMPs, which favors the apical migration and lateral extension of the pocket epithelium i.e. attachment loss.

Studies have shown that high concentrations of the natural tissue inhibitor of MMPs (TIMPs) are found in the gingival crevicular fluid of healthy gingiva 20. As we know that Type I collagen is the main ECM component in the soft (gingiva and periodontal ligament) and hard (alveolar bone) periodontal tissues and thus, collagen Type I degradation is regarded as one of the key factors in the uncontrolled destructive lesion 21, 22. The major collagenolytic MMPs associated with the severity of periodontal inflammation and disease are collagenase-2 (MMP-8) and collagenase-3 (MMP-13), whereas collagenase-1 (MMP-1) is related to the physiological periodontal tissue turnover 23, 24. Experiments on MMP-8 deficient mice have confirmed the concept that MMP-8 is a central mediator in chronic infection-induced inflammatory conditions, and can also exert, in addition to the classical surrogate tissue destructive properties, the anti-inflammatory and defensive properties 25-29. MMP-8 is the major type of interstitial collagenase present in human periodontitis-affected gingival tissue, GCF and peri-implant sulcular fluid 30.

Matrix metalloproteinases and bone resorption

Although, the role of MMPs in bone resorption is not completely clear, but present information suggests that they do have an important role in this process. The mineralized bone matrix is covered by an osteoid layer because of which osteoclasts are unable to attach to the bone surface. Now, this osteoid is composed of collagen Type I, proteoglycans, glycoproteins, and native collagen Types IV and IX. Osteoblast-derived collagenase (MMP-13) seems to be mainly responsible for degrading the nonmineralized osteoid layer covering the bone surfaces, exposing the mineralized matrix to the osteoclasts 31-33. Cleavage of collagen Type I by MMP-13 seems to be the initial step of the entire bone resorption process. Subsequent, denatured collagen fragments are also degraded by gelatinases (MMP-2 and MMP-9) 33. Another important MMP causing bone resorption is MMP-9. It cleaves acid insoluble collagen Type I at 37°C and presents strong proteolytic activity against denatured collagen Type I and Type IV 34-36.

Further, research done on chemically modified tetracyclines clearly indicates that MMPs have an important role in periodontal bone resorption 37.


MMPs are important components in many biological and pathological processes because of their ability to degrade ECM components. Considerable advancements have been made in the understanding of biochemical and structural aspects of MMPs, including their activation and catalytic mechanisms, substrate specificity, and the mechanism of inhibition by TIMPs.

The role of MMP’s in the progression of periodontal diseases has been well investigated. They can act as biomarkers for active periodontal disease. Studies have shown that collagenase-2 or MMP-8 can be used as a clinically useful oral fluid (GCF, mouthrinse or saliva) biomarker in periodontal and peri-implant diseases 38, 39.

Modulation of the host response is an important field of research in the treatment of periodontal diseases. Modification of host response via the use of MMP inhibitors, along with the use of bisphosphonates as blockers of periodontal tissue destruction, has shown promises in the treatment of these diseased states. Further research work is required to understand the role of MMPs in periodontal diseases and how it can be controlled or modified to prevent connective tissue destruction.


References available in the hard copy of the website

Periobasics: A textbook of periodontics and implantology

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