Bone

Contents
• Introduction
• Definitions & classification
• Bone development
• Physiologic functions of bone
• Macroscopic features
• Microscopic features
• Composition
• Bone remodeling
• Factors regulating bone formation & resorption
• Regulation of bone by systemic hormones
• Sites of Haematopoiesis
• Estimation of age
• Conclusion
Introduction :

Bone is a connective tissue element comprising of

cells & an intercellular matrix, the greater majority of
its cell being the osteocytes. The matrix is composed
in part of organic materials mainly collagen fibers &
the rest made of inorganic salts rich in calcium &
phosphate. Together these give bone its unique
mechanical properties.

Remarkably strong but light weight, bone is a

dynamic, ever changing tissue. Throughout life which
is continuously being broken down & reformed.
Classification & Terminology:
1. Macroscopic appearance of cut surfaces:
a) compact bone: the ivory surface layer of mature bone.
b) trabecular bone/ spongy bone: the interior of mature bone.
2. Developmental origin:
a) intramembranous(mesenchymal/dermal): direct transformation of
condensed mesenchyme.
b) intracartilagenous (cartilage/ endochondral bone): replacing a
preformed cartilage model.
3. Type of osteon( haversian system):
a) primary osteon: first formed, lamellar or non lamellar osteon.
b) secondary osteon: concentric lamellae around vascular canals of
mature bone.
4. Disposition of lamellae:
a) circumferential lamellae: parallel to both periosteal & endosteal
surface.
b) osteonic lamellae: concentric lamellae around canals of mature bone.
c) interstitial lamellae: between osteons.
5. General microstructure:
a) non-lamellar bone: includes early woven bone & primary osteon.
b) lamellar bone: almost all mature bone.
6. General terms:
a) surface bone: circumferential lamellae but may include woven &
bundle bone (sharpeys/ extrinsic fiber)
b) interstitial bone: between osteon, often lamellar remnants of secondary
osteon but may include woven or primary osteon fragments.
Calcification: a process in which tissue or noncellular
material in the body becomes hardened as the result of
precipitates or large deposits of insoluble salts of
calcium especially calcium carbonate & phosphate.

Ossification: formation of bone substance or the

conversion of other tissue into bone.

Mineralization: normal or abnormal deposition of

minerals in the tissues.
 Main characteristics of bone
Bone is a dynamic material
• Self repair
• Changes with aging : becomes stiffer and less ductile
• Changes with immobilisation : becomes weaker
Functions of bone in general:
1. Support
2. Protection
3. Movement
4. Mineral homeostasis
5. Site of blood cell production
6. Storage of energy
 Structure of bone:

Articular cartilage
Epiphysis
Metaphysis

Endosteum

Diaphysis
Periosteum

Medullary cavity
Structure of mature bone:
1. Macroscopic feature:

a) Compact bone:
• It forms the external layer of all bone of the body &
the bulk of the diaphysis of long bone.
• They provide protection & support & resists the
stress of weight placed on them.
b) Spongy bone tissue:

In contrast to compact bone spongy bone does not

contain true osteons. It consists of lamellae arranged in
an irregular lattice work of thin plates of bone called
trabeculae
• Spongy bone makes up most of the bone tissue of
short, flat & irregular shaped bone & most of the
ephiphysis of long bone.
• Osteons are not usually present in the spongy bone
because osteocytes are not deeply buried & have
access to nutrients directly from the blood.
Microscopic tissue types:
1. Woven bone:
- highly cellular osseous tissue that is formed rapidly.
- low mineral content, more random fiber orientation
& minimal strength

