86 Fundamentals of Oral Histology and Physiology

Circumpulpal dentin
Enamel The bulk of dentin is constituted of circumpulpal dentin exclu-
sively produced by the odontoblasts (Fig.  5.5a). Odontoblasts
initially form predentin, which undergoes mineralization and
Primary
Mantle becomes dentin. The daily production of dentin decreases from
dentin
dentin 10 μm/day at the onset of dentin formation to 4 μm/day. This
generates a rhythmic formation, and at each 4 μm a daily incre-
Secondary
dentin mental (von Ebner’s) line appears. This circadian rhythm is
Pulp
interacting with another longer-period rhythm forming Owen’s
Predentin
lines, appearing each 16–20 μm, due to unknown mechanisms.
The continual centripetal formation of primary dentin results in
Cementum
gradual reduction of the pulp volume (Figs. 5.2 and 5.3).
As each odontoblast deposits dentin matrix and retreats
toward the central pulp, it extends an elongated process from the
distal end of the cell body. Continual deposition and mineraliza-
tion of dentin matrix around the odontoblast process creates a
tubule within the dentin, from near the dentinoenamel junction
to the pulpal surface of the dentin. Dentin contains about 20,000
dentinal tubules per mm2, with variations between the outer
Figure 5.1 General organization of a tooth. Beneath the coronal and inner parts of the dentin layer. Due to space restriction,
enamel, the different layers of dentin are found. Mantle dentin is the more tubules are present in the inner third than in the outer
first dentin deposited; circumpulpal or primary dentin constitutes the third (between 18,000 and 21,000/mm2). The tubules are curved,
bulk of the tooth. Secondary dentin is deposited at a slow rate after displaying a gradual S shape from the dentinoenamel junction to
primary dentin formation is complete. A layer of predentin separates
the pulp. The primary dentin of the crown and most of the root
the dentin from the pulp. The root is covered by cementum.

Dentin location, types and structures

Enamel DEJ
Primary dentin Metadentin
Neonatal line Secondary
dentin Odontoblast
processes pulp
Hoehl' s cells

Odontoblast
cell bodies
predentin
Circumpulpal dentin

Mantle dentin
Hopewell-Smith & Tomes' granular layer

Figure 5.2 Schematic distribution of the different dentin layers located between the dentinoenamel junction (DEJ) and the dental pulp. Two
atubular peripheral layers are located beneath the enamel or cementum: the mantle dentin in the crown and the peripheral layers in the root,
including the atubular hyaline Hopewell-Smith layer and/or the Tomes’ granular layer. Primary dentin includes the atubular peripheral layers and
the tubular circumpulpal dentin of the crown and root. The neonatal line separates dentin formed before birth from dentin formed after birth.
Secondary dentin is formed continuously after the completion of primary dentin formation. The mineralization front, or metadentin, is the border
where dentin mineralization is initiated. Odontoblast processes cross the predentin (15-20 μm in width) and enter the dentinal tubules. Hoehl’s cells
may differentiate into odontoblasts, the dental pulp being located beneath.
Chapter 5 Dentin, Pulp, and Tooth Pain 87

(a) (b)

cp pd

e a
odontoblasts
pd

cp
d
odontoblasts

Hoehl' s
cells pulp

pulp
10 μm

Figure 5.3 Light micrographs of odontoblasts and forming dentin. (a) One-μm-thick section of a rat incisor. On the right, enamel (e) is formed by
secretory ameloblasts (a) The mineralization front (arrows) located at the edge of the dentin layer (d) is covered by silver grains (radioautography
using (3H) serine as phosphoprotein precursor). Odontoblast processes (arrowheads) extend through the predentin (pd). Dentin extracellular
matrix molecules are synthesized in odontoblast cell bodies. Hoehl’s cells may differentiate into odontoblasts. The odontoblast and Hoehl’s cell
layers include endothelial cells forming capillaries (cp). The dental pulp is located more centrally. Bar = 10 μm. (b) Hematoxylin- and eosin-stained
section of a human molar. Labels as in 5.3(a). Arrowheads indicate odontoblast processes, and arrows indicate Hoehl’s cells.

