THE SKELETAL SYSTEM

Cartilage and Bone

Prof Raymond Coleman

Department of Anatomy & Cell Biology
Rappaport Faculty of Medicine
Technion-Israel Institute of Technology
Haifa, Israel

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The Skeletal System
 The skeletal tissues
• Skeletal tissues consist mainly of
cartilage and bone
• These skeletal tissues are specialized
forms of connective tissue
• Like all connective tissue they
develop in embryos from embryonic
mesenchyme
 Cartilage
• Cartilage is mainly a tissue of embryos
and fetuses
• In postnatal life and in adults cartilage
has a very limited distribution, but serves
very important functions
• The slow-turnover of cartilage, limited
growth and repair processes in adults is
problematic
 Functions of cartilage
• Skeletal support in embryos prior to
development of bony skeleton
• Long bone elongation (endochondral
ossification)
• Articulating joints
• Flexible support and protection (ears,
trachea, bronchi)
 Cartilage
• Cells (chondrocytes) constitute only 2-5% of
tissue volume
• The main bulk of cartilage is the matrix (95-98%)
• Cartilage cells are located in lacunae in the
matrix
• Cartilage is avascular and lacks intrinsic nerves
• Cartilage receives nutrients from blood vessels
of the perichondrium
• Metabolites and nutrients diffuse via the matrix
(low metabolic activity)
 Types of cartilage
There are three morphological types of
cartilage:
Hyaline cartilage
Elastic cartilage
Fibrocartilage
The differences depend on the matrix
especially the type and amount of the
embedded fibers
 Hyaline cartilage
• This is the most common type of cartilage
• The name is derived from the Greek
“hyalos”=glass
• This is mainly found in developing
embryos where it forms the model for
developing bones
• In adults hyaline cartilage is found in the
respiratory tract, ventral part of ribs, and
articulating joints (articular cartilage)
Hyaline cartilage

Hyaline cartilage of the trachea

Hyaline cartilage
Hyaline cartilage
 Cartilage cells
• Chondroprogenitor cells (found in
the perichondrium). These develop
into chondroblasts
• Chondroblasts (actively secrete
matrix)
• Chondrocytes (mature cells derived
from the chondroblasts).
 Chondrogenesis
There are two different types of
cartilage growth
• Appositional growth (adding new
cells from the perichondrium)
• Interstitial growth (division of older
cells to form isogenous or nest cells
located in the same lacuna deeper in
the matrix)
 )Nest cells )isogenous cells

Interstitial growth of matrix occurs in deeper areas of cartilage and involves

division of chrondrocytes within a single lacuna
 Hyaline cartilage cells
Chondroprogenitor
cells

Chondroblasts

Chondrocyte

Nest or
isogenous cells
 Hyaline cartilage matrix
The main components of the matrix are:
• Water (“solvation water”) (72-75%)
• Proteoglycans (10%)
• Collagen )type II) (16%)
• Glycoproteins (e.g.chondronectin) 1.6%
• Minerals (0.5%)
 Proteoglycans of matrix
The proteoglycans are a complex of
protein and sulfated glycosaminoglycans
(GAGs) and in particular:
• chondroitin-4-sulfate
• chondroitin-6-sulfate
• keratan sulfate
• (non-sulfated) hyaluronic acid
 Cartilage matrix

HA, hyaluronic acid

LP, link protein
PC, core protein
CS, chondroitin sulfate
 Matrix staining properties
• Basophilic after H & E staining (mainly
due to sulfated glycosaminoglycans)
• Periodic acid-Schiff (PAS) positive
• Metachromatic
The matrix surrounding lacunae stains a
deeper color (territorial matrix) owing to
more GAGs than in the interterritorial
matrix
 Territorial matrix

The territorial matrix surrounding lacunae stains deeper than the interterritorial matrix
(owing to the greater amount of glycosaminoglycans (GAGs
Hyaline cartilage metachromasia
If stained with toluidine blue or Azur II the GAGs stain a purple
)color )metachromasia
Metachromasia of matrix
 Elastic cartilage
• Elastic cartilage has a very limited
distribution. It is present in the external ear
and epiglottis
• It contains large quantities of elastic fibers in
the matrix providing flexibility and elasticity
• It has a yellowish color in the fresh state
• Elastic fibers can be stained with orcein
• The numbers and quantity of elastic fibers
are greater in older matrix
 Elastic cartilage
Perichondrium

