Adhesives Symposium

Dentin: Microstructure and characterization

Grayson W. Marshall Ji*

Demin consists of several idetitifiahle structures: lubuies wilh cell processes andßuicl;
highly mineraliied periluhulai- dentin; and inierlubular deniin consisting mainly of col-
lagen and deposited apatite. The structural organization and microslructural variations
i-eßeet formadve influetices, such as taoth ii::e and shape, and alieralions caused by age,
insult, and disease. However, details of struciwe-chemistry-properiy relationships for
this anisotropie biologie composite are limited. Materials scientists are focusing on an
array of new spectroscopic, analytical, and imaging techniques thai are yielding im-
proved understanding of structural variations and their dependence on specimen prepa-
ration, tooth type, location, storage eonditions, and chemical and physical modifica-
tions. Such studies should lead lo the major refinements in structure-properties models
for dentin that are required for continued advances in dental composite resins and adhe-
sives. (QiiinLessence InL ¡993:24:606-6J7.)

Introduction restorations for the future, because durable bonding,

Dentin is the fuiidaraenLal subslrale of restorative
dentistry, and its properties and characteristics are key
determinants of nearly all restorative, preventive, and
disease processes for teeth. Thus its characteristics
hmit today's treatments and will also shape the treat-
ments of the 21st century. Improved understanding
of its nature should have important consequences for
today's dental procedures and should lead to new
methods to preserve and protect teeth, and repair de-
fects brought on by disease or trauma.
An important current area of research is focused
on new methods to produce a bond between restor-
ative materials and dentin. Research progress is being
made to improve such methods, which alter dental
practice, and new approaches and methods are be-
coming available at a rapid rate. Thus better bonding
to tooth structure implies improved composite resin
with adequate bond strength and the prevention of
microleakage, is fundamental to the success of such
restorations.
However, before we turn our attention to the struc-
tural characteristics of dentin, it is worth noting that
the same characteristics have important influences in
many other aspects of current and future dental prac-
tice. Dentina! structure and its physical and chemical
modifications are important in carious processes, in-
cluding primary, secondary, and Class V lesion de-
velopment and progress, as well as preventive thera-
pies and restorations, dental pain and sensitivity, and
tooth fracture. Thus it is a common factor in virtually
all of dentistry and demands detailed attention from
many viewpoints.
Approaches to dentinal bonding

Efforts to bond to dentin have been far ranging and

Professor and Chair. Division of Biomaterliils, Deparimeiil of
Restorative Dentisiry, University of California. San Francisco,
707 Parnassus Avenue. San Francisco. California 94143-0758.
can be broadly classified into methods designed to
chemically or meehanically bond to the various com-
ponents of the dentin. An excellent series of reviews
of the various approaches has been recently pub-
hshed.' Tbese approaches include efforts to bond
chemically to the mmeral phase, collagen, or prtcip-

606 Quintessence International Volume 24, Number


itates following chemical modification.' In addition,
a variety of micromechanical bonding approaches
have been investigated: initially through formation of
tags in the tubules, and more recently by modification
oí intertubular areas through partial demineraliza-
tion and attachment of polymer to precipitaies.- for-
mation of a hybrid dcntin-polymer structure by im-
pregnation of monomers into partiaiiy demineralized
dentin and subsequent polymerization."' and penetra-
tion into laser-modified dentin.^'' The broad array of
approaches to solve the dentinal bonding problem
highlights the need for a comprehensive mode! of the
dentinal structure.
Structural elements
Dentin can be considered as a complex hydrated com-
posite of four elements: (1) oriented tubules sur-
rounded by (2) a highly mineralized peritubular zone,
embedded in an intertubular matrix consisting largely
of (3) type i collagen with embedded apatite crystals,
and (4) dentinal fluid (Fig I). Other non-collagenous
proteins are present in smaller amounts and have spe-
cific functions.' These basic components are well
known.* It is an unusual composite because the den-
tinal tubules converge on the predentin from the den-
tinoenamel junction (DEJ) in the crown and from the
cementum in the root. Overall, dentin is composed of
about 50 vol% mineral, 30 vol% organic matter, and
about 20 vol% fluid.'-'»
The structural arrangement left by the formative
cells leads to variation in tubule size and number and
quantity of intertubular matrix; these vary with po-
sition. The tubule course, number, and size are gen-
erally known from developmental and morphologic
studies such as those of Garberoglio and Bränn-
ström." Pashley'- calculated the fractional areas oc-
cupied by tubules, peritubular matrix, and intertu-
bular matrix for occlusal dentin as a function of po-
sition from these data. Thus tubule area and diameter
vary from about 22% and 2.5 fim. respectively, near
the pulp to 1% and 0.8 |im. respectively, at the DEJ.
The intertubular matrix area reportedly varies from
12% at the predentin to 96% near the DEJ, whereas
peritubular dentin decreases from more than 60% to
2.9% at the DEJ.'- Other more widely varying esti-
mates of tubule size and density have been reported
and would lead to larger differences in areas of each
component-^ Additional work has recently been re-
ported by Suzuki and Finger,'^ fiom studies of 15
teeth in which the area percentage of solid dentin ver-
Quintessence international Volume 24, Number 9/t993
Adhesives Symposium
Fjg 1 Scanning electron mjerograpii of fractured dentin
showing iongitudinal and cross-sect i o na i views ol tubuies,
peritubuiar zone, and intertubuiar matrix.
sus remaining dentinal thickness was determined.
Their technique may have overestimated tubule size
somewhat, because the denlin was treated with 0.5 M
of ethylenediaminetetraacetic acid for 60 seconds, and
tubules of lesi; than 0.7 ¡im in diameter could not be
measured. The relationship found by Suzuki and Fin-
ger'- is compared with other available information
from Pashley,'- and additional estimates from Olsson
and Oilo.'* in Fig 2. which illustrates the disagree-
ments in the available information for the important
parameter representing the available surface area for
bonding.
Additional work needs to be carried out to define
more closely the structure to be encountered in bond-
ing at different locations and for differing tooth sizes
and types. The marked changes in the amount of each
structural element with location imply that the den-
tinal properties and the nature of the suhstrate pre-
sented for bonding will vary with location.
The dentinal tubules are not smooth pipes, but rath-
er have irregular walls with many lateral branches
and microchannels that connect neighboring
tubules'^'' (Fig 3). They are normally filled with fluid
or partially filled with odontoblastic processes. The
tubules permit rapid movement of fluid, which is
thought to be related to pain and sensitivity, and has
led to extensive studies of fluid flow and permeability
by Pashley and coworkers.'* Dentinal permeability is
highly variable and is dependent on location in the
tooth; permeabihty is greater near the pulp and pulp
horns than in adjacent areas. "*•'' This is to be expected
as the consequence of the convergence and increasing
diameter of the tubules closer to the pulp.'*
607
 AcJhesives Symposium

