Impact of Different Acid Etching Time On Microtensile Bond Streng

University of Iowa
Iowa Research Online
Theses and Dissertations
Summer 2009
Impact of different acid etching time on microtensile bond

strength to vital dentin
Aadarsh Gopalakrishna
University of Iowa
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Part of the Other Dentistry Commons
Copyright © 2009 Aadarsh Gopalakrishna
This thesis is available at Iowa Research Online: https://ir.uiowa.edu/etd/291
Recommended Citation
Gopalakrishna, Aadarsh. "Impact of different acid etching time on microtensile bond strength to vital
dentin." MS (Master of Science) thesis, University of Iowa, 2009.
https://doi.org/10.17077/etd.2not5n6h
Follow this and additional works at: https://ir.uiowa.edu/etd

Part of the Other Dentistry Commons
IMPACT OF DIFFERENT ACID ETCHING TIMES ON
MICROTENSILE BOND STRENGTH TO VITAL DENTIN
by
A thesis submitted in partial fulfillment

of the requirements for the
Master of Science degree in Operative Dentistry
in the Graduate College of
The University of Iowa
July 2009
Thesis Supervisor: Assistant Professor Saulo Geraldeli

Graduate College
The University of Iowa
Iowa City, Iowa
CERTIFICATE OF APPROVAL
________________________
MASTER’S THESIS
_______________
This is to certify that the Master’s thesis of
has been approved by the Examining Committee

for the thesis requirement for the Master of Science
degree in Operative Dentistry at the July 2009 graduation.
Thesis Committee: ____________________________________

Saulo Geraldeli, Thesis Supervisor
____________________________________
Steve Armstrong
____________________________________
Deborah Cobb
____________________________________
Fang Qian
To my parents, for their motivation, constant support and love.
To my wife, who is always there for me.
To all my family and friends, who are my well wishers.
To my mentors, for their willingness to teach.
ii
ACKNOWLEDGMENTS
I would like to express my gratitude to Dr. Saulo Geraldeli, my thesis advisor,
who has constantly supported me throughout this thesis project. I greatly appreciate his
passion in teaching and encouragement. His continued support helped me finish this
project on time.
I would like to acknowledge Dr. Steve Armstrong for being a part of my research
committee member and sharing his research experience in my thesis project. I would like
to thank him for letting me use his biomaterial lab for this thesis. I would also like to
thank him for all the extended support in literature development.
It is my great honor to have Dr. Deborah Cobb as my graduate program director
and my committee member. I thank her for her encouragement during my program and
my thesis project. She made me feel home away from home.
I would like to acknowledge Dr. Fang Qian for her support for analyzing the
statistics of this thesis and her valuable suggestions.
I would like to thank Dr. Ricardo Atui from Guarulhos University, Brazil, for all
his clinical support in this project with patient selection and placement of the restorations
for the in vivo aspect of this research.
I would like to thank John Laffon for his help with the Scanning Electron
Microscopy procedures.
I would like to acknowledge and thank Dr. Gerald Denehy for all his support and
help to make me a better clinician and for his motivation to teach.
I would also like to thank all the friends and faculty members of the Operative
Dentistry department.
iii
TABLE OF CONTENTS
LIST OF TABLES -------------------------------------------------------------------------------- vi
LIST OF FIGURE -------------------------------------------------------------------------------- vii
CHAPTER
1. INTRODUCTION----------------------------------------------------------------------- 1
2. LITERATURE REVIEW-------------------------------------------------------------- 4
2.1 Statement of problem----------------------------------------------------------4

2.2 Background of tooth structures and adhesion------------------------------4
2.2.1 Enamel--------------------------------------------------------------------5
2.2.2 Dentin- pulp complex-------------------------------------------------- 5
2.2.3 Resin based composite (RBC)-----------------------------------------6
2.2.4 Adhesion/Bonding------------------------------------------------------7
2.3 Definitions and characteristics----------------------------------------------10
2.3.1 Acid etch and acids used in dentistry-------------------------------10
2.3.2 Acid etching and its effects on enamel-----------------------------10
2.3.3 Acid etching and its effects on dentin-pulp complex------------ 11
2.3.4 Smear layer-------------------------------------------------------------13
2.3.5 Effect of acid on dentin-----------------------------------------------13
2.3.7 Effect of etching time on enamel------------------------------------14
2.3.6 Effect of etching time on dentin-------------------------------------14
2.3.7 Microtensile bond strength as a method for evaluation
of resin- dentin interface----------------------------------------------15
2.3.8 Scanning electron microscopy---------------------------------------15
2.4 Studies supporting extended etching times on dentin-------------------15
3. MATERIALS AND METHODS-----------------------------------------------------22
3.1 Overview----------------------------------------------------------------------22
3.2 Research question------------------------------------------------------------22
3.3 Hypotheses------------------------------------------------------------------- 22
3.4 Outcome of interest--------------------------------------------------------- 23
3.5 Operational definitions------------------------------------------------------ 23
3.6 Variables---------------------------------------------------------------------- 23
3.7 IRB approval----------------------------------------------------------------- 23
3.8 Teeth samples---------------------------------------------------------------- 24
3.9 Scanning electron microscopy--------------------------------------------- 33
3.10 Statistical methods--------------------------------------------------------- 33
4 RESULTS------------------------------------------------------------------------------- 34
4.1 Microtenslie bond strength evaluation------------------------------------ 34
iv
4.2 Statistical analysis ---------------------------------------------------------- 43
4.2.1 Statistical results without the pretest failure data----------------- 43
4.2.2 Statistical results with the pretest failure data--------------------- 43
4.3 Results from Scanning electron microscopy---------------------------- 44
5 DISCUSSION---------------------------------------------------------------------- 56
6 CONCULSIONS------------------------------------------------------------------ 62
APPENDIX-------------------------------------------------------------------------------------- 63
BIBILOGRAPHY--------------------------------------------------------------------------------- 66
v
LIST OF TABLES
Table 1: Materials, compositions and manufacturer------------------------------------------ 27
Table 2: Data of microtensile bond strength in MPa from for each group------------------35
Table 3: Mean microtensile bond strength by surfaces and etching times without
including the failure mode-------------------------------------------------------------- 36
Table 4: Mean microtensile bond strength by surfaces and etching times with
the pretest failure as 1 MPa-------------------------------------------------------------37
Table 5: Comparison of mean tensile bond strength with and without pre test failure--- 38
Table 6: Failure mode results--------------------------------------------------------------------- 46
vi
LIST OF FIGURES
Figure 1: Procedure of tooth preparation------------------------------------------------------- 28
Figure 2: Procedure of restoration--------------------------------------------------------------- 29
Figure 3: Tooth sectioning------------------------------------------------------------------------ 31
Figure 4: Dumbell sample on Dirks device----------------------------------------------------- 32
Figure 5: Graph showing data comparison of mean microtensile bond strength