.Elastic cartilage stained with Weigert’s elastic stain

The elastic fibers are present in greater numbers in older matrix rather than
in the newer matrix below the perichondrium
 Fibrocartilage
Fibrocartilage is found in areas subject to
high mechanical stress or weightbearing
such as:
• intervertebral disks
• pubic symphysis
• temporo-mandibular joints
• ligament connections to bones
(e.g. Ligamentum teres femoris)
• tendon insertions
 Fibrocartilage
• high tensile inelastic properties
• possesses large amounts of collagen
fibers (little amorphous matrix)
• white appearance because of large
quantities of collagen fibers
• acidophilic in H&E staining
• lacks perichondrium
 Fibrocartilage

The main feature of fibrocartilage is the dominance of large bundles of orderly-

arranged collagen fibers that provide great tensile strength to the tissue
Intervertebral disks
Nucleus
pulposus

Anulus fibrosus

Theanulus fibrosus is a form of fibrocartilage. Thenucleus

pulposus . represents remains of the embryonic notochord
Intervertebral disk
 Secondary cartilage
• This refers to cartilage that develops in
association with specific bones formed by
intramembranous ossification after the
bones are already present (unlike cartilage
associated with endochondral ossification)
• The temporo-mandibular joint (TMJ), for
example, has secondary cartilage
 Cartilage regeneration
• Cartilage in adults has very limited
regenerative ability if injured or after wear-
and-tear of aging
• This is a result of limited numbers of
cartilage cells with low metabolic activity
and minimal cell turnover
• Absence of an integral blood supply
 Bone Tissue

• Bone tissue is the

main skeletal tissue of
the body and is the
main component of
bones

(Cartilage-ligament skeleton (van Hagen

 Bone tissue
• Bone is a specialized form of
connective tissue and the main
element of skeletal tissues
• It is composed of cells and
extracellular matrix
• Unlike other connective tissues the
extracellular matrix becomes hard
and calcified
Bone
 Functions of bone
• Skeletal support
• Site of attachment of tendons and
muscles
• Protection for vital organs (skull protects
brain; rib cage protects heart and lungs)
• Hematopoietic tissue (bone marrow) is
enclosed and protected by bone
• Calcium homeostasis (main store of
calcium and phosphate)
Muscle man and his
skeleton
(Gunther van Hagen)

The skeleton provides the

support system for skeletal
muscle
 Bone characteristics
• Hard, brittle, light-weight
• Dynamic tissue, continuous formation
and resorption throughout life
• Remodelling is result of several
factors including: mechanical stimuli,
metabolic causes (diet, illness,
aging), endocrine changes, use of
drugs
 Macroscopic structure

Compact bone

Spongy bone

There are two main

categories of bone:
• Spongy bone
(trabecular, cancellous
bone)
• Compact bone
(cortical bone)
 Compact bone

Compact bone appears as a mass of bony

tissue lacking spaces visible to the unaided
eye. All bones (long bones, flat bones and
irregular bones) are composed of both
.compact and spongy bone
 Spongy bone

Spongy bone is composed of a lattice or

network of branching bone spicules or
trabeculae. The spaces between the bone
.spicules contain the bone marrow
 Spongy bone
)Osteoid )prebone

Bone trabeculae have a

lamellar structure.
Osteoblasts are active
at sites of bone Endosteum is a thin
formation.Osteoclasts layer of cells
are active at sites of between the
bone resorption trabecula and bone
((Howship’s lacunae marrow and is asite
of osteoprogenitor
.cells
 Preparation of histological
sections of bone tissue
• Decalcification (acids or chelating
agents such as EDTA)
• Ground sections
• Embed in hard resins and cut with
tungsten-carbide knives
• For electron microscopy small blocks
(under 1mm3) of undecalcified bone can
be cut with diamond knives
 Microscopic structure
• Cells form only a small
part of bone tissue
• The bulk of the tissue
is the calcified bone
matrix
• The matrix has two
main components:
)d) organic matrix
)e) inorganic matrix
 Bone matrix
Organic matrix
• Type 1 collagen fibers (95%)
• Amorphous ground substance: sulfated
GAG’s (chondroitin-4-sulfate, chondroitin-
6-sulfate, keratan sulfate); bone proteins
(sialoprotein, osteocalcin)
Inorganic matrix
• Mainly hydroxyapatite Ca10(PO4)6.(OH)2
Bone matrix
)Organic matrix )30%
Cells )2%)
)Matrix )98%

