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Tooth Development Oral Histology


Initiation of tooth development
Stages of tooth development:
Bud stage
Cap stage
Bell stage
Advanced Bell stage
Root formation
Histophysiology and clinical consideration
Molecular aspects of tooth development
Role of genes in tooth development
Tooth Type Determination


Early Embryogenesis
Primitive oral cavity called the stomodeum, is
lined stratified squamous epithelium called oral
ectoderm. This oral ectoderm contacts endoderm
of the foregut to form the buccopharyngeal
At the 27th day of gestation this membrane
ruptures and the primitive oral cavity establishes a
connection with the foregut

Underlying the oral ectoderm is an embryonic
connective tissue that originates from the neural
crest and is called ectomesenchyme

Primary Epithelium Band
Around 37
day of intrauterine life, a continuous
horse shoe shaped band of thickened epithelium
forms at the site of future maxillary and
mandibular arches called the primary epithelial
bands. They are divided into two parts:
1. Vestibular lamina
2. Dental lamina

This increase in thickness of primary epithelial
band occurs because of change in orientation of
mitotic spindle and cleavage plane of epithelial cell
1. Vestibular Lamina
Vestibular lamina/ lip furrow band is
responsible for the formation of future
vestibule. The cells proliferate into the
ectomesenchyme and the band enlarges
It then subsequently degenerates and hollows
out to form a cavity/ cleft/ vestibule between
the alveolar portion of the jaws and the lips
and cheeks.
2. Dental Lamina
This lamina leads to development of future
teeth. Dental lamina is seen as a localized
proliferation of the epithelium into the
ectomesenchyme. Formation of this dental
lamina begins in the distal (molar) region and
then proceeds towards the midline

What are neural crest cells?
Neural crest cells are derived from the dorsal most
region of the neural tube.
These are highly pluripotent cells which migrate
into the 1
branchial arch and assume a
mesenchymal phenotype.
Then a series of reciprocal interactions take place
with the oral epithelium leading to the formation of
tooth primordial
If the neural cells fail to migrate then they may lead
to anodontia (absence of teeth) and microdontia
(underdeveloped jaws)
All tooth structures except enamel are derived from
neural crest cells; enamel is ectodermal in origin

Tooth development (odontogenesis) takes
place by specific and reciprocal epithelium
mesenchymal interactions.
This involves cross-talk between the
epithelium derived dental lamina and the
neural crest cell derived underlying
Oral ectoderm gives rise to- enamel
Oral ectomesenchyme gives rise to- dentin,
pulp, cementum and PDL of the fully formed

Tooth Development Oral Histology

Derivatives of dental lamina:
1) Direct extension of dental lamina (lateral
lamina)leads to formation of deciduous
2) Lingual extension of free end of dental lamina
(successional lamina) opposite to the enamel
organ of each deciduous tooth gives rise to
the permanent teeth or the successors of
deciduous teeth
3) Distal extension (parental lamina) in dental
arches provides primordia for all teeth
lacking primary predecessors. e.g. permanent
Fate of dental lamina Rudimental lamina:
Total activity of dental lamina lasts for 5 years.
However, the dental lamina can still be active in
third molar region after it has disappeared
elsewhere. After tooth germ formation, dental
lamina disintegrates by autolysis. If it persists, it is
called as cell rests of serre. (May give rise to
eruption cysts, odontoma, or may be activated to
form supernumerary teeth)

At some points along the dental lamina localized
proliferation of ectodermal cells take place leading
to formation of knob like structures called the
enamel organ
Based on the morphology and histophysiology of
the enamel organ, the development of teeth can be
divided into the following stages:
Morphologic Stages Histophysiologic Processes
Dental Lamina Initiation
Bud stage
Cap stage
Bell stage (early) Histodifferentiation
Bell stage ( advanced) Morphodifferentiation
Formation of enamel
and dentin matrix

I. BUD STAGE (Initiation)
The oral epithelial cells and the adjacent
mesenchymal cells proliferate resulting in a bud
shaped enamel organ.
Histology: The enamel organ consists of
peripherally located low columnar cells and
centrally located polygonal cells
The ectomesenchymal cells surrounding the tooth
bud show condensation due to increased mitotic

