Lec.( 1 ) Oral histology dr.

lubna al khafaji

Development of the Tooth and Its Supporting Tissues

This discusses the histologic aspect of tooth development and the coming together of the different
tissues that form the tooth and its surrounding tissues.

What are the signals mediating the initial steps in tooth development?

A signaling molecule originating from the oral epithelium of the first branchial arch results in
the expression of both of (transcription factors) Lhx-6 and Lhx-7 genes in the neural crest–
derived ectomesenchyme of the oral portion of the first branchial arch as early as day 9 of
gestation ,

The earliest histologic indication of tooth development is at day 11 of gestation, which is marked
by a thickening of the epithelium where tooth formation will occur on the oral surface of the
first branchial arch

After about 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 horseshoe-shaped and
correspond in position to the future dental arches of the upper and lower jaw. Each band of
epithelium, called the primary epithelial band, quickly gives rise to two subdivisions which in
grow into the underlying mesenchyme colonized by neural crest cells. These are:
1- The dental lamina, which forms first, on the anterior aspect of the dental lamina,
continued and localized proliferative activity leads to the formation of a series of epithelial
outgrowths into the mesenchyme at sites corresponding to the positions of the future

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 deciduous teeth. Ectomesenchymal cells accumulate around these outgrowths. From this
point, tooth development proceeds in three stages: the bud, cap, and bell.
2- The vestibular lamina, which forms shortly afterward and is positioned just in front of the
dental lamina (Labial and buccal) . The vestibule forms as a result of the proliferation of
the vestibular lamina into the ectomesenchyme soon after formation of the dental lamina.
The cells of the vestibular lamina rapidly enlarge and then degenerate to form hollows or
cleft and forms the oral vestibule between the alveolar portion of the jaws (tooth-
bearing area) and the lips and cheeks. also termed the lip furrow band.

Fate of dental lamina

It is evident that the total activity of the dental lamina extends over a period of at least 5
years. Any particular portion of the dental lamina functions for a much briefer period since
only a relatively short time elapses after initiation of tooth development before the dental
lamina begins to degenerate at that particular location. However, the dental lamina may still
be active in the third molar region after it has disappeared elsewhere, except for occasional
epithelial remnants. As the teeth continue to develop, they lose their connection with the
dental lamina. They later break up by mesenchymal invasion, which is at first incomplete and
does not perforate the total thickness of the lamina.
Remnants of the dental lamina persist as epithelial pearls or islands within the jaw as well as
in the gingiva. These are referred to as cell rest of Serres.

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 Histophysiology of tooth developmet
1- Initiation: The dental laminae and associated tooth buds represent those parts of the oral
epithelium that have the potential for tooth formation. Different teeth are initiated at definite
times. Initiation induction requires ectomesenchymal–epithelial interaction.

2- Proliferation: Enhanced proliferative activity ensues at the points of initiation and results
successively in the bud, cap, and bell stages of the odontogenic organ. Proliferative growth
causes regular changes in the size and proportions of the growing tooth germ.
3- Histodifferentiation: The formative cells of the tooth germs developing during the proliferative
stage undergo definite morphologic as well as functional changes and acquire their functional
assignment (the appositional growth potential).
4- Morphodifferentiation: The morphologic pattern, or basic form and relative size of the future
tooth, is established by morphodifferentiation, that is, by differential growth.
Morphodifferentiation therefore is impossible without proliferation. The advanced bell stage
marks not only active histodifferentiation but also an important stage of morphodifferentiation in
the crown, outlining the future dentinoenamel junction.
5- Apposition: is the deposition of the matrix of the hard dental structures.

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 Developmental stages of tooth are:
1- BUD STAGE
 The epithelium of the dental laminae is separated from the underlying ectomesenchyme by a
basement membrane
 The bud stage is represented by the first epithelial incursion into the ectomesenchyme of the jaw
 The epithelial cells show little if any change in shape or function.
 The supporting ectomesenchymal cells are packed closely beneath and around the epithelial bud
 As the epithelial bud continues to proliferate into the ectomesenchyme, cellular density increases
immediately adjacent to the epithelial outgrowth .
This process is classically referred to as a condensation of the ectomesenchyme.

