Histology of Cartilage and Bone

Cartilage and bone are specialized connective tissues. (It may be useful to review the connective
tissue section of Fundamentals of Histology at this point). Like all connective tissues, cartilage
and bone consist of cells and extracellular matrix. The matrix of all CTs consists of fibers
(collagen, reticular, and elastic) and amorphous ground substance, which contains proteoglycans
and hyaluronic acid. Different connective tissues vary in the abundance and types of cells, in the
proportion of cells to matrix, and in the properties of the matrix itself, including the numbers and
types of fibres and the nature of the ground substance. The matrix is secreted by some of the cells
in connective tissues, fibroblasts in most CTs. In cartilage, it is chondroblasts and chondrocytes
that produce the matrix, while in bone, it is osteoblasts and osteocytes.

 Cartilage
 Hyaline Cartilage
 Low power view of hyaline cartilage
 Higher power view of hyaline cartilage
 High power view of the isogenous
groups in the cartilage
 High power view of the boundary
between the cartilage and the
perichondrium
 Low power view of cartilage from the
human trachea
 Elastic Cartilage
 High power view of elastic cartilage
 Fibrocartilage
 High power view of fibrocartilage from
a ligament
 Fibrocartilage from the cruciate ligament
of the knee
 Embryology of Cartilage
 Bone
 Compact Bone
 Low magnification of ground bone
 Higher power view of compact bone
 Compact bone that has been
demineralized and stained
 Spongy Bone
 Low power view of spongy bone from
the dog jaw
 Junction of compact and spongy bone
 Bone Cells
 Sharpey’s Fibres
 Sharpey’s fibres in jaw bone of dog
 Embryology and Formation of Bone
 Low power view of intramembranous ossification in the
head of the pig embryo
 A high power view of one of the spicules
 High power view of bone spicules with osteoclasts
 Cartilaginous rib of pig embryo
 Cartilage cells undergoing hypertrophy under bone collar
in rib
 High power view of bone collar and hypertrophic
cartilage cells
 High power view of blood vessels in hypertrophic
cartilage cells.
 Endochondral ossification in a long bone
 Epiphyseal plate of a long bone
 Low power view of vertebrae and disk
 High power view of intervertebral disk
 Cartilage–ossification boundary in a vertebra
 Tendon and muscle
 The Synovium
 Low power view of synovial joint
 High power view of synovial membrane
 High power view of the articular cartilage
 Cartilage:

Cartilage is a solid connective tissue that is to a certain extent pliable, making it resilient. These
characteristics of cartilage are due to the nature of its matrix. The ground substance of cartilage is
rich in proteoglycans consisting of a core protein with numerous- about 100- glycosaminoglycans
(GAGs) attached bottle-brush fashion around it. GAGs are made of repeating units of
disaccharides, one of which is always a glycosamine (hence the name) such as glucosamine or
galactosamine. (Glycosamines are also called hexosamines.) In cartilage, the GAGs attached to
the core proteins are chondroitin sulfate and keratan sulfate.

The proteoglycans themselves are attached, by special linker proteins to long, rigid molecules of
hyaluronic acid (HA). HA itself is a GAG, but is composed of several thousand disaccharide
units, rather than several hundred or less, as are other GAGs. About eighty proteoglycans are
attached to one molecule of HA.

[For those who thirst for knowledge: The repeating units of chondroitin sufate are D-glucoronic
acid and N-acetylgalactosamine-(4 or 6)-sulfate. The repeating units of keratan sulfate are
galactose or galactose 6-sulfate and N-acetylglucosamine 6-sulfate. The repeating units of
hyaluronic acid are D-glucuronic acid and N-acetylglucosamine].

In addition to the giant HA-proteoglycan macromolecules, there are other proteoglycans as well
as glycoproteins in the matrix. The matrix also has collagen fibers, but these are of a finer nature
(collagen Type II vs. Collagen Type I) than the collagen fibers in most other CTs. The
macromolecules are bound to the thin collagen fibres by electrostatic interactions and cross-
linking glycoproteins.

Between 60 and 80 percent of the net weight of (hyaline) cartilage is water, and this large
component of water accounts for the resilient nature of cartilage. Water is attracted to the
negative charges in the abundant sulfate and carboxyl groups on the GAGS. This hydration
permits diffusion of water-soluble molecules in the ground substance. However the movement of
large molecules and bacteria is inhibited. Cartilage is poorly vascularized, and gets most of its
nutrients through diffusion. In the adult, repair is poor.

