Seminar no

Collagen
SHASHI KANT CHAUDHARY
JR
DEPT OF PERIODONTOLOGY & ORAL IMPLANTOLOGY
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Content

History
Introduction
Structure of collagen
Synthesis
Types
Collagen in periodontium
Degradation and Remodeling
Clinical significance
Summary
References 4/6/17
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History

Collagen - Greek word kolla meaning glue and gen denoting

producing

The first evidence that it possesses a regular structure at the molecular

level was presented in the mid-1930s

Since that time, many prominent scholars, including Nobel laureates

Crick, Pauling, Rich and Yonath, and others, including Brodsky, Berman,
and Ramachandran, concentrated on the conformation of the collagen
monomer

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Introduction

Tissues are not made up solely of cells.

A substantial part of their volume is

extracellular space which is largely filled
by an intricate network of
macromolecules constituting the
extracellular matrix (ECM)

Often referred to as "connective

tissue."

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The ECM contains three major classes of biomolecules:

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Collagen

The major structural protein found in connective tissue.

Most abundant about 25- 30% of the total weight of protein

in the body is collagen.

It serves to hold together the cells in the tissues.

It gives strength support and shape to the tissues

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At least 28 distinct types of collagen made up of over 30 distinct

polypeptide chains have been identified in human tissues

In addition, a number of proteins (eg the C1q component of the

complement system, pulmonary surfactant proteins SPA and SPD)that
are not classified as collagens have collagen-like domains in their
structures

These proteins are sometimes referred to as "noncollagen collagens.

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Structure of collagen

Astbury was the first to suggest a structure for collagen in 1938, which
consisted of a mixture of Trans and Cis peptide units

The same feature was incorporated by Pauling and Corey in the model
proposed by them in 1951 that had three coaxial helices.

However, neither of these structures was in agreement with the observed

Xray diffraction pattern of collagen fibers.

It was Ramachandrans group from Madras, India, who first postulated a

triple helical structure for collagen.

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Structure

The collagen is a rod like structure

The tropocollagen, has a molecular weight of 285 kDa.

The tropocollagen is made up of three polypeptide alpha chains.

About 30 genes are responsible for collagen synthesis, and the

enzymes necessary for collagen synthesis.

Each polypeptide chain of collagen has about 1000 amino acid

residues.

The amino acid composition of collagen is quite unique

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 The repetitive amino acid sequence may be represented as 10
Gly - X - Y- Gly -X - Y - Gly - X - Y- Gly -X - Y -

Every third residue is glycine, where X is frequently proline and Y is often

hydroxyproline (but can be hydroxylysine).

Thus, most of the chain can be regarded as a polytripeptide whose

sequence can be represented as (GlyProHyp).

Proline helps in the formation of helical orientation of each alpha chain

Glycine, the smallest amino acid, It fits into the restricted spaces where
the three chains of the helix come together.

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 Three polypeptide alpha chains coiled around each other 11
to form the typical collagen triple-helix configuration

The three polypeptide chains are held together by

hydrogen bonds between the chains

Each polypeptide subunit or alpha chain is twisted into a

left-handed polyproline helix of three residues per turn

Three of these alpha chains are then wound into a right-

handed superhelix, forming a rodlike molecule 1.4 nm in
diameter and about 300 nm long.

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In type I collagen there are 338 [GlyX Y]

triplets repeated in a sequence per chain
accounting for 95% of the total structure in
the form of uninterrupted triple helix.
However the N- &C- termini of the chains
do not have the [Gly-X-Y] motif.
This produces a non helical conformation at
the ends of the molecule. These regions are
referred to as TELOPEPETIDES .
the telopeptides are particularly prone to
proteolytic attack
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 Band Spacing
D=670
FIBRIL
13
Overlap Zone (0.4D) Hole Zone (0.6D)

MICROFIBRIL

3000 (4.4D)
COLLAGEN
MOLECULE 15 Dia

104

TRIPLE
HELIX

PRIMARY Glycine 8.7 Glycine

STRUCTURE
IN -CHAIN
Y X Y X 4/6/17
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Quarter-Staggered Arrangement

The tropocollagen molecules are arranged in a 'quarter

staggered array' to form the collagen fibers (Molecules in each
row separated by 400 and adjacent rows by 680 ).

