CONNECTIVE TISSUE

NICANOR B. LACUESTA JR., MD, DPBO-HNS

• Provides and maintains the form of the body
• Major constituent is extracellular matrix –
protein fibers, ground substance, tissue fluid
– Tissue fluid:
• Simmilar to blood plasma in ion contents and diffusible
substances
• Contains a small percentage of plasma proteins of low
molecular weight
• Insignificant quantity under normal circumstances
 Function
• Structural
– Encapsulate and provides internal structure to
organs
– Tendons, ligaments, and areolar tissue around
organs
– Bone and cartilage – specialized types
• Defense:
– Due to content of phagocytic and
immunocompetent cells
– Cell that produce intermediates of inflammation
– A physical barrier – prevents dispersion of bacteria
in the body
• Nutrition:
– Due to the close relation with blood vessels
– Serves a medium for the exchange of wastes and
nutrients between blood vessels and cells
 origin
• Mesoderm
• Mesodermal cells migrate and surround and
invade developing cells to become
mesenchymal cells
– oval nucleus, prominent nucleoli, fine chromatin,
sparse cytolplasm –extends as thin processes
away from the nucleus
– Point of origin of all connective tissue cells, and
blood and blood vessels
 3 Classes of components

1. Ground Substance
2. Fibers
3. Cells
 Ground Substance
• Viscous, colorless, transparent, and
homogenous
• Fills space in between cells and fibers
• Acts as a lubricant
• Barrier to foreign particles
• Difficult to examine in fresh and fixed samples
due to high water content
• 3 classes of components:
–Glycosaminoglycans

–proteoglycans

–Structural proteins
 Glycosaminoglycans
• Linear polysaccharides
• Repeating disaccharide units - uronic and a
hexosamine
– Hexosamine: glucoseamine or galactosamine
– Uronic : glucoronic or iduronic acid
• largest, almost unique, most ubiquitous -
heparan sulfate (100s-1000s kDa)
• Intensely hydrophilic and act as polyanions
(hydoxyl, carboxyl, and sulfate groups)

• Binds a great number of cations (sodium),

intensely hydrated structures
 Glycosaminoglycans Repeating dissacharide Distribution Electrostatic interaction with
Hexuronic acid Hexosamine collagen

Hyaluronic acid D-Glucoronic acid D-Glucosamine Umbilical cord, synovial fluid,

vitreous humor, cartilage

Chondroitin 4-sulfate D-glucoronic acid D-Galactosamine Cartilage, bone, cornea, skin, High levels of interaction, mainly
notochord, aorta with collagen type II

Chondroitin 6- sulfate D-Glucoronic acid D-Galactosamine Cartilage, umbilical cord, skin, aorta High levels of interaction, mainly
(media) with collagen type II

Dermatan Sulfate L-iduronic acid or D- D-Galactosamine Skin, tendon, aorta (adventitia) Low levels of interaction, mainly
Glucoronic acid with collagen type I

Heparan Sulfate D-Glucoronic Acid or D- Galactosamine Aorta, lung, l iver, basal laminae Intermediate levels of interaction,
L-iduronic acid mainly with collagen types III and IV

Keratan Sulfate (cornea) D-Galactose D-Galactosamine Cornea

Keratan Sulfate D-Galactose D-Glucosamine Cartilage, nucleus pulposus, annulu

(skeleton)
 Proteoglycans
• composed of a protein core to which various
numbers and combinations of GAGs are
covalently attached

• carbohydrate portion constitutes 80-90% of

the weight
• Proteoglycans bind to collagen owing to
electrostatic interaction between their acid
groups and the basic amino acid residues of
collagen
• Synthesis of protein moiety begins in the RER
• Glycosylation initiated in the RER and
completed in the Golgi complex where
sulfation also occurs
 Multiladhesive Glycoproteins
• have multiple binding sites for cell surface
receptors (integrins) and other matrix
macromolecules
• very large molecules with branched
oligosaccharide chains
• important roles for the adhesion of cells to
their substrates
• Fibronectin
– (MW 235 - 270 kDa), dimeric
– Synthesized by fibroblasts and epithelial cells
– Has binding sites for cells, collagen, and
glycosaminoglycans
– Helps mediate normal cell adhesion and migration
• Laminin
– large (200-400 kDa), trimeric, cross-shaped
glycoprotein
– Detected in the basal laminae
– Partially responsible for the adhesion of epithelial
cells to basal lamina
• integrin
– act as matrix receptors for specifc sequences on
laminin, fbronectin, some collagens, and certain
other ECM proteins
– heterodimers of two transmembrane
polypeptides: the α and β chain
 3 Classes of components

• Ground substance
• Fibers
• Cells
 Fibers
3 main types
1. Collagen fibers
2. Reticular fibers
3. Elastic fibers