The sulfate groups of the GAGs that attract water also attract molecules of basic dye. Differences
in the intensity of staining within the matrix reflect differences in the abundance of
proteoglycans. Staining is most intense around the cartilage cells. The collagen fibres are not
visible in standard histological preparations as their refractive index is not markedly different
from that of the ground substance.

Some basic dyes used to stain cartilage can undergo a color change called metachromasia. In the
presence of the negatively charged groups in the GAGs, the dye molecules form aggregates of
dimers or polymers which changes their absorptive properties. Rather than staining blue, the
tissue will stain purple or reddish.

Their are three kinds of cartilage, hyaline cartilage, elastic cartilage and fibrocartilage.
 Hyaline Cartilage:

Hyaline cartilage is the most abundant type of cartilage. Most of the skeleton of the fetus is laid
down in cartilage before being replaced by bone. Hyaline cartilage in the adult is found in the
nose, parts of the respiratory tract, at the ends of ribs and at the articular surfaces of bones.
Low power view of hyaline cartilage

Figure 1 shows a low power view of cartilage in the nasal passage

of a cat. It is taken from your slide 51. In this slide, the cartilage
goes on a winding path, surrounded by the glands and blood
vessels of the nasal passageway. Glands and blood vessels can
also be seen in the section shown. The section of cartilage visible
in Figure 1 is in the shape of an upside down Y. (The two big
folds in the stem of the Y are an artifact of preparation.) The
things that appear as dots at this magnification are the
chondrocytes, and it can be seen that the matrix stains more
intensely in the middle of the cartilage (more purple) than at the
periphery (more yellow). The outer part of the cartilage is surrounded by the perichondrium
(red), made of dense connective tissue.
High power view of hyaline cartilage

Figure 2 shows a higher power view of the cartilage at the middle

of the Y. The chondrocytes are more evident and can be seen to
be clustered in groups, called isogenous groups. Such isogenous
groups reflect the last mitotic division of a chondrocyte whose
daughter cells have not secreted much more matrix around
themselves. The matrix in the middle part of the cartilage is
mottled, with areas of darker staining around the isogenous
groups. This darker staining reflects the greater abundance of
GAGs and is called the territorial matrix. The lighter-staining
areas between clusters of cells are called the interterritorial
matrix. Perichondrium can be seen around all of the cartilage shown. The border between the
perichondrium and cartilage is particularly evident at the lower part of the cartilage, where the
perichondrium stains quite red, while the outer part of the cartilage is more orange. The cells at
the outer part of the cartilage are also smaller than those farther inside (a bit difficult to see at this
magnification). The cartilage and its perichondrium lie in dense irregular CT in which blood
vessels, hair follicles and glands can be seen. (You don’t have to identify glands and hair
follicles.)
High power view of isogenous groups

Figure 3 shows a high power view of the isogenous groups in the

cartilage. The territorial and interterritorial matrices can be clearly
distinguished. Within the matrix, cartilage cells sit in spaces called
lacunae. An intensely staining ring is sometimes seen around the
edge of a lacuna. It is called the capsule and again reflects a higher
density of GAGs. In life, chondrocytes completely fill the lacunae, but during tissue preparation,
the cells frequently shrink and even fall out. Thus, in cartilage tissue, you sometimes see only
empty spaces where the chondrocytes once sat. Some of the cells in Figure 3 can be seen to have
shrunk a bit.

Although it cannot be differentiated with the light microscope, there is an inner and an outer part
of the perichondrium. The cells in the outer (or fibrous) part are regular fibroblasts that secrete
the collgen type I of the perichondrium. The cells of the inner (or cellular) part, however, are
capable of differentiating into chondroblasts. These chondroblasts then begin to secrete the
matrix of cartilage: collagen type II fibrils and the cartilage-specific ground substance. These
chondroblasts lie directly apposed to the perichondrium, when they have secreted enough matrix
to be completely surrounded by it, they are referred to as chondrocytes. Thus cartilage can grow
through two processes. Growth resulting from the secretion of matrix by chondroblasts of the
perichondrium is called appositional growth. Growth resulting from the secretion of matrix by
chondrocytes which are already embedded in matrix is called interstitial growth.
High power view of cartilage and perichondrium

Figure 4 shows a high power view of the boundary between the

cartilage and the perichondrium (which is torn a bit). The
flattened cells lying right along the boundary of the inner
perichondrium are chondroblasts. Cells completely surrounded by
cartilage are chondrocytes. Chondrocytes closer to the periphery
are more flattened than those deeper inside the cartilage.

