Guided By:- Presented By:-

Dr. SHASHIT SHETTY . Dr. Hitesh Chopra

Dr. VIPIN ARORA . M.D.S. 1
ST
Year
Dr. MANOJ HANS.
Deptt. Of Conservative Dentistry & Endodontics
INDEX
Introduction
Odontogenesis
Clip of odontogenesis
Signaling Molecules that determine Odontoblasts
differentiation & Pulp Formation.
Techniques to study ODONTOGENESIS
Structure of dentin
Structure of pulp



INTRODUCTION
The interrelationship between pulp & dentin is a major
theme/aspect in the field of dentistry
In fact interaction of pulp with various tissues including dentin
serves as a rationale for specialty of endodontics
Also from many perspectives dental health is related to the
health of unique tissue, that is dental pulp
However, the study of dental pulp, is not restricted to this
tissue alone , but extends to its interactions with many other
tissues in dental health &disease. For ex as dentin &pulp are
anatomically & functionally related they are commonly
referred to as pulp dentin complex
WHAT IS IT?...................
The dentin & pulp tissue remain closely
associated during development & throughout
life of an adult & hence are referred to as
Pulpdentin Complex.
ODONTOGENESIS
first signs of formation day 11
thickening of the epithelium where tooth formation will occur
on the 1
st
branchial arch
signals earliest mesenchymal markers for tooth formation
are the Lim-homeobox genes (Lhx-6 and Lhx-7) expressed as
early as day 9 in the neural crest cells of the tooth region
positions of the teeth are controlled by signals from the oral
epithelium role of FGF-8 and Pax-9 determine the position
of the tooth germs
more than 90 genes have been identified in the oral
epithelium, dental epithelium and dental mesenchyme!! so
exact signaling mechanisms remain unclear
Tooth formation: Initial stages
involves the physiologic process of induction
The primitive oral cavity or stomadeum is lined by
stratified squamous epithelium called the oral ectoderm
The oral ectoderm contacts the endoderm of the
foregut to form the buccopharyngeal membrane
At about 37
th
day /6
th
week of gestation this
membrane ruptures and the primitive oral cavity
establishes a connection with the foregut.
Most of the connective tissues cells underlying the oral
ectoderm are neural crest or ectomesenchyme in origin
which are thought to instruct or induce the overlying
epithelium to start tooth development.

the oral ectoderm gives rise to the primary epithelial
bands
a basement membrane separates the developing oral
epithelium and mesenchyme



















Primary epithelial bands: Horseshoe-shaped bands that appear
approximately around the 37th day of development, one for
each jaw.
-there are two subdivisions: vestibular lamina and dental
lamina
1) the dental lamina -develops a series of epithelial outgrowths
- grow deep into the mesenchyme
-develops in the future spot for the dental arches
-will form the midline for these arches
-arches then form posteriorly from this
point
-the ingrowths represent the future sites
for each deciduous tooth
-
Primary epithelial bands: Horseshoe-shaped bands that
appear approximately around the 37th day of development,
one for each jaw.
-there are two subdivisions: vestibular lamina and dental
lamina
-the dental lamina develops a series of epithelial outgrowths
- grow deep into the mesenchyme
-develops in the future spot for the dental arches
-will form the midline for these arches
-arches then form posteriorly from this
point
-the ingrowths represent the future sites
for each deciduous tooth
-
2) the vestibular lamina cells rapidly enlarge and then
degenerate forms a cleft that becomes the vestibule of
the oral cavity
Bud Stage

marked by the incursion of
epithelium into the mesenchyme
period of extensive proliferation and
growth of the dental lamina
forms into buds or oral masses that
penetrate into the mesenchyme
each tooth bud is surrounded by the
mesenchyme
buds + mesenchyme develop into the
tooth germ and the associated
tissues of the tooth
this developing tooth forms from
both the ectoderm and mesenchyme
and from neural crest cells that have
migrated into the mesenchyme 1. Tooth bud
2. Oral epithelium
3. Mesenchyme
Cap Stage
characterized by continuation of the
ingrowth of the oral epithelium into
the mesenchyme.
tooth bud of the dental lamina
proliferates unequally in different
parts of the bud
forms a cap shaped tissue attached
to the remaining dental lamina
looks like a cap sitting on a ball of
condensing mesenchyme
occurs for the primary dentition
(during the fetal period)
this stage marks the beginning of histodifferentiation
(differentiation of tissues)
the tooth germ also begins to take on form start of
morphodifferentiation

a depression forms in the deepest
part of each tooth bud and forms the
cap or enamel organ (or dental
organ) produces the future enamel
(ectodermal origin)
below this cap is a condensing mass
of mesenchyme dental papilla
produces the future dentin and pulp
tissue (mesenchymal origin)
the basement membrane separating
the dental organ and the dental
papilla becomes the future site for
the dentinoenamel junction (DEJ)
remaining mesenchyme surrounds
the dental/enamel organ and
condenses to form the dental sac or
the dental follicle
Hence,
-together the enamel
organ + dental papilla +
dental follicle is
considered the
developing tooth germ
or tooth primordium
-these primordium will
be housed in the
developing dental
arches and will develop
into the primary
dentition
Bell Stage
Continuation of
histodifferentiation and
morphodifferentiation
cap shape then assumes a more
bell-like shape
differentiation produces four
types of cells within the
enamel/dental organ
1. inner enamel epithelium
2. outer enamel epithelium
3. stellate reticulum
4. stratum intermedium

during the bell stage the
dental lamina is separated
from the dental organ
the dental papilla undergoes
differentiation and produces
two types of cells
1. outer cells of the DP forms
the dentin-secreting cells
(odontoblasts)
2. central cells of the DP
forms the primordium of the
pulp
dental sac increases its
collagen content and
differentiates at a later stage
than the EO and DP

Differentiation of the Enamel/Dental organ
outer enamel epithelium (OEE)
cuboidal shape
protective barrier during enamel
production
may also be called the outer
dental epithelium
very little cytoplasm
cells are separated from the
dental follicle by a basement
membrane
cervical loop
IEE
OEE
inner enamel epithelium (IEE)
short, columnar cells
differentiates into the enamel
secreting cells = ameloblasts
separated from the dental
papilla below it by a basement
membrane also
cells accumulate large
amounts of glycogen
may also be called the inner
dental epithelium
the IEE and OEE are
continuous
region where they connect
curved rim of the EO = cervical
loop

IEE
stellate reticulum
star-shaped cells in many
layers
center of the enamel
organ
forms a network =
reticulum
supports production of
enamel
stratum intermedium
inner layer of compressed
flat to cuboidal cells
very high levels of the
enzyme alkaline
phosphatase
supports production of
enamel
Bell Stage
-the cells in the center of
the enamel organ begin to
synthesize and secrete
GAGs
-this pulls water into the EO

-increasing amount of fluid
in the EO forces the central
cells apart
-however, they remain
connected via cellular
processes which makes them
star shaped = stellate ret.
B = inner dental epithelium (inner enamel epithelium)
Bell stage early crown formation
the dental papilla is separated from
the enamel organ by a basement
membrane
immediately below this BM is a region
called the acellular zone
this is where the first enamel proteins
will be laid down
the dental lamina begins to break up
into discrete islands of epithelial cells
(epithelial pearls) separates the oral
epithelium from the developing tooth
these pearls may form cysts and delay
eruption or they may develop into
supernumerary teeth
the IEE completes its folding and you
can begin to identify the shape of the
future crown pattern

Tooth development so far
Cap and Bell stages & Permanent teeth
during the cap stage the
development of the permanent
dentition begins anterior teeth
the primordia for these teeth
appears as an extension off the
developing dental lamina
penetrates into the mesenchyme
lingual to the primary primordium
its site of origin is called the
succesional dental lamina
these permanent teeth are called
succedaneous teeth (anterior teeth
and the premolars)
teeth that form with the primary
tooth buds (primary
predecessors)
permanent molars are non-
succedaneous - they are formed
by posterior proliferation of the
dental lamina.

Ameloblasts and Odontoblasts
ameloblasts
the cells of the IEE assume a
more columnar shape or they
elongate
differentiate into pre-
ameloblasts
this differentiation is
characterized by the
repolarization of these PAs
movement of the nucleus away
from the basement membrane
this repolarization is critical to
the differentiation of the PAs
continued differentiation and
maturation results in the
formation of ameloblasts
the pre-ABs induce the cells of
the dental papilla to differentiate
also
odontoblasts
differentiation by the mesenchyme of
the dental papilla
occurs after differentiation of pre-ABs
begins
results because the pre-ABs induce
differentiation of the mesenchymal cells
also
also undergo repolarization mirror
image of the pre-ABs (see Figure 6-12
and 6-13)
after differentiation the ODs then start
dentinogenesis
begin to deposit predentin on the
side of their basement membrane
forms a layer immediately below the
BM and above the cells (figure 6-13)
therefore dentin formation begins
before enamel synthesis
explains why dentin is thicker than
enamel

At 1 the epithelium is separated
from the dental papilla by an
acellular zone.
At 2 the cells of the inner dental
epithelium have elongated, and the
acellular zone begins to be
eliminated as odontoblasts
differentiate from
ectomesenchymal cells in the tooth
pulp.
At 3 the odontoblasts retreat
toward the center of the pulp,
leaving behind formed dentin.
At 4 the cells of the inner dental
epithelium, now ameloblasts, begin
to migrate outward and leave
behind formed enamel.
before dentin forms cells of
the EO receive blood supply
from vessels of the dental
lamina
as dentin forms, it cuts of this
papillary source of
blood/nutrients
this causes a drastic reduction
in the amount of nutrients
that reach the EO
but the ABs require extensive
nutrients to form enamel
stellate reticulum collapses
and invagination of the OEE
this brings in blood supply
from peripheral vessels found
outside the tooth

