THE ROLE OF BIOLOGY IN

ORTHODONTICS &
THE EVOLUTION OF
BIOLOGICALCONCEPTS IN
ORTHODONTICS.
 CONTENTS
INTRODUCTION
TOOTH SUPPORTING STRUCTURES
CELL BIOLOGY
TISSUE RESPONSE TO ORTHODONTIC
FORCE
CHEMICAL MEDIATORS
SECOND MESSENGERS
THIRD MESSENGERS &

DEVOLOPMENT OF BIOLOGICAL CONCEPTS IN

ORTHODONTICS
 INTRODUCTION
Orthodontic tooth movement results from forces
applied to the teeth that evokes cellular responses
in the teeth and their surrounding tissues
including PDL alveolar bone and gingiva.

These are series of biological cell occurrences that

turn biomechanical signals into chemical and
electrical messages to which cell responses.
It is advantageous for the orthodontist to
know the details of the biological events that
unfold during tooth movement, because
some of these details may differ from one
person to another, due to variables such as
gender, age, psychological status, nutritional
habits, and drug consumption. .
 Biological variations may be the foundation of the
differences that are frequently observed in the
outcomes of orthodontic treatment between
patients with similar malocclusions, treated
identically. The movements result from the
development of strains in dental and paradental
tissues, followed by modeling and remodeling of

these tissues .
 In some patients systemic conditions may exist,
evoking complications such as root resorption,
dehiscence and fenestrations of the alveolar bone.
Hence ,clinical orthodontics must be viewed as a
specialty staunchly entrenched in biology, all the
way to the molecular level.

As a clinical profession, it must be based on a

commanding knowledge of mechanics, biology,
physiology, and pathology.
 TOOTH SUPPORTING
STRUCTURES
PDL
1. IT IS A DENSE FIBROUS CONNECTIVE
TISSUE THAT OCCUPIES SPACE BETWEEN
ROOTS OF TOOTH AND ALVEOLUS.
2. THE MOST IMPORTANT ELEMENTS ARE
THE PRINCIPAL FIBRES, THAT ARE MORE
OF COLLAGEN TYPE-I AND LESS OF TYPE-
III
PROTEOGLYCANS
GROUND SUBSTANCE GLYCOS AMINO GLYCANS
GLYCOPROTEIN
 CELLS OF PDL

A-CEMENTOBLAST LINING THE CEMENTUM B-FIBROBLAST

B-OSTEOBLAST LINING THE BONE
CELLS OF PDL
FIBROBLAST
CEMENTOBLAST
CEMENTOCLAST
OSTEOBLAST
OSTEOCLAST
OTHER CELLS ( UNDIFFERENTIATED
MESENCHYMAL CELLS, DEFENCE
CELLS AND EPITHELIAL CELLS)
ALVEOLAR BONE

A-A thin layer of compact

bone lines the socket, and
gives attachment to some
of the principle fibres of
PDL.
B-Thicker layer of
compact bone forming
the external and internal
alveolar plates. Between
these plates of compact
bone are variable
amounts of spongy bone.
C-Represents arbitrary
boundary between
alveolar bone and jaw.
 ALVEOLAR BONE
1. MINERALISED CONNECTIVE TISSUE
2. 60% INORGANIC MATTER
3. 25% ORGANIC MATTER
4. 10% WATER
5. ABOUT 90% OF ORGANIC MATTER
CONSISTS OF TYPE-I COLLAGEN
 Cell Types in bone
Osteoblasts

Osteoclasts

Osteocytes

Osteoprogenitor cells

Bone lining Cells

 Osteoblasts

A layer of osteoblasts (arrowed) lining the

alveolar bone.
 Osteoblast, which are of mesenchymal
origin are the bone forming cells.
Many factors are shown to influence the
development of osteoblast from
mesenchymal pluripotent progenitors.
Certain growth factors like BMPs, gene
transcription factors such as core binding
factor alpha-1 etc.
Osteoblast synthesize and secrete, the extra
cellular organic matrix of bone including
type I collagen, osteoclacin, osteopontin,
osteonectin, alkalinephosphatase,
proteoglycans, growth factors.
 A recent advancement have been
identification of core binding factor
alpha I (Cbfa I ) gene during
osteoblast development and
function. Cbfa I is a transcription
factor expressed by the cells of
osteoblastic lineage and is necessary
for osteoblast differentiation.
 Osteoclasts