00 -

so -
ir

60 -
/ u Suzuki & Finger'=
•/ * Pashley,'^ based on
Garberogiro 8 Brännström"
40 -
a Olsson S 0ilo"
/

20 .
/

0 -
Fig 2 (ASD) Area of solid dentin
versus (RDTj remaining dentinal
thickness from various studies."""

Fig 3 Scanning electron micrograph showing micropores Fig 4 Scanning electron micrograph ot coilagen in lumen.
in peritubular dentin.

Variations in permeability tnay arise from regional
differences,'*'^ tubular irregularities associated with
mineral deposits, organic tromponents of the odon-
toblastic processes, or recently classified intratubular
deposits of collagen-" (Fig 4). These factors make the
functional size less than the apparent size of the tu-
bules, as seen in cross section in the scanning electron
microscope (SEM).''
In addition, an outward flow of dentinal fluid oc-
curs because of a small, but positive, pulpal pressure,
estimated to be approximately 10 mmHg or 15 cm
H;0.-- From a bonding perspective, the hydrated state
of the dentin and variability in its pemieability char-
acteristics cause substantial difficulties that will be
dependent on location in the dentin. Unless the bond-
ing agent can displace or is miscible in the flu^j ¡^

608 Quintessence International Volume 24, Number 9/1993


areas where the dentin is most permeable, it will be
ditficult to achieve a stable and long-lasting bond.
The normal mineral content of dentin is found in
two areas: between the tubules in the intertubular den-
tin in intimate association with the collagen fibrils and
concentrated in the peritubiilar dentin. The apatite
crystals are much smaller ihan Ihe apatite found in
enamel and therefore more diffieutt to study. Consid-
erable information is available from various sources
that demonstrates that the very small apatite crystals
(approximately 5 x 30 x 100 nm) have less calcium
than hydroxyapatite and contain 4% to 5% carbon-
ajç 9.IO,I3,;Î,:4 yj^g small crystallite size, which is com-
parable to that of bone, has chemical and biome-
chanieal consequences.-•• The small crystallite size
composed of a defective apatite with a lower calcium
content and higher carbonate content leads to higher
solubility and the potential for ionic substitutions,
such as fluoride, that have important consequences
for preventive and restorative treatments. As a poten-
tial bonding substrate, these characteristics suggest a
large and active surface area to which bonds could be
formed.
Crystallite orientation would logically be assumed
to have some relationship with tubule direction; this
could offer advantageous mechanical properties.
Studies in the laboratory tend support to this
idea.-'"-' However, it is widely held that the crystals
are randomly oriented.-" Sorting out any orientation
dependence has significance, because the overall me-
chanical properties and deminerahzation process of
the material are expected to he dependent on any ori-
entation relationships. It is now thought that the role
of collagen does not include control of the nucleation
and size of the crystals, however, because noncoUag-
enous proteins have been proposed for this role.-''
The collagen fibrils between the tubules constitute
the major remaining structural component of the den-
tinal intertubular matrix. The organic matrix is ap-
proximately 90% type I collagen' and has generally
been assumed to be uniform in composition and con-
formation throughout the tissue.
Bond strength testing
Because dentin is removed from its natural environ-
ment for structural study and bond strength evalua-
tion of new bonding treatments, the effects of various
storage and analytical procedures need clarification.
Bond strength tests have notoriously high variabihty,
Quintessence International Volume 24, Number 9/1993
Adhesives Symposium
and results in one laboratory are often difficult to
reproduce in another. Rueggeberg-' reviewed many of
the problems associated with establishing procedures
for testing bonding agents to tooth structure and of-
fered some recommendations for standardizing pro-
cedures. Variations in species need consideration be-
cause the use of animal dentin may have advantages
in certain instances, eg, greater availability and the
ability to make direct in vivo and in vitro compari-
sons, but the structure may differ from human dentin.
In most studies, the effects of postextraction time
on bond strength seem to be relatively minor for pe-
riods between 20 minutes and several years, and a 6-
month period has been recommended.'^ However,
several studies have reported a time dependence-"" that
may depend on the bonding agent.''
Storage solutions and conditions also vary widely.
Three solutions appear to be the most common: water
in various forms (distilled, deionized, and tap, with
or without antibacterial agents), saline, and formalin.
Kimura et aP- reported that formahn increased bond
strength, whereas Mitehem and Gronas^^ could fmd
no effect of initial storage in formalin. Permeability
studies have revealed a marked variation with time
that depends on the solution,^'' but this has only a
minor influence on bond strength.
Rueggeberg-' also noted that much more research
is needed on storage temperature, drying, and ther-
moeycling conditions; that surface preparation pro-
cedures prior to bonding vary widely; and that more
attention is needed on the influence of sterilization
and infection control procedures. Although ignored
in the past, extracted human teeth are now considered
potentially infectious, and the Centers for Disease
Control has issued guidelines for the United States.^^
Experience suggests that gamma irradiation may pro-
vide a method for sterilization with minimal altera-
tions of the dentin.^^"
The morphologic and structural variation of the
dentin with location also affects bond strength. Gen-
erally it has been shown to be higher in superficial
than in deep dentin."•^"•"•^'*-* This factor could help
explain variability in the bond strength results. How-
ever, there is contrary evidence, and the dependence
is hkely to be related to the specific bonding agent or
mechanism." It has been suggested that bond strength
testing be carried out after the tooth is sectioned 1
mm above the pulp horns or 1 mm below the DEJ to
control variables for comparative tesling.^^'^'
Structural and chemical studies also require speci-
men storage and often require desiccation and drying
609
Adhesives Symposium