The structure repeats after each row.

This arrangement is responsible for the banded appearance of

these fibers in connective tissues

Responsible for its tensile strength.

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Quarter-staggered arrangement in collagen fiber; each

row moves one-fourth length over the last row; the 5th
row repeats the position of the first row

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Synthesis of collagen

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Sites for synthesis of collagen

Mesenchymal Cells & Other Cells

Fibroblast (major
Their Derivatives Epithelial cells
cells)
Chondroblasts Endothelial cells

Osteoblasts Muscle cells

Odontoblasts Schwann cells

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Cementoblasts
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Synthesis of collagen

Formation of pro- chains

Hydroxylation
Intracellul
Glycosylation ar
Assembly and secretion

Extracellular cleavage of procollagen

Formation of collagen fibrils Extracellular
Cross-link formation
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Gene expression
Type Genes Tissue
I COL1A1, COL1A2 Most connective
tissues, including bone

II COL2A1 Cartilage, vitreous

humor
III COL3A1 Extensible connective
tissues such as skin,
lung, and the vascular
system
IV COL4A1COL4A6 Basement membranes
V COL5A1COL5A3 Minor component in
tissues containing
collagen I
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VI COL6A1COL6A3 Most connective
tissues 26

VII COL7A1 Anchoring fibrils

VIII COL8A1COL8A2 Endothelium
IX COL9A1COL9A3 Tissues containing
collagen II
X COL10A1 Hypertrophic
cartilage
XI COL11A1, Tissues containing
COL11A2, COL2A1 collagen II
XII COL12A1 Tissues containing
collagen I
XIII COL13A1 Many tissues
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XIV COL14A1 Tissues containing 27
collagen I

XV COL15A1 Many tissues

XVI COL16A1 Many tissues

XVII COL17A1 Skin hemidesmosomes

XVIII COL18A1 Many tissues (eg, liver,

kidney)

XIX COL19A1 Rhabdomyosarcoma

cells

Source: Adapted slightly from Prockop DJ, Kivirrikko KI: Collagens:

molecular biology, diseases, and potentials for
therapy. Annu Rev Biochem 1995;64:403. Copyright 1995 by Annual
Reviews, www.annualreviews.org. 4/6/17
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Types

Based on their location and functions in the body -

LIPPINCOTT

Fibril-forming collagens: Types I, II, and III

Network-forming collagens: Types IV and VII

Fibril-associated collagens: Types IX and XII

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Based Primarily on the Structures That They Form -

HARPER
Class Type
Fibril-forming I, II, III, V, and XI
Network-like IV, VIII, X
FACITs IX, XII, XIV, XVI, XIX
Beaded filaments VI
Anchoring fibrils VII
Transmembrane domain XIII, XVII
Others XV, XVIII 4/6/17
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Based on their supramolecular assemblies collagen

superfamily is subdivided into the nine subfamilies -
TENCATE
Subfamilies Type
Fibrillar collagens I, II, III, V, XI, XXIV, and XXVII
Basal lamina collagens IV
FACIT collagen IX, XII, XIV, XVI, XIX, XX, XXI, and
XXII
Network forming collagens VIII and X
Anchoring fibril collagens VII
Microfibril forming collagen VI
Transmembrane collagen XIII, XVII, XXIII, and XXV
Multiplexin collagen XV and XVIII
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Other collagen XXVI and XXVIII
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Fibrillar collagens

Types I, II, III, V, XI, XXIV, and XXVII

Aggregate in a highly organized manner in the extracellular

compartment to form fibrils with a typical 64-nm banding pattern.

Type I collagen is the most abundant in most connective collagen

tissues.

Collagen fibrils often are composed of more than one type of collagen.

Type V Collagen is believed to regulate fibril diameter.