with and without including pretest failures------------------------------------------ 41
Figure 6: Graph showing data comparison of mean microtensile bond strength---------- 42

Figure 7: Graph showing the fracture modes in 5 seconds group--------------------------- 47
Figure 8: Graph showing the fracture modes in 20 seconds group-------------------------- 48
Figure 9: Graph showing the failure modes in 80 seconds group--------------------------- 49
Figure 10: Graph showing a comparison of the fracture modes from each group-------- 50
Figure11: Cohesive failure in dentin------------------------------------------------------------ 51
Figure 12: Cohesive failure in composite resin------------------------------------------------ 52
Figure 13: Joint failures--------------------------------------------------------------------------- 51
Figure 14: Mixed failure-------------------------------------------------------------------------- 54
Figure 15: Adhesive failure in the hybrid layer------------------------------------------------ 55
vii
1
CHAPTER 1: INTRODUCTION
Currently there is an increase in demand for “tooth colored restorations’’ in
dentistry. With this growing demand for esthetic dentistry and decreasing demand for
amalgam restorations, these restorations are very popular as they resemble the natural
tooth in terms of color and translucency. Resins based composites (RBC) are the most
widely used tooth colored restorative materials. This material not only mimics tooth
structure in color, translucency and texture, but also exhibits adequate strength,
durability, good marginal adaptation and sealing and excellent biocompatibility compared
to other tooth colored materials. The evolution in both the physical and esthetic properties
of resin restorative materials has led to greater longevity and more esthetic restorations.
Another advantage of RBC is that these restorations can be bonded to the tooth
structure instead of the traditional mechanical way of obtaining retention at the cost of
removing healthy tooth structure. This bonding to tooth structure can be obtained by
dental adhesives. These adhesives, decrease the microleakage in the tooth-restoration
interface, decrease sensitivity and improve marginal sealing. Adhesives have allowed for
esthetically restoring the teeth with minimal or no tooth preparation.
RBC adheres to enamel and dentin by adhesives. However a major shortcoming
of today’s adhesive restoration is their limited durability in the mouth (Van Meerbeek et
al. 1998, 1-20) as bonding to dentin has been shown to be less reliable than bonding to
enamel and is considered to be a major cause for adhesive failure. Bonding to dentin is a
less reliable technique due to the intrinsic characteristics and composition of this
substrate, especially when compared to enamel bonding.
Whenever dentin is cut for placement of restorations, considerable quantities of
debris cover the surface of the dentin forming a smear layer. This layer is advantageous
to protect the pulp-dentin complex when non adhesive restorative materials are indicated.
However, adhesive materials such as resins require a more porous enamel and dentin
before its application in order to increase material retention.