Table ! Major inorganic constituents of dentin mation provided may often be complementary, a"^
use of .several methods often is necessary to provide
Constituent Mean dry wl% a comprehensive imderstanding of the surface. One
of the preferred methods is x-ray photoeiectrun spec-
Calcium 26.9 troscopy (XPS), also known as ESCA, which allows
Phosphorus 13.2 analysis of the top atomic layers of the surf.'^e and
Carbonate 4.6 provides information on the chemical bonding in that
Sodium 0.6 layer, but does not have high spatial resolution and
Magnesium 0.8 cannot discriminate between peritubular and inter-
Adapted from Dnesstns Lind Vcrbeeck.' tubular areas of the exposed surface. Ruse and Smith '
used XPS to determine the composition of various
surfaces of bovine dentin (Table 2).
Table 2 Bovine dentin composifion (atomic%) Fach of these surface conditions is of considerable
interest from the bonding standpoint. Fractured sur-
Surface C N O Ca P Si faces are generally thought to represent the undis-
turbed dentin, because fracturing exposes the surface
Fractured 51.5 8.0 25.9 6.7 6.5 — without contamination or artifact introduction
Ground 600 54.4 4.7 24.7 7.2 6.9 — through surface preparation or instrumentation. Den-
Peroxide 55.2 2.7 25.1 8.0 6,6 — tal instrumentation, grinding, and most other prepa-
1% HCl, 1 min 67.3 13.2 16.7 0.8 0.8 — ration methods introduce alteration of the surface
Phosph, 1 min 34.9 4.7 39.8 0.2 — 20.5 morphology and formation of a smear layer, which
Phosph, 2 niin 41.6 7.3 35.4 0.3 — 14.9 then covers the normal structural components of the
Calculated* 51.0 9.0 26.0 7.0 6.0 — dentin.
Adapted from Ruse and Smith.'" Although recognized in early optical studies in the
' Based on 40 wt% polyprolitie (NC5H,O)„-60% HA mixture.
17th century by Leeuwenhoek,''" the smear layer has
assumed increasing importance since concentrated ef-
forts have been made to bond to dentin. Characteri-
in very high vacuum for many analytical techniques; zations have been conducted by Eick et al^-' and Pash-
these considerations are discussed later in this article. ley et al,^ Advantages and disadvantages to keeping
or removing the layer prior to bonding have heen
discussed by Pashley.'* A major disadvantage of re-
Composition taining the smear layer is its inherent weak bond to
Most work on dentin has dealt with its overall com- the underlying dentin. Thus many adhesive systems
position, and composition has been described based use some form of treatment to remove it, and this
on organic and mineral components. The major in- trend is likely to continue.
organic constituents are shown in Table 1, which is The peroxide surface treatment evaluated by Ruse
basedonacompilationof studies from the literature."' and Smith"" represents a method for removing loose
These values suggest a calcium-phosphorous molar debris from the ground surface that seems not to alter
ratio of 1.57, which compares to 1.67 for hydroxy- the composition. The acid treatments remove smear
apatite, Caiii(PO4)e(OH):. However, an average of 1.97 layer and partially demineralize and potentially open
from literature sources has been suggested based on the structure for bonding. Ruse and Smith"s'" calcu-
the best estimate of the calcium-deficient, carbonate- lations (hydrogen-free basis), which assume dentin to
rich form of dentin apatite.'" be a mixture of 40% polyproline (approximation for
Because bonding involves reaction with a specific collagen) and 60% hydroxyapatite, are in good agree-
surface layer of the dentin, and the structural fluc- ment with the fractured surface results. The compo-
tuations noted previously imply regional composition sition of the smear layer (ground surface) and cleaned
differences, many workers are attempting to study the smear layer is similar to that of the fractured surface,
composition and structure of such surfaces. A host of whereas the acid treatments removed virtually all min-
sophisticated and surface-sensitive instruments are eral from the surface. Several other workers have con-
available, each of which has specific advantages and ducted similar studies on human den tin,'*'•''* but they
limitations in certain situations. However, the infor- report compositional values based on ratios of t.xper-