Dentinoenamel junction
after OD differentiation and the initiation of
dentinogenesis the BM between the pre-ABs and
ODs disintegrates
this allows direct contact between the pre-ABs and
ODs results in the completion of pre-AB
differentiation to mature ABs
ABs then begin amelogenesis apposition of
enamel matrix
replaces the disintegrating BM
each ameloblast forms a tapered portion that faces
the disintegrating BM - called a tome or Tomes
process
upon contact of the enamel matrix and dentin the
disintegrating BM begins to mineralize forms the
dentinoenamel junction or DEJ
the ODs form cellular process as they retreat toward
the dental papilla (figure 6-14) that penetrate the
forming predentin = dentinal tubules
mineralization of the developing dentin and enamel
is distinct for each type of tissue
the cell bodies of the ODs remain in the pulp tissue
the cell bodies of the ABs participate in tooth
eruption and will disappear shortly after
Pulp formation


while the cementum is forming - the central cells of the
dental papilla form the pulp
HISTOLOGICALLY
HISTOLOGICALLY
STAGES ANOMALIES
Initiation Anodontia , Supernumerary teeth
Proliferation
Histodifferentiation Dentinogenesis imperfecta, Atypical
Dentin
Morphodifferentiation Abnormality in form & size of tooth, ex
peg laterals , hutchinsons incisor ,
twinning
Appoposition Enamel hypoplasia, hypocalcification
,intrinsic staining
video
Signaling Molecules that determine
Odontogenesis
or
Initiation of tooth
Primary epithelial band - FgF- 8
Dental lamina FgF-8 + BMP-4
Tooth bud formation
Epith. BMP 4 Epith. FgF8
Msx1
Ectomesen. BMP 4 + activin A


Tooth bud formation
Both FGF and BMP activate Msx1 and Dlx2, although Msx2 is
only activated by BMP and Dlx1 by FGF (Vainio; Bei and
Kettunen).

Jukka Jernvall

, and Irma Thesleff

. Reiterative signaling and
patterning during mammalian tooth morphogenesis . Mech
Dev2000 ; 92 :19-29
Tucker, A.S., Yamada, G., Grigoriou, M., Pachnis, V. and Sharpe,
P., 1999. Fgf-8 determines rostral-caudal polarity in the first
branchial arch. Development 126, pp. 5161.
Vainio, S., Karavanova, I., Jowett, A. and Thesleff, I., 1993.
Identification of BMP-4 as a signal mediating secondary
induction between epithelial and mesenchymal tissues during
early tooth development. Cell 75, pp. 4558
Bei, M. and Maas, R., 1998. FGFs and BMP4 induce both
Msx1-independent and Msx1-dependent signaling pathways
in early tooth development. Development 125, pp. 43254333


Ferguson, C.A., Tucker, A.S., Christensen, L., Lau, A.L., Matzuk,
M.M. and Sharpe, P.T., 1998. Activin is an essential early
mesenchymal signal in tooth development that is required for
patterning of the murine dentition. Genes Dev. 12, pp. 2636
2649.
Kettunen, P. and Thesleff, I., 1998. Expression and function of
FGFs-4, -8, and -9 suggests functional redundancy and
repetitive use as epithelial signals during tooth
morphogenesis. Dev. Dyn. 211, pp. 256268.

Tooth bud cap stage

enamel knot ( total 10 signals belongs to 4
families BMP,FGF,WnT,HG)

No BMP4 + MSX1 No Enamel knot
BMP4 + MSX1 Enamel knot
. Restricted expression of altogether 10 signals belonging to
the BMP, FGF, Hh and Wnt families has so far been reported in
the enamel knot.
The enamel knot has been suggested to be an important
regulator of tooth shape, and its induction conceivably is the
prerequisite for the tooth to develop into the cap stage.
Indeed, tooth development is arrested at the bud stage in the
knockouts of Lef1, Msx1, and Pax9 (Kratochwil; Bei and Peters
As these transcription factors are targets for BMP, FGF, and
Wnt signaling, the formation of the tooth cap seems to
depend again on these signaling pathways.
No enamel knots develop in the arrested tooth buds in the
mutant mouse embryos and, interestingly, a common
feature in the three mutants is that Bmp4 expression is
absent from the dental mesenchyme.
BMP4 therefore is a good candidate for a mesenchymal
signal inducing the transition from the bud to the cap stage.
This is supported by experiments in which the arrested
tooth buds from Msx1 mutant embryos continued
morphogenesis to the cap stage in vitro when BMP4 protein
was added to the culture medium (Chen et al., 1996).
A role for BMP4 specifically in the induction of the enamel
knot was supported by in vitro studies where BMP4 beads
placed on isolated dental epithelium induced the expression
of p21 and Msx2, two early markers of the enamel knot
(Jernvall et al., 1998).

Jukka Jernvall

, and Irma Thesleff

. Reiterative signaling
and patterning during mammalian tooth morphogenesis .
Mech Dev2000 ; 92 :19-29
kratochwil, K., Dull, M., Farinas, I., Galceran, J. and
Grosschedl, R., 1996. Lef1 expression is activated by BMP-4
and regulates inductive tissue interactions in tooth and
hair development. Genes Dev. 10, pp. 13821394.
Bei, M. and Maas, R., 1998. FGFs and BMP4 induce both
Msx1-independent and Msx1-dependent signaling
pathways in early tooth development. Development 125,
pp. 43254333
Peters, H. and Balling, R., 1999. Teeth where and how to
make them. Trends Genet. 15, pp. 5965.
Chen, Y., Bei, M., Woo, I., Satokata, I. and Maas, R., 1996.
Msx1 controls inductive signaling in mammalian tooth
morphogenesis. Development 122, pp. 30353044.
Jernvall, J., berg, T., Kettunen, P., Kernen, S. and Thesleff, I.,
1998. The life history of an embryonic signaling center: BMP-4
induces p21 and is associated with apoptosis in the mouse
tooth enamel knot. Development 125, pp. 161169.

Anterior teeth BMP4
Inhibit
BarX 1 gene

Proof ;- NOGGIN + BMP4 + BarX 1 gene = MOLAR( Tucker).

Posterior teeth BarX 1 gene Molar

Jukka Jernvall

, and Irma Thesleff

. Reiterative signaling and
patterning during mammalian tooth morphogenesis . Mech
Dev2000 ; 92 :19-29
Tucker, A.S., Matthews, K.L. and Sharpe, P., 1998.
Transformation of tooth type induced by inhibition of BMP
signaling. Science 82, pp. 11361138.
Cusp development FgF4 & P 21

Initiation of cusps

FgF4 Related to cusp tips
P21 rest
Jukka Jernvall

, and Irma Thesleff

. Reiterative signaling and
patterning during mammalian tooth morphogenesis . Mech
Dev2000 ; 92 :19-29

GENES EXPRESSED DURING TOOTH DEVELOPMENT
Gli Glioma associated oncogene homologue (zinc
finger protein ) (TF)
Lef Lymhoid enhancer-binding factor 1 (TF)
Pax Paired box homeotic gene (TF)
Fgf Fibroblast growth factor (SP)
Msx Msh-like genes in vertebrates (TF)
Dlx Distaless omologue in vertebrtes (TF)
Wnt Wingless homologue in vertebrates (SP)
Lhx lim-homeobox domain gene (TF)
Bmp Bone morphogenic proteins (SP)
Shh Sonic hedgehog (SP)
Hgf Hepatic growth factor (SP)
Ptc Patched cell surface receptor for SHH
Smo Smoothed PTC co-receptor for SHH
Pitx Transcription factor named for its
expression in the pituitary gland
Slit Homologous to Drosophila slit protein (SP)
Barx BarH1 homologue in vertebrates (TF)
Otlx Otx-related homeobox gene (TF)

Techniques to study
ODONTOGENESIS
1) Tooth organ culture systems
2) Odontoblasts & dental pulp cell cultures
3) Transgenic & knock out mice
4) Laser capture microdissection
1) Tooth organ culture systems

In vitro approaches include;-

1) Tooth organ culture by means of Trowel type
system
In this either whole mandibular & maxillary implants or
individually dissected molar organs can be cultured in
enriched serum by means of Trowel type system which
involves placing the tooth organ in the correct orientation on
a filter that is supported by a metal grid at the gas liquid
interface

2) Functional tooth organ recombination assays
Dental epithelium is separated from papilla mesenchyme by
means of enzymes that degrade the basement membrane at
the interface.
Isolated epithelium & mesenchyme can be cultured separately or
recombined & then transplanted in vivo to study the effects
on tooth development









3) Bead implantation assays

Briefly, either heparin or agarose beads that are soaked in known
concentration of growth factor are placed on separated
dental mesenchyme . After approximately 24 hrs in culture,
the mesenchyme is analyzed for changes in gene & protein
expressin in the region surrounding the bead.

2) Odontoblasts & dental pulp cell cultures

Tooth organ culture system facilitates the studies of early
tooth development but this facilitates the study of late stage
of tooth development that involve cell differentiation &
matrix synthesis
Two approaches can be used -
1) To utilize hemisectioned human teeth from which dental
pulp has been carefully extirpated. The remaining layer of
intact odontoblasts then can be cultured with the native pulp
chamber , to which nutrient media & various growth factors
or cytokines are added.
2) Thick slices of human teeth with the odontoblast layer left
intact offer another useful approach to study the behaviour
of odontoblasts under conditions that stimulate dentinal
caries
Cell line of odontoblast resp. MO6 -G3 & MDPC-23 & Pulpal
cell lines RPC-C2A &RDP 4-1 have been successfully obtained
which are used to study various characteristics.
3) Transgenic & knock out mice
The modern era of recombinant DNA technology & genetic
engineering has made it possible to alter or mutate a gene of
interest in vitro & then inject into the pronucleous of a
fertilized mouse egg
Transgenic mice generated through conventional technology
can be designed to over express the gene of interest in cells
or tissues where it is normally expressed
When it is study the behavior of gene at an ectopic site , the
transgene of interest is placed behind the promoter of
another gene that will drive expression in tissues where it is
not normally expressed
In the case of a gene that is expressed inn multiple tissue or
organs its now possible to study activity at one particular site,
by driving the expression of a transgene with a tissue or cell
specific promoter
4) Laser capture microdissection
This is one of the most innovative techniques which is now
being used in Cancer Genomic Anatomy Project to catalog the
genes that are expressed during solid tumor progression & to
construct complementary DNA (C DNA) libraries from normal
and premalignant cell populations . Microarray panels
containing these index genes are being used to obtain gene
expression pattern in human tissue biopsies .
The fluctuation of expressed genes that correlate with
particular stage of disease is compared within or between
individual patients ,
Such a finger print of gene expression patterns will provide
important clue regarding etiology & contribute to diagnostic
decisions & therapy
Application in this area will be particularly useful for dental
pulp research , in which individual cell populations are
difficult to access
The progress in understanding odontoblast differentiation has
been slow because of serious limitations inherent to both in
vivo & vitro approaches.
Initial studies of dentin ECM gene expression in differentiating
odontoblasts have been promising. Data from the future use
of laser capture micro dissection will provide a correlation
between the morphologic changes and the expression of
known ECM genes during odontoblast differentiation.
Information generated from this approach will also be
valuable in developing a nomenclature that can be
consistently used by researchers.

