Connective Tissue

Dr. Jan Tambayong, PHK

Histologi FK-UPH
Connective Tissue
Connective tissue is formed by 3 classes
of components:
1. cells
2. fibers
3. ground substance

The major constituent of connective tissue
is the extracellular matrix.
Extracellular Matrix
Consist of different combinations of protein
fibers (collagen, reticular, elastic) and ground
substance.
Ground substance is a hydrophilic complex of
anionic macromolecules (glycosaminoglycans
and proteoglycans), and multiadhesive
glycoproteins (laminin, fibronectin, ect) that
imparts strength and rigidity to the matrix by
binding to receptor proteins (integrins) on the
surface of cells
Cells of the connective tissue
Fibroblasts
Synthesize fibers and ground substance
The most common cell in connective tissue
Two stages of activity: active & quiescent
Active fibroblast: abundant and irregularly branched
cytoplasm. Its nucleus is ovoid, large, and pale-
staining, with fine chromatin and a prominent
nucleolus.
Quiescent fibroblast (Fibrocyte) is smaller, tends to be
spindle-shaped, a smaller, darker, elongated nucleus.

Fibroblast dan fibrocyte
Fibroblast
Branched cytoplasm
Large, oval nucleus,
with fine chromatin

Fibrocyte
Spindle-shaped
Smaller, darker,
elongated nucleus
Connective cell lineage

Simplified representation of the
connective tissue cell lineage
derived from the multipotential
embryonic mesenchyme cell.
Dotted arrows indicate that
intermediate cell types exist
between the examples
illustrated. Note that the cells
are not drawn in proportion to
actual sizes, eg, adipocyte,
megakaryocyte, and osteoclast
cells are significantly larger
than the other cells illustrated
Macrophages and Mononuclear Phagocyte
System
Phagocytic ability
Morphologic feature: correspond to their
state of functional activity and to the tissue
they inhabit.
Oval, kidney-shaped, or indented nucleus,
with coarser chromatin granules than that
of fibroblasts
Cytoplasm: vacuolated or granulated
Macrophages and Mononuclear Phagocyte
System
Macrophages derive from bone marrow
precursor cells that divide, producing monocytes
that circulate in the blood. Monocytes can cross
the walls of venules and capillaries to penetrate
the connective tissue, where they mature and
become macrophages.
Macrophages, which are distributed throughout
the body, are present in most organs and
constitute the mononuclear phagocyte system
(Kupffer cells, microglial cells, Langerhans cells,
osteoclasts).
Mast cells
Oval to round connective tissue cells
Cytoplasm is filled with basophilic secretory
granules.
Small, spherical nucleus is centrally situated,
frequently obscured by the cytoplasmic
granules.These granules contain pre-formed
mediators such as histamine and proteoglycans.
Function: storage of chemical mediators of the
inflammatory response.
Mats cell granules are metachromatic

Mast Cell

Section of rat tongue.
Several mast cells in
the connective tissue
surround muscle cells
and blood vessels. PT
stain. Medium
magnification.
Plasma Cells

Plasma cell
Large ovoid cell, with
basophilic cytoplasm
The nucleus is spherical,
eccentrically placed
Coarse heterochromatin,
alternating with lighter
areas (resembles the face
of a clock)
Average life: 10-20 days

Adipose Cells
Photomicrograph of unilocular
adipose tissue of a young
mammal. Arrows show nuclei
of adipocytes (fat cells)
compressed against the cell
membrane. Note that, although
most cells are unilocular, there
are several cells (asterisks)
with small lipid droplets in their
cytoplasm, an indication that
their differentiation is not yet
complete. Pararosaniline
toluidine blue (PT) stain.
Medium magnification.
Connective Tissue Fibers
The 3 main types of fibers are:
Collagen
Reticular
Elastic

The predominant fiber type is responsible
for conferring specific properties on the
tissue.
Collagen
Representing 30% of the bodys dry weight
Fibrillar
Birefringent
640 (64 nm) periodicity
Specific stain: Mallory : purple blue
Masson : green
van Gieson : red

Collagen
Consist of tropocollagen molecules
Length : 280 nm
Width : 1.5 nm
Triple helix
consists of 3 polypeptide chain (= chain) make
up of 3 amino acids
1. other than (2) and (3)
2. proline
3. always glycine
Differences in the chemical structure of these
polypeptide chains are responsible for the various
types of collagen
Collagen Types
Collagen type I
Location
skin (loose connective tissue)
tendon
bone, dentin
Function
Resistance to tension
Collagen Types
Collagen type II
Location
Cartilage
Vitreous body