610 Quintessence International Volurrie 24, Number 9/1993

 Adhesives Symposium

imentally determined atomic percentages. A compar- Table 3 Atomic% ratios of elements

ison of values is shown in Table 3.
These studies show good agreement on the basic Surface C-0 C-N Ca-P Ca-C
composition of the various untreated and treated sur-
faces, and the values for calcium-phosphorous ratio Fractured* 1,99 6.43 1.03 0.130
are in general agreement. Furthermore, it is apparent Fractured^ 1.4 7.6 1.38 0-2
that varying surface treatments alter the mineral con- Ground* 2.20 11.57 1.04 0.132
tent of the specimen to varying extents. The effects of Ground^ 3.0 9,7 1,59 0.09
demineralÍ2ation on structure will be considered later Ground* 1.68 11.71 1.04 0,56
in this paper.
Peroxide* 2,19 20,4 1.21 0.145
1% HCl* 4.03 5.1 1,0 0.012
Mechanical properties
Phosph A* 0.87 4.7 0.005
The ultimate bond strength to dentin is limited by the Phosph A* 1.2 10.3 0/0 0
inherent strength of the material. Table 4 presents
typical values for dentin and enamel as reported by Gluma (EDTA)' 2.64 5.72 0.99 0.06
Waters'*^ and Craig and Peyton."'^ Gluma (EDTA)' 4.3 6.3 1.21 0.03
It would be extremely helpful to have a model of Calculated* 1.96 5.67 1.16 0.137
properties that depended on the components and their
relative distribution, and such models have been con-
' Adapted from Ruse und Smiih
' Adiipled from Thompson et ai.1
sidered for calcified tissues by Katz."" Microhardness ' Adapted from Eliades et al,''^
measurements and the other nonnal mechanical prop-
erty measurements produce values that represent a
large volumetric average for the tissue. Thus individ-
ual structural components, such as peritubular or in- Table 4 Mechanical properties of human enamel
tertubular dentin regions, are not sampled by tbese and dentin
techniques.
The problem associated with understanding the me- Enamel* Dentin* Dentin'
chanical properties of dentin is related to the presence
of two phases with very different properties. The Proportional limit 70-350 100-190 —
hardness and strength of the dentin stems from the (MPa)
degree of mineralisation, but tbe exact relationships Compressive 95^50 230-370 297
are unclear and complex. In a two-phase mixture such strength (MPa)
as dentin, composed of collagen and apatite, it is pos- Young's modulus 9-90 7-30 18,3
sible to establish upper and lower boundaries to a (GPa)
material's elastic modulus. The upper boundary is Tensile strength 8-35 31-65 51.7
predicted by the rule of mixtures, or the Voight model, (MPa)
where, for example, the Young's modulus of elasticity Shear strength (MPa) 90,2 138
is given by the equation: Flexural strength 60-90 245-280
(MPa)
E,, - E,V, + EcVc Hardness (KHN) 330-450 68-80

where E¿ is Young's modulus for dentin, £'.,K,j is the * From Waters."''

product of the Young's modulus for the pure apatite
phase and the volume fraction of the apatite phase in
the dentin, and ¿V^c is the similar product for the
collagen phase. The lower boundary is given by the
Reuss model:
where the variables carry the same meaning.
' Craig and Peyton."**
For many materials, the upper and lower bounda-
ries are nearly equivalent, and it is possible to use
these simple models to describe the material's elastic
response to applied loads. Unfortunately, the large
difference between the moduli for collagen and apatite

Quintessence International Volume 24. Number 9/1993 611

Adhesives Symposium
D— E-Voight
• • E-Reuss
100 - Modulus data compiled
D by Waters'" and variation
m m nerai fract on
80 -
60 -
40 -
20 -,
0 -
02 0.4 0.6
Volume fraction hydroxyapatile
leads to large differences between the upper and lower
boundaries for dentin, thereby making the simple
models of limited value for prediction of properties.
The moduli for apatite and collagen cited by Katz,''^
were used to calculate the upper and lower boundaries
for Young's modulus of elasticity in dentin versus the
volume fraction of apatite phase with the above
models (Fig 5).
It is clear from these calculations that there is a
large difference between the upper and lower bound-
aries. Dentin has a volume fraction of approximately
40% apatite, and the measured Young's modulus (see
Table 4J also ranges widely within the boundaries.
The problem, then, is establishing the relationship
between modulus and volume fraction of the apatite.
Neither of the above-mentioned models is adequate.
Therefore, a much more careful analysis must be per-
formed based on improved methods for determina-
tion of mineral density and measurement of properties
at the component structural level. Many of the tech-
niques needed to obtain the necessary information are
now available. Mineral density variations can be ob-
tained from backscattcred electron microscopy be-
cause the backscatter signal coefficient is related to
local atomic number,*""" and quantitation of the sig-
612
•
^
rf
/^
m
0.8
J Fig 5 Young's modulus of elastic-
ity versus volume fraction ot apatite
for the Voight and Reuss models ot
mechanical properties (Henshall et
al, unpublished data, 1992).
nal should be possible."'** Furthermore, nanoinden-
tation methods should permit the evaluation of hard-
ness and modulus at the local level. Combining in-
formation from these methods should lead to im-
proved models that permit incorporation of details on
the properties and distribution of the basic structural
elements of the dentin.
Standard microstructural techniques require speci-
men preparation, and potential benefits to a nonin-
vasive technique are great. Such a technique would
permit detailed evaluation of mineral variations as
well as time-dependent studies of de mineralization
processes or other dynamic processes. Our laboratory
group has been working to apply one such technique,
known as x-ray tomographic micro.scopv (XTM).
X-ray tomographic microscopy of dentin
This new technique images materials in three dimen-
sions noninvasively.^5 By usmg synchrotron radiation
it is possible to achieve three-dimensional spatial res-
olution of several micrometers. X-ray tomographic
microscopy was initially developed for nondestructive
characterization of processing and failure in advanced
composite materials. It differs from conventional op-
Quintessence internationai Voiume 24, Number
 Adhesives Symposium

