Development of

tooth
 Contents
•Introduction
•How the tooth development begins
•Phases of tooth development
•Primary epithelial band
•Dental lamina
•Vestibular lamina
•Successional lamina
•Parent lamina
•Stages of tooth development
•Bud stage
•Cap stage
•Early Cap stage
•Enamel nitch
•Lateral lamina
•Late Cap stage
•Enamel knot & cord
•Bell stage
•Crown pattern formation
CONTENTS
•Root formation
•Epithelial mesenchymal
interactions
•Homeobox genes
•Developmental disorders of
teeth

CONTENTS
 Introduction

Development of the tooth involves many complex

biological processes, including epithelial
mesenchymal interactions, morphogenesis and
mineralisation. In the human beings, 20 deciduous
and 32 permanent teeth develop from the interaction
between the oral epithelium cells and the underlying
mesenchymal cells. The basic developmental process
is similar for all teeth but each developing tooth
develops as an anatomically distinct unit.
 How does the tooth development begin?

On the 27th day of intra uterine life the

buccopharngeal membrane ruptures and the
POC comes in contact with the foregut.

Connective tissue underlying the oral

ectoderm is Neural Crest Cells or
Ectomesenchymal in origin, which induces
the overlying ectoderm to start tooth
development.
 Phases of tooth development
•Initiation
•Proliferation
•Histodifferentiation
•Morphodifferentiation
•Apposition
 Induction and Proliferation and Proliferation,
initiation stage 6 bud stage 8 week differentiation,morphoge
to 7 week nesis and cap stage 9 to
10 week

Induction,proliferati
Proliferation Maturation,varies
on and apposition
differentiation,morphoge per tooth
stage varies per
nesis bell stage 11 to 12
tooth
week
 INITIATION
The epithelial ectomesenchymal interaction is
essential for initiation induction.
If there is a lack of initiation ,it results in the absence
of either a single tooth or many teeth.
Abnormal initiation may result in the development of
single or supernumerary teeth.
PROLIFERATION
It is characterized by regular changes in the size and
shape of the developing tooth germ.In this stage the
tooth germ has a strong affinity to differentiate into
various structures.
 Histodifferentiation
Histodifferentiation shows maximum
development in the early bell stage of tooth
development .It is during this stage that the
formative cells of the tooth germ undergo
definite morphologic as well as functional
changes and acquire their functional roles.
The cells differentiate and give up their
capacity to multiply.
 Morphodifferentiation

Proliferation is essential for morphodifferentiation.

The basic anatomic form of tooth is established in the
late bell stage after active histodifferentiation .
In this stage the dentinoenamel and the
cementoenamel junctions are developed by the
continuous deposition of enamel,dentin and
cementum from ameloblasts,odontoblasts and
cementoblasts respectively,resulting in the
establishment of a complete morphologic pattern
,size,and shape of a tooth.
If any disturbance occurs in the
morphodifferentiation , it affects the form and
size of the tooth , but the functions of the
ameloblasts and the odontoblasts are not
affected

Morphodifferentiation
 Apposition
After the morphologic pattern of a tooth is
established, an additive growth of the hard dental
tissues occurs.apposition is characterized by the
rhythemic,layer-like deposition of an extracellular
matrix of enamel and dentin.
 PRIMARY EPITHELIAL BAND

After 37 days of development, a continuous band of

thickened epithelium forms around the mouth in the
presumptive upper and lower jaws.
These bands are roughly horse shoe shaped and
correspond in position to the future dental arches of
the upper and the lower jaws.
Each band of epithelium called the PEB gives rise to 2
subdivisions
•The buccaly located vestibular lamina
•The lingually located dental lamina
Primary Epithelial Band
A = Vestibular lamina B = Dental lamina

Oral vestibule Tooth bud

 DENTAL LAMINA

Dental lamina is the band of epithelium which

has invaded the underlying ectmesenchyme
along both the horse shoe shaped future dental
arches
It serves as the primordium for the ectodermal
portion of the deciduous teeth.
 VESTIBULAR LAMINA
The vestibule forms as a result of the proliferation of
the vestibular lamina into the ectomesenchyme.