2
Resin adhesives classified as etch-and-rinse adhesives require the removal of the
smear layer. To achieve that, the bur prepared dentin surface should be treated with an
inorganic acid such as ortho phosphoric acid. When dentin is etched, the acid removes the
smear layer and modifies the outer most surface of the dentin morphologically by
dissolution of mineral content, which are predominately hydroxyapatite crystals. This
leads to opening of the dentinal tubules of dentin and the exposure of non mineral layer
of dentin made of collagen fibrils and a depleted mineral content. This is desirable with
the use of etch-and-rinse adhesives as they bridge the restoration to the tooth by forming
resin tags in the micropores created by etching and they micromechanically interlock
with the exposed collagen of the dentin to form the hybrid layer, thereby providing
retention. Most of the adhesive systems presently on the market use an acidic conditioner,
generally 30 to 40% phosphoric acid, to prepare the dentin surface to receive the bonding
components. Although the interaction of the etching agents with dentin is limited by the
mineral and non-mineral phases, there is often a discrepancy between the depths of dentin
demineralization versus monomer penetration of the adhesive, which is greatly
influenced by the etching time. This remaining unprotected mineral depleted layer at the
interface permits leakage, degradation and alters the integrity of the bond that may lead to
bonding failure. It has been suggested that the degree of resin infiltration of the exposed
collagen fibrils within the demineralized dentin has a profound influence on bond
integrity. Many in vitro studies (Abu-Hanna and Gordan 2004, 105-110; Abu-Hanna,
Gordan, and Mjor 2004, 28-33; Bolanos-Carmona et al. 2006, 1121-1129;Hashimoto et
al. 2002, 99-105; Jacques and Hebling 2005, 103-109; Sardella et al. 2005, 355-362) have
been carried out to evaluate the effect of different etching times on the dentin prior to
bonding and placing restorations, the relationship between tensile bond strength in these
restorations and the effect of etching times influence on the demineralized dentin.
Based on previous studies that excessive etching creates a deep demineralized
zone, this research will evaluate the effect of different etch times on microtensile bond
3
strength of teeth which were treated in vivo. All teeth used for this research were on
patients who were scheduled for orthodontic treatment and who needed those teeth to be
extracted solely for orthodontic treatment. These teeth were treated with three different
etch times, bonded and restored with RBC. After extraction, these teeth were evaluated
for their microtensile bond strength. The bond strengths were compared between groups.
Therefore, the current study evaluates the effect of etch times on tensile bond strength in
a clinical situation versus evaluating in vitro where an ideal clinical environment is hard
to mimic. In addition to comparing bond strengths, the predominant failure mode was
determined for the different etching times. The results of this study may provide
information as to the clinical importance of avoiding extended phosphoric acid
application. On the other hand, 5 seconds application might result in equal values for
bond strength which will point out that shorter times could be used. This study could be a
valuable source of information in terms of improving the clinical longevity of the
restoration and better success of treatment with tooth colored restorations.

4
CHAPTER 2: LITERATURE REVIEW
2.1 Statement of problem
The durability of the bond between adhesive materials and tooth structure is most
desired in restorative dentistry. The integrity of this interface between tooth structure and
adhesive dental materials is considered as the key to the longevity of the restoration.
However there is a broad spectrum of factors affecting the adhesion of the materials to
the tooth structure. This includes; the tooth as a substrate and its intrinsic qualities, the
type of adhesive systems, the properties of the adhesive systems, types of application
procedures, pretreatment of tooth structures and much more. The focus of this research is
to understand and evaluate the effect of acid conditioning time on dentin bond strength
using microtensile bond strength as a measure and to analyze the fracture modes with
scanning electron microscopy. Many in vitro studies (Hashimoto et al. 2002, 99-105)
(Abu-Hanna, Gordan, and Mjor 2004, 28-33) have demonstrated that increase in etching
times can reduce the bond strength; however this research project focuses on comparing
the microtensile bond strength on the vital dentin when treated with different etching
times.
2.2 Background of tooth structures and adhesion
Understanding different tooth structures is important for a successful adhesive
restorative dentistry as different tooth structures as substrate behave differently to dental
adhesive bonding systems. It would be important to understand the adhesion principal as
well.
5
2.2.1 Enamel
Tooth enamel is the hardest substance in the human body. It is of ectodermal
origin and chemically composed of a highly mineralized crystalline structure with 95% to
98% inorganic matter by weight and 86% inorganic matter by volume, 2% organic matter
and 12% water by volume. Enamel is formed by ameloblasts. There is formation of
organic matrix and then calcium and phosphate in the form of hydroxyapitite are
crystallized and these crystals enlarge. The majority of enamel is in the form of
Ca10(Po4)6(OH)2, although there are other minerals present as trace elements.
Structurally, enamel is composed of millions of enamel rods or prisms. They are closely
packed crystal forms of enamel made from small elongated apatite crystals arranged in a
distinctive pattern which gives strength and structural identity to enamel rods. These
enamel rods can be described as a keyhole with a circular core and are about 5µm in
diameter. Enamel is homogenous in structure except for the outer surface where the
crystals are prismless and run parallel to each other and perpendicular to the surface. The
hardness and density of enamel vary on different locations of the tooth. Enamel is very
brittle structure with low tensile strength and high modulus of elasticity making it a rigid
structure but dentin below the enamel acts as a cushion and withstands the masticatory
forces (Sturdevant 1995, 18-24)
2.2.2 Dentin-pulp complex:
Dentin and pulp tissues, in spite of the differences in structure and composition,
they are related in many physiologic and pathologic reactions. They have the same
embryonic origin and are formed from the dental papilla and maintain this relationship
throughout the life of a vital tooth. The cells of the dentin-pulp complex are the
6
odontoblasts. These cells are considered as part of dentin and pulp tissues as their cell
bodies are in the peripheral part of the pulp but the cytoplasmic process called the tomes
process are in the dentinal tubules of dentin. Dentin is considered as a living tissue as it
lodges the odontoblastic cell process and can react to physiologic and pathologic stimuli.
The odontoblastic process plays a primary role in the formation of dentin and continues
to slowly form dentin even after the tooth has erupted in what is termed secondary dentin
(Sturdevant 1995, 24). After eruption localized stimuli such as caries, wear process or
restorative procedures affect the formation of dentin and this type of dentin is called the
tertiary dentin. Tertiary dentin varies in structure and components and represents the
defense mechanism of the dentin-pulp complex (Mjor, Sveen, and Heyeraas 2001, 427-
446).
The composition of human dentin is 70% inorganic, 18% organic material and
12% water by weight. Dentin is less mineralized than enamel but more than cementum or
bone. The mineral content is hydroxyapatite arranged in a less systemic manner than
enamel. The hardness of dentin is less compared to enamel and even within the dentin the
hardness decreases from superficial dentin to circumpulpal dentin. The morphologic
characteristic of dentin is the dentinal tubules which extend from pulp to dentin enamel
junction (DEJ). The dentinal tubules are filled with odontoblastic process and dentinal
fluid which is a transudate of plasma (Sturdevant 1995, 28-29). The odontoblastic
processes are extensions of odontoblasts which are present the peripheral layer of the
pulp which is responsible of dentin formation. The tubules have a highly mineralized
lining along the tubular wall termed as peritubular dentin. Dentinal tubules are separated
by hydroxyapatite embedded collagen matrix called intertubular dentin. However,