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Basal lamina collagen

Collagen type IV is similar in size to type I but does not

assemble as fibrils.

It contains frequent nonhelical sequences and aggregates in a

sheetlike, chicken-wire configuration.

Type IV collagen is a major component of the basal lamina and is

a product of epithelial cells.

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Fibril-associated collagens with
interrupted triple helices (FACIT)

Collagens IX, XII, XIV, XVI, XIX, XX, XXI, and XXII consist of
chains that have different lengths and contain a variety of
noncollagenous domains.

They exhibit several interruptions in the triple helix and are

found in various locations in different tissues.

Several of the FACIT collagens associate with fibrillar collagens

and other extracellular matrix components.

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Network-forming collagens

Type VIII and X collagen

Type VIII collagen assembles into a hexagonal lattice, which is

believed to impart compressive strength while providing an
open, porous meshwork.

Type X collagen has a similar size and structure and is largely

restricted to the hypertrophic zone of the epiphyseal cartilage
growth plate.

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Anchoring-fibril collagen

Collagen VII has unusually large nonhelical ends making

up two thirds of the size of the molecule.

The C-terminal ends associate to form dimers that

subsequently are assembled into the anchoring fibrils
that extend from the basal lamina into the underlying
connective tissue

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Microfibril-forming collagen

Type VI collagen, which has large N- and C-terminal

globular domains that associate in an end-to-end
fashion, forms beaded filaments.

Present in most connective tissues.

Has binding properties for cells, proteoglycans, and

type I collagen and matrix

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Transmembrane collagen

Types XIII, XVII, XXIII, and XXV

These collagens are transmembrane proteins with extracellular collagenous domains and a
C-terminal noncollagenous domain that functions in cell adhesion.

Type XVII collagen is found in hemidesmosomes of basal epidermal cells and attaches the
cells to the basal lamina.

Type XIII collagen is present in focal adhesion sites of fibroblasts and at cell-matrix interfaces
in some epithelia, muscle, and nerves, also present in the cell-cell adhesive specializations.

These collagens may interact with other cell surface or extracellular matrix molecules to
alter cell behavior.
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Multiplexin (endostatin-forming)
collagens

Type XVIII collagen is a component of basal lamina of epithelial and endothelial cells
and stabilize structures of the basal lamina.

Type XVIII collagen has multiple interruptions in the central helical domain and a
large, unique C-terminal nonhelical domain.

This C-terminal domain can be cleaved by extracellular proteases to form endostatin,

a potent inhibitor of endothelial cell migration and angiogenesis..

Type XV collagen has a similar structure and a wider distribution, including the
papillary dermis. However has less potent antiangiogenic activity than type XVIII.
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Other collagens

There are other collagens and proteins containing helical collagenous

domains that cannot be classified into other category.

Type XXVI is found in the extracellular matrix of the testis and ovary

The structure of type XXVIII has some similarities with type IV but the
triple helical domain is longer

Type XXVIII is predominantly expressed in the basement membranes

around Schwann cells of the peripheral nervous system and dorsal root
ganglia.
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Collagen in periodontium and

tooth structures

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DENTIN COLLAGEN

The organic matrix of dentin contains collagen , noncollagenous proteins (proteoglycans,

phosphophoryns, and glycoproteins), phospholipids, and growth factors.

Type I collagen is the major protein of dentin matrix. Lesser amounts of types III, V, and
VI collagen are also found in dentine and predentine

Demineralized dentine and predentine show closely packed collagen fibers of 20-50nm

Dentinal collagen contains 2-3 fold increase of hydroxylysine compared to that of soft
tissues

Dentinal collagen is relatively insoluble in acid and neutral solutions

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PULPAL COLLAGEN

The dental pulp is a connective tissue

In its developmental stage, it contain sparsely distributed, fine

collagen fibrils (types I and 111).

Initially rich in type III, as matures type I collagen bundles are formed

Approximately 34% dry weight of pulp is collagen

After a small increase in collagen synthesis which occurs at the time

of eruption and root closure, there is no change in collagen content
of the tooth for rest of the life
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Gingival collagen

Gingival connective tissue contains collagen types I, III, IV,

V, VI, and VII.