Its cells rapidly enlarge and then degenerate to form a

cleft that becomes the vestibule between the cheek and
the tooth- bearing area.
 Successional lamina
The free terminal of the dental lamina begins to proliferate in
the 4th month of fetal growth. (Crown Rump Length 55-100
mm). This newly formed growth center is known the
successional lamina and is destined to form the permanent
teeth.
 Parent dental lamina (lamina for permanent
molars)

The dental lamina

providing for the
formation of the 1st, 2nd, &
3rd permanent molars may
be referred to as the parent
lamina
Morphologic Stages of tooth development
 Bud Stage (proliferation stage)

CR Length= 17.0 mm)

 Bud stage is the initial stage of tooth

development, represented by the first epithelial
incursion into the ectomesenchyme of the jaw
forming tooth buds

 Simultaneous with the differentiation of each

dental lamina,round or ovoid swellings arise at
the basement membrane at 10 different points.
•The epithelial cells show little, if any change, in shape.

•The epithelium of the tooth bud forms the enamel organ.

•The supporting ectomesenchymal cells are packed closely

beneath and around the epithelial bud.

The enamel organ in the bud stage

•appears as a simple,spherical to ovoid, epithelial
condensation surrounded by mesenchyme.
•It contains 2 types of cells
1.Polygonal,which are centrally situated
2.Low columnar,which are peripherally situated.

Bud stage
• The cells of the tooth bud have a higher RNA
content than those of the overlying oral
epithelium,a lower glycogen content and an
increased oxidative enzyme activity.

• The area of ectomesenchymal condensation

immediately subadjacent to the enamel organ is
the dental papilla

• The condensed ectomesenchyme that surround the

tooth bud and the dental papilla is the dental sac.

Bud stage
Bud stage
Bud stage
 CAP STAGE
WHY IS IT CALLED CAP STAGE &
HOW IS IT FORMED?
A = Bud stage
B = Dental papilla
 Cap stage
During the ninth week of embryonic development,
the tooth bud differentiates into a cap-shaped
enamel organ extending from the dental lamina

A, Enamel organ
B, Dental lamina
C, Vestibular
lamina
D, Dental Papilla
E, Dental sac
A, Stellate reticulum; B, Outer enamel epithelium;
C, Inner enamel epithelium; D, Dental papilla; E, Dental sac
 EARLY CAP STAGE
(PROLIFERATIVE)
 The epithelial bud continues to proliferate into the
ectomesenchyme.
 Condensation of ectomesenchyme
 At this stage, the enamel organ, dental papilla and
dental follicle that constitute the dental organ/tooth
germ can be identified.
 The Enamel Niche is an apparent structure in
histologic sections, created because the dental lamina
is a sheet rather than a single strand and often
contains a concavity filled with connective tissue.
 Mesenchyme

Dental epithelium

ENAMEL NICHE is seen where the tooth germ appears to

have a double attachment to the dental epithelium. EN
appears as a funnel-shaped depression containing
connective tissue. The functional significance of EN is
unknown.
 LATERAL LAMINA

•As the tooth bud grows larger, it drags

along with it a part of the Dental lamina;
from that point on, the developing tooth is
attached to the Dental lamina by an
extension called as Lateral lamina.
Early Cap stage
 LATE CAP STAGE

Important developmental changes begin late in the cap

stage and continue during the transition of the tooth
germ from cap to bell. Through these changes ,termed
histodifferentiation , a mass of similar epithelial cells
transform itself into morphologically and functionally
distinct components.
The cells in the center of the enamel organ synthesize and
secrete glycosoaminoglycans into the extracellular
compartments between the epithelial cells.

Glycosaminoglycans are hydrophillic and so pull water

into the enamel organ because of which the cells loose
water and are forced apart but they retain connections
with each other through their desmosomal contacts, and
hence become star shaped.
The center of the enamel organ thus is termed Stellate
Reticulum
Late cap stage
•At the periphery of the enamel organ the cells
assume a low cuboidal shape and form the
outer(external)dental epithelium.

•The cells bordering on the dental papilla assume a

short columnar shape and are characterized by high
glycogen content. They form the internal dental
epithelium.
•Enamel knot & enamel cord can be seen.
Late cap stage
Enamel knot or Ahren’s knot

 It is a localized
mass of, or cluster
of non dividing
cells.
 The E K forms
a bulge into the
dental papilla, at
the center of the
enamel organ.
Late cap stage
THE ENAMEL CORD
Strand of cells seen at the early bell stage of
development, extending from the stratum
intermedium into the stellate reticulum.
When it completely divides the stellate reticulum into 2
parts,reaching the ODE. It is termed the Enamel
septum.
Where the enamel cord meets the OEE a small
invagination called the Enamel Navel may be seen.
EC may be involved in the process by which the cap
stage is transformed into the bell stage.
It is a focus for the origin of the Stellate Reticulum cells.
Late cap stage
Late cap stage
 BELL STAGE

•Transition from cap to bell stage occurs in the

embryo which is between 100-160 mm long(
CR length)

Why is this stage called the bell stage?