7
anastamosis between tubules have been described (Mjor and Nordahl 1996, 401-412).
The number and diameter of the dentinal tubules decreases towards the dentinoenamel
junction. Superficial dentin contains about 20,000/mm2 of dentinal tubules, which are
each about 0.8µm in diameter and deep dentin contains about 76,000/mm2 of dentinal
tubules which are about 2.5-3µm in diameter (Fosse, Saele, and Eide 1992, 201-210);
(Pashley 1996, 104-133). This translates to more dentinal tubules close to the pulp where
are greater in diameter than the superficial dentin close to the DEJ.
2.2.3 Resin based composite (RBC)
A major problem in restorative dentistry is that dental materials do not adhere
efficiently to natural tooth structure. Classic restorative materials such as amalgam do not
bond to the tooth structure and provide little or no reinforcement of the weakened tooth
structure (Swift, Perdigao, and Heymann 1995a, 95-110). Conversely, resin bonded
composites can be adhered to the tooth surface. The adhesion of RBC to tooth structure
also has been shown to increase resistance to caries (Grogono and Mayo 1994, 89-90).
Adhesion can also reduce marginal leakage of bacterial and salivary components
at the tooth/restoration interface (Asmussen 1985, 61-73). Research suggests that bonded
resin composite restorations provide substantial reinforcement (McCullock and Smith
1986, 405-409). RBC has a wide variety of use such as direct anterior and posterior
restorations, composite veneers, and pit and fissure sealants.
2.2.4 Adhesion/Bonding
Adhesion or bonding can be described as attachment or intimate contact of two
materials. RBC can be bonded to the tooth structure by dental adhesives. Adhesion of
restorative materials to the hard components of the tooth structure has been a goal
8
pursued by many researchers ever since Buonocore pioneered adhesive dentistry in 1955
(Buonocore 1955, 849-853). The basic mechanism of bonding to enamel and dentin is
essentially an exchange process involving replacement of minerals removed from the
hard dental tissue by resin monomers which, upon setting become micro mechanically
interlocked in the porosities created. This was first described by Nakabayashi, in 1992
(Nakabayashi and Takarada 1992, 125-130) and commonly referred to as ‘hybridization’
or the formation of hybrid layer.
Based upon the adhesion strategy, three mechanisms of adhesion are currently in
use. (Van Meerbeek et al. 2003a, 215-235). A review by De Munck described the
different types of adhesives (De Munck et al. 2005, 118-132). First type is the etch-and-
rinse adhesives which involve separate etch and rinse phases where acid is applied and
rinsed off followed by a application of primer and application of adhesive step or a
simplified procedure where in prime and adhesive are combined in one application
preceded by etch and rinse.
Second type is the self etch adhesives which are based on the use of non-rinse
acidic monomers that simultaneously condition and prime dentin. Regarding user-
friendliness and technique-sensitivity, this approach seems clinically most promising.
This approach eliminates the rinsing phase, which not only lessens the clinical
application time, but also significantly reduces the technique-sensitivity or the risk of
making errors during application. There are two types of ‘self-etch’ adhesives: ‘mild’ and
‘strong’ (Van Meerbeek et al. 2003b, 215-235; Van Meerbeek et al. 2003b, 215-235).
‘Strong’ self-etch adhesives have a very low pH of 1 and exhibit a bonding mechanism
and interfacial ultra-morphology in dentin resembling that produced by etch-and-rinse

9
adhesives. ‘Mild’ self-etch adhesives have a pH of around 2 and dissolve the dentin
surface only partially, so that a substantial number of hydroxyapatite crystals remain
within the hybrid layer. Specific carboxyl or phosphate groups of functional monomers
can then chemically interact with this residual hydroxyapatite (Yoshida et al. 2004, 454-
458). This two-fold bonding mechanism of mild self etch i.e., micro-mechanical and
chemical bonding is believed to be advantageous in terms of restoration durability. It has
a micro-mechanical bonding component that may provide particular resistance to
debonding stress. The chemical interaction may result in bonds that better resist
hydrolytic break-down and thus keep the restoration margins sealed for a longer period.
The third type is the Glass ionomers and glass ionomer adhesives which are
considered to self-adhere to tooth tissue (Yoshida et al. 2000, 709-714). A short
polyalkenoic acid pre-treatment cleans the tooth surface; it removes the smear layer and
exposes collagen fibrils up to about 0.5-1 µm deep (Inoue et al. 2001, 237-245); therein,
glass-ionomer components inter-diffuse and establish a micro-mechanical bond following
the principle of hybridization (Lin, McIntyre, and Davidson 1992, 1836-1841); Van
Meerbeek et al., 2001). In addition to this, chemical bonding is obtained by ionic
interaction of the carboxyl groups of the polyalkenoic acid with calcium ions of
hydroxyapatite that remained attached to the collagen fibrils (Yoshida et al. 2000, 709-
714). This additional chemical adhesion may be beneficial in terms of resistance to
hydrolytic degradation. Consequently, a two-fold bonding mechanism is established,
similar to that mentioned above for mild self-etch adhesives. The basic difference with
the resin based self-etch approach is that glass ionomers are self-etching through the use
of a relatively high-molecular-weight polycarboxyl-base polymer. This limits their