2.Lamellar bone:
- principal load bearing tissue of the adult skeleton.
- predominant component of mature cortical &
trabecular bone .
- formed slowly, highly organized matrix & densely
mineralized.
3. Bundle bone:
- characteristic of ligament & tendon attachment
along bone forming surfaces.
- striations are extensions of sharpeys fibers
composed of collagen bundles from adjecent
connective tissue that insert directly into bone
4. Composite bone:
- lamellar bone deposited on a woven bone matrix.
- During growth woven bone lattice grows out &
captures blood vessels along an endosteal or periosteal
surface.
- then fills paravascular space with high quality
lamellae, resulting in composite bone of high strength.
2. Microscopic structure of bone:
a) Cells of bone:
the cells of bone are of number of types ,
• Osteoprogenitor stromal cells ( give rise to various other forms of
bone cells)
• Osteoblasts ( lay down bone)
• Osteocytes (mature bone cells )
• Bone lining cells ( on surface of quiescent bone)
• Osteoclasts ( erodes the bone)
i) Osteoprogenitor cells:

• Derived from plueripotent stromal stem cells

(mesenchymal origin) which can proliferate &
differentiate into osteoblasts prior to bone
formation.
• Stromal stem cells resemble young fibroblasts are
responsible for bone formation during early
development.
• two types of osteoprogenitor cells,
- one totally committed to bone formation (found within bone).
- The other ( inducible osteoprogenitors) widely present in connective
tissue & are able to differentiate into various connective tissue cells (eg:
fibroblasts, myoblasts, pericytes, adipose cells, chondroblasts, osteoblast)
depending on nature of inducer.
ii) Osteoblasts :

• Are basophilic, roughly cuboidal mononuclear

cells about 15-30 µm across.
• Ultra structurally, osteoblasts have feature typical of
protein secreting cells i.e., a pale oval nucleus
placed away from the secreting surface, an
extensive granular ER, large golgi complex &
numerous secretary vesicles.
• Found on the forming surface of growing or
remodeling bone where they constitute a covering
monolayer, can also appear deep in compact bone
where remodeling is occurring.
• Osteoblasts are, differentiated nonmotile cells which
arise from osteoprogenitor cells & de differentiation of
osteocytes (released during resorption).

• The surface of osteoblasts are rich in alkaline

phosphatase activity, located at the plasma membrane
(detected in condition of rapid bone formation /
turnover).
• Major activity of osteoblast is,

- important role in mineralization of osteoid via

alkaline phosphatase

- to synthesize & secrete the organic matrix of bone

i.e. type I collage, small amount of type V collage &
various other macromolecules including osteonectin,
gamma carboxyglutamic acid which binds strongly to
mineral & collagen ( may act as cell adhesion factor)
- also considered to play an important though indirect
role in hormonal regulation of bone resorption since
they bear receptors for PTH & 1 -25,(OH)2 D3 &
other stimulants of bone resorption.
iii) Osteocytes:
• Constitutes the major cell type of mature bone, lying scattered within its
matrix & interconnected by numerous cellular extension to form a
complex cellular network.
• derived from osteoblasts which have reduced or ceased matrix formation
& become enclosed in matrix but retaining contact with each other &
cells at surface of bone throughout
the life span.
• A mature osteocyte is ellipsoidal in shape, basophilic cytoplasm
containing few organelles (little granular ER, a few free ribosomes small
golgi apparatus & a narrow rim of cytoplasm surrounding an oval
nucleus).
• Average life span of an osteocyte varies with the metabolic activity of
the bone & the likelihood that it will remodel.
• Exact functions unclear, however widely assumed to play an essential
role in maintenance of bone & their death leads to resorption of the
matrix by osteoclastic activity.
iv) Bone lining cells:
• Are flattened epithelium like cells, particularly evident
in the adult skeleton found on resting surface of bone
• They form continuous layers & are in contact with
neighboring osteocytes via gap junctions.
• Their exact role is unclear but they may have the
capacity to revert back to active state when stimulated
suitably.
• They may play an important role in regulating the
differentiation of osteoprogenitor cells, control the
access of osteoclast to bone surface & regulate mineral
homeostasis.
v) Osteoclasts:
• Are large polymorphous cells with an variable number
of oval closely packed nuclei often 15-20 or more.
• Found at sites of active removal of bone & lie in close
contact with the bone surface in pits , resorption bays
or lacunae of howships.
• Although exact mechanism of removal of bone is not
known, but thought
- to cause demineralization by proton release
- organic matrix destruction by lysosomal & non
lysosomal enzymes
• Various factors such as PTH & 1-25, vit D3 are also
involved in bone resorption.
• Their life span of an osteoclast is estimated to be 7
weeks, however knowledge of osteoclast longevity in
humans of different ages is scanty.
• the sequence of resorption events is considered as
follows,
- attachment of osteoclast to the mineralized surface of
bone
- creation of a scaled acidic microenvironment through
action of proton pump which demeneralizes bone &
exposes the organic matrix.
- Degradation of the exposed matrix by the action of
released enzymes such as acid phosphatase &
cathepsin B.
• Endocytosis at the ruffled border by inorganic &
organic bone degradation products.
• Translocation of degradation products in transport
vesicles & extracellular release along the membrane
opposite the ruffled border.
• The organic part of matrix is mainly Type I collagen.
This is made intracellularly as tropocollagen and then
exported. It then associates into fibrils.
• Also making up the organic part of matrix include
various growth factors, the functions of which are not
fully known.
• Other factors present include glycosaminoglycans,
osteocalcin, osteonectin, bone sialo protein and Cell
Attachment Factor. One of the main things that
distinguishes the matrix of a bone from that of another
cell is that the matrix in bone is hard.
Bone development:
Bone formation occurs by three main
mechanism
a) Endochondral bone formation
b) Intra membranous bone formation
c) Sutural bone growth
Endochondral bone formation
mesenchymal cells differentiates into chondroblasts,
produce cartilage matrix model of hyaline cartilage