tical and electron-beam microscopy in that the spec-

imen need not be harmed prior to characterization of
internal microstructure. There Is no requirement for
tlat optical surfaces or thin sections, and it is possible
to examine materials in their unaltered state.
X-ray tomographic microscopy is based on the
same principle as computerized tomography (CT), in
which the x-ray attenuation coefficient at a point in
a material can be determined from a finite set of
x-ray attenuation measurements (projection data)
taken at different angles. The transmitted x-ray in-
tensities reach a position-sensitive detector after pass-
ing through the specimen, and tomographic software
converts x-ray absorption profile data into two-di-
mensional reconstructions of the linear attenuation Fig 6 X-ray tomographic microscopic images of a natural
carious lesion: (top) a singie slice, 6.5 pm thick, taken ap-
eoefTicients in the sample interior. These values can proximately 130 um inward from the front surtace; (bottom)
be rendered into a three-dunensional view by high- parallel view, but 26 pm farther into the specimen.
speed digital image processing computers. In addition,
when elemental or phase-mapping details are desired,
excellent chemical contrast can be obtained by re-
cording data at two x-ray wavelengths and perform-
ing image subtraction to enhance chemical or phase-
specific information.-^ Conventional CT technology
bonding. A variety of decalcifying agents are utihzed
typically uses a polychromatic beam from conven-
with different bonding systems, which induce various
tional tube sources; this methodology effectively pre-
morphologic changes.^*
cludes quantitative chemical analysis.
Acid treatments and demineralization caused by
The XTM has been used to image the mineral den- carious attack preferentially remove the peritubular
sity variations associated with natural and artificial dentin, causing a funnel-shaped structure at the ex-
carious lesions. Figure 6 shows several reconstructed posed surface. The acid also demineralizes the adja-
sections from several hundred contiguous XTM sec- cent intertubular dentin; differences in the condition-
tions through a lesion (Kinney TH, et al, unpublished ing agent yield differences in extent of demineraliza-
data, 1992).'^ The complexity of the carious process tion and shrinkage of the demineralized dentin as a
is apparent, because some areas are hyp ermine rali zed depression or concavity forms.^* Furthennore, total
and others are demineralized along the direction of demineralization of dentin results in substantial
the tubules. shrinkage, estimated at about 18% by GarberogHo
In addition, the three-dimensional morphology of and Brännström," especially when specimens are
the deminerahzed area can be reconstructed to show dried. Thus shrinkage or deformity of the surface of
the demineralization along individual dentinal tu- dried and partially or totally demineralized dentin oc-
bules.'* Such details cannot be seen with conventional curs; it has been proposed that the extent of such
techniques such as microradiography. shrinkage may depend hnearly on the volume fraction
of mineral."' Drying itself could cause structural
changes.^" I have recently attempted to study several
DemineralizatioD processes and dentin of these problems using the relatively new method of
atomic force microscopy (AFM).
Alterations of the structure of dentin by deminerali-
zation processes are of great interest in restorative Atomic force microscopy offers the opportunity to
dentistry because of caries and because of various ap- image specimens in a variety of environments (eg, ex-
proaches to bonding to dentin. Many dentinal bond- posed to water, storage solutions, deminerahzation
ing methods rely on acidic treatments.=' It may be and re mineralization solutions, or bonding agents)
necessary to condition the dentin for either chemical with high spatial resolution. It is one member of the
bonding or formation of a resin-reinforced hybrid family of new scanning probe microscopes.*'•*'
structure''; this might be considered nanomechanical Tbe microscope consists of a microfabricated tip

Quintessence International Volume 24, Number 9/1993 613

Adhesives Symposium

Fig 7 Atomic force microscopic ¡mages from a drying ex-

periment: (top) initial image taken of dentin in deionized
water: (bottom) image taken after specimen has air dried
for 23

Fig 8b Same area after 60 seconds.

Figs 8a to 8c Atomic force microscopic images taken dur-

ing demineralization of dentin in dilute nitric acid.

Fig 8c Same area after 100 seconds.