Table 5.1 Schematic distribution of the successive layers and types of sound dentin in longitudinal section between the dentinoenamel (DEJ)
and the pulp

Location Dentin Type Structure

Peripheral dentin Crown: Atubular dentin, 30–150 µm thick.

● Mantle dentin Initial dentin formed in the crown.
Non-phosphorylated proteins.
Resilient border.
Root: Initial atubular dentin.
● Hopewell-Smith layer: 8–15 µm Calcospherites or globular structures and interglobular spaces.
and/or A few bent minute tubules.
● Tomes’ Granular layer: 8–15 µm
Circumpulpal dentin Primary dentin The dentin formed between the creation of initial peripheral
dentin and early tooth functioning. The tubules display a
longitudinal S-shaped curvature. The formation of primary dentin
is limited by the outer DEJ and the initiation of secondary dentin
deposition.
Secondary dentin Formed normally and continuously after completion of the crown
and root. Results from the daily 4-µm dentin formation. The S-shape
of the tubules is more accentuated, and the tubules are less
numerous, due to space restriction.
Peripheral dentin Atubular dentin ● Mantle dentin
● Hopewell-Smith & Tomes’ granular layers
● Fibrodentin formation in the mesial and distal surfaces of
the root canal
Circumpulpal dentin Tubular dentin Intertubular dentin: a collagen-rich dentin associated with non-
collagenous proteins along and between the collagen fibrils. The
hydroxyapatite crystallites are 3–5 nm thick and 60 nm long.
Peritubular dentin: 25–30 nm crystallites, and an amorphous
network without collagen fibrils lining the lumina of dental
tubules.
88 Fundamentals of Oral Histology and Physiology

Table 5.2 Pathologic dentin

Causative Event Response Structural Alteration

Genetic alterations: Gene deletion of collagen I, or phosphorylated Abnormal dentin properties and tooth structure
● dentinogenesis imperfecta proteins (SIBLINGs) (DSPP, DSP, DMP1)
● dentin dysplasia
● X-linked hypophosphatemia
Environmental effects: Odontoblast gene expression changes Hyper/hypomineralization, structural alterations
● fluorosis
● dioxin
● other toxic agents
Loss of tooth structure: Reactionary dentin Ortho- or osteodentin formed beneath a
● caries calciotraumatic line
● abrasion
● restorative dental material
Pulp exposure Reparative dentin Dentinal bridge, partial or total pulp mineralization

(a) (b)

IG

DEJ MD CD GT
E
E

10 μm CA 10 μm

Figure 5.4 Electron micrographs of a rat molar. (a) Atubular mantle dentin (MD) in the coronal part of the forming molar. DEJ: dentinoenamel
junction. E: enamel. CD: circumpulpal dentin. (b) Beneath the forming enamel (E) and cervical ameloblasts (CA), calcospherites contribute to the
formation of the superficial Tomes’ granular layer (GT). An interglobular space (IG) is located between the granules. Bar = 10 μm.