Low power view of cartilage from human trachea

Figure 5 shows a low power view of cartilage from the human

trachea. The characteristics are the same as those described in the
previous four figures. A few glands can be seen at the bottom
right of the figure.

Elastic Cartilage:

The structure of elastic cartilage is very similar to that of hyaline cartilage, but in addition to the
other components, its matrix has elastic fibres and interconnecting sheets of elastic material. This
gives elastic cartilage an elasticity not present in hyaline cartilage. Elastic cartilage is found in
the external ear, the walls of the external auditory canal, the Eustachian tube, the epiglottis and
the larynx.
 High power view of elastic cartilage

Figure 6 shows a high power view of elastic cartilage. The

density of elastic material, especially toward the middle of the
matrix, has made it appear quite red, similar in color to the
perichondrium. Toward the periphery of the cartilage, wisp-like
ends of the elastic fibres can be seen. With other stains, such as
orcein used to stain elastin, the elastic fibres would stand out
more clearly as black lines and bands.

Fibrocartilage:

Fibrocartilage has characteristics intermediate between those of hyaline cartilage and dense
connective tissue. Its presence indicates the need for resistance to compression and shear forces.
It is found in the intervertebral disks, the symphysis pubis, the articular discs of the
sternoclavicular and temperomandibular joints, the menisci of the knee joint and some places
where ligaments or tendons attach to bones. The amount of cartilage in fibrocartilage is variable,
it generally occupies a smaller amount of the tissue. There is no perichondrium.
High power view of fibrocartilage

Figure 7 shows a high power view of fibrocartilage from a

ligament. The isogenous groups of cartilage cells are in a line,
surrounded with a bit of purple matrix, the rest of the tissue is
dense connective tissue (stains pink).

Low power view of cruciate ligament

Figure 8 shows fibrocartilage from the cruciate ligament of the

knee. Many rows of isogenous groups are seen. There is a
relatively greater amount of matrix in the fibrocartilage shown
here than was present in the previous figure.

Embryology of Cartilage:

Cartilage arises from mesenchyme. Some mesenchyme cells aggregate to form a blastema. The
cells of the blastema begin to secrete cartilage matrix and are then called chondroblasts. They
move apart as they deposit matrix, and when they are completely surrounded by matrix they are
called chondrocytes. The mesenchymal tissue surrounding the blastema gives rise to the
perichondrium.
 Bone:

Bone is a connective tissue distinguished by the fact that its matrix is mineralized by calcium
phosphate in the form of crystals very similar to hydroxyapatite. The minerals are both in the
(Type I) collagen fibers which constitute about 95% of the matrix, and in the ground substance.
Bone is both resilient and hard. Its resilience is due to the organic matter (collagen), its hardness
due to the inorganic minerals. Bone serves as a storage site for calcium and phosphate. Blood
calcium levels are regulated by the hormones parathormone (parathyroid hormone), which raises
blood calcium levels by stimulating bone resorption, and calcitonin, which reduces blood calcium
by suppressing bone resorption and increasing osteoid calcification. (Osteoid is the matrix
secreted by osteoblasts and osteocytes prior to mineralization).

There are two kinds of mature bone: compact bone and spongy bone. Compact bone is also
called dense bone and cortical bone. Spongy bone is also called cancellous bone, trabecular bone
and medullary bone. [Another term, lamellar bone, is sometimes used to mean only compact
bone, and sometime used to denote mature bone, whether spongy or compact, as opposed to
immature bone (see below). Therefore, the term lamellar bone is a bit ambiguous].

Compact bone and spongy bone are found in specific locations. In long bones, most of the
thickness of the diaphysis is made of compact bone, with a small amount of spongy bone facing
the marrow cavity. The ends (epiphyses) of long bones, however, consist mostly of spongy bone
covered with a shell of compact bone. The flat bones of the skull have a middle layer of spongy
bone sandwiched between two relatively thick layes of compact bone.
Compact Bone:

Compact bone is composed of cylindrical structures called osteons or Haversian systems. An

osteon consists of concentric lamellae of bone matrix, mainly collagen fibres, surrounding a
central canal called the Haversian canal, which contains small blood vessels and nerves. The long
axis of osteons is usually parallel to the long axis of the bone. The collagen fibres within any one
lamella are generally parallel with one another, but the collagen fibres in the different lamellae of
an osteon are oriented at different angles. This increases the strength of the osteon. Canals called
Volkmann’s canals link the Haversian canals of different osteons with one another and with the
marrow cavity. They provide the major route for blood vessels from the marrow cavity to the
Haversian canals of osteons.