Function
Resistance to pressure
Collagen Types
Collagen type III
Location
Skin
Muscle
Blood vessels
Function
Structural maintenance in expansible
organs
Collagen Types
Collagen type IV
Location
All basement membranes

Function
Support of delicate structures
Filtration
Collagen Types
Collagen type V
Location and function
Fetal tissues
Skin
Bone
Placenta
Most interstitial tissues
Function: Participates in type I collagen
function

Elastic Fiber
Elastic
Homogenous
Can branch
Resistant to boiling, acid and alkali
extractions
Specific stains:
Orcein brown
Resorcin fuchsin purple blue
Reticular Fibers
Are extremely thin, collagen III fibers
Not birefringent
64 nm periodicity
Form extensive network in certain organs,
particular abundant in smooth muscle,
endoneurium, and the framework of
hematopoietic organs, and constitute a network
around the cells of parenchymal organs.
Specific stain
Silver Impregnation (eg Bielschowsky method)
PAS (periodic acid Schiff)
Ground Substance
The intercellular ground substance is a highly
hydrated, complex mixture of
glycosaminoglycans, proteoglycans, and
multiadhesive glycoproteins.
Glycosaminoglicans (originally called acid
mucopolysaccharides) are linear
polysaccharides formed by repeating
disaccharide units, usually a uronic acid and a
hexosamine.
The proteoglycans are composed of a core
protein with 4 main glycosaminoglycans,

Types of Connective Tissue
Connective tissue proper
Loose connective tissue (areolar)
Dense connective tissue
Regular
Irregular
Collagen
Elastic

Connective Tissue with special
properties
Adipose tissue
common (univacuolar/unilocular/yellow)
brown (multilocular)
Elastic tissue
Hematopoietic tissue (lymphatic and
myeloid)
Mucous tissue


Supporting Connective Tissue
Supporting connective tissue:
Cartilage
Hyaline
Elastic
Fibrocartilago
Bone
Compact bone
Cancellous bone

Connective Tissue Proper
Loose connective tissue
Fills spaces between muscle cells, supports
epithelial tissue, and forms a layer that
sheathes the lymphatic and blood vessels.
The most numerous cells are fibroblasts and
macrophages
Dense connective tissue
Is adapted to offer resistance and protection.
Fewer cells and predominance of collagen
fibers. Dense regular and dense irregular.
Common Adipose Tissue
Color: white to yellow
One large lipid droplet
(unilocular/univacuolar)
Lipid removed, each
cell appears as a thin
ring of cytoplasm
surrounding the
vacuole, called signet
ring cell
Eccentric and flattened
nuclei
Common Adipose Tissue
Storage of energy (triglycerides)
Thermal insulation
Shock absorbers (soles and palms)
Shape the body (subcutaneous fat)
Found throughout the human body, except
the eyelids, the penis, the scrotum, auricle
Developed from lipoblast (mesenchym)
Multilocular Adipose Tissue
Smaller cell
Multiple lipid droplets
and numerous
mitochondria (colored
cytochromes)
Limited distribution
Nucleus at the center
Function: heat
production

Supporting Tissue
Cartilage
Avascular, no lymphatics or nerves
Nourished by diffusion of nutrients from
capillaries in perichondrium or by synovial
fluid from joint cavities.
Perichondrium: dense connective tissue. Joint
cartilage is devoid of perichondrium (nutrients
from synovial fluid).
Growth: interstitial + appositional
Hyalin Cartilage
Chondroblasts at the
periphery
Chondrocytes in lacunae
Homogenous matrix
Territorium: matrix
surrounding chondrocytes
is metachromatic due to
glycosaminoglycans
Nest/isogenous cells
A perichondrium present
Location
Elastic Cartilage
Location: auricle,
eustachian tube,
epiglottis, walls of
external auditory canal,
the cuneiform cartilage of
the larynx
Matrix appear dirty
(elastic fibers)
A perichondrium present

