COURSES > HUMAN ANATOMY II, DDS09, AUT06 > CONCISE ORAL HISTOLOGY > AMELOGENESIS, DENTINOGENESIS AND

ROOT FORMATION

Amelogenesis, Dentinogenesis and Root Formation

Enamel Formation
Dental enamel is the most mineralized biological substance found in vertebrate animals, and the
inorganic material is present in a highly ordered, crystalline state. Despite the fact that mature enamel is
acellular and largely devoid of organic material, it is far from being inert. On the contrary, enamel is an
active chemical system that participates in a variety of reactions, including solute and ion transport from
saliva to dentin and back, ion-exchange reactions with saliva and demineralization-remineralization
processes. Although a sound knowledge of the crystalline structure of enamel is vital to understanding
dental caries and many aspects of dental therapeutics.

Amelogenesis

Amelogenesis actually begins after the start of dentin formation, but it is easier to consider building the
enamel contours of the tooth first. There are five stages in the life cycle of the ameloblast that are
important:

1. Prior to the differentiation of odontoblasts the cells of the inner enamel epithelium proliferate to
form the the basic shape of the tooth, i.e. they form the contours of the dentinoenamel junction.
At the end of this stage they become post-mitotic, terminally differentiated cells (ameloblasts).
2. Ameloblast differentiation begins with the elongation of the the inner enamel epithelial cells, and
a reorientation of their intracellular organelles. Most epithelial cells are polarized, they have a
basal end that sits on the basement membrane and an apical end that is involved in secretion,
absorption etc. As the inner enamel epithelial cells differentiate in to preameloblasts, the
'secretory pole' is reoriented towards the basement membrane, i.e. towards the DEJ. This
process is called 'repolarization'.
3. With the formation and mineralization of of dentin, the preameloblasts become fully differentiated
secretory ameloblasts and they begin to secrete enamel matrix. The differentiation of
ameloblasts begins at the cusp tips and incisal edges, and proceeds down the cusp slopes to the
cervix of the crown. Enamel matrix formation occurs on a diurnal cycle, with approximately 4 μm
of enamel formed every day, and the matrix is immediately mineralized to about 30% mineral by
weight. Each secretory ameloblast continues to synthesize matrix until the entire thickness of
enamel is it programmed to secrete is formed.
4. After matrix synthesis and secretion has been completed the secretory ameloblast differentiates
to become a 'maturation ameloblast'. The maturation ameloblast is responsible for the continued
mineralization of the enamel to it finally hardness, approximately 96% mineral by weight. The
maturation ameloblast actually cycles between two distinct morphological forms, called ruffle-
ended ameloblasts and smooth-ended ameloblasts.
5. When maturation is complete the ameloblasts 'de-differentiate', and together with the remnants
of the stratum intermedium, the stellate reticulum and the outer enamel epithelium, they form the
reduced enamel epithelium. The role of the reduced enamel epithelium in eruption and formation
of the junctional epithelium with be discussed in another lecture.

Disturbances in either the secretion or maturation of the enamel matrix can lead to defects in
enamel structure.

z Enamel hypoplasia is due to a decrease in the amount of matrix synthesized by the ameloblasts
z Enamel hypomineralization/maturation is caused by a lack of sufficient mineral incorporated

The biochemistry of the enamel matrix

The mineralization of the enamel matrix with a carbonated hydroxyapatite mineral generates the most
dense of all vertebrate mineralized tissues, and it is the unique proteins of the enamel matrix that are

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responsible for this. The principal proteins in the enamel matrix are:

1. Amelogenins are the major protein (~90%) secreted into the enamel matrix. They are a group of
heterogeneous proteins (20-30 kDa) that are hydrophobic and rich in proline, histidine and
glutamine. Amelogenins are thought to play a role in the organization and regulation of crystal
growth.
2. Tuftelin is a 45 kDa acidic, phosphorylated glycoprotein whose secretion is limited to the initial
formation of enamel at the DEJ. It's function is not known.
3. Ameloblastin (Amelin, Sheathelin)constitutes 5-10% of the enamel matrix. It is thought to
promote mineralization and crystal elongation.
4. Enamelins (60-80 kDa) are a heterogeneous group of proteins that may be involved in cyrstal
nucleation.
5. Proteinases are important components of the enamel matrix. They are responsible for the
progressive proteolytic cleavage of amelogenins. The processing of amelogenins to smaller
peptides is necessary for the regulation of crystal organization and growth.

Mineralization of the enamel matrix

The mineralization of the enamel matrix is described as a two-step process.

1. The ameloblasts secrete an organic matrix that is immediately mineralized to about 30% by
weight.
2. When the full thickness of enamel has been secreted by an ameloblast, a progressive increase in
mineral content begins. Smooth-ended ameloblasts remove of water and proteins from the
enamel matrix, whereas ruffle-ended ameloblasts participate in the active transport of calcium
and phosphate into the matrix. The cells of the stratum intermedium assist in the maturation
phase of amelogenesis.

Dentin Formation
Dentin makes up the bulk of the hard tissue in the tooth, and the organic and inorganic components
provides an appropriate supporting material for enamel. Despite the fact that mature enamel is highly
mineralized, and therefore hard and fairly brittle, enamel rarely shears off of the underlying dentin core
when subjected to stresses developed during mastication or bruxing. On the contrary, enamel and
dentin form a cohesive unit that successfully transmits forces through the enamel, and spreads them
through the more deformable dentin. Those stresses are subsequently transmitted through the root to
the mandibular and maxillary bone.