is formed before eruption. The curvature is more accentuated in hydroxyapatite crystallites. The needle-like crystallites are 3 nm
tubules formed after the eruption of the tooth. in thickness and 60 nm in length. Each crystallite is formed by
Secondary dentin formation begins after completion of pri- the association of two plates (each 1 nm thick) separated by a
mary dentin formation. The structure and composition of sec- thin electron-lucent layer (Fig. 5.8a).
ondary dentin are similar to those of primary dentin. A slight The lumen of the dentinal tubules contains non-mineralized
change in the direction of the dentinal tubules occurs when sec- collagen fibrils and some amorphous proteins forming a perio-
ondary dentin formation begins. The formation of secondary dontoblastic structure around the odontoblast process. Peritubular
dentin continues at a slow rate throughout life, but is gradually dentin forms a highly mineralized ring around the tubules but
reduced in later years (Fig. 5.2 and Table 5.1). never completely fills the lumen. Isodiametric crystals about
Between the tubules, intertubular dentin is found (Figs. 5.5a, 25 nm in width and 9 to 10 nm in length resist abrasive forces and
5.6 and Table 5.1). This dentin specifically results from the con- occlusal pressures (Fig. 5.5b and Table 5.1). These crystallites have
tinuous apposition of predentin components at the mineraliza- high magnesium and carbonate content. They display high solu-
tion front (Fig.  5.3). It is a collagen-rich structure, containing bility in acid and chelating solutions. Peritubular dentin formation
fibrils about 80 to 100 nm in diameter (Fig. 5.6). Non-collagenous begins within the tubules, a few micrometers from the predentin-
proteins are associated either with the surface of collagen fibrils dentin junction. Present mostly in the inner two-thirds of the den-
and holes due to the quarter-stagger transversal bands of colla- tin, it is reduced in the outer one-third and almost lacking in the
gen fibrils, or they fill the spaces between collagen fibrils mantle dentin. Peritubular dentin does not contain collagen fibrils,
(Fig. 5.7). The dentin organic matrix plays a crucial role in pro- but is comprised of non-collagenous proteins, namely phospholip-
moting crystal nucleation and allowing limited growth to ids or lipoproteins and proteoglycans (Fig. 5.5b).
 (a) (b)

Lu

100 nm 100 nm

Figure 5.5 Electron micrographs of unstained ultrathin sections of human dentin. (a) Intertubular dentin. Crystallites are located along and
between collagen fibrils. The periodic banding of collagen is apparent. Barâ•›=â•›100â•›nm. (b) Peritubular dentin displays isodiametric crystallites about
35â•›nm in diameter. Lumen of dentinal tubule (Lu). Barâ•›=â•›100â•›nm.

co

100 nm

Figure 5.7 Electron micrograph of phosphorylated extracellular

0.5 µm
matrix€proteins stained with phosphotungstic acid–chromic acid (PTA)
in intertubular dentin. The electron-dense staining is located along and
Figure 5.6 Electron micrograph of type I collagen fibrils (co) in between the collagen fibrils. Barâ•›=â•›100â•›nm.
intertubular dentin. Demineralized section. Barâ•›=â•›0.5â•›μm.

(a) (b)

PD PD

MD

D 0.5 µm 0.1µm
D

Figure 5.8 Electron micrographs of rat molar dentin. (a) Unstained section. Needle-like crystallites located at the mineralization front between the
predentin (PD) and dentin (D) are seen along the collagen fibrils. Barâ•›=â•›0.5â•›μm. (b) The distal border (metadentin: MD) located between the predentin
(PD) and intertubular dentin (D) is densely stained by the phosphotungstic acid–chromic acid mixture. Barâ•›=â•›0.1â•›μm.
90 Fundamentals of Oral Histology and Physiology