Between the lamellae of an osteon are lacunae containing bone cells called osteocytes. Canaliculi
connect the lacunae with one another and with the Haversian canal. The canaliculi contain the
processes of the osteocytes, which communicate with one another via gap junctions. Thus
nutrients and other substances, such as hormones, can pass from blood vessels in the Haversian
canal to distant osteocytes via a sort of bucket brigade. Osteocytes can be involved in both bone
deposition and bone resorption.

In addition to the lamellae of osteons, lamellae not belonging to any osteon can be seen in
compact bone. These are called interstitial lamellae and are the remnants of previous osteons.
They reflect the fact that bone is not static but is constantly being remodelled. In addition, the
inner and outer surfaces of long bone have lamellae that run the length of the shaft. They are
called, respectively, the inner and outer circumferential lamellae.
Low power view of ground compact bone

Figure 9 is a low magnification of ground bone. Ground bone is

bone that has been dried, cut (with a saw, not a microtome) and
ground to a sufficient thinness to be viewed with a microscope.
Ground bone is used to observe the architecture of bone. This
section is not stained.

Note the numerous osteons, of variable sizes and shapes. The size
of their Haversian canals is also quite variable. Many interstitial
lamellae are seen among the osteons. A Volkmann’s canal is seen
arising from one Haversian canal. (Sometimes you will see a
Volkmann’s canal making a "textbook" connection between two Haversian canals. This one is
less perfect.) The lacunae can be seen as black spots lying between the lamellae of osteons. This
section does not show inner or outer circumferential lamellae.
High power view of ground compact bone

Figure 10 shows a higher power view of compact bone. A

complete osteon is shown at the centre of the figure with parts of
several osteons surrounding it. The lacunae can be clearly seen,
and the canaliculi are discernible as fine black lines.

  Stained, demineralized compact bone

Figure 11 shows compact bone that has been demineralized and stained (taken from slide 88, of
the dog jaw). The two lines at the right are an artifact of the tissue
folding. The osteons appear as red circles with the Haversian canal
(blue) in the centre. Lacunae appear as dots. Many interstitial
lamellae are visible. Several Volkmann’s canals are evident, even
though the section did not go right through the canals themselves.
Even at higher magnifications, the canaliculi would not be
distinguishable.

Spongy Bone:

Spongy bone is composed of bone spicules, also called trabeculae, of varying shapes and sizes.
The spaces between the spicules are filled with marrow. The composition of spongy bone (cells
and matrix) is the same as that of compact bone. In spongy bone, however, the lamellae of
collagen are not arranged concentrically around a central canal, but run parallel to one another.
Osteocytes sit in lacunae between lamellae.
Low power view of spongy bone

Figure 12 shows a low power view of spongy bone from the dog
jaw (slide 88). The variability of the bone spicules is evident. In
the spaces between the spicules, the myeloid elements of bone
marrow can be seen as bluish patches.

Junction of compact and spongy bone

Figure 13 shows the meeting of compact bone (above) with

spongy bone (below) in the jaw (slide 88).

Bone Cells:

Osteoprogenitor cells (periosteal and endosteal), osteoblasts, osteocytes, osteoclasts

Just as cartilage is surrounded by a perichondrium, bone is surrounded by a periosteum of dense

connective tissue. As in the perichondrium, the periosteum has two layers: an outer fibrous layer
with typical fibroblasts, and an inner cellular layer, which contains osteoprogenitor cells. The
osteoprogenitor cells in this location are called periosteal cells. They are capable of giving rise to
osteoblasts, which secrete the extracellular matrix of bone.

The marrow surface of compact bone, and the spicules of spongy bone, are lined by an (often
single) layer of cells called the endosteum (endosteal cells). Like the periosteal cells, these
endosteal cells are also osteoprogenitor cells, capable of becoming osteoblasts. (The two names
periosteal cells and endosteal cells refer to their different locations, both function as
osteoprogenitor cells).

Osteoblasts secrete the collagen fibres and ground substance (matrix) of bone and are responsible
for the calcification of the matrix. They retain the ability to divide and communicate by thin
cytoplasmic processes which form gap junctions.