Moreover , known and unknown genes will be identified from
the developmentally staged, odontoblast specific cDNA
libraries.
Genes that are defined for each stage of primary dentin
formation will provide important clues about temporal
patterns of genes expression and the potential functions of
encoded protein products in dentin mineralization .
Such fundamental information will be useful in characterizing
cells within the cell-rich zone of dental pulp, identification of
replacement population of pulpal cells involved in reparative
dentin formation , and in development of vital pulp therapies
aimed at hastening the healing of the injured pulpo - dentin
complex.
Dentin
STRUCTURE

Dentin consists of microscopic channels, called dentinal
tubules, which radiate outward through the dentin from the
pulp to the exterior cementum or enamel border.
From the outer surface of the dentin to the area nearest the
pulp, these tubules follow an S-shaped path.
The diameter and density of the tubules are greatest near the
pulp.
Tapering from the inner to the outermost surface, they have a
diameter of 2.5 m near the pulp, 1.2 m in the middle of the
dentin, and 0.9 m at the D.E.j.
Their density is 65,000 per square millimeter near the pulp,
whereas the density is 15,000 at the DEJ.

Changes in dentin structure with depth
The area occupied by the lumina of dentinal tubules can be
calculated as the product of the cross sectional area of a
single tubule ,r2 & N , the no. of tubules per cmsquare .
As both the radius of dentinal tubules & their no. per unit
area increases as one moves from DEJ to pulp , the area
occupied by dentinal tubules also increases.
The water content of dentin near the DEJ is about 1% & while
that of dentin near the pulp is about 22% , a 22 fold variation.
This is important with respect to bonding with composite
restorative materials in deed cavities in a vital tooth but not in
endodontically treated tooth


STRUCTURE
Cont.
Made up of CHEMICAL COMPOSITION ;-
70% inorganic materials (mainly hydroxylapatite and some
non-crystalline amorphous calcium phosphate),
20% organic materials (90% of which is collagen type 1 and
the remaining 10% ground substance, which includes dentine-
specific proteins),
and10 % water (which is absorbed on the surface of the
minerals or between the crystals).
Types
A) Based on time of formation
Mantle Dentin
1) Primary dentin
Circumpulpal dentin
2) Secondary dentin
3) Tertiary dentin Reactionary Dentin
Reparative Dentin
B) Based on age changes
1) Dead tracts
2) Sclerotic dentin
3) Reparative dentin
mantle dentin
tertiary dentin
primary dentin
secundary dentin
predentin
Section of the tooth types of dentin
1) Primary dentin
Mantle dentin
Its the name of the first formed dentin in the crown
underlying D.E.J
Its thus the most peripheral part of the primary dentin &
is about 20 m thick.
Circumpulpal dentin-
It forms the remaining primary dentin or bulk of the
tooth.
Its the Circumpulpal dentin that represents all of dentin
before root completion

Mantle dentin Circumpulpal dentin-
Collagen fibers large small
Lightly packed Tightly packed
Less mineralized More mineralized
Has fewer defects Has more defects
2) Secondary dentin
Secondary dentin is formed after root formation is complete,
normally after the tooth has erupted and is functional.
It grows much slower than primary dentin, but maintains its
incremental aspect of growth.
It has a similar structure to primary dentin, although its
deposition is not always even around the pulp chamber. It is
the growth of this dentin that causes the decrease in the size
of the pulp chamber with age; by depositing more on the roof
& floor of the coronal pulp chamber.
3) Tertiary dentin
Tertiary dentin is dentin formed as a reaction to external insult such
as caries. It is of two types, either Reactionary, where dentin is
formed from a pre-existing odontoblast or is it Reparative, where
newly differented odontoblast-like cells are formed due to the
death of the original odontoblasts, from a pulpal progenitor cell.
Tertiary dentin is only formed by an odontoblast directly affected
by stimulus, therefore the architecture and structure depends on
the intensity and duration of the stimulus e.g. if the stimulus is a
carious lesion, there would be extensive destruction of dentin and
damage to the pulp, due to the differentiation of bacterial
metabolites and toxins.
Thus tertiary dentin is deposited rapidly, with a sparse and irregular
tubular pattern and some cellular inclusions known as osteodentin.
However if the stimulus is less active, it would be laid down less
rapidly with a more regular tubular pattern and hardly any cellular
inclusions.

Reactionary dentinogenesis
Its by definition dentin secreted by surviving primary
odontoblasts
So its a less complex formation as compared to that of
reparative dentinogenesis in which dentin is secreted by
odontoblasts like cells
Factors infleuencing Reactionary dentinogenesis
1) R.D.T
2) Etchant
3) Restorative material

1) R.D.T
It was significantly the most
important factor in
determining the Secretion of
reactionary dentin , which is
increased in area by 1.187
mm2 for every 1mm
decrease in the R.D.T
beneath the cavity

2) Etchant
A. Time of etching
Cavity etching can positively influence the secretion of
reactionary dentin .
Treatment wit EDTA for 0 sec. ., 60 sec. , & 120 sec. lead to a
ranking of 60 sec. > 120 sec. > 0sec. for reactionary
dentin secretion
Hence reduced reactionary dentinogenesis after 0 & 120 sec.
of treatment was associated with decreased odontoblast
survival
B . Type of etchant
Phosphoric acid has less stimulatory effect as compared to EDTA
on reactionary dentinogenesis
3) Restorative materials
Restorative materials that are capable stimulting reactionary
dentinogeneis , such as calcium hydroxide , hv similar action
on dentin-matrix releasing growth factors , which then diffuse
to the odontoblasts & prompt their upregulation
Reparative dentinogenesis
Its by definition dentin secreted by odontoblasts like cells
In non exposed pulps it may be sequel to reactionary
dentinogenesis or it may occur independently in the absence
of reactionary dentin if the injury is of sufficient intensity (e.g.
an active carious lesion)
The reparative response of tertiary dentinogenesis always
takes place at sits of pulp exposure because of loss of
odontoblasts & need for dentin bridge formation
Its a more complex formation as compared to that of
reactionary dentinogenesis in that progenitor cells from the
pulp must be recruited & induced to differentiate into
odontoblasts like cells before their secretions may be up
regulated to form the reparative dentin


B) Based on Age changes

1) Dead tracts
Disintegration of odontoblastic processes due to various stimuli
leads to formation of dead tracts
They are called so because tubules get filled with air which appear
black in transmitted & white in reflected light

2) Sclerotic dentin
Deposition of apatite crystals leads to formation of Sclerotic or
transparent dentin
They are called so because refractive indices of intertubular dentin
after crystal deposition becomes equal to that of peritubular
dentin.
It appear light or transparent in transmitted & dark in reflected
light

3) Reparative dentin

The empty ( normal) dentin tubules
Mineral deposits narrow down dentin
tubules

Smear layer is debris after
instrumentation
Its about 1-3m

Smear plug in coronal or
radicular dentinal tubules
are 1-3m long , smear
plugs in instrumented
root canals reach upto 40
times longer i.e.. 40m .
SMEAR LAYER & SMEAR PLUGS
Pulp
ANATOMY OF PULP

Each person can have a total of up to 52 pulp organs, 32 in the
permanent and 20 in the primary teeth.
The total volumes of all the permanent teeth organs is 0.38cc
The mean volume of a single adult human pulp is 0.02cc
Consists of two parts Coronal Pulp & Radicular Pulp.
Apical foramen is the opening of the Radicular Pulp into the
periapical connective tissue.
The average size is 0.3 to 0.4 mm in diameter.
STRUCTURE OF PULP
Has three layers (Outerrmost to
innermost)
1. Odontoblastic Layer; outermost layer
which contains odontoblasts and lies
next to the predentin and mature
dentin
2. Cell Free Zone (Zone Of Weil ) which
is rich in both capillaries and nerve
networks. The nerve plexus of
Rashkow is located in here

3. Cell Rich Zone; innermost pulp layer
which contains fibroblasts and
undifferentiated mesenchymal cells


Nerve fibers Pulp and Dentin entering
the teeth have been identified
histologically as myelinated A-
fibers and unmyelinated C-fibers
which enter through the apical
foramina of the teeth, passing
through the radicular to the coronal
pulp where they fan out and diverge
into smaller bundles.As divergence
continues; individuals-fibers within
small bundles lose their myelin
sheath and divide repeatedly before
finally ramifying into a plexus of
single axons known as the
SUBODONTOBLASTIC PLEXUS OR
PLEXUS OF RASCHKOW in Cell Free
Zone
Neuroanatomy of Pulp and Dentin
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." From this plexus nerve fibers are
distributed toward the pulp-dentin
border with terminals showing a
characteristic bead-like structureThis has
been summarized by Trowbridge
1) MARGINAL FIBERS : S.O.P O.L
They do not reach the predentin.
2) SIMPLE PREDENTINAL FIBERS
S.O.P PREDENTIN
They donot branch
3) COMPLEX PREDENTINAL FIBERS
S.O.P PREDENTIN
multiple branches and multiple ending-
like enlargements on each branch.
4) DENTINAL FIBERS
S.O.P DENTIN
pass through the predentin without
branching and enter the dentin through
the dentinal tubule. The penetration is
limited to approximately 100 um.