Quintessence International Voiume 24, Number 9/1993

614

assembly (a sharp tip, cantilever, and cantilever hold-
er), and a laser beam and photo detector to sense
deflection. The specimen is located on a Piezo tube
that can provide three-dimensional motion to the
specimen with subnanometer accuracy. The specimen
is brought in contact with the tip and is scanned so
that no change in cantilever deflection (as recorded
by the laser beam) is detected. Thus the three-dimen-
sional motion of the Piezo corresponds to the surface
morphology of the scanned area. The conventional
AEM tip, made of Si^Nj, has a typical radius of cur-
vature of 20 nm and exerts a force of about 10"" N
on the specimen. The in-plane resolution of the mi-
croscope is dictated by the radius of the curvature of
the tip, but the vertical resolution is about 0.1 nm.
Reeent work with the AEM suggests that localized
changes in tubule diameter and spacing are insignif-
icant during drying of mineralized dentin. although
microstrains result in buckling of the surface.'*-' Fig-
iire 7 shows AEM images of dentin, initially taken
under deionized water and taken again after 23 hours
of air drying. There was no detectable change in the
localized structural components.
The initial stages ofdemineralization have also been
evaluated with the AEM. Figures 8a to 8c show a
portion of a demineralization experiment with dentin
in dilute nitric acid. The development of the funnel-
shaped demineralization zone in the peritubular ma-
trix is clearly evident, in addition, the image shows
that the intertubular matrix begins to contract or col-
lapse during the initial stages of deminerahzation.
Thus it is difficult to demineralize the surface locally
without introducing a change in the surface geometry.
This suggests that new treatments might have advan-
tages in adding to bond strength if they can stabilize
the structure as demineralization occurs.
Along this line, Nakabayashi"" reported that ferric
chloride additions to citric acid stabilize the collagen
during demineralization processes. Sugizaki" also re-
ported that some primers appear to restore the de-
mineralized region to nearly its original position. Van
Meerbeek et al*^-*' have also recently reported on the
tnorphologic variations in this ¿one. Transmission
electron microscopy was used to identify three layers
in the zone associated with varying degrees of collapse
and resin entrapment.
It is clear that dentin and structural changes in-
duced by local or partial demineralization are com-
plex and difficult to study, but understanding their
nature is vital to improving dentinal adhesives. Eur-
thermore, understanding such processes is also im-
Quintessence International Volume 24, Number 9/1993
AdhesJves Symposium
portant in improving knowledge about deminerali-
zation and remineralization processes associated with
physiologic or pathologic changes in the structure of
dentin. As pointed out by Brickson'"* and Duke and
Lindemuth,^' formation of caries and sclerotic dentin
may induce regions of hypermineralization and oc-
clusion of the tubule lumen by crystalline deposits,
which appear to alter the ability to bond effectively
to the structure. Eurthermore, alterations of the min-
eral density of the dentin are the norm when the den-
tin is altered by caries or formation of erosive lesions.
Therefore, details of the structure and composition of
these altered forms of dentin are vital to the future
success of bonding systems.
Acknowledgment
This work was partialty supporled by NIH-NIDR grant No. POt
DE 09859; I would like (o (hank Dr S. J. Marshall. Dr J, H, Kinney.
Dr M. ßalooch. Dr G A. Henshall, L. Watanabe, S. Strawn. and
M. Ashton for their as,íislatice in this work.
References
1, Proceedings of the International Symposium on Adhesives in
Dentistry. Oper Dent 1992; (suppi 5).
2. Bowen RL, Marjenhoff WA. Development of an adhesive
bonding system, Oper Dent t992;(suppl 5);75-SO.
J, Fusayama T. New Concepts in Operative Dentistry. Chicago:
Quintessence, 1980.
4. Nakabayashi N. Adhesive bonding with 4-META, Oper Dent
1992;(suppl j ¡ : 125-130.
5. Cooper LF, Myers ML, Nelson DGA, Mowery AS. Shear
strength of composite bonded lo laser-pretreated dentin J
Prosthet Denl 1988;60:45^9,
6. White JM, Goodis HE, Rose CM, Khosrovi P.M. Hornberger
B, Shear bond strength of Nd:YAG laser treated dentin [ab-
stract 1048], J Dent Res 1991:70:397.
7. Butler WT. Dentin extracellular matrix and denlinogenesis.
Oper Dent ]992;(suppl 5);18-23.
8. Scott JH, Symons NBB. Introduction to Dental Anatomy, cd
Ö. Edinburgh; Churchill Livinstone, 1971.
9. Mjor IA. Fejerskov 0 . Histology of the Human Tooth, ed 2,
Copenhagen. Munksgaard, 1979.
10. Dtiessens FCM, Verbeeck RMH, Biominerals. Boca Raton,
FL: CRC Press, 1990:153-178,
11. Garbcroglio R, Bránnstróm M. Scanning electron microscopic
investigation of human deutinal tubules. Arch Oral Biol
1976;21.355-362.
12. Pashley DH. Dentin: A dynamic substrate—A review. Scan
Microscl989;3:161-'176.
13. Suzuki T, Finger WJ. Dentin adhesives: Site of dentin vs. bond-
ing of composite resins. Dent Mater 1988;4:379-3S3
14. Olsson S, 0ilo G. The structure of dentin surfaces exposed for
bonding [abstract 732]. J Dent Res 1989;68:958.
15. Jones SJ, BoydeA, Ultrastructure of dentin and dentinogenesis.
In: Linde A (ed). Dentin and Denlinogenesis, vol I. Boca Raton,
FL: CRC Press, 1984:81-134.
615
Adhesives Symposium
16. Piishley DH. Andringa HJ. Derkson GD, Derkson ME. Ka-
lathoor SR. Regional variability in Ihe permeability uf human
dentine. Arch Oral Biol l987;32:3lii-523,
17. Fogel HM, Marshall FJ, Pashley DH, EFfects of dislance from
the pulp and thickness on the hydraulic inductance of human
reticular dentin. J Dent Res I9SS;67:]381-1385.
18. Pashley DH. CHnica! correlations of dentin structure and func-
tion, J Prosthet Dent 1991 ;66:777-781.
ly. Sturdevant J, Pashley D. Regional dentin permeability of class
I and class II cavity preparations [abstract 173]. J Denl Res
19S9;68:203.
20. Dai XF, Ten Cate AR, Limeback H, The extent and distribu-
tion of intra tubular collagen fibrils in human dentine. Arch
Oral Biol 1991:36:775-778.
21. Michelich V, Pashley DH, Whitford GM. Dentin permeability:
A comparison of functional versus anatomical tubular radii. J
Dent Res 1978:57:1019-1024.
22. Vongsavan N, Matthews B, Fluid flow through cat detitine in
vivo. Arch Oral Biol 1992;37:175-185.
23. LeGeros R2, Apatites in biological systems. Prog Crystal
Growth Charact 1981:4:1^5.
24. Posner AS, Tannenbaum PJ. The mineral phase of dentin. In:
Linde A (cd|. Dentin and Dentinogesis, vol II, Boca Raton.