When an osteoblast is completely surrounded by matrix, it is called an osteocyte. Osteocytes are

responsible for maintaining the matrix, and can both secrete and resorb matrix. Thus
osteoprogenitor cells, osteoblasts and osteocytes are all part of a series derived from mesenchyme
cells. There is evidence that osteocytes and osteoblasts can revert to earlier stages.

Another type of cell called the osteoclast is responsible for the resorption of bone. These large,
multinucleated cells arise from monocytes. They release lysosomes, organic acids and hydorlytic
enzymes to break down bone matrix.
Sharpey’s Fibres

Collagen fibres from tendons and ligaments extend into bone

tissue at an angle, and become continuous with the collagen
fibres of the bone matrix. Figure 14 shows Sharpey’s fibres in the
jaw bone, extending downward from the connective tissue above.

Embryology and Formation of Bone:

Intramembranous ossification, endochondral ossification

In the embryo, bone tissue arises through two processes, intramembranous ossification and
endochondral ossification. In intramembranous ossification, bone is formed directly from
mesenchymal tissue. The flat bones of the skull and face, the mandible and the clavicle develop
in this manner. In endochondral ossification, a cartilage model of the bone is formed first, and is
later replaced by bone. The weight-bearing bones of the axial skeleton and the bones of the
extremities (= most of the skeleton) develop in this manner.

The first bone to arise, whether from mesenchyme or from cartilage (or in fracture repair
postnatally), is in the form of spicules. These first spicules are made of immature bone, also
called woven bone. In immature bone, the collagenous lamellae are not arranged in parallel or
concentric arrays (as in mature spongy and compact bone, respectively), but are randomly
oriented and loosely intertwined (hence woven). Immature bone also has more ground substance
than mature bone. Consequently, immature and mature bone show different staining
characteristics, immature bone stains more with hematoxylin and mature bone more with eosin.

The spicules of immature bone are remodelled. The remodelling process can eventually give rise
to more spongy bone or to compact bone. The remodelling of bone continues throughout life
(remember those interstitial lamellae in compact bone represent former osteons). Immature bone
is the predominant bone in the developing fetus. In the adult, most immature bone is replaced by
mature bone, but immature bone is seen where bone is being remodelled or repaired, and in
certain specific areas, such as the alveolar sockets of the oral cavity. Because most of the spongy
bone (= in form of spicules) you look at in the lab is from the embryo, you may erroneously
come to think of spongy bone as synonymous with immature bone.

When bone matrix is first secreted, it is not yet mineralized and is called osteoid. As mentioned
above, osteoblasts also bring about the mineralization of bone.

Intramembranous Ossification:

The first step in intramembranous ossification is the aggregation of mesenchyme cells in the area
where bone is to be formed. The tissue in this area becomes more vascularized, and the
mesenchyme cells begin to differentiate into osteoblasts, which secrete the collagen and ground
substance (proteoglycans) of bone matrix (collectively called osteoid). The osteoblasts maintain
contact with one another via cell process. The osteoid becomes calcified with time, and the
processes of the cells (called osteocytes when they are surrounded with matrix) become enclosed
in canaliculi. Some of the mesenchymal cells surrounding the developing bone spicules
proliferate and differentiate into osteoprogenitor cells. Osteoprogenitor cells in contact with the
bone spicule become osteoblasts, and secrete matrix, resulting in appositional growth of the
spicule. Intramembranous ossification begins at about the eighth week in the human embryo.
Low power view of intramembranous ossification

Figure 15 shows a low power view of intramembranous

ossification in the head of the pig embryo. A number of bony
spicules are seen lying in the mesenchyme from which they
developed. A band of cartilage (not involved with these spicules)
is seen lying below them. Osteocytes can be seen within the
spicules, and each spicule is surrounded by osteoblasts.

High power view of bone spicule

A high power view of one of the spicules from Figure 15 (the

seahorse-shaped one at the far right) is shown in Figure 16.
Numerous osteocytes are seen enclosed within the spicule, and
most of the periphery is lined by osteoblasts. (A characteristic
feature of immature bone is a greater abundance of cells than in
mature bone). Several developing blood vessels are seen in the
mesenchyme.