10
infiltration capacity, so that only shallow hybrid layers are formed. In addition, because
of this high molecular weight, they cannot infiltrate phosphoric acid decalcified dentin.
Consequently, such aggressive conditioners should not be used with glass ionomers (De
Munck et al. 2004, 73-83).
2.3 Definitions/Characteristics
2.3.1 Acid etch and acids used in dentistry
Buonocore in 1955 proposed the use of acids in dentistry. He stated industries
used acids like phosphoric acids to obtain better adhesion of paints and resin coating to
metal surface, so he proposed that acids could be used to change the surface of the
enamel and render it more receptive to adhesion. (Buonocore 1955, 849-853). He started
the use of acids on enamel. Further work suggested that a tag like extension of resins into
enamel after the acid use (Gwinnett and Matsui 1967, 1615-1620). Later it was actually
known as acid etch technique (Swift, Perdigao, and Heymann 1995b, 95-110). Ever since
then different acids have been tried, they include polyacrylic acid, citric acid, nitric acid,
with phosphoric acid most commonly used.
2.3.2 Acid etching and its effects on enamel
Acidic solutions are normally used to etch enamel and dentin in commercial
dentin adhesive systems. Bonding to enamel is a reliable technique due to the
composition of enamel. The goal of enamel etching is to increase the surface free energy
for better monomer infiltration (Nakabayashi et al., 1998) and form resin tags. Peumans
described two types of resin tags. Macro tags are formed between prism periphery in a
circular manner and micro tags which is much finer network at the core of the prisms
where hydroxyapitite crystals are been removed by the effect of the acid. Micro tags are
11
responsible for most of the bond strength because of their large surface area. Generally
acid etchants remove about 10µm of the enamel surface and dissolve the rod core and
periphery to form microporisities from 5 to 50 µm in depth. The effect of acid etching on
enamel depends on many factors like type of acid, acid concentration, the time of etching,
rinsing time etc. There are three enamel etching patterns described. Type 1 is
predominantly dissolution of prism cores, type 2 is predominantly dissolution of prism
periphery and Type 3 in which no prism structure remain evident. Enamel surface treated
with acid has a high surface energy that allows resin monomer flow by capillary
attraction before polymerization to form resin tags. Enamel consists of mainly inorganic
hydroxyapatite which has high surface energy. Thus, bonding to enamel is easier than to
other tooth tissues and has been proven to be successful and predictable (De Munck et al.
2003, 136-140). However the solubility of enamel when exposed to acid many vary from
enamel surface to DEJ or the presence of fluoride (Sturdevant 1995, 23).
2.3.3 Acid etching and its effects on dentin-pulp complex
The composition of dentin differs markedly from that of enamel. Dentin is a
dynamic and a heterogeneous substrate which makes it more difficult to bond . Most of
the commercially available dentin bonding system use acid conditioners which remove
the smear layer and partially demineralize the intertubular and peritubular dentin. Dentin
has a higher amount of organic content than enamel and when acid demineralizes dentin,
protein rich collagen is exposed. This process changes the surface free energy of dentin.
The amount of demineralization depends on many factors like application time,
concentration and ph of acid, modifiers, surfactants and thickeners in acid.
The depth of dentin also plays a role in acid demineralization. The deeper dentin
where the dentinal tubules are closer show more demineralization than the superficial
dentin as the distance between the tubules are less with less intertubular dentin. The
deeper dentin show more dentinal tubules and the diameter of the dentinal tubules
increase with lesser intertubular dentin (Sturdenant 1995, 24). The degree of
12
mineralization also decreases with deeper dentin thus resulting in a more
demineralization when compared to superficial dentin when acid is applied.
Dentin is a dynamic structure which undergoes continuous physiologic and
pathologic changes. With age, dentin undergoes physiologic dentin sclerosis or reactive
sclerosis in response to mild irritation like abrasion and erosion. The result of dental
sclerosis is the formation of a precipitation of mineral crystals into the peritubular dentin
which leads to obstruction of the dentinal tubules. This process leads to less receptiveness
to adhesive treatments. With response to caries or attrition, there is formation of
hyperminerization leading to obstruction of dentinal tubules by crystalline deposits and
these dentin would respond to longer etching time for adhesive treatments and have
shown better bond strengths with extended duration of acid exposure in caries affected
dentin (Arrais et al. 2004, 458-464).
Numerous dentinal tubules are the present in the dentin which are filled with
dentinal fluid. The fuild in the dentinal tubules are under positive pressure. (Ciucchi et al.
1995, 191-194). There is no outward fluid movement from these tubules when they are
sealed with enamel and cementum but the fluid can show an outward movement when
this external seal is lost due to dental caries, tooth preparation, or tooth wear which could
interfere with adhesive procedures. This fluid due to dentin permeability caused by tooth
preparation makes dentin more challenging for bonding compared to enamel since the
fluid in these tubules can interfere with monomer infiltration of the adhesive system
(Pashley 1991, 777-781).
2.3.4 Smear Layer
The smear layer has been described as any surface debris, produced by grinding