Types I and III form the major collagen fibers , occupy

approximately 60% of the extracellular space.

The connective tissue of the marginal gingiva is densely

collagenous, and it contains a prominent system of
collagen fiber bundles called the gingival fibers.

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Collagen Location
I Lamina propria
III Lamina propria
IV Basement membrane
V Subepithelial basement
membrane
VI Microfibrils
VII Subepithelial basement
membrane
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PERIODONTAL COLLAGEN

Collagen is the most abundant protein of ECM of the periodontal

ligament

Type I collagen accounts for approximately 80% of the total collagen

content major component of the principal fibers.

Type III collagen, accounting for about 15% is preferentially localized

in reticular fibers located around blood vessels and peripheral nerves.

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Type IV collagen forms the major

fraction of basal lamina protein of the
blood vessels and nerves in the PDL.

Collagen types V and VI form a minor

fraction of the PDL collagen.

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The most important elements of the

periodontal ligament are the
principal fibers, which are
collagenous and arranged in bundles

When viewed in longitudinal section

follow a wavy course

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CEMENTAL COLLAGEN

Consist of a calcified interfibrillar matrix and collagen fibrils.

The two main sources of collagen fibers

Sharpey fibers (extrinsic)- fibroblast and

fibers that belong to the cementum matrix (intrinsic)-

cementoblast

The major proportion is type I (90%) and type III (about 5%)
collagens
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SHARPEYS FIBERS

The terminal ends of the collagenous principal

fibers are inserted in to bones to form Sharpeys
fibers

At their insertion the collagen bundles of PDL are

embedded into cementum and alveolar bone in
a manner similar to tendon inserting in to the
bone

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They usually tend to be concentrated in the crestal

region

Sharpey fibers are composed of mainly type I collagen

Type III collagen appears to coat the type I collagen of

the Sharpey fibers

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BONE COLLAGEN

Bone consists of two thirds inorganic matter and one third

organic matrix.

The organic matrix consists mainly of collagen type I (90%)and

type V (5%)

Alveolar bone consists type I,V,III and XII

Initial mineral depostis in the organic matrix of bone appears to

occur at discrete sites, in or on, the collagen fibrils

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Functional adaptation of
collagen

The periodontal ligament is very important to the maintenance of the

periodontal apparatus

This is proved by the atrophy of the alveolar bone that often follows
the removal of the teeth and associated PDL

Under the scanning electron microscope it is revealed that the point

where the fibers insert obliquely , the bone surface exhibits a stepped
appearance that indicates mineralization occurs approximately at right
angles to the axis of the fiber.
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These mineralized points provide mechanical advantages for transmitting

axially directed forces

The mineral interface also appears concave and this maximum strength to
the mineral-collagen interface

In case of periodontal diseases there is destruction of PDL , sharpeys

fibers

Due to this teeth is not able to withstand the varying types of mechanical
forces that ultimately leads to tooth mobility
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Age changes in collagen

As age advances qualitative and quantitative changes are seen

Higher conversion of soluble to insoluble collagen

Increased mechanical strength

More organized and thicker

Areas of hyalinization

Increased denaturing temperature

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Apparent decrease in the number of collagen & PDL fibers due to

Increased Fibrosis and decreased cellularity

Gradual decrease in the number of synthesizing CT cells

A gradual recession of alveolar bone

Irregular Alveolar Bone & cementum surface

These changes are due to higher cross linking and stabilized forms of
collagen

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COLLAGEN DEGRADATION 57
EXTRACELLULAR

Break down of the collagen matrix element is a key component of any normal tissue
that is undergoing morphogenesis and growth

But it is vital that this process is kept under rigid control

Collagen degradation and synthesis should be in equilibrium

Although several enzymes are involved in the destruction of matrix components

collagen breakdown is mediated primarily by the COLLAGENASES ( Type of MMP)