II. CAP STAGE (Proliferation)
The tooth bud continues to proliferate and exhibits
unequal growth in different parts leading to cap
shaped enamel organ
It is characterized by a shallow invagination on the
deep surface of the bud
Tooth germ: enamel organ+ dental papilla + dental
follicle are seen in the cap stage

Histologically, the enamel organ in the cap stage
consists of:
1. Outer and Inner Enamel Epithelium
The peripheral cells of the cap covering the
convexity are cuboidal and are called the outer
enamel epithelium (OEE). The cells lining the
concavity of the cap become tall columnar and is
the inner enamel epithelium (IEE)
2. Stellate Reticulum
The central area of the enamel organ between the
outer and inner epithelium consist of polygonal
These cells in the centre synthesize and secrete
GAGS (Glycosaminogylcans) which gives cushion
like consistency, supports and protects the delicate
Tooth Development Oral Histology

enamel forming cells. As GAGs are hydrophilic in
nature, water is pulled into the enamel organ. This
increases the volume of extracellular compartment
and cells are forced apart thus becoming star
shaped and are called stellate reticulum.
3. Dental Papilla: ectomesenchymal condensation
immediately subjacent to enamel organ
The ectomesenchymal cells are partly covered by
the invaginated portion of the inner enamel
epithelium. It condenses to form the dental papilla
which is the formative organ of dentin and pulp
The dental papilla shows mitotic cell division
along with proliferation of new blood capillaries
The peripheral cells near the inner enamel
epithelium increase in size and differentiate to
form odontoblasts
4. Dental Sac (Dental follicle): ectomesenchymal
condensation surrounding the tooth bud and the
dental papilla
The cells of dental sac form the cementum and
periodontal ligament
Blood supply for ameloblast for most of its life
cycle is from the dental sac

Transient structures seen in enamel organ
during cap stage:

Enamel knot / Ahrens knot: Dense cluster of non
dividing epithelial cells detected first at the tip of
the tooth bud
Enamel cord: Vertical extension of the enamel knot
between the IEE and OEE is the enamel cord
Enamel septum: When the enamel cord extends to
meet the OEE, it is termed as enamel septum.
Enamel navel: The OEE at the point of meeting
shows a small depression called enamel navel
Enamel niche: An apparent structure seen in
histologic sections due to funnel shaped
depression of the dental lamina. It creates an
impression that tooth germ has double attachment
to the oral epithelium by two separate strands of
dental lamina enclosing the ectomesenchyme
called as enamel niche between them

It is the 3
stage of enamel organ formation in
which the crown form is established characterized
by histodifferentiation and morphodifferentiation.
As the invagination of the epithelium deepens and
its margins continue to grow, the enamel organ
assumes a bell shape
On light microscopic examination of bell stage
four different types of epithelial cells can be seen:
1. Inner Enamel Epithelium
Consists of a single layer of cells that
differentiate prior to amelogenesis to form tall
columnar cells called ameloblasts
The cells of the inner enamel epithelium exert
an organizing influence on the underlying
mesenchymal cells in the dental papilla, which
later differentiate into odontoblasts
2. Stratum Intermedium
Formed by a few layers of squamous cells
between the IEE and stellate reticulum.
They have a high activity of enzyme alkaline
phosphatase and increased glycogen deposits
which are essential to enamel formation
3. Stellate Reticulum
It expands further by increase in intracellular
fluid. The cells are star shaped with processes
that anastomose with those of adjacent cells
Enamel knot and cord act as reservoir of cells for
growing enamel organ
Enamel knot expresses various signalling
molecules like Bmp-2, 4, 7; Fgf-4, 9; Wnt-10b, Slit-
1 and Shh
FGF-4 expressed in the enamel knot, stimulates
cell division in adjacent enamel epithelium and
dental papilla
However, enamel knot cells themselves do not
express FGF-4 receptors and thus are unable to
respond to the proliferative mitogenic stimuli of
FGF and remain as nondividing cells
Thus they regulate the epithelial folding and
help in transformation from bud to cap stage
They also regulate the expression of Cbfa1 gene
in the mesenchyme which induces odontoblast
differentiation and thus play a role in tooth