Increase in length (interstitial growth) & thickness

(appositional growth)

chondrocytes in the mid region hypertrophy, accumulate

glycogen for energy & produce enzymes to catalyze chemical
reaction, which trigger calcification
 once cartilage gets calcified

nutrients required for cartilage cells no longer diffuse

leading to death of cartilage cells

lacunae of the cells killed are empty

a nutrient artery penetrates the perichondrium & along with it

the mesenchymal cells accompany which differentiates into
osteoblasts & begin to deposit osteoid

near the middle of the model capillaries of the periosteum

grow into distintegrating calified cartilage the vessels associated
with osteoblasts, osteoclasts & red marrow cells are known as
periosteal bud (primary ossification center).
 Secondary ossification center develops around the time of
birth, when the epiphyseal arteries enter the epiphyses.
different from primary ossification center,
- spongy bone retained in interior of the ephiphyses
- no medullary cavities are formed
- hyaline cartilage remains covering the epiphyses as
articular cartilage & between epiphyses & diaphysis as
epiphyseal plate
Endochondral bone formation
 Close to the zone of ossification, the cartilage can
usually be divided into a number of distinct zones:
• Reserve cartilage, furthest away from the zone of
ossification, looks like immature hyaline cartilage.
• A zone of chondrocyte proliferation contains
longitudinal columns of mitotically active
chondrocytes, which grow in size in
• the zone of cartilage maturation and hypertrophy.
• A zone of cartilage calcification forms the border
between cartilage and the zone of bone deposition.
Intramembranous ossification:
mesenchymal cells become vascularized, cluster
& differentiates into osteoprogenitor cell
then into osteoblasts
( center of ossification )

osteoblasts secrete the organic matrix of bone

stop secretion once completely surrounded by the matrix,

cells now are called the osteocytes

later calcium & mineral salts are deposited

on outside of bone, vascularized mesenchyme

develops into periosteum
Sutural bone growth:

• Sutures play an important role in the growing face &

skull.
• Understanding the structure of a suture is based on the
knowledge of the periosteum of bone which consists of
two layers an outer fibrous & an inner cellular layer.
• At the suture the outer layer splits, the outermost leaf
run across the gap of the suture to form a uniting layer
with the outermost leaf from the outer side.
• The inner most leaf together with the osteogenic layer
of the periosteum runs through the suture along with
the corresponding layer from the outer bone.
• The osteogenic layer of the suture called, the cambium
& the inner leaf, the capsule.
• Between the two layers is a loose cellular & vascular
tissue.
• Suture are best regarded as having the same osteogenic
potential as periosteum. When two bones are
separated, for example the skull bones forced apart by
growing brain, bone forms at sutural margins with
successive waves of new bone cells differentiating
from the cambium.
Bone remodelling:
 Modeling
1. The process by which the
overall size & shape of the
bone is established is
referred to as bone
modeling, which extends
from embryonic bone
development to preadult
period of human growth.
2. Formation & growth of
bone in childhood &
adolescence.
3. Primarily of processes at
the periosteum &
endosteum
4. Spatially related & proceed
in uncoupled manner
Remodeling
1. The replacement of old bone
by new bone tissue is called
remodeling.
2. Remodeling usually start once
the adult bone has been
formed
3. It can occur at any region no
specific site.
4. Remodeling is cyclical &
usually covers only a small
area.
• The current concept of bone remodeling is based on
the hypothesis that osteclastic precursors become
activated & differentiates into osteoclasts which begin
the process of bone resorption which is followed by a
bone –formation phase or activation frequency is
determines the rate of tissue turnover.

• The termination of bone resorption & initiation of

bone formation in the resorption lacunae occurs
through a coupling mechanism.
• The coupling process ensures that the amount of bone
removed is similar to the amount of bone laid down
during the subsequent bone formation phase.

• Nature of activation & coupling mechanism is still

unknown, although some growth factors, such as
various lymphokines, fibroblast growth factor beta ,
PG have been proposed.
• As the osteoclasts move through bone, the leading
edge of resorption is termed “ cutting cone” which is
characterized in cross section by scalloped array of
howships lacunae each housing osteoclasts.

• behind the cutting cone is a migration of

mononucleated cells on the roughened cylinder, as
these cells differentiate into osteoblasts,
• they produce coating termed the cement or reversal
line, thin layer of glycoprotien comprising of
sialoprotein & osteopontin that acts as a cohesive,
mineralized layer between the old bone & the new
bone to be secreted
• On top of the cement line osteoblasts begin to lay
down new bone matrix & mineralizing it from outside
in.
• the entire area of the osteon where active formation
occurs is termed the filling cone

• once formation is complete, haversian canal contains a

central blood vessel & a layer of inactive osteoblasts,
the lining cells that communicate by means of cell
processes with the embedded osteocytes.
Factors regulating bone formation:
it involves two major steps ;
a) The production of new organic matrix by osteoblast
b) Mineralization of that matrix
agents that regulate bone formation act on the
osteoblast to either increase or decrease replication
of cell in the osteoblastic lineage or to modify the
differentiated function of the osteoblast.
 The local regulators of bone formation are growth
factors that act directly on cells of osteoblastic
lineage.

Systemic hormones can regulate growth factors

activity by one or more of 4 different mechanism,
i) By regulating factors synthesis, as well as release
from cells
ii) If factors are released in a latent form, its
physiologic effect can be regulated by controlling ,
when the factor become activated.
iii) By regulating receptors binding
iv) By regulating the production of the binding protein
that stabilizes the factors & promotes its binding to
receptors.
Growth factors that regulate bone formation have some common features:
- They are polypeptides
- exert their activity by binding to specific receptors on cell surface
- they primarily act locally
- they natural products
- they are multi functional in that they can stimulate a wide variety of
cellular activity
a) Platelet derived growth factor:
- is cationic, heparin binding polypeptide with a
molecular weight of approximately 30,000d.
- 3 isoforms of PDGF exists: PDGF-AA, PDGF-AB,
PDGF-BB
- PDGF-BB is the most biologically active produced
by osteoblasts but much of the PDGF found in bone
is derived from serum & platelets.
functions:
- stimulates DNA synthesis & cell replication in
osteoblasts
- increases bone collagen synthesis & the rate of
matrix apposition
- also reported to increase bone resorption &
collagen degradation
b) Heparin binding growth factor:
- acidic fibroblasts growth factor & basic fibroblast
growth are the two better known forms of HBGF &
were the first to be purified, sequenced & cloned.

- bone matrix is rich in FGF

- in bone cells, FGF interacts with heparin which
enhances the effect of FGF on bone cell replication.
but their specific function in bone cell
biology & their role as therapeutic agents require
further study.
c) Insulin like growth factor (IGF)
are non glycosylated polypeptide with molecular
weight of approximately 7,500d
IGF are of two types IGF-1
IGF-2

• IGF-1 , principle growth regulator in bone & cartilage.