FL: CRC Press, 1984:17-36,
25. Marshall SJ, Marshall GW, Jendresen MD, Tomsia AP, Kinney
JH. Preferred orientation of apatite crystals in dentin [abstract
1288]. J Dent Res I99O;69:369,
26. Kinney JH, Balooch M, Marshall GW, Marshall SJ, White JM,
Goodis HG. AFM of drying induced changes in mineralized
human dentin [abstract 1732]. J Dent Res 1993;72:320.
27. Kinney JH, Balooch M, Marshall GW, Marshall SJ. Atomic
force microscope study of dimensional changes in dentin during
drying. Arch Oral Biol (in press).
28. LeGeros RZ. Calcium phosphates in Oral Biology and Medi-
cine. Basel: Karger, 1991,
29. Rueggeberg FA. Substrate for adhesion testing to tooth struc-
ture— Review of the literature. Report of ASC MDI56 Task
Group. Dent Mater lí'9!;7:2-lü.
30. Causton BE, Improved bonding of composite restorative to
dentine. Br Dent J 19B4;156:93-95.
31. Beech DR, Tyas MJ, Solotnon A. Bond strength of restorative
materials to human dentin: Influence of post-ex traction time.
Dent Mater 199I;7:I5-17.
32. Kimura S, Shimizu T, Fujii B. Influence of dentin on bonding
of composite resin. Part 1. Effect of fresh dentin and storage
conditions. Dent Mater J 1985:4:68-80.
33. Mitchem JC, Gronas DG. Effects of time after extraction and
depth of dentin on resin dentin adhesives, J Atn Dent Assoc
1986;113:285-287.
34. Goodis HE, Marshall GW Jr, White JM. The effects of storage
after extraction of tbe teeth on human dentine permeability in
vitro. Arch Oral Biol 1991:36:561-566.
35. Centers for Disease Control. CDC Recommendations for tbe
Decontamination of Fxtracted Teeth for Use in Dental Edu-
cational Settings, document reference No. lC-2, Division of
Oral Health, National Center for Prevention Services, Centers
for Disease Control, US Public Health Service. 1992,
36. White JM, Goodis HH, Marshall SJ, Marshall GW, Steriliza-
tion of teeth by gamma irradiation. Trans Acad Dent Mater
I99O;3(1):141-142,
37 White JM, Goodis HE, Marsball SJ, Marshall GW. Steriliza-
tion of teeth by gamma irradiation. J Dent Res (in press),
38. Nakamichi I, Iwaku M, Fusayama T, Bovine teeth as possible
substitutes in tbe adhesion test. J Dent Res 1983;62;1076-1081.
616
39. Causton BE. In viiro assessment of dentin bonding ag ^^
Today's Dentist, Proceedings of International ^ï^'P jjgtry
Adhesion, its Theory and Practice m Restorative
London; Curretit Medical Literature, 1987:18-27.
40. Tagami J, Tao L, Pashley DH, Correlation among •i^-^"" ^P '
permeability and bond strength of adhesive resms. DenL Mater
1990:6:45-50.
41. Ruse ND, Smith DC. Adhesion to bovine dentm-sunace char-
acterization, J Dent Res ,991:70.1002-1008.
42. O-Sullivan R, Flannelly M. Leeuwenhoek and the structure of
dentine, J Ir Dent Assoc 1990:36:129-133,
43. Eick JD, Wilko RA, Anderson CH, Sorenson S. Scanmng elec-
tron microscopy and electron m.croprobe analysis of cut tooth
surfaces. J Dent Res ¡970:49:1359-1368,
44 Pashley DH, Tao L, Boyd D, King GE, Horner JA, Scanning
electron microscopy of the substructure of smear layers in hu-
man dentine. Arch Oral Biol 1988;33:265-27Ü.
45 EUades G Palaghias G, Vouglouklakis. Surface reactions cf
adhesives on dentin. Dent Mater 1990;6:208-216.
46, Thompson VP, Edler TL, Davis G. XPS characterisation of
dentin treated with bonding primers [abstract 733], J Dent Res
1989:68:958.
47 Waters NE. Some mechanical and physical properties of teeth.
In: Vincent JFV, Currey JD (eds]. The Mechanical Properties
of Biological Materials. Cambridge, England: Cambridge Uni-
versity Press, 1980:99-136.
48. Craig RG, Peyton FA. Elastic and mechanical properties cf
human dentin, J Dent Res 1958;37:710-718.
49. Katz JL, Haid tissue as a composite material. I. Bounds on the
elastic behavior, J Biomech 1971:4:455^73,
50. Jones SJ, Boyde A. Scanning microscopic observations on den-
tal caries. Scan Microsc. 1987:1:1991-2002.
51. Pearce EIF, Nelson DGA. Micro structural features of carious
humnn enamel imaged with backscattered electrons. J Dent Res
1989;68:113-118,
52. Marshall GW Jr, Staninec M, Torii Y, Marshall SJ. Compar-
ison of backscattered scanning electron microscopy and micro-
radiograpby of secondary caries. Scan Microsc 1989:3:1043-
1050,
53. Nelson DGA, Backscattered electron imaging of partially-de-
mineralized enamel. Scan Microsc 1990;4:31-42,
54. Kinney JH. Marshall GW, Marshall S J. Quantitative BSEM of
mineral phases in dentin [abstract 828|. J Dent Res 1992;72:619.
55. Kinney JH, Nichols MC. X-ray tomographic microscopy using
synchrotron radiation. Ann Rev Mater Sei 1992;22:l21-152.
56. Kinney JH, Marshall SJ, Marshall GW, X-ray lomographic
microscopy of carious dentin [abstract 526]. J DenL Res
1992:71:171
57. Bertolotti RL, Conditioning of tbe dentin substrate, Oper Dent
1992;(suppl 5):131-136.
58. Inokosbi S, Fujitani T, Negisbi T, Hirasawa K. Study on resin
impregnated layer of varions dentin bonding techniques. J ipn
Soc Dent Mater Dev 1989;8(special issue 14):95-96.
59. Ten Cate JM, Nyvad B, van de Plassche-Simons YM, Fejerskov
O. A quantitative analysis of mineral loss and shrinkage of in
vitro demineralized human root surfaces. J Dent Res 1991:
70:1371-1374,
60. Van der Graff ER, ten Bosch JJ Changes in dimensions and
weight of human deninte after different drying procedures and
during subsequent rehydration. Arch Oral Biol 1993;38:97-99.
61. Jahanmir J, Haggar BG, Haynes JB, The scanning probe mi-
croscope. Scan Microsc 1992:6:625-660.
Quintessence International Volume 24, Number 9/1993
62. RLidinuther M, Tillmmin R\V. Frilz M. Giiub HE. From mol- ISBN1-B5D97-020-3
eciiles to cells: Imaging soft samples witii tiie alomic force mi- 122pp;66iilus(2eincûlor):
croscope. Science 1992;2S7:1900-1905. US $48
63. Sugizaki J. Tlie efTect of the various primers on ilie dentiii
adhesion of resin composites-SEM and TEM o be rvai ions of the
resin-impregnated layer and adhesion promoting effect of tiie
primers. Jpn .1 Conserv Dent 199t;34:228-265.
64. Van Meerbeck B, Inokoshi S, Braem M, Lanibrechts P, Van-
herie G. Morphologicül aspects of the resin-dentin interdiffu-
sion zone with different adiicsivo systems. J Dont Res
1992:71:1530-1540.
65. Van Mecrbcek B, Dhem A, Goret-Nicaisc M, Braem M, Lam-
brechts P. Vanherle G. Comparative SEM and TEM exami-
nation of Ihe ul I rast rue ture of the resin-iJenlin interdifTusion
zone. J Dent Res 1993;72:495-501.
66. Erickson RL. Surface interactions of dentin adhesive materials.
OperDenl I992:(suppl 51:S[-94,
Endodontic Surgery
67. Duke ES. Lindemuth J. Variability of clinical dentin substrates. C.R. Stockdale
Am J Dent 1991:4:241-246. " D