or instrumentation of enamel, dentin, cementum or as contaminant that precludes
interaction with the underlying substrate (Ishioka and Caputo 1989, 180-185). The
thickness of the smear layer varies from 0.5- 5 µm (Pashley 1992, 215-224). A smear
13
plug is debris that occludes dentinal tubules. The surface smear layer and smear plugs are
porous with submicron channels but are reported to reduce dentin permeability by 86%
(Pashley 1992, 215-224). Removal of the smear layer or plug, as well as the mineral
content of intertubular dentin will increase dentin permeability.
An in vitro study by Spencer closely examined the smear layer, created by carbide
and diamond burs and the effect of acid etch on this smear layer (Spencer et al. 2001,
1802-1807). In this study, use of acid showed incomplete removal of the smear layer.
2.3.5 Effect of acid on dentin
Smear layer is not a stable layer and during its formation, the smear layer gets
incorporated into the underlying tissue which makes it impossible to remove by
scrubbing. On the contrary these particles are small giving them a high surface area to
mass ratio facilitating its rapid demineralization in acids (Pashley et al. 1988, 265-270).
So application of acids such as phosphoric, maleic, nitric or citric acid to dentin surface
results in removal of smear layer and demineralizing the underlying dentin (Eliades 1994,
73-81;Eliades 1994, 73-81). This acid demineralizes intertubular and peritubular dentin
and exposes the collagen along with increasing the microporosity of intertubular dentin
(Pashley 1992, 215-224; Perdigao et al. 1995, 1111-1120). Dentin is demineralized up to
7.5µm depending on the type of acid used and its concentration (Chiba, Itoh, and
Wakumoto 1989, 76-85; Van Meerbeek et al. 1992, 1530-1540). The changes produced
in the mineral content of the substrate also change the surface free energy of dentin
making it more receptive to adhesive (Erickson 1992, 81-94).
2.3.6 Effect of etching time on Enamel
With the use of acids, there is microporosities created within the enamel which
promote micromechanical retention of adhesive resin material into this porosity,
however, this depends on several parameters of which etching time is one. Traditionally
14
enamel was etched up to 60 seconds to produce an optimum surface for bonding but
etching times with lower etching times of 30 seconds is considered as an ideal time for
the peak quality of the etched enamel (Gardner and Hobson 2001, 64-67).
2.3.7 Effect of etching time on dentin
Acids, such as phosphoric acid, demineralize dentin, but the depth of
demineralization depends on the concentration of the acid and the duration (Chiba, Itoh,
and Wakumoto 1989, 76-85). Excessive acid conditioning for dentin pretreatment would
form a deep demineralized dentin zone within the bonded structure (Hashimoto et al.
2002, 99-105;Hashimoto et al. 2000, 406-411); (Nakabayashi, Watanabe, and Arao 1998,
379-385). There is deeper demineralization of both intertubular and peritubular dentin,
which would lead to incomplete infiltration by resin monomers.(Hashimoto et al. 2002,
99-105; Hashimoto et al. 2000, 406-411) These studies suggested that when failure was
initiated, this weaker zone created decreased bond strength. There was a direct correlation
between etching time and depth of demineralized zone. The hybrid layer thickness is
correlated directly to the etching time. Increased etching time demineralizes the dentin
surface to a depth greater than resin monomers could penetrate, producing a thick, poorly
infiltrated hybrid layer. Reducing etching time reduces the depth of the demineralized
zone and may be effective for achieving complete penetration and for sealing the dentin
surface (Abu-Hanna, Gordan, and Mjor 2004, 28-33);(Abu-Hanna and Gordan 2004,
105-110). On the contrary, the shorter etching times less than 15 seconds did not affect
the shear bond strengths of dentin as much as the longer etching times (Abu-Hanna and
Gordan 2004, 105-110).
2.3.8 Microtensile bond strength as a method for evaluation
of resin-dentin interface
Microtensile bond strength is considered as a suitable measure for the dentin
adhesive restoration joint evaluation (Pashley and Carvalho 1997, 355-372; Strang et al.
1998, 191-207; Tam and Pilliar 1993, 953-959; Eick et al. 1997, 306-335). More over
15
many investigators have suggested that shear bond strength is less used as they do not
measure the adhesive properties of dentin-resin interface (Nakabayashi, Watanabe, and
Arao 1998, 379-385; Pashley and Carvalho 1997a, 355-372; Strang et al. 1998, 191-
207;Eick et al. 1997, 306-335). The dentin adhesive restoration joint is made of dissimilar
materials and when force is applied there is non-uniform distribution of stress with
adhesive showing higher strain than the adherends (Armstrong, Keller, and Boyer 2001,
201-210). This would lead to joint failure that can be measured. Microtensile bond testing
is being known to produce distinctive debonding in the joint but if the samples are tested
after a very short storage it may not produce joint failures consistently.
2.3.9 Scanning electron microscopy
Scanning electron microscopy (SEM) can demonstrate the different layers of the
dentin-adhesive-resin joint and the dentinal tubules showing resin tags formed by the
bonding systems after demineralization. SEM could evaluate the depth of dentin
demineralization and adhesive penetration into them.(Van Meerbeek et al. 1993, 1423-
1428) The specimens for SEM should be air-dried or vacuum dried prior to fixation and
examination with the SEM. Sometimes artifacts can develop generally during this process
which may provide distortion in the images (Perdigao et al. 1995, 1111-1120; Perdigao et
al. 1995, 1111-1120).
2.4 Studies supporting extended etching times on dentin