2- CAP STAGE
 As the tooth bud grows larger, it drags along with it part of the dental lamina; so from that point
on, the developing tooth is tethered to the dental lamina by an extension called the lateral
lamina
 The epithelial outgrowth, which superficially resembles a cap sitting on a ball of condensed
ectomesenchyme, is referred to widely as the dental organ but actually should be called the
enamel organ (epithelial part of the tooth germ) , because it eventually will form the enamel
of the tooth.
 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. A section through this arrangement creates the impression that the tooth germ
has a double attachment to the oral epithelium by two separate strands .
 The ball of condensed ectomesenchymal cells, called the dental papilla, will form the dentin
and pulp.

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 The condensed ectomesenchyme limiting the dental papilla and encapsulating the enamel
organ—the dental follicle or sac—gives rise to the supporting tissues of the tooth.
 Because the enamel organ sits over the dental papilla like a cap, this stage of tooth development
is known as the cap stage.
 The enamel organ, dental papilla, and dental follicle together constitute the dental organ or
tooth germ.
 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 transforms itself into morphologically and functionally distinct
components.
 Outer enamel epithelium:
consists of a single layer of cuboidal cells, separated from the surrounding connective tissue of
the dental sac by a delicate basement membrane.
Prior to the formation of hard structures, this regular arrangement is maintained only in the
cervical parts of the enamel organ. At the highest convexity of the organ become irregular in
shape and cannot be distinguished easily from the outer portion of the stellate reticulum.
The capillaries in the connective tissue surrounding the epithelial enamel organ proliferate and
protrude toward it.
Immediately before enamel formation commences, capillaries may even indent the stellate
reticulum. This increased vascularity ensures a rich metabolism when a plentiful supply of
substances from the bloodstream to the inner enamel epithelium is required .
During enamel formation, cells of the outer enamel epithelium develop villi and cytoplasmic
vesicles and large numbers of mitochondria, all indicating cell specialization for the active
transport of materials.
The capillaries in contact with the outer enamel epithelium show areas with very thin walls, a
structural modification also commonly found in areas of active transport

 The cells in the center of the enamel organ synthesize and secrete glycosaminoglycans into the
extracellular compartment between the epithelial cells. Glycosaminoglycans are hydrophilic
and so pull water into the enamel organ. The increasing amount of fluid increases the volume of
the extracellular compartment of the enamel organ, and the central cells are forced apart.
Because they retain connections with each other through their desmosomal contacts, they
become star-shaped .The center of the enamel organ thus is termed the stellate reticulum.

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 Enamel knot Enamel knots are clusters of nondividing epithelial cells visible in sections of
molar cap stage tooth germs.
 Enamel cord: In some planes of section, one can see cells extending from the enamel knot
across the stellate reticulum to the outer enamel epithelium .This structure is referred to as the
enamel cord; while it could be:
1- part of the enamel knot organizational center,
2-It could also be related anatomically to the site where the lateral lamina attaches to the
enamel organ cap.

This gives the stellate reticulum a cushion like consistency and acts as a shock absorber that may
support and protect the delicate enamel-forming cells.
There are temporary structures (transitory structures) that disappear before enamel formation
begins:
The cells in the center of the enamel organ are densely packed and form the enamel knot This
knot projects in part toward the underlying dental papilla, so that the center of the epithelial
invagination shows a slightly knob like enlargement that is bordered by the labial and lingual
enamel grooves .
At the same time a vertical extension of the enamel knot, called the enamel cord occurs. When
the enamel cord extends to meet the outer enamel epithelium it is termed as enamel septum, for
it would divide the stellate reticulum into two parts. The outer enamel epithelium at the point of
meeting shows a small depression and this is termed enamel navel as it resembles the umbilicus.
The function of the enamel knot and cord:
1- May act as a reservoir of dividing cells for the growing enamel organ.
2- Recent studies have shown that enamel knot acts as a signaling center as many important
growth factors are expressed by the cells of the enamel knot and thus they play an important part
in determining the shape of the tooth.

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 Lec.( 2 ) Oral histology dr. lubna al khafaji

3- BELL STAGE
 As the invagination of the epithelium deepens and its margins continue to grow, the enamel
organ assumes a bell shape, so called because the enamel organ comes to resemble a bell as
the undersurface of the epithelial cap deepens.(early ell stage)

 In the bell stage crown shape (tooth morphogenesis) is determine assumes its final shape
(morphodifferentiation), is under the control of genes and their signaling molecules and
growth factors.