This bookse«ki to simplify the subject and spells out the

"'art of the possible." Perhaps of more importance, it
enables the practitioner to maximize the task of long-
term success from the patient's point of view. It recog-
nizes the problems and failures of etidodontic surgery
and recotnmetids techniques to minimize them. The
scope is such that it will appeal to sttidents, general
practitioners, and endodontists alike.

Contents include:
Introduction, Classification and Indications. Applied
Anatomy, Pathology of Periapical Lesions, Ciinical Di-
agnosis, Diagnostic Radiulogy, Patient Assessment and
Preparation. Anaesthesia, Design of Mucoperiosteal
Flaps, Surgical Technique of Apical Resección, Surgical
Management, Postoperative Care, Failures and Their
Management, and Medico-Legal Considerations.

CALL OR FAX TO ORDER: Tet: mm 621-0387 or

(7081 682-3223: Fax: 1708) 682-32S8
or 1111 out Ihs order lonn ^nd ísnú lo: OuinlËascnci: Publishing Cn,
Inc, 551 N. Kimberly Drive, Carol Stream, IL 6Ü188-188t

I Send me copies of (8SO4) Endcidontit Surgery I

al US î4S/copy.
Name
Streel
City . Slale .Zip
Teiephone _

[ 1 General Practitioner I 1 Endodontist

[ ] Bill me, including shipping & handling
I 1 Charge to my credit card plus shipping & handling
[ I Visa/MasterCard [ I American Espress

. Esp.

Signalure

L to change wiihoul nolice. All ^ale« sie fln^l. Shippirig [ui

ge.f will 1« adúcil i,> nil orders. For tllirmis nnil Caniulian
s las will be .tdclcd :rupplii:abl<:. P^ymem niusLlie niuli: i

Quintessence internationai Voiume 24, Number 9/1993