There are many reported studies comparing the effects and alterations in etch
times to microtensile bond strengths of dentin-adhesive-resin interface. Many parameters
are looked into using scanning electron microscopy, transmission electron microscopy,
optical microscopy and dye staining to better understand the interface itself.
An in vitro study done by Hashimoto et al (Hashimoto et al. 2002, 99-105);
compared the over etching effects on tensile bond strength in two dentin bonding
systems. Their aim was to determine the weakest zone of resin-dentin bonds and the
relation between bond strength and failure mode to clarify the effect of demineralized
16
dentin. They used human premolars which were sectioned to expose the dentin surfaces,
and the dentin surfaces were conditioned with phosphoric acid for 15, 60, 120 or 180
seconds. Resin-dentin bonded specimens were produced using two adhesives, One-Step
and OptiBond Solo and tested for microtensile bond tests. Mean bond strengths were
statistically compared using two-way ANOVA and Duncan's multiple-range test and p
value was set to 0.05. The fractured surfaces of all specimens were examined using SEM,
and the areas of failure were measured using an image analyzer. They found that for One-
Step, the bond strength decreased with increase in acid conditioning time; 15 seconds
showed bond strengths measuring 50.7+/-9.7 MPa, 60 seconds = 40.8+/-11.0 MPa, 120s
= 23.6+/-4.9 MPa and 180 seconds = 12.1+/-4.6MPa. For OptiBond Solo, the bond
strength in the case of 15 seconds acid conditioning time = 42.6+/-7.9MPa which was
significantly greater than that for the other times of 60 seconds = 31.9+/-10.3 MPa, 120
seconds = 31.8+/-14.4 MPa and 180 seconds had 31.8+/-7.4MPa. The rationale for their
study was excess etching of dentin has been shown to reduce in vitro bond strength as
adhesive may fail to penetrate the over etched demineralized collagen network. The
purpose of the present research is similar, but evaluates over etching with vital dentin
using the micro tensile bond strength as a measure. The hypothesis for their study was
that increasing acid etching time in dentin will reduce the microtensile bond strength and
a reduction in etching time may produce more functional hybrid layers. The hypothesis in
the above study was tested in vitro whereas the current research may provide evidence of
clinical relevance regarding the relationship of bond strength to vital dentin.
Extending etching times has shown similar results on primary dentin as well. In
contrast to the permanent teeth, primary teeth have less mineral content (Sanchez-
Quevedo et al. 2001, 827-832; Sanchez-Quevedo et al. 2001, 827-832). An in vitro study
(Bolanos-Carmona et al. 2006b, 1121-1129) compared the tensile bond strength of
primary teeth etched at 3 different intervals and compared with a case control study. The
results revealed that 5 seconds etching time of primary dentin with 37% orthophosphoric
17
acid produced a visibly demineralized layer when specimens were stained with Masson’s
technique using optical microscopy. Tensile bond strength was significantly lower in
primary dentin etched for 5 s compared to 15 or 30 seconds of etching. They concluded
that 15 seconds and 30 seconds etch times produced better tensile bond strength
nevertheless 5 seconds still produced some demineralization. Although their primary aim
was to evaluate etch times and bond strength, they also evaluated the demineralized zone
thickness in the dentin after etching by using a dye which could stain the demineralized
dentin and compare their thickness to etch times. They used interface morphology along
with tensile bond strength determination. The interface morphology was used to
determine the associated interdiffusion zones by scanning electron microscopy and
optical microscopy. They treated the fractured /failure areas after testing for tensile bond
strength with Masson’s trichromic acid staining technique which stains the mineralized
type I collagen, resulting in staining collagen green. Etching of dentin with
orthophosphoric acid removes collagen, resulting in generally red stain. Specimens were
then examined in an optical microscope for presence or absence of a red band. They used
image analysis software. In each slide, three measurements of the depth of the
demineralized dentin layer were taken and 5 seconds etching time of primary dentin with
37% orthophosphoric acid produced a visibly demineralized layer when specimens were
stained but the thickness of demineralization increased with increase in etching time.
Most of the studies carried out are based on the in vitro hypothesis testing which
is specific to sound dentin. A few studies comparing etch time with caries affected dentin
were done. A study (Arrais et al. 2004, 458-464) concluded that extended etching can
improve bonding to caries affected dentin; however the adhesives applied on sclerotic
showed the best results for bonding. The authors evaluated the effects of additional and
extended acid etching on microtensile bond strength (µTBS) of two adhesive systems to
sound and caries-affected dentin. Additional samples were prepared for scanning electron
microscopy observations. They found that extended etching significantly increased