Bell stage
 EARLY BELL STAGE (CR LENGTH
=120MM)

•Further morphodifferentiation and

histodifferentiation of the tooth germ leads to the
early bell stage.
•The configuration of the internal enamel
epithelium broadly maps out the occlusal pattern of
the crown of the tooth.

•A high degree of histodifferentiation is achieved in

the early bell stage.
•The enamel organ shows 4 distinct layers:
External enamel epithelium, Stellate Reticulum,
Stratum Intermedium, and internal enamel
epithelium.

•During this stage,the crown assumes its final

shape(morphodifferentiation)and the cells ameloblast
and odontoblasts acquire their distinctive
shape.(histodifferentiation)

Early bell stage

 OUTER OR EXTERNAL DENTAL
EPITHELIUM :

 The ODE cells flatten to a low cuboidal form.

 At the end of the bell stage preparatory to and
during the formation of enamel, the formerly
smooth surface of the outer enamel epithelium is
laid in folds. Between the folds the adjacent
mesenchyme of the dental sac forms papillae that
contain capillary loops and thus provide rich
nutrition supply for the intense metabolic activity
of the avascular enamel organ.

Early bell stage

 INNER DENTAL EPITHELIUM
•IDE consists of a single layer of tall columnar
cells,which differentiate into specialised cells called
Ameloblasts ,before amelogenesis.

•The IDE cells are rich in RNA but, unlike the SI & SR
do not contain alkaline phosphatase.

•During enamel formation there is reversal of polarity

in the ameloblasts in order to derive nutrition through
the outer enamel epithelium.
Early bell stage
•The cells of the IDE exert an organising influence
on the underlying mesenchymal cells in the dental
papilla which later differentiates into Odontoblasts

Inner dental epithelium

(Early bell stage)
 STELLATE RETICULUM (ENAMEL PULP)
 It is fully developed at the bell stage.
 Cells of this layer possess little endoplasmic reticulum
and few mitochondria. There is a relatively well-
developed golgi complex.
 Numerous tonofilaments are present within the
cytoplasm, and desmosomes and gap junctions are
present between the cells.
 The proteinaceous fluid containing albumen gives a
cushion- like consistency to the SR that supports and
protects the delicate enamel-forming cells against
physical disturbance and maintenance of tooth shape.
Early bell stage
CERVICAL LOOP/ZONE OF REFLECTION

The outer and inner dental epithelia are

continuous. The region where they meet at the
rim of the enamel organ is known as Cervical
Loop

Early bell stage

 STRATUM INTERMEDIUM

In the bell stage ,some epithelial cells between the

inner dental epithelium and the stellate reticulum
differentiate into a layer called the Stratum
Intermedium.

 Consists of 2 or 3 layers of flattened cells lying over

the IEE. The cells of the SI resemble the cells of the
SR but the intercellular spaces are smaller and the
cells contain increased alkaline phosphatase levels.

Early bell stage

•Well developed cytoplasmic organelles,acid
mucopolysaccharides & glycogen deposits indicate
high degree of metabolic activity.

•SI is concerned with the synthesis of proteins

•Although these cells are histologically distinct from

the cells of the IDE, both layers have been
considered as a single functional unit responsible for
the formation of enamel.

STRATUM INTERMEDIUM

(early bell stage)

 Early bell stage of tooth development .
EO seems to be divided by the enamel septum
or cord
 DENTAL PAPILLA
•1st the odontoblasts are cuboidal – shaped and are
then elongated to become columnar in shape and
laying down of the dentin starts before the inner
enamel epithelium lays down the first layer of enamel
matrix.
•The dental papilla is referred to as the “tooth pulp”
when the 1st calcified matrix appears at the cuspal tip
of the bell stage tooth germ.
•Membrana preformativa seperates the enamel organ and
the dental papilla before the dentin develops.
Early bell stage
Ectomesenchymal cells closely aggregated to form
the dental papilla
Early bell stage
 DENTAL SAC

Before formation of dental tissues begins, the dental sac

shows a circular arrangement of fibers and resembles a
capsular structure.

With the development of the root, the fibers of the

dental sac differentiate into the peridontal fibers and
become embedded in the developing cementum and
alveolar bone.