In the sclerotic zone of carious decay, or after pathologic abra- dentin. These non-collagenous components have important
sion, or as a consequence of the aging process, the tubules may functions in mineral nucleation and crystal growth.
be filled by intratubular mineralization. Such heterogeneous
mineralization results from the occurrence of pathologic pro- Dentin formation
cesses that favor the precipitation of calcium and phosphate ions
into non-apatitic forms within the lumen of the tubules. Pre-odontoblast differentiation and tooth
The reparative process occurs in arrested carious lesions, or in morphogenesis
response to the preparation of cavities. It also may be due to the
effects of the release of resin monomers by restorative dental As described in Chapter 3, at early stages of tooth formation, neural
materials. These events lead to the formation of tertiary dentin, crest-derived mesenchymal cells migrate toward the first branchial
classified as either reactionary dentin or reparative dentin. arch. Interaction between these cells (condensing mesenchyme)
Hoehl’s cells (Fig.  5.3) and surviving secretory odontoblasts and epithelial cells located specifically at placode sites leads to the
form reactionary dentin, which appears either as orthodentin or proliferation of the epithelial cells and formation of a dental lamina.
osteodentin (Table  5.2). If both the odontoblastic and Hoehl’s The two tissues form the rudiment of the embryonic tooth, epithe-
cell layers are irreversibly altered, pulp progenitor cells contrib- lial cells forming the enamel organ and mesenchymal cells forming
ute to the formation of reparative dentin. the dental papilla, or embryonic pulp (Fig. 5.9).
Crown morphogenesis starts with the formation of dental buds,
followed by the cap stage, and later by early and late bell stages. The
Dentin includes atubular peripheral dentin (the mantle lateral proliferation of epithelial cells occurring at the periphery con-
dentin in the crown and Hopewell-Smith and/or Tomes’ tributes to increase the diameter of the enamel organ. Four epithelial
granular layers in the root) and tubular circumpulpal den- layers form the enamel organ: the outer enamel epithelium, the stel-
tin. After the formation of primary dentin, secondary dentin late reticulum, the stratum intermedium, and the inner enamel
is formed during the entire life of the functioning tooth. epithelium, the latter eventually differentiating to ameloblasts
Between the tubules, there is a network of intertubular (Figs. 5.9–5.12). The cells undergo a series of divisions, leading to the
dentin, whereas around the lumen of the tubules peritubu- formation of folds, the onset of shaping grooves and cusps. In the
lar dentin increases resistance to occlusal pressures. After
mouse molar, cell division in the embryonic mesenchymal pulp stops
a mild lesion (caries, abrasion, and/or restorative material),
before it does in the enamel organ, leading to a larger surface of inner
odontoblasts and Hoehl’s cells contribute to the formation
of reactionary dentin (also named tertiary dentin) beneath enamel epithelium compared with pre-odontoblasts. Folds begin to
a calciotraumatic line. After irreversible alteration of these form, leading to cusp formation. In the central area of the enamel
cells, or a pulp exposure, pulp cells contribute to the forma- organ, the enamel knot regulates the morphogenetic process, influ-
tion of reparative dentin. encing the process through cell proliferation and apoptosis.
Epithelial-mesenchymal interactions are instrumental in the
transition from mesenchymal embryonic pulp cells to a pre-
odontoblastic stage. The cells at the periphery of the embryonic
Dentin composition pulp undergo asymmetric division. After the last mitosis, the
Dentin consists of an organic matrix composed largely of Type I larger cells in contact with the basement membrane (BM)
collagen, within which hydroxyapatite mineral crystals are (basal lamina) become early pre-polarized odontoblasts
deposited. Mature dentin contains more organic substance than (Figs.  5.10 and 5.11). These cells ultimately differentiate into
enamel, but less mineral (Table 5.3). In addition to Type I colla- polarized secretory odontoblasts (Figs. 5.13a and b). The cells
gen, small amounts of Type III and Type V collagen are present. some distance away from the BM are smaller and retain their
Non-collagenous components, including phosphorylated and potential as stem cells. They are grouped in the sub-odontoblas-
non-phosphorylated proteins, proteoglycans, lipids, growth tic cell layer of Hoehl (Figs. 5.2 and 5.3), which may differentiate
factors, and enzymes, account for about 10% of the organic ultimately toward the odontoblastic lineage. It is crucial to high-
matrix (Table 5.4). While many of the non-collagenous compo- light that odontoblasts are post-mitotic cells. In case of carious
nents also are present in bone and other tissue, products of the decay or odontoblast injury, differentiation of Hoehl’s cells is
dentin sialophosphoprotein gene are specific to odontoblasts and reactivated and polarized secretory odontoblasts are implicated
in reactionary dentin formation (Figs. 5.14–5.16b and 5.16c). If
both the odontoblastic and Hoehl’s cell layers are irreversibly
Table 5.3 Summary of the global composition of dentin.

Component By Weight By Volume

Early tooth development involves reciprocal interactions
Mineral content 68–70% 50% between neural crest-derived mesenchymal cells and oral
ectoderm-derived cells of the enamel organ. The peripheral
Organic content 21% 30% mesenchymal cells of the dental papilla proliferate and
Water (free and bound) 11% 20% become pre-polarized odontoblasts, subsequently differen-
tiating into odontoblasts. Cells in the sub-odontoblastic
Note: Substantial differences occur between the different types of dentin layer retain their potential as stem cells.
described in the text.