– Distributed unevenly throughout the body

– Predominant type determine the property
of connective tissue
 Collagen
• Most abundant protein in the human body
(30% dry weight)
• 28 types have been described but the most
important, best studied, and most common
types are types I, II, III, IV, V, VII, IX, XII, XIV
 Categories of Collagen
• Fibrillar Collagen (types I, II, III)
– have subunits that aggregate to form large fibrils
clearly visible in the electron or light microscope
– Collagen type I, the most abundant and widely
distributed collagen, forms large, eosinophilic
bundles usually called collagen fibers.
– densely fll the connective tissue, forming
structures such as tendons, organ capsules, and
dermis
• Sheet-forming collagens (type IV)
– subunits produced by epithelial cells and are the
major structural proteins of external laminae and
the basal lamina in all epithelia

• Linking/anchoring collagens (type VII)

– short collagens that link fbrillar collagens to one
another (forming larger fbers) and to other
components of the ECM
• Synthesis of collagen is very widespread
among many cells though originally thought to
be restricted to fibroblasts, chondroblasts,
osteoblasts, and odontoblasts
• Principal amino acids composing collagen
– Glycine (33.5%)
– Proline (12%)
– Hydroxyproline (10%)
• Amount of collagen in a tissue can be
determined by measuring the hydroxyproline
content
• 2 amino acids characteristic of collagen
protein: hydroxyproline and hydroxylysine
– Not incorporated in the protein itself but results
from the hydroxylation of the proline and lysine in
the nascent collagen polypeptide in the RER
during synthesis
 Structure of Collagen
• TROPOCOLLAGEN
– Protein unit that polymerizes to form collagen
fibrils
– Elongated (280 nm in length and 1.5 nm in width)
– Consists of 3 unit polypeptide chains intertwined
in a triple helix
– Difference of chemical structures of these
polypepetide chains account for the different
types of collagen
• Tropocollagen molecules aggregate into
microfibrillar subunits packed together to
form FIBRILS
• Aggregation and packing is achieved by
hydrogen and hydrophobic interactions
• Structure is reinforced by covalent cross-links,
catalyzed by lysyl oxidase
• FIBRILS – thin and elongated structures
– Variable diameter (20 to 90 nm)
– Traverse striations (periodicity of 64 nm)
• Determined by the overlapping arrangement of subunit
tropocollagen mollecules.
• Dark band retain more stain due to more free chemical
groups
• In types I and III – fibrils associate to form
fibers
• Type I – fibers can associate to form bundles
• Type II – can occur as fibrils does not form fibers
• Type IV – does not form fibrils or fibers but occur as
unpolymerized or sparsely polymerized procollagen
molecules
• Type I, II, and III – (form fibrils) are called
interstitial collagens
• type IV and V – do not form fibrils
 Synthesis
1. Polypeptide alpha chains are assembled on
the polyribosomesin RER and injected into
the cisternae
2. Hydroxylation of proline and lysine occurs
– Hydroxylation begins after the peptide chain has
reached a certain minimum length and is still
bound to the ribosomes
– Enzymes:peptidyl proline hydroxylase and
peptidyl lysine hydroxylase
3. Glycosylation of hydroxylysine
– Different collagen – different amounts of
carbohydrate ( galactose or glycosylgalactose
linked to hydroxylysine)
4. Alpha chain is synthesized with registration
peptides (NH- and COOH- terminal ends)
assemble to form the procollagen which is
transported into the extracellular
environment
– Registration peptides ensure proper positioning
of the alpha chains, makes the procollagen
soluble and prevents premature assembly and
precipitation in the cell
5. Registration peptides are removed by
procollagen peptidases to turn procollagen
into tropocollagen
6. Fibrils aggregate to spontaneously form
fibers
7. Fibrilllar structures are reinforced by covalent
crosslinks , catalyzed by lysl oxidases
• Degradation is acheved by collagenases
 Collagen fibers
• Most abundant fibers in connective tissues
• Causes tissue to become white
• Bifringent
• Inelastic and have a tensile strength greater than
steel
• Consist of closely packed thick fibrils (75 nm
thick)
• Can be organized in parallel array forming
bundles
• Better studied in spread preparations and
appear as elongated and tortous cylindric
structures – indefinite length; diameter of 1-
20 µm
• Acidophilic
– Pink – eosin
– Blue – Mallory’s trichrome stain
– Green – Masson’s trichrome stain
– Red - Sirrius red
 Reticular fibers
• Extremely thin (0.5 to 2 µm)
• Extensive network in some organs
• Non visible in H&E preparations
• Stained black by silver salts (agyrophilic)
• PAS positive
– Staining due to high content of glycoproteins
• Have 6-12% hexoses compared to 1% in
collagen
• Composed mainly of type III collagen
• Formed by loosely packed, thin (45nm) fibrils
bound together by small interfibrillar bridges
(proteoglycans and glycoproteins)
• Weak bifringency
• Abundant in smooth muscle, endoneurium
and the framework of hematopoeitic cells
• Constitute a network around cells of
parenchymal organs
• Abundance of reticular fibers in
embryogenesis, inflammatory processes,
wound healing but is replaced eventually by
regular collagen fibers
• Create a flexible network in organs subjected
to increase volume
 Elastic fibers
• Fibers branch and unite with one another into
an irregular network
• Characteristic yellow color in fresh samples
• Capable of stretching to 150% of their length
and yields very easily to traction but readily
returns into original shape when relaxed
• Methods to demonstrate
– Resorcin-fuschin: purple
– Aldehyde-fuschin: black
– Orcein:dark blue
• 2 components:
– Amorphous central region containing elastin
– Sheath of 14 nm tubular microfibrils
• Development:
– Microfibrils appear first
– Amorphous material fills the space in tube
• Elastin
– Secreted as proelastin( MW 70,000) which
polymerizes to form elastin
– Produced by fibroblasts in skin, tendon, smooth
muscle
– Resistant to boiling, extraction with dilute acids
and alkali and not digested by trypsin
– Has a tertiary and quaternary structure, stabilized
by hydrophobic interactions between the non
polar peptide chains
• Amino acid composition resembles collagen
but has greater quantity of valine and alanine
• Two unusual amino acids: desmosine and
isodesmosine, (formed by covalent reactions
among 4 lysine residues) thus crosslinks
elastin and is thought to give its elastic
properties
• Occurs in a nonfibrillar form as fenestrated
membranes in some walls of blood vessels
 3 Classes of components