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Neurophysiology ( General )
TYPE OF NERVE
FIBRE
SIZE VELOCITY OF
CONDUCTION
FUNCTION
A
( Myelinated )
12-20 m 70-120 m/sec proprioception.
A
( Myelinated )
5-12 m 30-70 m/sec transmission of touch and
pressure.
A
( Myelinated )
3-6 m 15-30 m/s ec for motor function to the
spinal nerves
A
( Myelinated )
2-5 m 12-30 m/sec transmission of pain,
temperature, and touch.
B
( Myelinated )
1-3 m 3-15 m/sec preganglionic autonomic
function
C
( Non- Myelinated )
0.2-2 m 0.5-2 m/sec postganglionic
sympathetic pain and
possibly heat, cold, and
pressure

Neurophysiology of Pulp and Dentin
Dental pulp is innervated by both
1) Myelinated and
2) Nonmyelinated nerve fibers

Recent electrophysiological investigations on intradental nerves
of experimental animals confirm histological evidence that
two fiber groups exist ;-

1) A
2) C nerve fibres









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A Nerve fibres
C Nerve fibres
Myelinated Non-Myelinated
Fast conduction velocity Slow conduction velocity
Initial response Late response
Periphery Center
Heat/Cold sensitive Heat sensitive
Sharp localised pain Dull poorly localised pain
Get necrosed easily More resistant to necrosis
Mechanisms of Stimulus Transmission Across
Dentin

Four theories are given
1) DIRECT STIMULATION OF NERVE FIBERS
2) THE DENTINAL RECEPTOR MECHANISM HYPOTHESIS
3) THE HYDRODYNAMIC THEORY
4) ALTERNATIVE MECHANISM (MODIFIED HYDRODYNAMIC
THEORY)



Anderson and Naylor proposed two possible explanations:
There were no nerve elements in dentin. When pain was evoked it
was due to stimulation of receptor mechanisms in the pulp by a
disturbance transmitted through the tubules by non-neural means.
There are receptor mechanisms in dentin that could be stimulated
indirectly, but cannot be reached by direct stimulation from
chemical agents because of some barrier to diffusion in the tubules.

3) Newly erupted toot does not have Plexus Of Raschkow & yet its
sensitive
Conclusion ;-
Autoradiography studies have demonstrated that nerve fibers in the
dentin is not a necessary prerequisite for its sensitivity, which also
supports the evidence of Lilja that root dentin contains no
intratubular nerves, but nevertheless is very sensitive.

DISCARDED
1) DIRECT STIMULATION OF NERVE FIBERS
Given by - Anderson and co-workers
According to this theory dentin was directly innervated by
nerve fiber as a result stimuli to exposed dentin would cause
pain.

Contraindications ;-
1. a) Application of algogenic (pain-inducing) substances such
as potassium chloride, acetylcholine, 5-hydroxytryptamine,
and histamine failed to elicit a response; whereas when
applied directly to exposed pulpal tissue, an immediate
response was elicited.
b) Similarly, topical anaesthetic solution when applied to
the exposed sensitive dentin did not decrease sensitivity.
2) THE DENTINAL RECEPTOR MECHANISM
HYPOTHESIS


Acc. to this hypothesis odontoblast has a special sensory
function , the processes of odontoblast in close contact with
terminal sensory nerve endings forms so-called specialized
junctional complexes reffered to as NEUROSENSITIVE
COMPLEX which is than responsible for pain/ sensivity
transmission through exposed dentin
Points against this theory

Dentinal sensitivity persists following the degeneration of
both odontoblasts and intratubular nerve fibers in the inner
third of dentin. Such studies appear to contradict the
hypothesis that odontoblasts act as a dentinal receptor
mechanism.
Morphological evidence of a synaptic relationship between
odontoblast and sensory nerve endings is lacking.
Impulse is too low.
Application of local anaesthetics does not remove pain/
sensivity


Points in favour of this theory

Odontoblast is of neural crest origin & it retains an ability to
transduce & propogate an impulse.

EVIDENCE FROM HISTOCHEMICAL AND ELECTROPHYSIOLOGICAL
STUDIES

1. Impulse transmission via Neurotransmitter

Avery and co-workers have shown that odontoblast
protoplasmic extensions were cholinesterase positive.
Ten-Cate and Shelton demonstrated cholinesterase
activity in both myelinated and non-myelinated nerve fibers of the
pulp, but not close to, or in, odontoblasts or their processes. They
concluded that if the transmission of impulses associated with
dentinal sensitivity was via a dentinal receptor mechanism, then
there was no evidence to suggest that these impulses were
mediated by cholinergic activity


2) Impulse transmission by polypeptides, such as plasma kinins
and Substance P (modulation theory).
However it also fails to prove nerve transmission.


DISCARDED



3) THE HYDRODYNAMIC THEORY


Proposed by BRAINSTORM
Most widely accepted theory.
Acc. to this theory- The movement of fluid in dentinal
tubules , the functional unit of dentin is responsible for
transmission of pain/sensitivity

Hagen Poiseuielle equation
- fluid movement -
basis of the hydrodynamic theory
V=Pr
4
/ 8L

V: fluid movement
P: pressure difference
r: radius
: viscosity
L: length


Points in favour of this theory
1) Newly erupted toot does not have Plexus Of Raschkow & yet
its sensitive
2) Experimental evidence ;-

1) The effect of pressure:
Both increase & decrease of pressure results in rapid fluid shift
which elicits pain e.g...Following a decrease in pressure, an
immediate pain response was elicited which persisted for as
long as there was decreased pressure. Histological examination
showed odontoblasts nuclei in the tubules. Brainstorm
concluded that these effects were probably due to intense
evaporation from the dentinal surface. The dislocation is
probably due to aspiration of the odontoblasts into the dentinal
tubules in connection with the capillary fluid flow.
2) The effect of dehydration:
It also elicits painful response because withdrawing of fluid
contents of dentinal tubules
Hypertonic solutions such as sugar and calcium chloride also
elicit pain by the same effect of dehydration of the dentinal
surface
This effect, too, can be explained by dentin tubule fluid
movements, since fluids of a relatively low osmolarity (e.g.,
dentinal tubule fluid) will tend to flow toward solutions of
higher osmolarity.
When isosmotic solutions are applied, no stimulus is
perceived.
3) The effect of thermal changes

Cold stimulus - causes contraction of tubule contents, which in
turn a rapid outward movement of fluid away from the pulp.
Hot stimulus- expansion of tubule contents occurred with a
subsequent increase in pressure, which resulted in a rapid
inward movement of fluid toward the pulp.

4) The effect of mechanical probing

These procedures could cause the removal of dentinal fluid from
the exposed dentin surface and by capillary action elicit an
outward flow of tubule contents from the pulp, stimulating the
odontoblast structure and causing pain.


3) It also explains why application of local anesthetics does
not remove pain/ sensitivity


ACCEPTED


4) ALTERNATIVE MECHANISM
(MODIFIED HYDRODYNAMIC THEORY)
Desensitization of dentin by blocking nerve activity
(direct ionic diffusion)

Several investigators have used a neurophysiological model to
evaluate dentin sensitivity. The results from these studies,
together with the work of Kim and other Theory)
investigators, would suggest that application of various
chemical solutions (in particular K+-containing compounds)
to dentin resulted in raising the intratubular K+ content
which in turn rendered the intradental nerves less excitable
to further stimuli by depolarizing the nerve fiber(s)
membrane.

On the basis of these studies, Kim and Markowitz and Kim
proposed an alternative mechanism, namely desensitization
of dentin by blocking nerve activity (direct ionic diffusion).This
recent hypothesis has, however, been criticized

. According to Sena, Kim's work was based on deep-cut cavity
preparations, with only a very thin slice of dentin between the
exposed dentin surface and the pulp. In consequence K+ had
only a short distance to traverse the length of the tubule. In
the normal clinical situation, however, the incoming K+ (i.e., if
applied in a toothpaste on exposed cervical dentin) would
have to overcome the opposing pulpal pressure that produces
an outward flow of dentinal fluid. Such an outward flow can
prevent the inward diffusion of substances from the oral
cavity.
While the alternative or modified hypothesis of stimulus
transmission across dentin as proposed by Kim and Markowitz
and Kim appears to be an attractive alternative to the
hydrodynamic theory, this hypothesis nevertheless requires
further investigation.


Reference
3) Western Society of Periodontics
Abstract Index,
Volume Number 2, 1995,
D.G. GILLAM
Mechanisms of Stimulus Transmission Across Dentin

Dentin is the only innervated hard tissue of the tooth.

As dentin is basically made of dentinal tubules exposure to
oral environment invariably results in fluid shift.

These fluid shift can directly stimulate odontoblasts , pulpal
nerves and subodontogenic blood vessels by applying large
shear forces on their surfaces as the fluid streams through
narrow spaces .

Fluid shift in dentinal tubules

Physiological changes in blood vessels

Increased flow of plasma fluid & plasma proteins from vessels
into pulpal tissue spaces & out into dentinal tubules

Extravasation leads to of plasma causes increase in local pulpal
tissue pressure

Increased firing of sensitized neurons

Increased sensitivity & pain
Increase of outward fluid movements from the
pulp during inflammation
DYNAMICS OF THE PULP DENTIN
COMPLEX
D.H. Pashley
Department of Oral Biology, School of
Dentistry, Medical College of Georgia,
Augusta, Georgia 30912-1129, USA
Introduction

The purpose of this review is to integrate a great deal of
recent information from a wide variety of fields on how the
pulpo-dentin complex responds during nonphysiologic
conditions.
Indeed, it is during pathologic stresses that the full range of
biologic responses of the pulp to changes that occur in dentin
can be expressed.
These are the conditions with which most dental clinicians
must contend. It is hoped that insights gained by the study of
pulpal pathobiology will provide new therapies that will be of
use to dentistry in the future.