Early bell stage

 Advanced Bell Stage (Appositional Stage)
2 main features seen:

•Future dentinoenamel junction forms from the boundary

present between IDE & odontoblasts.

•Hertwig’s epithelial root sheath develops from the

cervical portion of the enamel organ.
•It is during the advanced bell stage that dental lamina breaks
down and the enamel organ loses connection with the oral
epithelium.

•At the same time the dental lamina between tooth germs
also degenerates.

Advanced bell stage

 Breakup of dental lamina :
•The Dental lamina and the Lateral lamina joining the tooth
germ to the oral epith. break up into discrete islands of
epithelial cells which normally degenerate;

If they persist, they are called as epithelial pearls.

•These may form Eruption cysts over the developing tooth

and delay eruption or may form supernumerary teeth, &
cysts (odontogenic tumours , cyst linings.)

 Remnants of the dental lamina may contain keratin and

can be involved in the etiology of cysts.
Advanced Bell stage
The dental lamina is
disintegrating ,tooth divorced
from the oral epithelium.
The crown pattern of the tooth
has been established by the
folding of the inner dental
epithelium.
This folding reduces the amount
of SR over the future cusp tip.
Dentin and enamel formation
have begun at the crest of the
folded IEE.
Advanced bell stage
 Reduced enamel epithelium
• The reduced enamel epithelium consists of the
mature/protective ameloblasts and remanants of the outer
layers of the enamel organ
• Numerous capillaries,which had formed to supply oxygen
and nutrients to the ameloblasts following dentin formation,
surround the reduced enamel epithelium.

A, Reduced enamel epithelium; B, Maturative/protective ameloblasts; C,

Capillary
 Crown pattern determination :

•The Inner dental epithelium lies between two opposing

pressures i.e. from the Stellate reticulum cells of the
Enamel organ and from the Dental papilla, and is
therefore in a state of equilibrium.

•The folding that occurs as the crown develops results

from the differential rates of mitotic division within the
Inner dental epithelium.
•When the tooth germ is growing rapidly during the cap to
bell stage, cell division occurs throughout the Inner dental
epithelium. As development continues, division ceases at a
particular point because the cells are beginning to
differentiate into Ameloblasts. This represents the site of
future cusp development.

•Because the Inner dental epithelium is constrained between

the Cervical loop and the cusp tip, continued cell
proliferation causes the Inner dental epithelium to buckle
and form the cuspal outline.
CROWN PATTERN FORMATION : Darkened area –zone of cell
division
White area –cells have
differentiated.
 Root formation
•The development of the roots begins after Enamel and Dentin
formation has reached the future Cemento-Enamel
junction.

•The Enamel organ plays an important role in root

development by forming Hertwig’s Epithelial Root Sheath.

•Hertwig’s root sheath consists of Outer and Inner Dental

epithelia.
•When the inner cells of this sheath have induced the
differentiation of radicular cells into Odontoblasts and first
layer of Dentin has been laid down, the root sheath loses its
structural continuity.

•Its remnants persist as Rests of Malassez. Apparently

functionless, they are the source of epithelial lining of dental
cysts that develop in reaction to inflammation of periodontal
ligament.
 Root formation in single rooted teeth
•Prior to the beginning of root formation, the root
sheath forms the Epithelial Diaphragm. The Outer
and Inner dental epithelia bend at the future CEJ
into a horizontal plane, narrowing the wide
cervical opening of the tooth germ.

•The epithelium proliferates coronal to the Epithelial

Diaphragm.
Root formation in single rooted teeth
•Differentiation of Odontoblasts and formation of Dentin
follow the lengthening of the root sheath.

•At the same time, the connective tissue cells of the Dental
Sac proliferate and invade the root sheath. These cells come
into contact with the outer surface of dentin and
differentiate into Cementoblasts that deposit Cementum
onto the surface of the Dentin.

ROOT
FORMATION
 Root formation in multi-rooted teeth :
•During the general growth of the Enamel organ, the
expansion of its cervical opening occurs in such a way
that long, tongue-like extensions of the horizontal
Diaphragm develop.

•The free ends of these horizontal epithelial flaps grow

towards each other and fuse. The single cervical
opening of the Enamel organ is then divided into
two/ three openings.
 ROOT
FORMATION
 ROOT
FORMATION
How is dental development initiated?
 Nature of the Inductive Message

During tooth development messages pass between the

epithelium and mesenchyme to produce changes of increasing
complexity (i.e. differentiation) with in the cell layers.