• Ground substance
• Fibers
• Cells
 Cells
FIBROBLASTS
• Most commonly found cell
• Synthesizes fibers and amorphous intercellular
substrate
• 2 different morphologic types
– Younger, more actively synthesizing cell
(fibroblast)
• Abundant and irregularly shaped cytoplasm; nucleus is
ovoid, large and pale staining, with fine chromatin,
prominent nucleolus; cytoplasm rich in RER, well
developed Golgi complex
– Mature, queiscent (fibrocyte)
• Found in the already formed tissue
• Smaller cell, spindle shaped, fewer processes; nucleus
is smaller, darker; acidophilic cytoplasm; less well
developed RER and Golgi complex
• When actively stimulated, mature cells may
revert to younger, active form (i.e. wound
healing)
• Synthesize collagen reticular and elastic fibers,
glycosaminoglycans, and glycoproteins
• Fibroblasts rarely undergo division
• Mitoses are only observed when the organism
requires additional fibroblasts (tissue damage)
 Macrophages
• Phagocytic capacity
• Derive mainly from precursor cells that
produce monocytes – transported where they
mature into macrophages
• Can proliferate locally to produce more cells
• Ingestion of particles and their digestion by
the lysosome
 Mast Cells
• Oval to round connective tissue cell (20-30
µm)
• Cytoplasm filled with basophilic granules
• Nucleus is small spherical and centrally
situated
• metachromatic
• Release leukotrienes
• 2 populations:
– connective tissue mast cells: proteoglycan is
heparin
– mucosal mast cells: proteoglycan is chondroitin
sulfate
• Contain specific receptor for IgE,
 Plasma cells
• Large ovoid cells, with a basophilic cytoplasm,
rich RER
• Nucleus is spherical and eccentrically placed
with compact coarse hetrochromatin
• Responsible for the synthesis of antibodies
 Adipose Cells
• Specialized for storage of neutral fats or heat
production

Leukocytes
 Types of connective tissues
Connective tissue proper
• Loose connective tissue:
– Areolar tissue
– More abundant type
– Fills space between fibers and muscle sheaths
– Supports epithelial tissue
– Composed of all mian compoonents of connective
tissue proper, most abundant cell is the fibroblast
and macrophages
Dense connective tissue
• Same component as loose but with a clear
predominance of collagen fibers
• Fewer cells – fibroblasts most common
• Less flexible and resistant to stress
• Can be
– Dense irregular – without definite orientation
– Dense regular tissue – 3D network and provide
resistance to stress from all directions (tendons)
Elastic tissue
• Composed of thick, parallel elastic fibers
• With a surrounding small amount of loose
connective tissue, flattened fibrocytes
• Occurs in the yellow ligament of the vertebral
column and in the suspensory ligament of the
penis
Reticular tissue
• Specialized loose connective tissue variation,
provides the architechtural framework of the
myeloid and lymphoid hemopoietic cells
• Reticular cells – fibroblasts specialized in the
production of reticular fiber constituents
Mucous tissue
• Abundance of amourphous ground substance
composed chiefly of hyaluronic acid
• Jelly like tissue containing collagen fibers and
few elastic or reticular fibers
• Mainly fibroblasts
• Principal component of the umbilical cord,
also found in the pulp of young teeth