This review will cover the
1) physiology of dentin, the mechanical properties of dentin,
2) how dentin structure can be modified for therapy,
3) changes in pulpal innervation in response to exposure of
dentin,
4) dentin sensitivity, the inter-relationships among pulpal
nerves, dentin, and
5) pulpal inflammation, and the reactions of the pulp to wound
healing.

(I) The Concept of the Pulpo-Dentin Complex

The physiologic concept of the "pulp-dentin complex has
recently been challenged as an oversimplication (Goldberg
and Lasfargues, 1995).
These authors correctly point out that there are a number of
differences between the chemistry of dentin and that of pulp.
For instance, the collagen matrix of dentin is composed
primarily of type I collagen, while that of the pulp contains
collagen types I, III, and V, among others. Type III collagen is
associated with three-dimensional fibrillar networks, while
type IV collagen is found in basement membranes.
Since peripheral dentin has no basement membrane,it is not
surprising to find that it has no type IV collagen.
The distribution of noncollagenous proteins is also different.
Ex. Phosphophoryn is found in mineralized dentin and seems
to modulate mineralization, but it is not located in non-
mineralizing pulpal soft tissue (Butler, 1995; Linde and
Lundgren, 1995).
These differences led Linde and Goldberg (1993) to conclude
that although pulp and dentin tissues share a common
ancestry, the dental papilla, and although the development of
the two is closely interrelated, there are no direct chemical
similarities between the pulp and dentin.
There is a great deal of evidence that dentin and pulp are
functionally coupled and hence are integrated as a tissue.
1) When normal intact teeth are stimulated thermally, dentinal
fluid expands or contracts faster than does the volume of the
tubules that contain the fluid. This causes hydrodynamic
activation of intradental nerves and is an example of the
functional coupling of pulp and dentin in an intact tooth.
2) As soon as the external tissues that seal dentin peripherally
(e.g., enamel and cementum) are lost for any reason, the
normal compartmentalization between the two tissues is
lost, and they become functionally continuous.
Under these pathologic conditions, there is a fluid-filled
continuum from the dentin surface to the pulp. It is through
this medium that external stimuli are transduced to physical
disturbances in the pulp.

It is through this fluid medium that bacterial substances can
diffuse across dentin to produce pulpal reaction(Bergenholtz,
1977, 1981, 1996; Bergenholtz and Warfvinge, 1982)

The pulp responds to these insults in the
1) short-term by mounting an inflammatory response which
produces an outward movement of fluid (Vongsavan and
Matthews, 1991, 1992a,b, 1994; Matthews, 1996) and
macromolecules (Raab, 1989; Maita etal, 1991; Byers, 1996).
2) In the long term, pulpal tissues produce tertiary dentin as a
biologic response to reduce the permeability of the pulpo-
dentin complex and to restore it to its original sequestration.
Radioactive tracer experiments clearly demonstrate the
continuity of the dentinal fluid-pulpal fluid-circulation in
exposed dentin and the importance of pulpal blood flow in
clearing pulpal interstitial fluids of exogenous material
(Pashley, 1979, 1985; Potts et al, 1985).


(II) Dentin
DENTIN MORPHOLOGY

Dentin can be regarded as a porous biologic composite made
up of apatite crystal filler particles in a collagen matrix (Fig. 1).
This mineralized matrix was formed developmentally by
odontoblasts which began secreting collagen at the DEJ and
then grew centripetally while trailing odontoblast processes.

The presence of these cellular processes makes primary and
secondary dentin tubular in nature.

Since the circumference of the most peripheral part of the
crown or root of a tooth is much larger than the
circumference of the final pulp chamber or root canal space,
the odontoblasts are forced closer together as they continue
to lay down dentin, ending up in a columnar layer in parts of
the coronal pulp, especially over pulp horns (Couve, 1986).
They are more cuboidal in the root canal and become flat
near the apex. The convergence of dentinal tubules toward
the pulp creates a unique structural organization to dentin
which has profound functional consequences, as will be
discussed below.
This convergence has been estimated to be
a) 5:1 (HoppeandStiiben, 1965),
b) 4:1 (Walton et a!., 1976), or
c) 3:1 (Fosse et al, 1992) in coronal dentin.
Each individual dentinal tubule is an inverted cone with the
smallest dimensions at the DEJ and the largest dimensions at
the pulp.
Originally, each tubule had a diameter of nearly 3 m.
However, within each tubule is a collagen-poor,
hypermineralized cuff of intertubular dentin which is called
"peritubular dentin" in many texts.
It is really periluminal dentin or, more accurately, intratubular
dentin (Linde and Goldberg, 1993). Its formation narrows the
lumen of the tubule from its original 3 m to as little as 0.6-
0.8 m in superficial dentin.
This larger amount of peritubular dentin in superficial dentin
near the DEJ is due, in part, to the fact that it is "older than
middle or deep dentin, which was more recently secreted and
mineralized.
Thus, the width of intratubular (peritubular) dentin decreases
as tubules are followed inward toward the pulp, with the
exception that there is no peritubular dentin in intraglobular
dentin (Blake,1958).
Very close to the pulp, there is no intratubular or peritubular
dentin, and the tubule (luminal) diameter is about 3 m
(Garberoglio and Brannstrom, 1976).
Thus, most of the narrowing of the tubule lumen as one
observes dentin more peripherally is due to deposition of
peritubular dentin (Frank and Nalbandian, 1989).
The composition of this dentin is different from that of
intertubular dentin, being collagen-poor and mineral-rich.
The mineral is in the form of small calcium-deficient
carbonate- rich hydroxyapatite crystals which have a higher
crystallinity and is almost 5 times harder than intertubular
dentin (Kinney et al, 1996).
Little is known about the biologic control of peritubular dentin
apposition.
Presumably, it is a very slow process, slower than the
incremental formation of secondary dentin in the pulp
chamber. This process can be accelerated by occlusal abrasion
(Mendis and Darling, 1979) and other forms of pulpal
irritation and may be more rapid in deciduous (Hirayama et
al, 1985) than in permanent teeth.
When dentin is exposed by attrition, there is a net outward
movement of dentinal fluid for the first time.
Although the complete composition of dentinal fluid is
unknown, it presumably has an ion product of calcium and
phosphate which is near or above the solubility product
constants for a number of forms of calcium phosphate.
This would tend to form mineral deposits in dentinal tubules
which can take on many forms (Mjor, 1985), since the
outward movement of dentinal fluid would present a larger
amount of mineral ions to the walls of the tubules than could
occur by diffusion in sealed tubules.
This principle has been used experimentally to slow thedepth
of demineralization of dentin in vivo under simulated caries-
forming conditions (Shellis, 1994).

(A) The extent of the odontoblast process

Controversy continues regarding the extent of the
odontoblast process.
Developmentally, odontoblast processes do indeed extend
from the odontoblast cell body, through mineralized dentin
matrix, to the dentin-enamel junction at the bell stage of
tooth development.
However, as dentin thickens, the cellular processes must
elongate
Since there are no supporting cells or blood vessels, the ability
of the odontoblast cell body to support a long cytoplasmic
process is in question.
In human teeth, the thickness of dentin is about 3-3.5 mm or
3000- 3500.
If the diameter of the process is (on average) 1 m, then the
volume of such a process would be (v =wr2 x length, or 3.14
[0.5 m x 3000 m = 2356 m).
The cell body of a columnar odontoblast is about 5 m in
diameter and 30 m long, or a volume of 589 m
Thus, the volume of the cellular process would be fourfold
larger than that of its cell body.
This difference is even larger in cuboidal or flattened
odontoblasts. It is not so much the disparity between the
volumes of the cell body and the cell process that matters, but
rather the diffusion distances involved.
An odontoblast process at the DEJ or CD) would be 3 mm or
3000 m from the nearest capillary, which is a very long
diffusion distance.
Although nerve axons are that long or longer, they are not 3
mm from supporting cells or capillaries. In the absence of any
evidence for cytoplasmic streaming analogous to axon
transport, it seems unlikely that the odontoblast could
maintain such a long process.
The length of the odontoblast processes of most
odontoblasts, regardless of dentin thickness, is between 0.1
and 1.0 mm (Holland, 1976; Byers and Sugaya, 1995).
The extent varies somewhat with species and position within
the tooth ( Holland, 1976, 1985).
The extent of the odontoblast process is important for a
number of reasons
1) If odontoblasts participate directly in the mechanism of
dentin sensitivity to surface stimuli, then these stimuli must
interact with a cytoplasmic process. If the process does not
extend to the surface, then the odontoblast cannot be
directly involved in dentin sensitivity.
2) When cavities are prepared in deep dentin in restorative
procedures,the odontoblast processes are amputated,
thereby irritating the cell body residing in the pulp.
The fact that many odontoblast processes are found to extend
only about one-third the distance from the tubule has been
explained as being due to a retraction of the process during
extraction (LaFleche et al, 1985).
Others have shown that the process does not extend more
than

Others have shown that the process does not extend more
than one-third the length of the tubule under normal
Conditions (Holland, 1976; Thomas, 1985; Weber and Zaki,
1986) and that LeFleche et al (1985) may have been confused
by fixation and tissue preparation artefacts.
(B) Changes in dentin structure with depth

The area occupied by the lumina of dentinal tubules can be
calculated as the product of the cross-sectional area of a
single tubule, r, and N, the number of tubules/cm2. The
term "r" is the radius of the tubule.
Since both the radius of dentinal tubules and their number
per unit area increase as one examines (Mjor and Fejerskov,
1979; Fosse et al, 1992;Olsson and 0ilo, 1993; Dourda et al,
1994) dentin from the DE) to the pulp, the area occupied by
tubule lumina also increases.
Garberoglio and Brannstrom (1976) measured the tubule
radius and number very carefully. They were the only authors
to correct for shrinkage artifact. Using their experimental
data, Pashley (1984) calculated the area occupied by tubule
lumina at the DEJ to be approximately 1% of the total surface
area at the DE] and 22% at the pulp (Fig. 2) (Table1).
Since this area is occupied by dentinal fluid, which is 95%
water, these areas are also approximately equal to the tubular
water content of these areas.
That is, the water content of dentin near the DEI is about 1%
(volume percent), while that of dentin near the pulp is about
22%. Textbooks list the water content of dentin at
approximately 10% by weight or 20% by volume (LeGeros,
1991), but that is an average value.