The surface of the alveolar bone shows resorption concavities

(Howships lacunae), in which lie the osteoclasts (arrowed).
 The osteoclasts which differentiate from
heamopoetic cells. Many chemical mediators
of macrophage family are known to influence
osteoclast differentiation, they are
Cytokines Ex: tumor necrosis
factor,interleukin-1 alpha & IL-6 alpha
Growth factors Ex: macrophage colony
stimulating factor, granulocyte, macrophage
colony stimulating factor & Prostaglandins
Osteoblasts itself seems to regulate the
differentiation of osteoclasts. OPG gene
secreted by osteoblasts and its ligand OPG-L
seems to moderate osteoclastic activity.
 An another molecule osteopotegerin
(OPG) and its binding ligand (OPGL)
as an essential regulators of
osteoclast formation and activity has
been discovered. OPG secreted by
osteoblast functions to block the
formation of osteoclast.
 The other cells include
a. Osteocytes which are trapped
osteoblassts in its own matrix.
b. Osteoprogenitor cells Are mesenchymal
fibroblasts like cells which are regarded
as stem cells to generate osteoblasts.
c. Bone lining cells Are the
undifferentiated flattened cells lining the
bone surface. They may represent active
osteoblasts.
 Cell Biology
When force system is applied to it. The first
structure of note is the cell membrane. This
lipid bilayer "double soap bubble skin"
separates the vast extra cellular space
outside of the cell from the closed
compartmentalized and highly organized
cytosol. Extending from this cell membrane
are a variety of discrete structures that
permit some sophisticated forms of
communication across this barrier. These
include mechanogated ion channels, ligands,
receptor sites, oligosaccharide chains,
surface globular proteins, cholesterol
molecules and miscellaneous protein in the
shape of alpha helix and beta sheets.
All of these complex structures on or in the
cell membrane carry electrical charges and
form regions of polarity. They form a
heterogeneous forest of sites that act as
stereometric molecular keyholes or
receptacles. They can also be depicted as
antennae for communication of signals from
the outside to the inside of the cell. These
signals that affect the cell may take the form
of mechanical, electrical, chemical, or
physical (heat or light) perturbations. Once
they activate a receptor or group of
receptors, a stimulatory or inhibitory signal
may be activated on the membrane.
This receptor may directly affect an event
on the other side of the cell membrane
that is in contact with the cytosol.
Alternately, it may communicate with one
or more intermediary receptors before
making a product that is seen inside the
cell. A signal must be translated into a
biochemical/electrochemical language
that can be understood by the control
mechanisms on the inner surface of the
cell membrane
The primary purpose is to activate or
deactivate the process of phosphorylation
and a change in adenosine triphosphate
producing energy, the driving force of a
chemical reaction. An early by-product is
a second messenger molecule that
produces an internal affector molecule (s)
at a more distal level. Once the signal is
understood, energized, and the specific
command executed, there must be a
critical mass of affector molecules
produced at a sufficient concentration or
concentration gradient at critical sites to
produce a biochemical cascade of events.
This means that one molecule may processes a
tenfold by-product. This by-product, in turn,
produces an amplification of molecules that
induce the next stage of events. This continues
in a biochemical cycle stepwise fashion that
stops when a final product or event is
achieved. It is regulated by a biofeedback
mechanism. Ultimately, a critical mass of cell
products may be necessary to push events in a
given direction.
Keep in mind that during the course of normal
cell-to-cell interaction, a cell membrane
continuously receives intermittent stimuli
telling the cell to respond in all sorts of ways.
The accumulation of crucial molecules in a
biochemical critical mass drives the reaction to
a programmed completion, a pathway cycle
This could range from the synthesis of a
specific type of collagen molecule for a
repair of tissue, regeneration of bone, to the
alteration of the cytoskeleton proteins and
actin molecules leading to a cell movement.
The most important lesson from the
transcellular membrane physiology
previously discussed is that some signals
stimulate processes, whereas others inhibit
or control critical biochemicals that are part
of some event cascade. The pathway is an
external signal that affects a receptor. This
produces a signal that is transduced
(transmitted to another point in a usable or
translated fashion) and ultimately amplified
to critical concentrations in the cytosol.
 Based on the current understanding when
we apply orthodontic force to the tooth,
the following changes occur
1. There is development of tissue strain