)Inorganic matrix )70%

)Hydroxyapatite )95%
 Bone cells

Osteoprogenitor Osteoblast Osteocyte Osteoclast

cell
in osteogenic tissue) on surfaces of) in lacuna of bone) in Howship’s lacuna of)
of periosteum and (developing bone (tissue (bone undergoing resorption
(endosteum
 Bone cells
• Osteoprogenitor cells are found in the
periosteum and endosteum and possess
“osteogenic potential”
• Osteoblasts are found on surfaces of
newly forming bone
• Osteocytes are present in lacunae of
bone matrix
• Osteoclasts are found in matrix being
resorbed (Howship’s lacunae)
 Osteoblasts

Osteoblasts

Osteoblasts are present on surfaces of new bone development. Osteoblasts are

characterized byhigh alkaline phosphatase activity. They secrete the components
.of osteoid (prebone) which undergoes calcification to form the calcified matrix of bone
 Osteocytes

Osteocytes in lamellar bone are located in lacunae. Lacunae are connected

to each other by means ofbone canaliculi. Cell processes from osteocytes
penetrate the bone canaliculi. Because the matrix is calcified nutrients can
.only reach osteocytes by diffusion via these bone canaliculi
 Osteocytes
Osteocytesare located in lacunae of bone matrix and
communicate with each other by means of processes via bone
canaliculi
 Osteoclasts
Osteoclasts are found inHowship’s lacunae at areas of
.bone resorption. Osteoclasts originate from monocytes

Osteoclasts are multinuclear, highly acidophilic cells. They secrete

lysosomal enzymes, which are acidic and lead to the breakdown and
.erosion of bone matrix
Osteoclasts
 Osteogenesis: woven bone
• The first bone to develop is woven bone
)immature bone, primary bone). This a
temporary form of bone in which the
lacunae are disordered and the matrix has
thick, irregularly-arranged collagen fibers
• Over the age of 14 woven bone is rare
and only found in sutures of flat bones,
tooth sockets, some tendon insertions and
temporarily during fracture repair.
 Woven bone
• Woven bone (b) is
temporary and is
rapidly remodelled
into lamellar bone (a)

IB , immature boneMB, mature bone

 Lamellar bone
There are 3 lamellar
arrangements in
compact bone:
• Osteons (Haversian
Systems)
• Circumferential
lamella (outer, inner)
• Interstitial systems

The periosteum is attached to

compact bone by Sharpey’s fibers
((perforating fibers
 )Osteons )Haversian systems
Osteons constitute the
morphofunctional units of
compact bone. They
consist of 4-20 concentric
lamellae surrounding a
central vascular Haversian
canal. Cement lines
surround each osteon.
The interstitial systems (2)
are remains of earlier
osteons after remodelling.

Interstitial System
(Osteons (Haversian systems
Osteons in compact bone
 Lamellar arrangements in compact bone

Osteons Outer circumferential

lamellae
Inner
circumferential
lamellae

Endosteum
(Lamellar arrangements (diaphysis of tibia
Bone lamellae: polarization microscopy

Alternate lamella are seen as bright when bone is examined using

polarization microscopy. This anisotropy )birefringence) results from the
orderly arrangement of the collagen fibers in secondary bone and their
alternating direction in adjacent lamella.
Ansiotropy of lamella
Anisotropy of lamella
Labelling bone growth

Tetracycline is incorporated into developing bone

matrix and can be used as a marker to determine rate
of lamellar formation. Tetracycline-labelling can be
.seen using fluorescence microscopy
 Remodelling of bone

During remodelling of bone there is erosion of osteons by osteoclasts

that results in connecting resorption cavities from adjacent osteons.
When sufficient resorption has occurred, osteoblasts appear in the
resorption cavities and start building a new generation of osteons.
The remnants of earlier osteons are seen as interstitial systems.
 Interstitial systems

Interstitial systems represent remains of osteons following remodelling

 Blood supply of compact bone

Blood supply to
compact bone is from
vessels in the
perichondrium leading
to transverse vessels
(Volkmann’s canals)
between the osteons
that connect with the
longitudinal
Haversian canals.
 Blood supply to osteons of compact bone

Haversian canal in center of osteon Volkmann canal )transverse) connects

periosteal blood with osteons
 Trabecular bone

Trabecular )spongy)
bone is also lamellar.
Unlike compact bone
the lamellae do not
show a concentric
arrangement.
 Ossification
Ossification is the process of bone
formation. There are two types of
ossification:
• Intramembranous ossification
(e.g. in flat bones)
• Endochondral ossification
(e.g. in long bones)
Intramembranous ossification