Tooth Development Oral Histology

Before enamel formation begins, the stellate
reticulum collapses reducing the distance
between the centrally situated ameloblasts
(IEE) and the nutrient blood capillaries
This change begins at the height of the cusp or
incisal edge and progresses cervically
4. Outer Enamel Epithelium
The OEE is made of low cuboidal cells and is
thrown into folds by the end of bell stage
Between the folds, the mesenchyme forms
papillae that contain capillary loops, thus
providing a source of nutrition to the
metabolically active avascular enamel organ

Zone of reflexion or cervical loop: The junction of
IEE and OEE is called the zone of reflexion or
cervical loop. It gives rise to the epithelial
component of root formation.
Changes in dental papilla: Before the inner
enamel epithelium begins to produce enamel, the
peripheral cells of the dental papilla differentiate
into odontoblasts which are initially cuboidal and
later become columnar and produce dentin.
Membrana Preformativa: It is the basement
membrane that separates the enamel organ and
dental papilla just prior to dentin formation
Changes in dental Sac: Initially it shows a circular
arrangement of its fibres and resembles a capsular
structure. With the development of root, the fibres
of dental sac become embedded in the cementum
and alveolar bone
Two important events occur during bell stage
Break-up of the dental lamina
Inner enamel epithelium folds
This stage is characterized by the commencement
of mineralization and root formation

Steps in hard tissue formation (reciprocal induction):
Initially, the IEE cells induce the underlying
ectomesenchymal cells of dental papilla to
differentiate into odontoblasts
Odontoblasts lay down the first layer of dentin
along the DEJ at the region of future cusps
After the first layer of dentin is formed the IEE
differentiate into ameloblasts ( tall columnar cells
with reversal of polarity) and lay down enamel
matrix over the dentin
Note: Enamel does not form in the absence of dentin


The development of roots begin after enamel and
dentin formation has reached the future
cementoenamel junction (CEJ)
The cervical portion of enamel organ gives rise to
HERS which is a double layered cell sheath formed
by the down growth of IEE and OEE
It outlines the future root and is thus responsible
for the shape, length, size and number of roots
It extends around the dental pulp and forms the
epithelial diaphragm which encloses primary
apical formation
In multirooted teeth, epithelial diaphragm exhibits
differential growth, causing division of root trunk
and development of multiple roots (AIIMS-89)

During expansion of pulp, the IEE cells induce the
ectomesenchymal cells of the dental papilla to
differentiate into odontoblasts which form dentin
After the root dentin forms; the HERS cells
disintegrate and move away from the surface of
dentin thereby exposing it to the connective tissue
Tooth Development Oral Histology

cells of the dental sac, which differentiate into
cementoblasts and deposit cementum
Remnants of HERS may persist as a strand of cells
found in the periodontal ligament (PDL) of
erupted teeth called as Rests of Malassez
The continuity of HERS may be broken or not
established prior to dentin formation, leading to
the development of accessory root canals

If the cells of the epithelial root sheath remain
adherent to the dentin surface they may
differentiate into fully functioning ameloblasts and
produce enamel. Such droplets of enamel are
called enamel pearls and are usually found in the
furcation area of roots of permanent molars

1. Initiation
2. Proliferation
3. Histodifferentiation
4. Morphodifferentiation
5. Apposition
1. Initiation
Specific cells within the dental lamina have the
potential to develop into enamel organ or induce
tooth development by epithelial mesenchymal
Lack of initiationAnodontia / Partial Anodontia
Abnormal initiation Supernumerary teeth
Initiation at abnormal locations(ectopic) Ovary
(dermoid tumors or cysts) / Hypophysis

2. Proliferation
It causes regular changes in the size and
proportions of the developing tooth germ
Disturbances has different effects depending on
the time of occurrence and stage of development