The term Induction is used to describe the effect that one cell
layer has on another.
 Three main hypothesis to explain how information
leading to induction may be transferred between
epithelium & mesenchyme:

1. A chemical substance (Short range hormone) is produced by

one cell layer and diffuses across the narrow intervening space
to be taken up & cause induction in the other cell layer.

2. Induction is triggered by direct cell to cell contact & does not

involve a diffusible molecule.
3. Induction is due to the presence of the initial extracellular
matrix, a thin layer situated between the Epithelium &
Mesenchyme and comprising the basal lamina & adjacent
region. The extracellular matrix has a complex composition,
consisting of collagen, proteoglycans & glycoproteins.
 Homeobox Genes

•Genes highly conserved throughout the evolution of diverse

organisms.

•All homeobox genes contain a similar region of 180 nucleotide

pairs and function by producing transcription factors that
bind to the DNA of other genes, thereby regulating their
expression.

•Examples of those involved in tooth development include MSX,

DLX, BARX, LHX
 What are the signals mediating initial steps in
tooth development?

•The earliest mesenchymal markers for tooth formation are

Lhx genes i.e. Lhx-6 and Lhx-7 expressed in the 1st
branchial arch.

 Expression of these genes results from a signaling molecule

originating from the epithelium of 1st branchial arch i.e. Fgf-
8.

1st branchial arch epith + 2nd arch mesenchyme= Lhx Induced

1st arch mesenchyme + 2nd arch epith=Lhx down regulated

 What controls the position and no. Of tooth
germs?
•Signals for these aspects appear to originate from the oral
epithelium.

•Pax-9 gene plays an imp. role in defining the localization of

tooth germs.

•A no. of other genes are also expressed in the oral epithelium

e.g. Dlx-2, Msx-1. In fact, to date, more than 90 genes have
been identified during initiation of tooth development.
 Tooth Type Determination

•The determination of specific tooth types at their correct

positions in the jaws is referred to as “ patterning of the
dentition”.

•Two hypothetical models—

Field model
Clone model
 Field Model (Homeobox Code Model)

•It proposes that the factors responsible for tooth shape i.e.
Homeobox genes reside within the ectomesenchyme in
distinct but graded fields for each family.

•The expression of Msx-1 and Msx-2 is restricted to regions

where incisors and canines develop while Dlx-1 and Dlx-2
are expressed where multi-cuspid teeth develop.

 Expression of Barx-1 overlaps with Dlx-1, Dlx-2.

 Clone Model

Each tooth class is derived from a clone of ectomesenchymal

cells programmed by epithelium to produce teeth of a
given pattern.
 Both models can be combined

For instance, the coded pattern of homeobox gene expression

in the ectomesenchyme might be expressed following an
epithelial signal.
Ectomesenchyme eventually assumes the dominant role in
crown pattern formation.
 Examples to proove ectomesenchyme eventually
assumes the dominant role in crown pattern
formation.

Murine 1st branchial arch epithelium combined with cranial

neural crest in the anterior chamber of the anterior chamber
of the eye leads to tooth formation.

Epithelium from other sources e.g. 2nd arch does not elicit
this response
After 12th day of development ,1st arch epithelium loses its
odontogenic potential which is assumed by the
ectomesenchyme e.g. 1st arch ectomesencyme recombined
with embryonic foot epithelium leads to formation of dental
organ

Examples to proove ectomesenchyme eventually

assumes the dominant role in crown pattern
formation.
Tooth that developed from the skin epithelium
 Recombination of dental epithelium and
ectomesenchyme

A.Incisor epithelium combined with molar papilla results in

a molariform tooth
B.Molar epithelium combined with incisor papilla results in
an incisiform tooth
Summary
Macrodontia
 Missing teeth

Anodontia hypodontia
Dilaceration
Dens in dente

Type I II III
Fusion
Germination
Taurodontism
Supernumerary roots
 Cusp of carabelli

CUSP OF
CARABELLI
Supernumerary teeth
Enamel pearl
Talons cusp
 References
•Oral histology –Ten Cate’s & advanced Ten
Cate
•Orban’s – Oral histology and embryology
•Satish Chandra - Oral histology and
embryology
•Provenza
•Textbook of oral pathology-Shafer
•Oral and maxillofacial pathology-Neville
JOUNAL OF DENTAL RESEARCH,Vol.79,1469-1475(2000)
JOUNAL OF DENTAL RESEARCH, Vol.81(4) : 274-278 (2002)
JOUNAL OF ARC ORAL BIOLOGY 15:1315,1970
CONCLUSION
Thank You
Thank you!