High power view of bone spicules with osteoclasts

Figure 17 shows a high power view of a bone spicule with three

osteoclasts, only one of which (the top one) is obvious. Although
this osteoclast will be multinucleate, only one nuceus, toward the
left of the cell, can be clearly seen. No osteoblasts are evident
around this spicule, but osteocytes can be seen within it. The
structure of the mesenchyme can also be seen quite well: small
spindle-shaped cells with processes. These processes contact the
processes of neighboring cells via gap junctions. In life, the extracellular space is occupied by a
viscous ground substance. It is lost during routine histological tissue preparation.

Endochondral Ossification:

Endochondral ossification also begins with the aggregation of mesemchyme cells, but these
differentiate into chondroblasts which secrete hyaline cartilage matrix. The cartilage is secreted
in the general shape of the bone that it will become, and grows by both interstitial (mostly in
length) and appositional (mostly in width) growth.

Sometime during the growth of this cartilage model (starting at about week 12 in the human
fetus), some of the inner perichondrial cells begin to give rise to osteoblasts instead of
chondroblasts. (As a result, the former perichondrium is now called the periosteum.) In long
bones, this process begins at the mid-region of the bone. The newly formed osteoblasts secrete
osteoid, forming a bone collar around the cartilage model. Therefore the very first bone that is
formed during endochondral ossification is considered to arise by intramembranous ossification.

With the formation of the bone collar, the cartilage cells in the underlying cartilage begin to
hypertrophy and secrete alkaline phosphatase. The surrounding cartilage matrix becomes
calcified. The bone collar and the calcified matrix inhibit diffusion of nutrients, and the
chondrocytes begin to die. The matrix breaks down, and the lacunae of the dying chondrocytes
become confluent making a large cavity. At the same time, blood vessels grow through the bone
collar, bringing osteoprogenitor (periosteal) cells with them. These differentiate into osteoblasts
when they come in contact with the calcifying cartilage matrix. Osteoid is deposited on the
calcifying matrix which is eventually removed. (This is the endochondral part of endochondral
ossification.) The bone spicules are remodelled by osteoclasts and osteoblasts. The invading
blood vessels also bring with them cells which dfferentiate into the hematopoietic cells of the
bone marrrow.

In long bones, the midregion where bone formation is initiated is called the primary ossification
centre. Secondary ossification centres will develop after birth at the ends (epiphyses) of the bone.
During the years of growth, the primary and secondary ossification centres are separated by a
cartilage plate called an epiphyseal (growth) plate. This plate allows the bone to grow in length.
Cartilaginous rib of pig embryo

Figure 18 shows part of a rib of the pig embryo that has not yet
begun the process of ossification. The cartilaginous rib is
surrounded by a perichondrium.
 Cartilage cells undergoing hypertrophy under
bone collar in rib

Figure 19 shows the other end of the rib, in which the first
step of endochondral ossification has begun (you will recall
this is actually a form of intramembranous ossification). A
bone collar lined with osteoblasts (which arose from what
used to be the perichondrium and is now the periosteum) can
be distinguished, but is most easily seen toward the right of
the figure. The cartilage cells are very obviously undergoing
hypertrophy, but the matrix is not yet calcified, and no
osteoid has yet been deposited.
High power view of bone collar and
hypertrophic cartilage cells

A high power view of the bone collar and the hypertrophic

cartilage cells is shown in Figure 20. The lacunae are
enlarged and becoming confluent and the matrix is reduced.
The bone collar is lined with osteoblasts.

High power view of blood vessels in

hypertrophic cartilage cells

Figure 21 (taken from slide 97 of the vertebral column of a

young cat) shows a high power view of some blood vessels
that have invaded the hypertrophic cartilage during
endochondral ossification. The lacunae can be seen to be
hypertrophic, the chondrocytes are shrunken due to tissue
preparation. Red blood cells can be seen in the vessels but
the osteoprogenitor cells cannot be distinguished. The
vertebrae ossify postnatally, in humans the process is not
completed until adulthood.
Endochondral ossification in a long bone

Figure 22 is taken from slide 21 (tibia and knee joint of rat). It

shows endochondral ossification at a later stage, in the primary
ossification centre of a long bone. Remnants of purple cartilage
matrix are surrounded by bone, staining pink. In between the bone
spicules are the myeloid elements of bone marrow. The bone
collar of the long bone is not visible in the field of view.
 Epiphyseal plate of a long bone