Nonetheless, the physical structure and biochemistry of dentin make it more susceptible to wear and
caries progression than the enamel. In particular, dentin is less mineralized (70% by weight), and
numerous tiny channels (dentinal tubules) make dentin quite porous.

Dentinogenesis

Dentinogenesis actually begins before the start of enamel formation, and unlike amelogenesis, it occurs
throughout the life of the individual:

1. Prior to the differentiation of odontoblasts the cells of the inner enamel epithelium proliferate to
form the the basic shape of the tooth, i.e. they form the contours of the dentinoenamel junction.
Odontoblast differentiation begins as the inner enamel epithelial cell undergo 'repolarization' to
become preameloblasts.
2. Each odontoblast secretes a collagen-rich organic matrix. This organic matrix is made up mainly
of type I collagen, but other proteins are also synthesized and secreted that play a role in
mineralization. Unlike the enamel matrix, predentin contains no mineral.
3. The organic matrix matures and is modified in order to regulate mineralization, and the
odontoblastic process plays an important role in mineralization. Certain proteoglycan species
inhibit mineralization, and as these are degraded they are endocytosed by the processes, which

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 concurrently secrete dentin phosphoproteins and other proteoglycans that appear to initiate
mineralization. The initial sites of mineralization resemble small globes in histological sections
and are called calcospheres.

Disturbances in either the secretion or maturation of the dentin matrix can lead to defects in dentin
structure, and as a consequence to the supportive function of dentin. Three different types of inherited
defects in dentin matrix are classied under the term dentinogenesis imperfecta. In these individuals, the
crowns are found to have a bulbous contour and the pulp chambers become obliterated by poor quality
dentin. Clinically, this results in a bluish or brownish cast to the teeth, and shortly after eruption the
enamel fractures away leaving the soft inner core of dentin exposed.

The biochemistry of the dentin matrix

The details of organic matrix interactions and their functional role in dentin mineralization are still under
investigation, but the principal proteins in the predentin are:

1. Collagen type I
2. Dentin phosphoprotein
3. Osteocalcin, Osteonectin and Osteopontin
4. Matrix Gla-protein (a γ-carboxyglutamate containing protein)

Types of dentin

Primary dentin makes up the bulk of the crown and root. The initial layer of dentin (closest to the DEJ) is
called mantle dentin and the remainder of primary dentin is called circumpulpal dentin. The outline of
dentinogensis described above is applicable to the formation of circumpulpal dentin. The synthesis and
mineralization of mantle, however, is somewhat different. During the formation of mantle dentin the
odontoblastic process is rudimentary. As a result, collagen fibers become oriented more perpendicular
to the DEJ. In addition, the initiation of mineral deposition occurs through a different mechnism, i.e. the
matrix vesicle. Matrix vesicles are small pieces of odontoblast membrane and cytoplasm that are
imbedded in the newly formed matrix. The crystallization of mineral takes place within these vesicles.

Primary dentin is formed at a rate of 4μm/day. Changes in the orientation of collagen fibers occur
approximately every five days, and this leads to the formation of lines of von Ebner. Longer term
changes in the rate of dentin formation cause the appearance of lines of Owen. The neonatal line in the
dentin of teeth whose crowns were forming at the time of birth is an example of a line of Owen.

After root formation is complete, the odontoblasts continue to form secondary dentin on the pulpal
surface of the dentin. Secondary dentin is added at a much slower rate than that of primary dentin, but
the basic structure of primary and secondary dentin do not differ.

In response to various types of external stimuli (chemical irritants, caries, restorative procedures,
attrition or trauma), tertiary dentin may be formed. Tertiary dentin is made very rapidly, and the
architecture of this type of dentin varies depending on the character of the stimulus. However, it is
generally far less organized than either primary or secondary dentin. The cells that form tertiary dentin
may be existing odontoblasts or activated mesenchymal cells from within the dental pulp. In the former
case, the dentin is often called reactive dentin, whereas the synthesis of tertiary dentin by newly
differentiated 'odontoblasts' is called reparative dentin.

Root Formation
After crown formation is complete, the cells of the inner and outer enamel epithelial continue to
proliferate (extending from the cervical loop) as the epithelial root sheath (of Hertwig). Unlike the
enamel organ in the crown, there is no intervening stratum intermedium or stellate reticulum. The
downward growth of root sheath determines both root number and form. Interactions between the inner
enamel epithelial cells and the cells of the dental papilla induces the formation of root dentin, and this is
follwed by the disruption of the root sheath. Gaps in the root sheath allow dental follicle cells to interact

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with the dentin, and they differentiate into cementoblasts. The cells of the root sheath remain as
epithelial islands within the periodontal ligament space.

In normal histological sections the interface between the newly formed cementum and root dentin has
been called the hyaline layer, or intermediate cementum. However, it is not cementum. Most
investigators now believe that the hyaline layer is a optical artifact caused by the structural
characteristics of this specialized interface. Specifically, the collagen fibers in dentin and in cementum
are interdigitated at right angles to one another, thus forming a very strong bond between the two
mineralized tissues.

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