2. There is movement of tissue fluids

3. Dilatation of blood vessels causes

decreased oxygen in the pressure side and
increased oxygen in the tension side
leading to the release of various chemical
mediators such as cytokines,
Prostaglandins etc.
4. Generation of piezo electricity (Bio
Electric potential)
5. Release of neurotransmitters from the
nerve endings of the damages tissues.
Advances signals to
Nucleus
Gene transcription factors released
Cfos mRNA, Cjun, egrI, AP-I, SP-I,
growth
differentiation factor 9B, Cytokinin,
 Chemical Mediators
Prostaglandins
Prostaglandins were introduced by Von Euler.
They are synthesizes from fatty acids by a
microsomal enzyme complex (PG Synthetase)
found in mammalian tissue. The most
abundant precursor of PG s is Arachidonic acid
which is present in membrane phospholipids.
Arachidonic acids can be released either by
phospholipidases activated by direct cellular
damage or by non destructive perturbation of
the membranes by physical or chemical or
neurohormonal stimuli.
 Prostaglandins can be termed as local
hormones functioning to co-ordinate effect of
those other hormones which induce
prostaglandin synthesis and prostaglandins
functioning through G-Protein linked
receptors to their cellular effect.
Classically PGs are one of the mediators of
inflammation cause an increase in
intracellular cyclic AMP and Calcium
accumulation by monocytic cells which then
modulate and activate osteoclastic activity.
Recent studies indicate PGs increase the
number of osteoclasts as well as stimulate
osteoblastic cell differentiation and new bone
formation.(Monomet et el.,)
 CYTOKINES & GROWTH FACTORS IN OTM
The early phase of OTM involves an acute
inflammatory response characterized by
periodontal vasodilatation and migration of
leukocytes out of PDL capillaries. The released
inflammatory mediators such as Prostaglandins
and interleukins (IL)1, Interact with bone cells.

Cytokines secreted by leukocytes may interact

directly with bone cells or indirectly, via
neighboring cells, such as
monocytes/macrophages, lymphocytes and
fibroblasts, through their production of cytokine,
or a variety of Growth factors. Cytokines released
have multiple activities, which include bone
remodeling, bone resorption, new bone deposition
 Growth Factors
Growth Factors are also released during
inflammation and repair by the cells of PDL
and bone. Another theory stated that the
growth factors may secreted by bone cells
and stored (bound to bound matrix). They
are likely to released and activated during
bone resorption. These Factors include :
Fibroblast Growth Factor (bFGF & a FGF ),
Insulin like Growth Factors (IGF I, IGF
II),
Transforming Growth Factor (TGF ),
Platelet Growth Factor (PDGFS),
Bone Morphogenic Proteins (BMP)
 How do cells detect the mechanical
signals?

Three theories has been suggested

1. Strain Released potential
Application of small bending force to
the bone produce flow of interstitial fluid and
generate streaming potential this leads to
the production of chemical mediators.
2. Activation of Ion Channels.
Ion channels are tunnel shaped proteins
that cross the width of the cell membrane as
well as membranes surrounding intercellular
organells. These are three types , Voltage
gated, Liagan Gated and stretch channels.
 EXTRACELLULAR MATRIX &
CYTOSKELETON REORGANIZATION.
The principle elements of ECM of either
PDL or the bone may be considered as
collagen fibrous network providing
structural support embedded in and
interacting with a non collagenous matrix
consist of proteoglycans & various
glycoprotein.
The macromolecules which make up the
ECM include collagen & glycose amino
glycans. These macro
molecules are secreted at local levels by
cells, particularly fibroblast in the PDL &
osteoblast in the bone.