Figure 23 shows a low power view of the epiphyseal plate. The

primary ossification centre that you saw in Figure 22 is toward the
right of Figure 23. The epiphyseal plate itself is the vertical purple
band, and a secondary ossification centre (at the epiphysis of the
bone) is seen at the left of the figure. The layers of the epiphyseal
plate extend from left to right. The rat from which this slide was
made was reaching maturity, and hence the layers of the plate are
not as large or as well-defined as you might see in textbooks.
Read the lecture notes or text for a description of events in each
zone.
Low power view of vertebrae and disk

Figure 24 is a low power view of parts of two vertebrae separated

by the intervertebral disk. Toward the disk side, both vertebrae
are still cartilaginous. (The cartilage stains more red than in the
previous pictures you have seen.) Toward the middle of each
vertebra, ossification is proceeding and bone spicules separated
by marrow spaces can be seen. The intervertebral disk consists of
a puffy-looking nucleus pulposus (a remnant of the notochord)
and a ring of fibrocartilage.
High power view of intervertebral disk

Figure 25 shows a higher power view of the central part of Figure

24. The fibrocartilage and nucleus pulposus of the disk can be
seen between the cartilage of two vertebrae. Because of the
staining on this slide, the fibrocartilage may be harder to identify
than that which you saw in the section on cartilage (Figures 7 and
8).

Cartilage–ossification boundary in a vertebra

Figure 26 shows a high power view of the boundary between the

cartilage and the ossification centre in a vertebra. In the spicules
(left), the bone is slightly more eosinophilic than the cartilage
matrix on which it is being deposited. The tear in the cartilage
(top right) could represent the start of a new ossification centre.
 Tendon and muscle

Figure 27 shows a tendon joining a muscle. The striations in the

muscle are barely visible. Tendon is made of regularly arranged
bundles of collagen secreted by fibroblasts (tendon cells). In this
section, the tendon appears a bit wavy. This is not the case if the
tissue is completely straight when sectioned. A few strands of the
tendon can be seen to penetrate into the muscle.

The Synovium

The synovium is the joint cavity between two movable bones. The bone surfaces in the synovium
are covered in cartilage, called the articular cartilage, which lacks a perichondrium. The ends of
the two bones are enclosed and united by an articular capsule, which has two parts. The outer
part is a sleeve of fibrous tissue (fibrous capsule) which extends well beyond the articular
cartilage of each bone. The inner part is called the synovial membrane, it lines the fibrous capsule
and is reflected onto the bone, which it covers right up to the articular cartilage. Thus the joint
cavity is lined everywhere with either articular cartilage or synovial membrane.

The synovial membrane is a thin sheet of connective tissue, with abundant blood vessels and
lymphatics. The surface facing the joint cavity is lined by epithelioid cells which secrete
hyaluronic acid and phagocytize debris. (They do not sit on a basement membrane so they cannot
be called an epithelium.) The hyaluronic acid and a dialysate of plasma from the blood vessels of
the membrane constitute the synovial fluid, a viscous substance which lubricates the joints.

Synovial membranes can be quite variable in appearance, they may be closely apposed to the
fibrous capsule or thrown into many folds (sometimes called villi), they may rest directly on
periosteum or on loose connective, dense fibrous or adipose tissue. If extensive folds in the
membrane are present, islets of cartilage may develop inside them, by metaplasia of the synovial
fibroblasts.

Inflammation of the synovial membrane is called rheumatoid arthritis, and results in the
stimulation of the pain ending of nerves. The formation of uric acid crystals in the joints is called
gout, and damages the articular cartilage. Various kinds of trauma to the joint can damage the
cartilage to the point that it begins to calcify and be replaced by bone. This can lead to a bony
fusion called ankylosis which results in a loss of mobility.
 Low power view of synovial joint

Figure 28 shows a low power view of part of a synovial joint

(from slide 21 of the rat tibia). The articular cartilage at the end of
one of the bones can be seen covering the bony spicules and
marrow spaces of its epiphysis. The fibrous capsule and the
synovial membrane lining it can be seen at the left. (These tissues
are very difficult to scan and in this computer image are barely
distinguishable from background, they will be a bit easier to see
through a real microscope.)
High power view of synovial membrane

Figure 29 shows a higher power view of the synovial membrane

(again slightly hard to see). The membrane is made of loose
connective tissue with some areas of adipose tissue, and is lined
with epithelioid cells. No blood vessels can be unequivocally
identified in the field of view.

High power view of the articular cartilage

Figure 30 shows a high power view of the articular cartilage

covering the epiphysis. Note the absence of a perichondrium. The
synovial membrane can be seen at the left.