It is clear that the water content or wetness of dentin is not
uniform, but varies 20- fold from superficial to deep dentin.
The high water content of deep dentin is responsible, in part,
for the difficulty in bonding to deep dentin (Prati and Pashley,
1992), where water competes with resin monomers for the
surfaces of collagen fibrils (Pashley and Carvalho, 1997; Eick
et al, 1996).
The true water content of various dentin regions can be
masked by the presence of a smear layer and smear plugs
(Pashley, 1984; Prati et al, 1991).
They physically occupy a significant fraction of the luminal
area that could be occupied by water in their absence
(Pashley et al, 1978).
It has been calculated that the presence of smear plugs and
the smear layer occupy 78.5% of the area that would normally
be occupied by water (Pashley,1984).

This means that these structures decrease the water content
of those surfaces by 78.5%. The presence of grinding debris in
the tubule orifices and on the dentin surface also lowers the
permeability of dentin (Pashley et al, 1978).
(C) Physical characteristics of dentin

Dentin has been described as a heterogeneous composite
material that contains micrometer-diameter tubules
surrounded by highly mineralized (ca. 95 vol% mineral phase)
peritubular dentin embedded within a partially mineralized
(ca. 30 vol% mineral phase) collagen matrix (intertubular
dentin) (Marshall, 1993; Marshall et al,1996)
The bulk of tooth structure is made up of dentin, which is the
vital part of the tooth. Dentin is much softer (Knoop hardness
number, KHN = 68) than enamel (KHN = 343; Craig, 1993) and
exhibits much faster wear.
The modulus of elasticity of enamel is about 84 GPa (Craig,
1993) compared with dentin, which has a modulus of about
13-17 GPa (Table 2). Due to its more elastic nature, dentin is
tougher than enamel and serves as a stress-breaker or shock
absorber for the overlying enamel.
Regional differences in the shear strength of dentin were
reported by Smith and Cooper (1971).
They made thin ground sections of teeth and then, using 100
m diameter punches, measured the shear strength of
enamel, the dentino-enamel junction (DEJ), and dentin
fromthe DE J to the pulp.
The shear strengths of these very small dentin specimens
varied from about 132 MPa in superficial dentin to 45 MPa
near the pulp.
Watanabe et al. (1996), 72.4 and 86.9 MPa, which is
intermediate between the extremes reported by Smith and
Cooper (1971).
Early measurements of the ultimate tensile stress (UTS) and
modulus of elasticity (E) of normal human dentin by
Bowen and Rodriguez (1962) gave values of 52 MPa and 19.3
GPa, respectively .
Lehman (1967) of 36.6 MPa and 11.0 GPa, respectively.
Recently, Sano et al. (1994), using smaller specimens,
obtained ultimate tensile strengths and E for human dentin of
100 MPa and 14 GPa, respectively. In that study, they also
measured these same tensile properties in demineralized
dentin, and reported values of 26 MPa and 0.26 GPa,
respectively.
In that study, they also measured these same tensile
properties in demineralized dentin, and reported values of 26
MPa and 0.26 GPa, respectively.
This means that dentin collagen is responsible for 26%of the
ultimate tensile strength of dentin but only 1.6% of its
stiffness.
The tensile strength of dentin collagen reported by Sano et al.
(1994) was similar for either human or bovine demineralized
dentin, which confirmed report by Akimoto (1991) that
demineralized bovine dentin had a tensile strength of 28 MP
a

Many D.B.A use acid conditioning agents to demineralize the
dentin surface and uncover collagen fibrils. Adhesive resins
are then applied to the demineralized surface in an attempt to
envelop the collagen fibrils, which then provide
micromechanical retention for the resins. If the resin-dentin
bond is stressed to failure, presumably the weakest link in the
adhesive assembly will break first.
Many investigators had thought that the weakest link was the
collagen fibrils.
However, if collagen fibrils can withstand a stress of 26-28
MPa, they may be stronger than many dentin bonds
.
It has been argued that since the cross-sectional area of
demineralized dentin that is occupied by collagen fibrils is
only 1/3 of the measured physical cross-sectional area, the
true modulus of elasticity and the yield stress of collagen
should be multiplied by 3 .This would give values of about 80
MPa. The infiltration of adhesive resins may reinforce the
strength of the demineralized dentin to, make it even
stronger.
Sano et al. (1995) reported that resin-infiltrated
demineralized dentin had an ultimate tensile strength of
between 60 and 120 MPa, depending upon the bonding
system, indicating that, in terms of tensile strength, resins can
increase the strength of demineralized dentin from 26 MPa to
120 MPa, which is not statistically significantly different from
the ultimate tensile strength of mineralized dentin .

Sano et al.(1995) proposed this as a model for the strength of
the resin-infiltrated hybrid layer that couplesstrength of
mineralized dentin strength of the resin-infiltrated hybrid
layer that couples many adhesive resins to dentin
(Nakabayashi, 1992).
When they measured the modulus of resin-infiltrated dentin,
they found that it increased from 0.26 GPa to about 3-5 GPa,
which is far below that of mineralized dentin, 13-17 Gpa.
Recently, modifications have been made to an atomic force
microscope (AFM) by use of a stainless steel cantilever and a
diamond stylus to permit an AFM to be used for
measurement of both nanohardness and the modulus of
elasticity of intertubular and peritubular dentin (Kinney et al,
1996).
Peritubular dentin had a Knoop hardness of 250, while that of
intertubular dentin was only 52 (KHN).


(D) Dentin permeability

It is the tubular structure of dentin which provides the
channels for the permeation of solutes and solvents across
dentin. The number of dentinal tubules per mm2 varies from
15,000 at the DEJ to 65,000 at the pulp (Garberoglio and
Brannstrom, 1976; Fosse et al, 1992; Dourda et al, 1994).
Since both the density and diameter of the tubules increase
with dentin depth from the DEI, the permeability of dentin is
lowest at the DEI and highest at the pulp.
However, at any depth, the permeability of dentin in vitro is
far below what would be predicted by the tubule density and
diameters (Koutsi etal., 1994), due to the presence of
intratubular material such as collagen fibrils, mineralized
constrictions of the tubules, etc.


Dentin permeability can be divided into two broad
categories:

(1) transdentinal movement of substances through dentinal
tubules (such as fluid shifts in response to hydrodynamic
stimuli), or
(2) intradentinal movement of exogenous substances into
intertubular dentin, as occurs during infiltration of
hydrophilic adhesive resins into demineralized dentin
surfaces during resin bonding or demineralization of
intertubular dentin by bacterially derived acids (Kinney et
a\., 1995).

The easiest method of measuring transdentinal permeability
is to quantitate its hydraulic conductance
This measures the ease with which fluid can filter across a
unit surface area of dentin in a unit time under a unit pressure
gradient (Pashley, 1990).
The hydraulic conductance of dentin is responsive to a
number of variables, the most important of which is the
fourth power of the radius (Pashley, 1990).
The presence of smear debris in tubules (or any intratubular
material) lowers the fluid conductance of dentin and hence its
permeability.
The hydraulic conductance of dentin increases as dentin
thickness decreases (Fogel et al, 1988; Koutsi et al, 1994) in
unobstructed dentin.
However, the presence of a smear layer is more important
than dentin thickness in reducing hydraulic conductance.
When smear layers are removed to facilitate dentin bonding
the hydraulic conductance increases, and the etched dentin
tends to become wet with pulpal/dentinal fluid, making it
more difficult for resin monomers to infiltrate wet dentin (Tao
and Pashley, 1989).
The severity of pulpal reactions to restorative procedures is
more severe when smear layers are removed and dentin is
made thinner (Fujitani etal., 1996).
Dentin can be regarded as both a barrier or a permeable
structure, depending upon its thickness, age, and other
variables (Pashley and Pashley, 1991).
Due to its tubular structure, dentin is very porous. The
minimum porosity of normal peripheral coronal dentin is
about 15,000 tubules per mm2.
Once uncovered by trauma or tooth preparation, these
tubules provide diffusion channels from the surface to the
pulp.
The rate of diffusional flux of exogenous material across the
dentin to the pulp is highly dependent upon dentin thickness
and upon the hydraulic conductance of dentin (Pashley, 1985,
1990).
Thin dentin permits much more diffusional flux than does
thick dentin. However, there is a competition between the
inward diffusional flux of materials and the rinsing action of
outward convective fluid transport (Pashley and Matthews,
1993)
This may serve a protective role in mitigating the inward flux
of potentially irritating bacterial products into exposed,
sensitive dentin.
The permeability of dentin is not uniform but varies widely,
especially on occlusal surfaces, where perhaps only 30% of
the tubules are in free communication with the pulp (Pashley
et al, 1987).
Scanning electron microscopic examination of acid-etched
occlusal dentin reveals that all the tubules are exposed, but
functional studies of the distribution of fluid movement
across the occlusal dentin reveal that the tubules that
communicate with the pulp are located over pulp horns and
that the central region is relatively impermeable.
Apparently, there are intratubular materials such as collagen
fibrils (Dai et ai, 1991) and mineralized deposits that restrict
fluid movement, even though the peripheral and central ends
of the tubules are patent. Even microscopically, within any
100 x 100 m field, only a few tubules are open (Hughes etal,
1996).
Axial dentin is much more permeable than occlusal dentin
(Richardson etal., 1991).
The gingival floor of proximal boxes or the gingival extension
of finish lines in crown preparations often ends in regions of
high dentin permeability (Garberoglio, 1994).
In a full-crown preparation on a posterior tooth with a
surface area of approximately 1 cm2, there are as many as 3-4
million exposed dentinal tubules (Pashley, 1991).
Although the tubules are occluded by smear layers and/or
cement following placement of castings, both cement and
smear layers have finite solubilities and may permit some
tubules to become exposed over time. This is most likely to
occur at the most peripheral extensions of restorations,
where diffusion distances to the external environment are
closest
The permeability of sclerotic dentin is very low (Tagami et ai,
1992), regardless of whether the sclerosis was due to caries or
was physiologic or pathologic, because the tubules become
filled with mineral deposits.