The matrix metallo proteins (MMPS)

represents major class of enzymes
responsible for ECM metabolism. The
growth and repair of connective tissue is
delicately balanced process of ECM removal
and replacement, with significant control by
MMP & Tissue inhibitors of metallo
proteinases(TIMPS)
Cytoskeleton represents a frame work
attaching cell to cell or cell to extra cellular
matrix there by presenting a possibility of
transducing mechanical forces acting on the
cells or on their adjacent matrices.
Cytoskeleton has three main components,
Microtubules, Microfilaments and
Intermediate filament.
Cell membrane integral proteins are also
identified as cell surface receptors termed as
Integrins.
These intgerins mediate cell to cell
attachment or cell attachments to extra
cellular molecules such as Fibrinonectin, It
also binds with microfilaments of
cytoskeleton.
Integrins have been found to regulate
signaling pathways by changing intracellular
calcium regulating insitol lipid turnover and
phosphorylation of intracellular proteins.
Integrins bind to the osteoblasts and
osteoclasts at the receptor binding sites ie.,
RGD ( Arginin Glycenase Phosphate)
RGES (Arginin Glucinacid-Glutamic acid).
 Production Of Second Messengers.
The first messengers alter
activity through cell membrane and
receptor sites and message goes to the cell
through the cytoskeleton framework. This
leads to increase in intercellular levels of
second messengers ie.,CAMP, CGMP, IP3,
Ca+ and K+.
This is demonstrated by Davidovitch. 1973,
74, 76.
 The intramembranous components that
have been shown to play a vital role in
production of CAMP are Ca+ ions and
cell membrane enzymes. CAMP is
known to activate Protein Kinase A, an
enzymes responsible for protein
phosphorelation. CAMP response is also
modulated by Prostaglandins. The
response of increase in intracellular
CAMP levels can be inhibited by
Ibuprofen (many other NSAIDS), a drug
commonly used as pain killer.
Production of Inositol phosphate, another
second messenger which was first
investigated by Hokin and Hokin, is similar
in its generation of CAMP. Inositol phosphate
in presence of phospholipase gets converted
to phosphotidylinositol biphosphate (IP2).
Phospolipase then cleaves IP2 into Inositol
Triphosphate (IP3) & Diacyl glycerol with
subsequent release of calcium from
intracellular stores. The calcium thus
released is responsible for Protein
Phosphorylation in nucleus. The Diglycerol
formed activates protein kinase C. This
protein Kinase C, is an enzyme responsible
again for Protein phosphorylation.
 Next how the nucleus responds for the
second messengers. Second messengers
send signals to the nuclei through a series
of kinases. In the nucleus different second
messengers account for differential
patterns, immediate early gene
expression ,IGE or also termed as third
messenger. This transcription of IGEs ie.,C
fox gene, C jun gene have shown to be
influenced when exposed to cytokines.
 Development of Biological
Concepts in Orthodontics
Recognition of malocclusions and variability
in features of morphology and function was
first noted in the ancient world by Hippocrates
(460-337 B.C) and Aristotle
(384-322B.C)
Hippocrates was among the first comment
about craniofacial deformity
Among those individuals with long shaped
heads, some have thick necks, strong parts
and bones, others have strongly arched palate,
their teeth are irregularly arranged crowding
one another and they are bothered by
headaches and ottrohea.
 Clsus was the first to recommend moving
of teeth by application of finger pressure.
Cajus Plinisns Secundus (23-79 A.D)
recommended tooth extraction for
orthodontic reasons.
Adamondos (5 centurs A.D) noted that
Those persons whose lips are pushed out
because of cuspid diplacement are ill
tempered abusive shouters and defamers.
Crude appliances that seemingly were
designed to regulate the teeth have been
found as archaelogical artifacts in tombs
of ancient Egypt,Greece and Mexico.
 Orthodontics as we think of it today however probably
has its roots in France in the 18 century when Pierre
Fauchard(1723) described an orthodontic appliance
that use silk or silver ligature to move malposed teeth
and a pelican pliers that was used to straighten
crowded incisors instantly by bending alveolar bone.
He also published an article in 1728 called Bandelette
now called as expansion arch.
Hunter (1778) explained that teeth may be
moved by applied force, because bones move
out of the way of pressure.
Delabarre (1815) reported that orthodontic
forces cause pain and swelling of paradental
tissues (inflammation), Farrar (1888) explained
that orthodontic forces move teeth mainly
because of bending of the alveolar bone
therefore he recommended to apply heavy
forces to the teeth to achieve rapid movement.
 He proposed the piezo electric theory of tooth
movement .
The first histological examination of tissues
surrounding orthodontically-treated teeth was
reported by Sandstedt (1904-5). He subjected
teeth in a dog to orthodontic forces for
increasing periods of time, and observed
stretching of the PDL in tension sites, and
narrowing of the tissue in compression sites.
New alveolar bone formation was seen in the
former sites, while necrosis (hyalinization) and
bone resorption, direct and undermining, was
noticed in the latter sites.
In 1911-2, Oppenheim reported that tooth
movement in one pre-adolescent baboon
resulted in complete transformation
(remodeling) of the entire alveolar process,
indicating that orthodontic force effects spread
beyond the constraints of the PDL
 The effects of orthodontic force magnitude on
the dogs paradental tissue responses was
examined by light microscopy by Schwarz (1932).
He concluded that an optimal force is smaller in
magnitude than that capable of occluding PDL
capillaries. Occlusion of these blood vessels, he
reasoned, would lead to necrosis of surrounding
tissues, which could harm the tissues and slow
down the rate of tooth movement.
This opinion was supported by Reitan, who
conducted comprehensive histological
examinations of paradental tissues incidental to
tooth movement. These studies were conducted
on a variety of species, including rodents,
canines, primates, and humans, and their results
were published during the period of the 1940s
to the 1970s.
 Reitan favored the use of light intermittent
forces, because they cause minimal
amounts of tissue damage and cell death.
He noted that the nature of tissue response
differs from species to species, limiting the
value of extrapolations. He also called
attention to factors such as gender, age,
and type of the alveolar bone, in
determining the nature of the response of
paradental tissues to orthodontic forces.