These are specialized enzymes that have evolved specifically to hydrolyze collagens,
because their triple helical collagen structure is resistant to most common proteinases

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MECHANISM OF DEGRADATION

The collagenases belong to a family of enzymes called Matrix Metallo

Proteinase(MMPs) that consists of at least 13 members with closely related
domain structures and discrete functions

MMPs are usually secreted by the connective tissue cells (predominantly

fibroblasts ) but are also produced by some leucocytes ( PMNs , Macrophages )

Based on their substrate specificity they have been classified as COLLAGENASES

, GELATINASES , STROMELYSINS , MATRILYSINS

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Collagenase-1 (Also called MMP-1) or fibroblasts type
collagenase

This can hydrolyses type I II III IV VIII X collagens and

gelatins

They cleave 1(I) and (I)2 chains at glycine-leucine bonds

Collagenase -2 (MMP-8) or PMN type collagenase

This can hydrolyses type I and type III

Only found in the granules of neutrophils.

Gelatinase Gelatinase-A (MMP-2) Gelatinase-B(MMP-9)

They degrade gelatin and type IV VII X XI collagen and 4/6/17

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Stromelysins

Stromelysins-1(MMP-3) Stromelysins -2(MMP-10)

Stromelysins -3

They degrade proteoglycans, basement membrane,

laminin and fibronectin and collagens

Matrilysin(MMP-7)

Metalloelastase(MMP-11)

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Regulation of MMP

Two key regulators are IL-1 and TGF-

Inhibitors of MMP are 2- macroglobulin and TIMP

(TIMP )Tissue inhibitor of MMPs are group of inhibitors that are

distributed in the body tissues and fluids.

TIMP is highly conserved glycoprotein that forms irreversible

complexes with MMPs via non covalent interactions. Its usually is
secreted by Fibroblasts and macrophages
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COLLAGEN DEGRADATION
INTRACELLULAR
Most important mechanism for the physiologic turnover and
remodeling of collagenous connective tissues

Done by fibroblasts Process includes:

A. Ingestion of extracellular collagen fibrils

B. Formation of phagosome

C. Initial fusion of lysosomes with collagen-containing phagosome

D. Advanced stages of collagen degradation in dense lysosomal

structures

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Clinical significance

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Collagen disorders

About 30 genes encode the collagens

Pathway of biosynthesis is complex, involving at least eight

enzyme-catalyzed posttranslational steps.

Due to mutations in collagen genes or in genes

encoding some of the enzymes involved in these
posttranslational modifications. The diseases affecting bone
(eg, osteogenesis imperfecta) and cartilage (eg, the
chondrodysplasias) 4/6/17
Gene Diseases
66
COL1A1, COL1A2 Osteogenesis imperfecta, type 1
Osteoporosis
Ehlers-Danlos syndrome type VII
autosomal dominant

COL2A1 Severe chondrodysplasias

Osteoarthritis

COL3A1 Ehlers-Danlos syndrome type IV

COL4A3COL4A6 Alport syndrome (including both
autosomal and X-linked forms)

COL7A1 Epidermolysis bullosa, dystrophic

COL10A1 Schmid metaphysial
chondrodysplasia
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Enzyme Disease
Lysyl hydroxylase Ehlers-Danlos syndrome type
VI
Procollagen N -proteinase Ehlers-Danlos syndrome type
VII autosomal recessive
Lysyl hydroxylase Menkes disease

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Scurvy vitamin C deficiency

Scurvy affects the structure of collagen.

Due to a deficiency of ascorbic acid not a genetic

disease.

Impaired synthesis of collagen due to deficiencies of

prolyl and lysyl hydroxylases, both of which require
ascorbic acid as a cofactor.
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Complete hydroxylation of Proline has a Tm of 39*C. However

underhydroxylated molecules melt at lower temperatures i.e the Tm is
around 25*C.

Therefore complete hydroxylation of proline and lysine is essential for

collagen thermal stability at physiologic temperature.