Indeed, this is a fortuitous reaction that slows the caries
process and tends to protect the pulp. Most pulpal reactions
to cavity preparations or restorative materials used on carious
dentin are due to changes that occur across adjacent normal
dentin rather than the almost-impermeable caries-affected
dentin.
(E) Balance between the permeation of noxious
substances across dentin and their clearance by the
pulpal circulation

There is a balance between the rate at which materials
permeate across exposed dentin to the pulp and the rate at
which they are cleared from interstitial fluid by the pulpal
microcirculation (Pashley, 1979).
Procedures which cause reductions in pulpal blood flow
(activation of pulp sympathetic nerves, administration of
vasoconstrictor agents with local anesthetics) upset this
balance, permitting higher interstitial fluid concentrations of
extrinsic material to exist than could occur at normal levels of
capillary flow (Pashley and Pashley,1991)
Electrical stimulation of the inferior alveolar nerve in cats
causes an increase in pulpal blood flow and an increase in
outward dentinal fluid movement (Vongsavan and Matthews,
1994).
This response is interpreted as being due to axon reflex
activity. That is, antidromic stimulation of the inferior alveolar
nerve causes simultaneous depolarization of all of the nerve
terminals providing sensory innervation of the teeth. This
release of neuropeptides from the terminals causes both
increased pulpal blood flow and extravasation of plasma
proteins from the microcirculation (Raab, 1989, 1992; Olgart
and Kerezoudis, 1994).
Vongsavan and Matthews (1994) speculated that exposed
dentin that was stimulated hydro dynamically (for instance
mechanically) causes release of neuropeptides, increased
local blood flow, increased tissuepressure, and increased
outward fluid flow. This outward fluid flow might rinse the
tubules free of inward-diffusing noxious substances by
"solvent drag".
Indeed, Vongsavan and Matthews (1991) demonstrated that
Evans blue dye would not diffuse into exposed dentin in vivo
but would do so in vitro.



They speculated that, in vivo, the outward fluid flow blocked
inward diffusion. This notion was tested in vitro by
quantitation of the decrease in the inward flux of radioactive
iodide when a simulated outwardly directed 15 cm H2O
pulpal pressure was applied to dentin disks. This outward fluid
movement produced a 50-60% reduction in the inward
diffusion of radioactive iodide across acid-etched dentin, in
vitro (Pashley and Matthews, 1993)
Thus, stimulated dentin with open tubules (i.e., hypersensitive
dentin) should have higher rates of outward fluid flow and less
inward diffusion of bacterial toxins than nonstimulated dentin
This protective effect is diminished in the presence of a smear
layer (Pashley and Matthews, 1993).



Pashley (1992) speculated that bacterial invasion of tubules
or formation of crystalline precipitates in tubules would
interfere more with outward fluid flow than with inward
diffusion of noxious materials, due to the higher sensitivity of
bulk fluid movement to changes in tubule radius, r (which
varies with r4), compared with diffusion (which varies with
r2).
Under these conditions, the inward flux of noxious substances
may increase and permit higher interstitial fluid
concentrations to be achieved (Pashley, 1979, 1985) than
would occur if there were more outward fluid movement.

In exposed dentin, the outward fluid flow through tubules is a
first line of defense against the inward diffusion of noxious
substances.
Dentinal fluid also contains plasma proteins (albumin,
globulins) which can bind or agglutinate some materials which
may also serve a protective role.
A second defensive reaction occurs in freshly exposed dentin
that causes the permeability of dentin to fall following cavity
preparation in vital dog teeth but not in nonvital teeth
(Pashley, 1985).
Further, in experiments in which there was a continuous
inward-directed bulk fluid movement, the rate of decrease in
dentin permeability was delayed, suggesting that the
reductions in permeability were due to the slow outward
movement of some unidentified substance.
These experiments were repeated on dogs which were
treated with an enzyme which depleted fibrinogen from their
blood. Under these conditions, cavity preparation produced a
much smaller reduction in dentin permeability.
Apparently, the outward movement of fluid is associated with
an outward flux of plasma proteins (Maita et ai, 1991),
including fib- rinogen.
Attempts to recover fibrinogen in dentinal fluid have been
unsuccessful (Pashley, unpublished observation), although
labeled fibrinogen has been seen in the dentin of rat molars
(Chiego, 1992)
This may be because the dentin thickness in rat molars
following cavity preparation is only about 100 m .
(Bergenholtz et al. (1993, 1996) found fibrinogen only in very
deep cavities prepared in monkey or human teeth.
Pashley et al. (1984) reported that dentin removed 61% of
fibrinogen as it passed through a 1.0-mm disk. Presumably, in
vivo, even larger amounts of fibrinogen would be removed,
because, the tubules contain odontoblast processes at their
terminations, which further reduces the sizes of the channels
through which this high-molecular-weight molecule must
move.
In Summary, This molecule may be responsible for reducing
dentin permeability, especially at the pulpal terminations of
the tubules, where it may polymerize into fibrin.

The permeability properties of dentinal tubules indicate that,
functionally, they have much smaller dimensions than their
actual microscopic dimensions (Pashley, 1990).
Although the microscopic diameter of dentinal tubules at the
DE) has been reported to be 0.5 to 0.9 m, they function as if
they are 0.1 m in diameter.
Dentin can remove 99.8% of a bacterial suspension of
streptococci that are approximately 0.5 m in
diameter(Pashley, 1985).
This tends to prevent infection of the pulp, even when
patients masticate on infected carious dentin.
Fluid shifts across dentin can occur and can cause sharp, brief
pain, but the fluid is virtually sterile.This is because of the
presence of intratubular deposits of mineral and collagen that
form multiple constrictions within the tubule to dimensions
less than those of most micro-organisms.
Ten Cate's group recently reported that 65% of the dentinal
tubules in occlusal coronal dentin contain large collagen fibrils
(Dai et al., 1991).
If we understand the biologic mechanisms controlling this
reaction, we may be able to reduce the permeability of all
exposed dentinal tubules by collagen secretion into tubules,
thereby decreasing dentin sensitivity and the potential for
pulpal irritation.

(F) Control of dentin permeability

(1) Peripheral Reductions in Dentin Permeability
Whenever dentin is exposed, the pulp is placed at risk
because of the relatively high permeability of normal dentin.
Depending on the magnitude of the permeability of the
exposed dentin (which varies with location, depth,age, and
time (Pashley and Pashley, 1991] and with whether it is
sclerotic, infected with bacteria, etc.), the types of pulpal
irritation would range from hydrodynamic (e.g., fluid shifts in
either direction in response to evaporation, thermal stimuli,
etc.) to immunologic (bacterial products).
The junctional complexes of the odontoblast layer are
disrupted (Turner et al, 1989; Ohshima,1990; Turner, 1992),
permitting relatively large molecules (e.g., albumin, globulins,
fibrinogen) to exit pulpal blood vessels, gain access to
extracellular spaces, and reach the dentinal tubules, where
they can be detected in dentinal fluid (Bergenholtz et al,
1993, 1996).
While it may be difficult to prevent these changes during
restorative dental procedures, they can be minimized by
avoidance of air drying of dentin.
The subsequent pulpal irritation due to the diffusion of
bacterial products across dentin (Bergenholtz, 1981) could be
avoided if the relatively high permeability of freshly exposed
dentin could be reduced.
Dentinal tubules can be occluded with various crystalline
materials following topical application (Pashley and Galloway,
1985; Suge et al, 1995).
The use of adhesive resins to occlude tubules continues to
improve in terms of their sealing ability (Pashley et al, 1992b;
Tay et al, 1994).
Ideally, the rapid formation of peritubular dentin would lower
dentin permeability to near zero.
Similar low degrees of dentin permeability have been shown
by Tagami et al (1992) in both aged and caries-affected
dentin, but it took years for sclerosis to develop by accretion
of peritubular dentin formation.
However, the biologic regulation and control of peritubular
dentin formation are unknown.
(2) Internal Reductions in Dentin Permeability
An alternative approach is to apply a biologic growth factor(s)
to the exposed dentin, allowing it to diffuse across the dentin
to reach the pulp, where it could either stimulate fully
differentiated, normal odontoblasts to secrete additional
dentin matrix more rapidly than normal, or promote the
migration and differentiation of mesenchymal cells into
odontoblasts if the original odontoblasts had been destroyed.