 It was thought that the shape of the crown is due to the pressure exerted by the growing
dental papilla cells on the inner enamel epithelium. This pressure however was shown to be
opposed equally by the pressure exerted by the fluid present in the stellate reticulum.

 During this stage, the tooth crown and the cells that will be making the hard tissues of the
crown (ameloblasts and odontoblasts) acquire their distinctive phenotype
(histodifferentiation).

 Cells begin to differentiate only when cells cease to divide. The inner enamel epithelial cells
which lie in the future cusp tip or incisor region stop dividing earlier and begin to differentiate
first. The pressure exerted by the continuous cell division on these differentiating cells from of
a cusp tip. The cells in another future cusp area begin to differentiate, and by a similar process
results in a cusp tip form.

 The area between two cusp tips, i.e. the cuspal slopes extent and therefore of cusp height are
due to cell proliferation and differentiation occurring gradually from cusp tips to the depth of
the sulcus. Cell differentiation also proceeds gradually cervically, those at the cervix are last
to differentiate.

 At the periphery of the enamel organ, the cells assume a low cuboidal shape and form the
outer enamel epithelium.

 The cells bordering on the dental papilla assume a short columnar shape and are
characterized by high glycogen content; they form the inner enamel epithelium.

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 The outer and inner enamel epithelia are continuous; the inner epithelium begins at the point
where the outer epithelium bends to form the concavity into which the cells of the dental
papilla accumulate.

 Four different types of epithelial cells can be distinguished on light microscopic

examination of the bell stage of the enamel organ. The cells form the inner enamel epithelium,
the stratum intermedium, the stellate reticulum, and the outer enamel epithelium.

 The junction between inner and outer enamel epithelium is called cervical loop and it is an
area of intense mitotic activity, this point is where the cells continue to divide until the tooth
crown attains its full size and which, after crown formation, gives rise to epithelial
component of root formation.

 In the bell stage, some epithelial cells between the inner enamel epithelium and the stellate
reticulum differentiate into a layer called the stratum intermedium. The cells of this layer
soon are characterized by an exceptionally high activity of the enzyme alkaline phosphatase .

 Although the stratum intermedium cells are histologically distinct from the cells of the
inner enamel epithelium, both layers work synergistically and have been considered as a
single functional unit responsible for the formation of Enamel.

 Two other important events occur during the bell stage.

1- The dental lamina (and the lateral lamina) joining the tooth germ to the oral epithelium
fragments, eventually separating the developing tooth from the oral epithelium.

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 The dental lamina is seen to extend lingually and is termed successional dental lamina as it
gives rise to enamel organs of permanent successors of deciduous teeth (permanent
incisors, canines and premolars). The enamel organs of deciduous teeth in the bell stage
show successional lamina and their permanent successor teeth in the bud stage.

2- The inner enamel epithelium completes its folding, making it possible to recognize the
shape of the future crown pattern of the tooth.

Fragmentation of the dental lamina results in the formation of discrete clusters of

epithelial cells that normally degenerate, but some may persist and are given the name
epithelial pearls.

These clusters of cells may form small cysts (eruption cysts) over the developing tooth
and delay eruption; may give rise to odontomes; or may be activated to form
supernumerary teeth.

An important consequence of the fragmentation of the dental lamina is that the tooth
continues its development within the tissues of the jaw divorced from the oral epithelium.
Thus, before the tooth can function, it must reestablish a connection with the oral
epithelium and penetrate it to reach the occlusal plane.

 When the tooth germ is growing rapidly during the cap-to-bell stage, cell division occurs
throughout the inner enamel epithelium. As development continues, division ceases at a
particular point because the cells are beginning to differentiate and assume their eventual
function of producing enamel.

 The folding that occurs as the crown develops results from intrinsic growth caused by
differential rates of mitotic division within the inner enamel epithelium. The cessation of
mitotic division within cells of the inner enamel epithelium determines the shape of a tooth.

 Before enamel formation begins, the stellate reticulum collapses, reducing the distance
between the centrally situated ameloblasts and the nutrient capillaries near the outer enamel
epithelium. Its cells then are hardly distinguishable from those of the stratum intermedium.
This change begins at the height of the cusp or the incisal edge and progresses cervically
 The cells of the outer enamel epithelium 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

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 dental sac forms papillae that contain capillary loops and thus provide a rich nutritional supply
for the intense metabolic activity of the avascular enamel organ.
 Before the inner enamel epithelium begins to produce enamel, the peripheral cells of the
mesenchymal dental papilla differentiate into odontoblasts under the organizing influence of
the epithelium. First, they assume a cuboidal form; later they assume a columnar form and
acquire the specific potential to produce dentin. The basement membrane that separates the
enamel organ and the dental papilla just prior to dentin formation is called the membrana
preformativa.