While under hydroxylated molecules are denatured and degraded

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Collagen as membrane

Collagen has shown to be particularly suitable for GTR applications

because it is chemotactic for periodontal ligament fibroblasts,

Acting as a barrier for migrating epithelial cells

Providing hemostasis,

Serving as a fibrillar scaffold for early vascular and tissue ingrowth

Membrane is semi-occlusive (effective pore size 0.004 pm)

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They are obtained from type I or type III collagen or both.

Some commonly available membranes are: BioMend

(Zimmer Dental,), Bio-Gide(Geistlich Pharma), OraMem
(Salvin, ), RCM6 Resorbable Collagen Membrane and
conFORM (ACE Surgical,), and Periogen (Collagen Corp.) -

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Hemostatic Collagen

Used for control of bleeding

CollaPlug, CollaCote, and CollaTape (Zimmer Dental) and Helistat

Soft , white, pliable, nonfriable, coherent, sponge-like structures

Fabricated from bovine collagen

Indications are for wound protection and for control of oozing or

bleeding from clean oralwounds

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Collagen as Bone Substitutes

Used as implantable carriers for bone-inducing proteins such as

bone morphogenetic protein-2 (rhBMP-2)

Demineralized bone collagen is used as a bone graft material for

the treatment of acquired and congenital defects either by itself
or in combination with hydroxyapatite

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Collagen for Local Drug Delivery
System

A degradable controlled-release device based on formaldehyde cross-linked

bycoprotein matrix containing chlorhexidine was described by Steinberg et al
1990.

PerioCol(Eucare Pharmaceuticals Private Limited,) is the most commonly used

collagenous membrane for local drug delivery system and has two contents:
chlorhexidine and collagen

Another material commonly used for local drug delivery is Periodontal Plus AB
(EnColl). This product comprises tetracycline fibers impregnated in collagen
fibers, which are available in vials. These brownish-colored fibers are resorbable

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Summary

All collagencontain a repeating Gly-Pro-X sequence and fold into a characteristic

triple-helical structure.

The various collagens are distinguished by the ability of their helical and nonhelical
regions to associate into fibrils, to form sheets, or to cross-link differentcollagen
types.

Mostcollagenis fibrillar and composed of type I molecules. A two-dimensional

network of type IV collagen is unique to thebasal lamina.

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Fibrous typecollagenmolecules (e.g., types I, II, and III) assemble into

fibrils that are stabilized by covalent aldol cross-links .

Procollagen chains are modified and assembled into a triple helix in

theER. Helix formation is aided by disulfide bonds between N- and C-
terminal propeptides, which align thepolypeptidechains in register.
Generally, the propeptides are removed after secretion, and then
collagenfibrils form in the extracellular space

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References

Oral Cells And Tissues , Garant P.R , Quintessence Publishing Co,inc ,2003.

Ten Cate`s Oral Histology ; Nanci A Et Al ;7th Edition ; Mosby ; 2008.

Carranza`s Clinical Periodontology; Newman M G.,Takei H H; Klokkevold P R 10th

Edition;saunder Elsevier;2006.

Orban`s Oral Histology And Embryology ,Kumar Gs , 12th Edition , 2008, Elsevier Pvt.

Ltd

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Harper's Illustrated Biochemistry, Robert K. Murray, David A Bender, Kathleen M. Botham ,

Peter J. Kennelly, Victor W. Rodwell , P. Anthony Weil, 28edition Lange

Lippincotts Illustrated Reviews: Biochemistry Richard A. Harvey, Denise R. Ferrier, Fifth Edition

2011 Lippincott Williams & Wilkins, A Wolters Kluwer Business

Textbook Of Biochemistry Dm Vasudevan, Sreekumari S, Kannan Vaidyanathan Sixth Edition

2011 Jaypee Brothers Medical Publishers (P) Ltd

Regeneration in Periodontics: CollagenA Review of Its Properties and Applications
in Dentistry Lanka Mahesh, BDS, MBA; Gregori M. Kurtzman, DDS, DICOI, DADIA;
and Sagrika Shukla, MDS

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Thank you

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