Fibrocartilago
Always between dense connective tissue
and hyaline cartilage.
The border areas is not clear-cut.
Chondrocytes usually in rows or singly
separated by coarse collagen type I fibers
Location: intervertebral disks and
symphysis pubis
No perichondrium
Photomicrograph of Fibrocartilago
Note the rows of
chondrocytes
separated by collagen
fibers.
Fibrocartilago is
frequently found in
the insertion of
tendons on the
epiphyseal hyaline
cartilage.
Bones
Main constituent of adult skeleton
Supports fleshy structures, protects vital organs
(thorax, cranium), harbors the bone marrow.
Reservoir of calcium, phosphate, and other ions
Bone matrix with 3 cell types: osteoblasts
(synthesize organic components), osteocytes (in
lacunae), and osteoclasts (resorption and
remodeling of bone tissues).
A periosteum present
Bone
Long bone diaphysis
Showing haversian
systems, outer and inner
circumferential lamellae.
The protruding haversian
system on the left shows
the orientation of collagen
fibers in each lamella. At
the right is a haversian
system showing lamellae,
a central blood capillary
and many osteocytes with
their processes.
Osteoblasts
Synthesize organic components of bone matrix.
Newly formed bone matrix: osteoid (not yet
calcified)
Located at the surfaces of bone tissue
Once surrounded by newly formed matrix, these
cells are now called osteocytes, located in
lacunae and connected to one another via
canaliculi, each housing cytoplasmic processes
of osteocytes.
Growth: appositional
Part of a haversian system
Note the numerous
canaliculi that permit
communication
between lacunae and
the haversian canal.
In adjacent lamellae,
the collagen fibers are
oriented in different
directions, which
provide the bone with
great strength.

Osteocytes
Osteocytes derived from osteoblasts, lie in
lacunae situated between lamellae.
One osteocyte is found in each lacuna.
The thin, cylindrical matrix canaliculi house
cytoplasmic processes of osteocytes.
Actively involved in the maintenance of the
bony matrix.
Osteoclasts
Multinuclear cells (5-50 nuclei in 1 cell)
Lie within enzymatically etched depression
in the matrix known as Howships lacuna
Secretes collagenase and other enzymes
and pumps protons into a subcellular
pocket, promoting digestion of collagen
and dissolving calcium salt crystals.
Its activity is controlled by cytokines and
hormones.
Bone Matrix
Inorganic matter represents 50% of the dry
weight of bone matrix.
Storage of calcium and phosphor
Organic matter: type I collagen and ground
substance.
After bone decalcification, its shape is preserved
but become as flexible as tendon
Removal of the organic part of the matrix, mainly
collagen, also leaves the bone with its original
shape; however, it becomes fragile, breaking
and crumbling easily when handled.
Types of Bone
Compact and cancellous (spongy) bone
Epiphysis consists of spongy bone
covered by a thin layer of compact bone
Diaphysis is almost totally compact bone
Flat bones have two layers of compact
bone called plates, separated by a layer of
spongy bone called diplo.

Histogenesis
Bone can be formed in 2 ways:
Intramembranous ossification
Endochondral ossification
Newly formed bone (osteoid) is primary bone
and soon replaced by the definitive lamellar,
secondary bone.
The combination of bone synthesis and removal
(remodeling) occurs not only in growing bones
but also throughout adult life.
Remodeling
Intramembranous Ossification
Also called desmal ossification
Is so called, because it takes place within
condensations of mesenchymal tissue.
Forming of flat bones
The starting point for ossification is called a
primary ossification center.
It begins with the differentiation of osteoblasts,
forming bone matrix which calcifies, forming
islands of developing bone. Several islands fuse
to form spongy bone.
Endochondral Ossification
Takes place within a piece of hyaline
cartilage whose shape resembles a small
version (model) of the bone to be formed.
Principally responsible for the formation of
short and long bones.
Endochondral ossification of a long bone
consists of the following sequence of
events.
Growing at the epiphyseal plate
Epiphyseal Plate
Resting zone : no morphologic changes
Proliferative zone: chondrocytes divide rapidly
and form columns of stacked cells parallel to the
long axis of bone
Hypertrophic zone: large chondrocytes whose
cytoplasm has accumulated glycogen.
Calcification zone: calcification, death of
chondrocytes,
Ossification zone: bone tissue appears
Fracture Repair
Bone fracture hemorrhage + clotting
Fibroblasts + blood capillaries enters blood
clot granulation tissue fibrous dense
connective tissue cartilage formation
endochondral ossification callus
formation (bone trabeculae reunite both
ends of fractured bone).
Spongy bone replaced by lamellar bone
Bone Repair