Experimenting in rodents, Storey (1973)

used light microscopy to determine that
these forces can be classified as bioelastic,
bioplastic, and biodisruptive. One common
denominator to all these force categories
was found to be inflammation.
 Storey reported on observing migration of
leukocytes out of PDL capillaries 20 minutes
after orthodontic force application to incisors
in rabbits. He concluded that inflammation is
an integral part of the tissue response in
orthodontics, a process which provides cells
essential for both the resorptive and depository
functions that facilitate tooth movement.
The participation of blood borne cells in the
remodeling of the mechanically-stressed PDL
was confirmed by Rygh (1973, 74, 76). He
utilized transmission electron microscopy in
studies on rodents, and detected macrophages
at the edge of the hyalinized zone, during the
early phases of treatment, invading the
necrotic PDL, phagocytizing its cellular debris
and strained matrix.
Identification of cellular and matrix changes in
paradental tissues following the application of
orthodontic forces led to histochemical studies
aimed at elucidating enzymes that might
contribute to the remodeling of these tissues.
In 1983, Lilja, Lindskog, and Hammarstrom
reported on the detection of various enzymes in
the mechanically-strained paradental tissues of
rodents, including acid and alkaline
phosphatases, b-galactosidase, aryl transferase,
and prostaglandin synthetase. Meikle et al
(1989) stretched rabbit coronal sutures in
vitro, and recorded increases in the tissue
concentrations of metalloproteinases, such as
collagenase and elastase, and a concomitant
decrease in the levels of tissue inhibitors of this
class of enzymes
 Davidovitch et al (1975, 78, 80, 92, 96,
2000) utilized immunohistochemistry to
localize a variety of first and second
messengers in cats mechanically-stressed
paradental tissues. These molecules
included cyclic nucleotides,
prostaglandins, neurotransmitters,
cytokines, and growth factors.
Is it Possible to enhance the rate of
orthodontic tooth movement and shorten
the time of this treatment?
The realisation that tissue remodelling in
orthodontics is mediated by a variety of cells
including fibroblasts, root and bone surface lining
cells endothelial cells, nerve cells and defence
cells promoted clinical investigators to apply
physical chemical agents concomitant with
orthodontic force in order to augment the effect
of mechanical force.
The actual rate of tooth movement may depend on
the rate of bone turnover. modifying this
turnover pharmacologically in rats by inducing
either hypothyroidism or hyperthyroidism (Verna
et al., 2000)
In rats with high bone turnover rate of tooth
movement was increased while it was reduced in
animals with low turnover.
 Histological examination of these rat jaws
showed root resorption in both groups but
it was more in low turnover group.
Affect of light on bone turnover is
examined by Miyoshi et al., in rats. Rats
exposed to light for 24 hours 12 hours per
day for 21 days and were subjected to
orthodontic force only during the light
period, presented double of the rate of
tooth movement and bone remodeling as
compared with animals that received force
during 12th hour daily darkness.
Application of physical agents
Tweedle (1965) used local application of
heat to paradental tissues surrounding
orthodontically-treated teeth in dogs;
Beeson et al (1975), Davidovitch et al
(1980, 82, 84), and Giovanelli et al (1994)
used minute electric currents; and
Blechman (1998) advocated the use of
static magnetic fields.
Beeson et al reported that their electric
device, which had been fabricated for use
in cats, and whose electrodes were placed
at a noticeable distance from the tooth,
failed to accelerate its movement
 In contrast, Davidovitch et al placed the
electrodes much closer to the cats canine,
resulting in a significant enhancement of
movement.
Blechman hypothesized that magnets
generate mechanical forces, as well as
magnetic fields, and that this combination
acts synergistically, causing the teeth to
move faster.
However, an experiment in rats (Tengku et
al, 2000) revealed that magnets do not
speed-up the mesial movement of maxillary
molars, and actually increase root
resorption in the early phases of treatment.
Utilization of chemical agents in attempts to
increase the pace of tissue remodeling and
tooth movement have been tested in various
laboratories and clinics.