Flat bones, such as those of the calvarium, develop by

intramembranous ossification directly from connective tissue
Intramembranous ossification

Early stages Advanced stages

Intramembranous ossification

mesenchyme ,2
blood vessel ,3 Developing calvarium in embryo
bone matrix ,4
osteoblasts ,5
osteoclast ,6
 Osteoblasts, osteoid and matrix vesicles

osteoblast

Matrix vesicles

osteoid Calcified matrix Thematrix secreted by osteoblasts is

at first non-calcified (osteoid or
prebone). Processes of osteoblasts are
nipped off to form membrane-bound
matrix vesicles. Calcification processes
occur on these matrix vesicles resulting in
deposition of hydroxyapatite crystals
. andmineralization of the matrix
 Endochondral Ossification

Endochondral ossification involves an

initial model of cartilage which provides
the basis for subsequent development of
bone tissue. This process is best
illustrated in long bones. Initially
endochondral ossification occurs at the
growth plate and primary center of
ossification of the diaphysis. At later
stages the process also occurs in the
epiphyses.
Endochondral ossification
Endochondral ossification
Endochondral ossification
 Growth plate of long bone
Resting zone

Zone of
proliferation

Zone of
hypertrophy

Zone of
calcification
and primary
spongiosa
formation

The cartilage matrix is stained with alcian blue. The bone develops on
the remains of the calcified cartilage.
Growth processes in long bones
Elongation in long bones
)Synovial joints )diarthroses
Diarthrosis
Synovial joint
 Synovial joint

a , capsular ligamentb , adipose tissuec, synovial membrane

 Synovial membrane

Two types of cells are found lining synovial joints: fibroblast-like cells
and macrophages
 Dynamic processes in bone

Activation Resorption Reversal Formation Bone multicellular unit (BMU)

Bone is continually undergoing change and the dynamic remodelling

processes involve coordination between several bone types. These are
sometimes described as bone multicellular units. These are regulated by
interaction of several endocrine factors
 Fracture repair

Repair of a fractured bone by formation of new bone through

proliferation of periosteal and endosteal cells
 Fracture repair
Bone heals more rapidly than cartilage because its blood supply is more
plentiful and there is rapid activation and turnover of bone cell types
 Growth factors and bone
• All stages of bone development and growth are
tightly regulated by a large complex system of
systemic and local growth factors.
• Homeostatic mechanisms of bone physiology
differ according age, gender and growth phase.
It is convenient to classify these stages as:
• Fetal
• Neonatal
• Juvenile
• Adult
• Senile
 Endocrine regulation of bone
Major systemic hormones involved in skeletal
growth and homeostasis include:
• Growth Hormone
• Insulin
• IGFs
• Glucocorticoids
• Thyroid Hormones )T3, T4)
• Androgens
• Estrogens
• Calcium-regulating hormones
)Parathyroid hormone, Calcitonin, Vitamin D3)
Published March 2000
pp, 110 Illustrations 512
 Systemic and local growth factors
• Normal skeletal growth results from a balance
between the processes of bone matrix synthesis
and resorption. These activities are regulated by
both systemic and local factors.
• Bone turnover is dynamic, and skeletal growth
must be maintained throughout life.
• Although many growth promoters are associated
with bone matrix, it is enriched particularly with
transforming growth factor beta (TGF-beta)
activity.
TGF-beta as a skeletal growth factor
• Experimental evidence indicates that TGF-beta
regulates replication and differentiation of
mesenchymal precursor cells, chondrocytes,
osteoblasts, and osteoclasts.
• Recent studies further suggest that TGF-beta
activity in skeletal tissue may be controlled at
multiple levels by other local and systemic
agents.
• The intricate mechanisms by which TGF-beta
regulates bone formation are likely to be
fundamental to understanding the processes of
skeletal growth during development,
maintenance of bone mass in adult life, and
healing subsequent to bone fracture.
 Growth factors and bone

Recent studies have indicated a wide range of

bone regulatory local growth factors including:
• Insulin-like growth factors (IGFs)
• Fibroblast growth factors (FGFs)
• Transforming growth factors (TGFs)
• Bone morphogenetic protein (BNP)
• IGF-binding proteins (IGFBPs)
Possible regulation and effects of bone-related growth factors