As dentin matrix is synthesized, secreted, and mineralized,
dentin formation appears to trap a number of growth factors
into its structure (Finkelman et al, 1990; Magloire et al,
1992).
That is, growth factors such as BMP-2, FGF, EGF, IGF-1, and
TGF-p may be incorporated into mineralized dentin matrix.
Magloire and his colleagues believe that this occurs during
carious invasion of dentin. The release of these preformed
growth factors may permit their diffusion into them, pulp,
where they can activate appropriate genes to initiate repair
processes.
The goal of being able to stimulate dentinogenesis through
intact dentin has recently been accomplished.
Smith et al. (1994) created class V cavities in ferret canines
using an atraumatic technique.
Control cavities were treated with either nothing or with
rabbit serum albumin. Experimental cavities were treated
with either lyophilized EDTA-extracted rabbit dentin matrix or
with collagenase-treated insoluble residue remaining after
EDTA extraction. These crude fractions were covered with a
Teflon film and restored with ZOE. In as few as 14 days, there
was significant deposition of reactionary dentin by normal
odontoblasts
In contrast, control cavities lacking treatment with dentin
matrix components showed no evidence of reactionary dentin
deposition. Presumably, the production of reactionary dentin
by normal, fully differentiated odontoblasts was due to one or
more soluble growth factors from the dentin matrix
They speculated that the combination of TGF-p and BMP-2,
plus some unknown factors in the crude extract, might have
been,responsible for the reactionary dentinogenesis.
Smith et al. (1990) reported that
EDTA-soluble matrix proteins from rabbit dentin induced
dentinogenic activity in vivo in pulp exposures in ferret canine
teeth. In young adult ferrets, the reparative dentin resembled
normal primary tubular dentin, while in older animals it
resembled osteodentin in that it was atubular and included
cellular inclusions
Smith et al. (1994) referred to the dentinogenesis that they
simulated in fully differentiated odontoblasts as "reactionary
dentinogenesis", to distinguish it from dentin formation that
occurs following destruction of the original odontoblasts,
which they termed "reparative dentinogenesis".
The response was less in cavities with thicker dentin between
the cavity floor and the pulp chamber than if the dentin was
thinner.
Dentin matrix formation was sometimes so rapid that cells
became trapped within the matrix. The rate of dentinogenesis
seemed to become slower as more dentin was formed. As
more dentin is formed, the amount of growth factors diffusing
from the floor of the cavity to the pulp would be expected to
decrease, which, in turn, should slow matrix secretion.
It would be advantageous to induce odontoblast-like cells to
produce atubular dentin following the topical application of
growth factors to dentin.
This would seal the pulp from hydrodynamic and bacteriologic
insult.
It would also interfere with further diffusion of growth factors
to the pulp and would, therefore, be self-limiting.
However, this may not be possible if the odontoblasts are
normal, since the odontoblast process phenotype requires the
production of tubular dentin.
Apparently, only less-differentiated odontoblastoid cells
without processes can form atubular dentin
(Ill) Dynamics of Dentin

Whenever dentin is exposed to the oral environment, it is
subjected to large chemical and mechanical stimuli, in
addition to thermal stimuli and smaller mechanical stimuli
which are effective even in intact teeth.
Fluid shift in dentinal tubules

Physiological changes in blood vessels

Increased flow of plasma fluid & plasma proteins from vessels
into pulpal tissue spaces & out into dentinal tubules

Extravasation leads to of plasma causes increase in local pulpal
tissue pressure

Increased firing of sensitized neurons ( Narhi 1978)

Increased sensitivity & pain
The outward fluid flow may have a protective, flushing action
which may reduce the inward diffusion of noxious bacterial
products in both exposed cervical dentin (Pashley and
Matthews, 1993) and perhaps even in leaking restorations
(Pashley and Pashley, 1991)

(A) REACTIONS TO CAVITY PREPARATIONS:
FLUID SHIFTS
Although most dentists regard cavity preparation as a minor,
routine restorative procedure, from the perspective of the
pulp ,its a crisis .
The use of cutting burs in handpieces produces vibrations,
inward fluid shifts (because of frictional heat generation on
the end of a poorly irrigated cutting bur), outward fluid shifts
due to evaporative water loss (if only air cooling is used), and
slight inward fluid shifts due to osmotic movement of cooling
water into dentin (Horiuchi and Matthews, 1973).
These fluid shifts occur in both directions at various stages of
cavity preparation.
Further outward fluid shifts accompany the application of
hypertonic conditioners,primers, varnishes, or bonding agents
(Pashley et al,1992b), and then, during light-curing of
adhesive resins, additional inward fluid shifts would occur due
to heat generated during polymerization of adhesive resins
and resin composites (Hussey etal., 1995)
All of these fluid shifts create a barrage of hydrodynamic
stimuli across dentin into the pulp. These will obviously cause
pain if the patient is unanesthetized, and will release
sufficient neurotransmitters to cause local pulpal neurogenic
inflammation under the irritated tubules (Olgart, 1992, 1996)
and changes in pulpal blood flow (Kim and Dorscher-Kim,
1996).
(B) DISRUPTION OF ODONTOBLAST LAYER
Deep cavity preparation in rat molars causes aspiration of
odontoblasts (Byers et al, 1988), while more shallow cavity
preparation causes disruption of junctional complexes
between odontoblasts (Ohsima, 1990), which decreases their
barrier properties (Bishop, 1987, 1992), allowing large
molecules (horseradish peroxidase) from the blood stream to
penetrate dentin (Turner, 1989; Turner et al, 1992).
The loss of gap junctions may interfere with the ability of the
odontoblasts to secrete a collagen matrix in a synchronous,
coordinated manner, due to the loss of cell signaling between
adjacent cells. The proteins that make up gap junctions are
called connexins (Pinero et al, 1994)
How long it takes odontoblasts to reestablish the continuity of
gap junctions and to revert to tissue rather than cellular
function is unknown and remains a fruitful area for research.
The commercial availability of antibodies to connexins
provides the opportunity for the study of their importance in
reparative dentin formation and in pulpal healing
followingcavity preparation.
Presumably, bacterial substances in plaque and saliva could
easily move from the oral cavity into the pulp after such
operative procedures (Warfvinge and Bergenholtz, 1986;
Pissiotis and Spangberg, 1994; Bergenholtz, 1996).
The mechanism(s) responsible for the disruption of rat
odontoblasts during cavity preparation has not been studied
but is probably due to rapid, outward movement of dentinal
fluid in response to evaporative water loss from dentin during
cavity preparation.
As sub-odontoblastic capillaries "loop up" into the
odontoblast layer, disruption of the layer will also cause
microhemorrhages and direct irritation to capillaries that
sustain the odontoblasts.
They can leak plasma proteins (Chiego, 1992) such as
fibrinogen out into tissue spaces which are in communication
with dentinal fluid.
The permeability of dog dentin fell in a time-dependent
manner following cavity preparation (Pashley, 1985). When
the animals were depleted of their plasma fibrinogen, this
reaction was greatly attenuated (Pashley, 1985).
Several investigators have measured the flux of plasma
proteins across dentin following cavity preparations (Pashley
et al, 1981; Maita et al, 1991; Knutsson et al, 1994).
They reported that several plasma proteins, including
albumin, could be detected in relatively high concentrations
immediately after cavity preparations in monkeys and humans
(Knutsson et al, 1994) rjut that their concentration fell over
the next few hours to days. These plasma proteins contain
immunoglobulins which may inactivate some bacterial
products.
There is the possibility that bacterial or salivary products may
activate complement, which would also contribute to pulpal
inflammation.
Many of these reactions to cavity preparation would be
immediate and short-term. Displacement of odontoblasts up
into tubules disrupts their internal cytoskeleton and causes
cell death (Eda and Saito, 1978; Ohsima, 1990). These cells
undergo autolysis over the next few days and are replaced by
mesenchymal reserve cells which begin to differentiate into
new odontoblasts
The cell signals that are responsible for this process are
beginning to be understood (D'Souza and Litz, 1996).
Displacement of odontoblasts and their replacement by new
cells (Fitzgerald, 1979) are associated with very little pulpal
inflammation if the environment is sterile and if the dentin is
well-sealed.
It is possible for cavities in teeth to be prepared with no
pulpal inflammation or formation of reparative dentin (Smith
et al, 1994). This is done with copious air-water spray, light,
intermittent cutting forces, and the use of sharp burs.
However, if the dentin is not sealed (i.e., cervical abrasion) or
if the restored cavity exhibits microleakage, the pulpo-dentin
complex will undergo long-term reactions which are the result
of continual leakage and permeation of bacterialsubstances
around gaps in restorations (Pashley and Pashley, 1991) and
through unsealed dentinal tubules into the pulp.
SUMMARY
There is abundant evidence that dentin and pulp function as an integrated
unit which is generally termed as pulpo-dentin complex. While dentin or
pulp can be discussed separately for purposes of instruction or to simplify
issues, it should be stressed that the two function together as a unit.

The pulpo-dentin complex is an important concept in understanding the
pathobiology of dentin & pulp .

Developmentally , pulpal cells produce dentin , nerves & blood vessels .

Although dentin & pulp have different structures & compositions , once
formed they react to stimuli as a functional unit



Toooften , for technical or experimental reasons .,the
individual components of the pulpo dentin complex are
studied independently.

However ,it is becoming clear that the individual components
are very interactive & that each modifies the activity of other.
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Jukka Jernvall

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contSignaling Molecules that determine
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Bei, M. and Maas, R., 1998. FGFs and BMP4 induce both
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contSignaling Molecules that determine
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kratochwil, K., Dull, M., Farinas, I., Galceran, J. and Grosschedl,
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make them. Trends Genet. 15, pp. 5965
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contSignaling Molecules that determine
Odontogenesis

Chen, Y., Bei, M., Woo, I., Satokata, I. and Maas, R., 1996.
Msx1 controls inductive signaling in mammalian tooth
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Jernvall, J., berg, T., Kettunen, P., Kernen, S. and Thesleff, I.,
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2) Mechanisms of Stimulus Transmission
Across Dentin

D.G. GILLAM, Mechanisms of Stimulus Transmission Across
Dentin,Western Society of Periodontics,Volume Number 2,
1995.

3) Pulp Dentin Complex

D.H. Pashley
Department of Oral Biology, School of Dentistry, Medical
College of Georgia, Augusta, Georgia 30912-1129, USA



Books

ORBANS Oral histology & Embryology S.N.Bhaskar
ORAL HISTOLOGY- Development , Structure & Function
Ten Cate
DENTAL PULP-Seltzer & Benders




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