Yamasaki, Miura, and Suda (1980), and

Yamasaki et al (1984) injected prostaglandin
E1 into the gingiva of moving teeth in human
subjects, resulting in rapid movement.
Systemic application of misoprostol, a PGE1
analog, to rats undergoing tooth movement for
2 wk, increased significantly the pace of
movement without enhancing root resorption
(Sekhavat et al, 2002). Similar results were
reported following
intraperitoneal injections of PGE2 in rats
(Seifi et al, 2003). Chumbley and Tuncay
(1986) administered systemically
indomethacin, a prostaglandin synthethase
inhibitor; Collins and Sinclair (1988) used
local applications of vitamin D, while
Engstrom, Granstrom, and Thilander
(1988) moved teeth in hypocalcemic,
vitamin D-deficient lactating rats.
The bone matrix component osteocalcin
was injected in rats, causing rapid tooth
movement due to the attraction of
numerous osteoclasts to this site
(Hashimoto et al, 2001).
The reports cited above suggest that the
extent of tissue remodeling and the rate of
tooth movement can be significantly
influenced by numerous factors capable of
interacting with paradental cells.
Selinkale et al., (2004) reported comparison of
effect of 1,25 dihydrocolcalciferol and PGE2 on
orthodontic tooth movement. In this
experimental group both enhanced the amount
tooth movement significantly.
Application of steroids in rats al.,
experimented by Colin K .L et al.,(2000)
suggest steroid treatment suppresses clastic
activity.
Injection of L Arginine (Nitric oxide precursor)
in rats reported by Mohsen Shiraziekal et al.,
(2002) suggest a definet role of nitric oxide in
orthodontic tooth movement ie., when there is
increase of nitric oxide there is increase in
bone remodeling and increased tooth tooth
movement.
 CONCLUSION
A THOROUGH KNOWLEDGE OF
COMPLEX EVENTS THAT ARE
TAKING PLACE IN THE CELL WILL
LEAD TO THE CLINICIANS ABILITY
TO MOVE TEETH MORE
EFFICIENTLY AND MORE FASTER BY
CHANGING THE HOST RESPONSE
WITHOUT DAMAGING THE TISSUES.