 The point at which inner enamel epithelial cell differentiation first occurs represents the site of
future cusp development. Because the inner enamel epithelium is constrained between the
cervical loop and cusp tip, continued cell proliferation causes the inner enamel epithelium to
buckle and form a cuspal outline. Thus the future cusp is pushed up toward the outer enamel
epithelium.
Eventually differentiation of inner enamel epithelium and papilla cells sweeps down along the
cusp slopes and is followed by the deposition of dentin and enamel first at the cusp tip. These
two matrices are deposited face to face, thereby defining the dentinoenamel junction. The
occurrence of a second zone of cell differentiation within the inner enamel epithelium leads to
the formation of a second cusp, a third zone leads to a third cusp, and so on until the final
cuspal pattern of the tooth is determined.

 Before formation of dental tissues begins, the dental sac shows a circular arrangement of its
fibers and resembles a capsular structure. With the development of the root, the fibers of the
dental sac differentiate into the periodontal fibers that become embedded in the developing
cementum and alveolar bone.

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 VASCULAR SUPPLY : The enamel organ is avascular, although a heavy concentration of
vessels in the follicle exists adjacent to the outer enamel epithelium. Clusters of blood vessels
are found ramifying around the tooth germ in the dental follicle and entering the dental papilla
during the cap stage. Their number in the papilla increases, reaching a maximum during the
bell stage when matrix deposition begins.

 NERVE SUPPLY :

 Pioneer nerve fibers approach the developing tooth during the bud-to-cap stage of
development.
 The target of these nerve fibers clearly is the dental follicle; nerve fibers ramify and
form a rich plexus around the tooth germ in that structure.
 Not until dentinogenesis begins, however, do the nerve fibers penetrate the dental
papilla (pulp).
 Although a possible relationship has been assumed between the developing nerve and
blood supplies (i.e., that the nerves might supply the vessels), the timing differs in
establishment of the papillary vascular and neural supplies.

Advanced bell stage

 This stage is characterized by the 1- commencement of mineralization and 2- root formation.
 During the advanced bell stage, the boundary between inner enamel epithelium and
odontoblasts outlines the future dentinoenamel junction.
 The formation of dentin occurs first as a layer along the future dentinoenamel junction in the
region of future cusps and proceeds pulpally and apically.
 After the first layer of dentin is formed, the ameloblast which has already differentiated from
inner enamel epithelial cells lay down enamel over the dentin in the future incisal and cuspal
areas.
 The enamel formation then proceeds coronally and cervically, in all regions from the
dentinoenamel junction (DEJ) towards the surface.
 In addition, the cervical portion of the enamel organ gives rise to the epithelial root sheath of
Hertwig. The Hertwig’s epithelial root sheath (HERS) outlines the future root and is thus
responsible for the shape, length, size, and number of roots.

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FORMATION OF THE PERMANENT DENTITION

 The permanent (secondary) dentition also arises from the dental lamina.

 The tooth germs that give rise to the permanent incisors, canines, and premolars form as
a result of further proliferative activity within the dental lamina at its deepest extremity. This
increased proliferative activity leads to the formation of another tooth bud on the lingual aspect of
the deciduous tooth germ .

 The molars of the permanent dentition have no deciduous predecessors, so their tooth
germs do not originate in the same way. Instead, when the jaws have grown long enough, the
dental lamina burrows posteriorly beneath the lining epithelium of the oral mucosa into the
ectomesenchyme. This backward extension successively gives off epithelial outgrowths that,
together with the associated ectomesenchymal response, form the tooth germs of the first,
second, and third molars.

 The teeth of the primary and secondary dentitions form in essentially the same manner,
although at different times.

 The entire primary dentition is initiated between 6 and 8 weeks of embryonic development;
the successional permanent teeth between week 20 in utero and 10 months after birth; and
the permanent molars between week 20 in utero (first molar) and 5 years of age (third molar).
Aberrations in this pattern of development result in missing teeth or the formation of extra teeth.

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