Produced mainly in liver
• Its systemic role unclear
• In connective tissue, IGF-1& IGF-2 are among the most
abundant growth factor present & synthesized by most cell
type in skeleton tissue including fibroblast & osteoblasts.
• Its production is increased by growth hormone,
estradiol & PGE2, where as its production inhibited by
cortisol.

• Act by- increasing preosteoblastic cell replication ,

- effect on osteoblastic collagen synthesis &
- bone matrix apposition &
- decreases the degradation of collagen & also
maintenance of bone mass.
d) Transforming growth factor β:
- molecular weight of 25,000 d synthesized by
skeletal matrix
- are polypeptide isolated from a variety of normal
& malignant cells.
- several isoforms identified with activities similar
to TGF –bone morphogenetic proteins
the activins
the inhibins.
- TGF β 1, found in the proliferating cell zone & pre
hypertrophic cell zone.
- TGF β 2, distributed through out the growth plates
& highest level in hypertrophic & calcifying cartilage.
- TGF β 3, is in proliferating cell zone &
hypertrophic cell zone most commonly found in
intramembranous bone.
e) Bone morphogenetic proteins:
- during repair & bone matrix destruction many
growth factors are released that affects the healing
process, one of them is BMP is a factor with
osteoinductive activity
- its implementation at ectopic sites as well as in
bone, induces production of new bone through an
endochondral pathway.
actions on bone cells:
- direct effect on osteoblasts
- stimulation of osteoblasts precursor cells into more
mature osteoblasts
- stimulates collagen production by mature
osteoblasts
- via endochondral pathway, induces chondrocyte
differentiation & matrix mineralization.
Actors regulating bone resorption:

1. Interleukin 1:
- it’s a powerful & potent bone resorbing cytokines.
IL-1 α & IL-1 β are equally potent in stimulating
bone resorption.
- they act by – stimulating proliferation of precursor
cells
- act indirectly on mature cells to stimulate bone
resorption.
- effect of IL-1 probably occurs by two mechanism,
a) stimulation of production & release of PGE2
which in turn causes resorption
b) direct action of IL-1 on the osteoclast, which is
independent of PG synthesis.
- in transient exposure IL-1 has been shown to
stimulate bone formation by osteoblasts.
2. Tumor necrosis factor & lymphotoxin:
- are two closely related cytokines that have similar
effects on bone cells
- they stimulate multifunctional cytokines produced
by activated lymphocytes
- the effect of TNF is mediated by PGE2 & also by
IL -6
- also effects cells with osteoblastic phenotype,
stimulating differentiated function & stimulating
cell proliferation.
3. Interleukin-6:
- it stimulates bone resorption
- they are responsible for formation of cell with an
osteoclastic phenotype

4. Gamma interferon :
- is an multifunctional cytokine i.e. most biologic
system has effects similar to those of TNF or IL-1
- its more effective in inhibiting IL-1beta or TNF
induced bone resorption than are systemic
hormones like PTH or vit D3.
5. Colony stimulating factor:
- they stimulate the differentiation of osteoclast
precursor into mature osteoclast.
- CSF s are found to be associated with tumor like
granulocytosis.
- Tumors , hypercalcemia are associated with
increased bone resorption
6) Prostaglandins & other arachidonic acid
metabolites:

- PGEs are slow acting but powerful mediators of

bone resorption & they affect both active mature
osteoclasts & differentiated osteoclast precursors.
- it mediates effects of other factors such as TGF-β

- PGE is produced by osteoblasts & affects not just

bone resorption but bone formation as well

- in vivo & in vitro studies shown that high levels

lead to resorption while low levels of these PGE
stimulate bone formation.
Regulation of bone by systemic hormones:

The Role of Parathyroid Hormone in Normal Calcium

Homeostasis:

• Major calcium-sensing glands in the body

•Secrete parathyroid hormone in response to low
circulating calcium
•Parathyroid hormone increases serum calcium
by:
1) Promoting bone resorption
2) Diminishing urinary calcium excretion
3) Indirectly promoting increased dietary calcium
absorption via increased renal production
of1,25(OH)2D
Role of 1, 25- dihydroxy vitamin D3:
• 1,25-(OH)2 & 24,25(OH)2 D3 are active metabolites
that have shown to directly affect bone cells, catabolic
& anabolic effects respectively.
• Both hormones promote absorption of Ca& P from the
gut.
• Effects on osteoblasts –to decrease cell proliferation
& stimulate osteoblast differentiation.
• excessive vit D3 , increases both osteoclast number &
an increase in ruffled border size & clear zone
volume.
Calcitonin:
• The effect of calcitonin in bone resorption are short
lived. However osteoclasts eventually lose their
responsiveness to calcitonin after continued exposure.
• Possible explanation for this phenomenon involves a
decrease in receptor number after long period of
exposure. Or emergence of a second population of
osteoclasts i.e. not responsive to calcitonin.
• Believed to inhibit bone resorption transiently when
bone turn over is not needed for homeostasis
Estrogen :
Inhibits the increase in bone resorption associated
with menopause.
Following estrogen withdrawal , an initial increase in
bone turnover can be observed , but later resorption
occurs faster than bone formation.
effects of estrogen on bone can be mediated both
directly by the action of hormone on bone cells &
indirectly by action of growth hormones (IGF-1&
TGF-β)
ALVEOLAR BONE:
…is the portion of the maxilla & mandible that forms &
supports the tooth sockets. It forms when tooth erupts to
provide attachment to the forming periodontal ligament, it
disappears gradually after the tooth is lost.

It comprises of the following

- External plate of cortical bone formed by haversian bone
& compact bone lamellae.
- Inner socket wall thin , compact bone called the alveolar
bone proper/cribriform plate /lamina dura in radiographs.
- Between the compact bone layers , the interdental septum
consists of cancellous supporting bone.
Lability of alveolar bone:
A balance between bone formation & resorption
maintains physiologic lability.
Cellular activity manifest in 3 areas
- Adjacent to PDL
- Periosteum of facial & lingual plates
- Endosteal surface of marrow spaces,
socket wall reflects responsiveness to occlusal
forces- trabeulae. Increased occlusal forces trabeculae
increased in number & thickness of bone added to
labial & lingual plates.
decreased occlusal forces , bone resorbed, bone
height diminished
Misch bone density classification:
D1 - dense cortical bone
D2 - thick dense to porous cortical bone on crest
&coarse trabecular bone within.
D3 - thin porous cortical bone on crest & fine trabecular
bone with in
D4 - fine trabecular bone
D5 - immature nonmineralized bone
Estimation of age :

The age at which the epiphysis of bone appears &

then fuses with the diaphysis provides the estimation
of age. Thus the ossification of bones is helpful in
determination of age till 25yrs of age.
above 25yrs of age the estimation of age is possible
by studying closure of cranial sutures.
Haematopoiesis:
In human embryo, the yolk sac is the main site of
haematopoiesis in the first few weeks of gestation.
Haematopoiesis commences in the bone marrow by
about 4th & 5th month & becomes fully active by 7th &
8th month so that at birth all the bones contain active
marrow. During normal childhood & adult life, the
marrow forms the source of new blood cells.
However there is progressive fatty replacement
throughout long bone so that by adult life the
haemotopoietic marrow is confined to central skeleton
(vertebrae, sternum, ribs, skull &pelvis)&proximal
ends of femur, tibia & humurus.
Conclusion

It is essential for a contemporary clinician to have

a firm grasp of the modern concepts of bone
physiology, metabolism & biomechanics. This is
often the only objective means for designing and
implementing an innovative treatment plan
References:

1. Principles of anatomy & physiology. Tortora

Gerard
2. Gray’s Anatomy. Peter L William & et al
3. Physiology. Robert M.R
4. Carranza’s Clinical periodontology.
5. Oral histology. Ten cate
6. Clinical periodontology and implant dentistry.
J.Lindhe