3. Histodifferentiation
In this stage, cells undergo definitive morphologic
and functional changes, they differentiate and
restrict multiplication to don their new functions
During histodifferentiation, the peripheral cells of
the dental papilla differentiate into odontoblasts to
produce dentin which further induces IEE cells to
differentiate into ameloblasts and form enamel
Eg. Disturbance: Vitamin A deficiency at this
stage leads to formation oosteodentin

4. Morphodifferentiation
Establishment of morphologic pattern, basic form
and size of future teeth takes place by differential
growth at this stage

Morphogenesis is not possible without proloiferation
In this stage the DEJ and CDJ are developed by the
continuous deposition of enamel, dentin and
cementum, giving the completed tooth its
characteristic form and size
Disturbances affect the form and shape of teeth:
a. Supenumerary cusps/ loss of cusps
b. Twining
c. Peg shaped tooth
d. Endocrine disturbances: hypopitutarism
/hypothyroidism small crowns

5. Apposition
It is the regular and rhythmic deposition of
extracellular matrix of enamel and dentin
Disturbances due to genetic or environmental
factors leads to hypoplasia and hypocalcification

Odontogenesis is initiated by factors resident in
the first arch epithelium influencing
ectomesenchyme, but with time, this potential is
assumed by ectomesenchyme

Role of Homeobox genes
The Homeobox gene was initially found in
Drosophila melanogaster and is called as the
master gene. It constitutes a large family of genes
that encode the transcription factors and regulate
the expression of downstream target genes
A set of coded patterning homeobox genes is
required to specify correct positioning and bring
about the development of cephalic structures

Subfamilies of Homobox genes:
Muscle segment (Msx)
Distal less (Dlx)
Orthodenticle (Otx)
Goosecoid (Gsc)
Bar class (Barx)
Paired related (Prx)
LIM homeobox

Tooth Development Oral Histology


Genes associated with tooth development
Gene Function
Lhx-6, Lhx-7
domain genes
Earliest mesenchymal
markers for tooth formation
Expressed in the neural crest
derived ectomesenchyme of
the oral half of the first
branchial arch by day 9
Fibroblast growth
factor 8
(Secreted protein)
Expressed in the first
branchial arch for induction
of the Lhx genes
Establishment of oral -
aboral axis
Determines the positions for
formation of tooth germs
Induction of Pax-9 gene
Paired box
homeotic gene
Its expression co localizes
with the exact sites where
the tooth germs appear
It is induced by Fgf-8 and
repressed by BMP-2 & BMP-
Sonic hedgehog
(Secreted protein)
Induces local epithelial cell
proliferation to produce
invaginations that are
reminiscent of tooth buds
Stimulation of epithelial cell
Signaling tooth initiation
enhancer binding
Initially expressed in dental
epithelial thickenings
During bud formation it
shifts to being expressed in
the condensing mesenchyme
Gene Abbreviation Function
Msh like genes
(Transcription factor)
Anterior teeth
Distaless homologue
(Transcription factor)
BarH1 homologue
(Transcription factor)

Humans have heterodont teeth (falling into 3
categories: Incisiform, caniniform and molariform)
The determination of the type of tooth which will
develop at a particular position is called as
patterning of dentition
This can be explained with the help of two
hypothetical models:
Field model
Clone model
1. Field model (Butler 1839)
It proposes that the factors responsible for tooth
shape reside within the ectomesenchyme in
distinct graded and overlapping fields for each
tooth family
This theory is supported by expression of different
homeobox genes at different position

2. Clone model (Osborn 1978)
It proposes that each tooth class is derived from a
clone of ectomesenchymal cells programmed by
epithelium to produce teeth of a given pattern. The
coded pattern of homeobox gene expression in the
ectomesenchyme are expressed following the
epithelial signal

The specific clone induces the dental lamina to
begin tooth development. At its posterior border
the clone and dental lamina grow by means of the
progress zone. When a clone reaches the critical
size, a tooth bud is initiated at its centre. A zone of
inhibition surrounds the tooth bud and the next
tooth bud is not initiated until the progress zone of
the clone has escaped its influence.