18
microtensile bond strength in caries affected dentin but they concluded that longevity
with these results were questionable and recommended further studies.
In relation to microtensile bond strength, (Paul et al. 1999, 181-188) tested the
microtensile bond strength on extracted teeth which was stored in dry storage, wet
storage and in a dye and the results showed no significance statistically in terms of
storage but showed a significant difference with etch time. With the increase in etch
time, the bond strength decreased. The etch times used were 15s, 30s and 60s. The aim
with the study was to evaluate the influence of etch times with microtensile bond
strength of Single Bond and to verify the leakage of silver ions within the hybrid layer.
After etch, bond and resin application, the teeth were sectioned and alternate slices were
either dried for 30 minutes in air, kept wet, or they were coated with fingernail varnish
except for 0.5 mm around the bonded area. Only the varnished samples were then
stained with 50% AgNO3. Microtensile bond strength was tested using a Vitrodyne V-
1000 universal tester. The samples of the stained group were embedded in self-curing
PMMA and polished. All samples were observed with an SEM. Nanoleakage of silver
ions was measured by exposure to laser ablation with an inductively connected plasma
mass spectrometer and by electron dispersive elemental analysis. Increasing etching
times seemed to have a negligible effect on bond strength of Single Bond, producing an
average value of ca 38 MPa. However, the silver uptake increased upon prolonged
etching times. Short-term results suggested that overetching has no detrimental effect on
bond strength values of Single Bond. Increased silver uptake, depending on the etching
time, raises concern about the long-term stability of the bond.
In another study (Spencer and Swafford 1999, 501-507), they stained the
exposed collagen in various adhesive group interfaces, found that exposed protein
stained red/orange in color using a light microscope which were indentified with all the
adhesives and were obscure with transmission electron microscopcy. Simple techniques
to evaluate the hybrid layers would lead to their improvements. Here the author
19
concluded that microtensile bond strength studies with One-Step as the adhesive
invariably showed most failures in the hybrid layer with the increase in etching time of
15 to 180 seconds. Their aim was to determine acid conditioning times associated with
various dentin bonding agents, that resulted in incomplete penetration of bonding areas
leaving a weak interface and to demonstrate this interface by a staining technique. Their
rationale was that other studies done to demonstrate the weak interface in bonding
created by the acid conditioning were not being effective, so developing a non
destructive staining technique to expose the collagen at this interface. Incomplete resin
infiltration of demineralized dentin can leave exposed collagen which may be penetrated
and degraded by some exogenous substances leading to weak interface. They used third
molars with various adhesive systems and the interface junction was cut for microscopic
sectioning and stained with Goldner’s trichrome. The exposed protein stained red/orange
in color using a light microscope was indentified with all the adhesives and were
obscure with transmission electron microscopy. They concluded that evaluation of
adhesive penetration in the decalcified dentin should be the first step in determining the
tooth resin-composite interface and simple techniques to evaluate the hybrid layers
would lead to their improvements.
On the other hand, a in vitro study (Abu-Hanna and Gordan 2004, 105-110)
compared etch times with lower and higher etching times than the recommended 15
seconds with 3 different 2 step etch and rinse adhesive system with shear bond strength
as a measure. The main aim of their study was to evaluate the effect of etching time by
lowering it to 5 seconds and increasing it to 30 seconds from the recommended etching
time of 15 seconds and evaluate their dentin bond strengths. 108 molars where distributed
equally among the 3 bonding agent groups which used a 2 step total etch system and each
group was further divided into 3 groups based on the etching times. Acetone based One-
Step (OS), ethonal based Single Bond (SB) and water based Syntac Single Component
(SSC) were the three different bonding agents. After etching the exposed flattened dentin
20
based on the etching times of the groups, restoration was done with resin based
composite (Z100) using one of the three adhesive systems based on the bonding agent
groups following the manufacturer’s instructions. After 300 cycles of thermocycling
between 5oC and 55oC, the teeth were tested for the shear bond strength and the fracture
mode was analyzed using SEM. The results analyzed using two way ANOVA showed no
statistical differences between the etching groups in OS and SB but the SSC groups
showed higher bond strengths with 5 seconds when compared to 15 and 30 seconds. This
results may indicate that lower etch times would create an area of demineralization
enough to facilitate complete infiltration of the adhesive monomer, which produces better
bond strengths.
A study evaluated the effect of acid concentration on dentin (Perdigao et al. 1996,
262-271). Six types of phosphoric acid etching agents were evaluated and the
independent variables were two acid concentrations of 10% and 32%-37% and three
thickener conditions. The hypothesis was that the use of different etchants with similar
concentrations of phosphoric acid would result in similar depths of dentin
demineralization. They obtained thirty dentin disks from extracted human teeth by
sectioning. The dentin surfaces were etched with one of the etching agents, fixed,
dehydrated and dried. The specimens were observed with SEM. The mean deepest
demineralization of intertubular dentin was measured from the fracture surfaces of the
disks. These values were analyzed by ANOVA and Duncan’s test. The morphological
appearance of the dentin surfaces was compared using the presence of a cuff of
peritubular dentin; relative thickness of the layer containing residual collagen or smear
layer particles and formation of a submicron hiatus at the bottom of the exposed collagen
network. The pH of each of the etching agents was measured. A correlation analysis was
made of the pH vs. the depth of dentin demineralization. The results indicated that silica-
thickened etchants did not demineralize dentin as deeply as the polymer-thickened
etchants and unthickened etchants. High magnifications revealed three distinct zones
21
within the demineralized dentin layer: An upper porous zone of residual smear layer or
denatured collagen and residual silica particles in silica thickened etchants, an
intermediate area with randomly oriented collagen fibers, and a lower zone with a
submicron hiatus, few collagen fibers, and scattered hydroxyapatite inclusions. The
results obtained suggested that similar concentrations of phosphoric acid etchants
containing distinct thickeners result in different demineralization depths as well as
different morphology of etched dentin.

22
CHAPTER 3: MATERIALS AND METHODS

3.1 Overview
The aim of this study was to evaluate the influence of the different etching times
on microtensile bond strength in vital dentin associated to a simplified two-step etch-and-
rinse adhesive. Twenty-six adults in the age range of 18-25 years and who needed
extractions of premolars for orthodontic reasons were selected in this study. The subjects
were randomly divided into three groups of 5, 20 and 80 seconds. Class I cavity
preparation were made in these teeth and restored with resin-based composite. After
extraction, the teeth were sectioned to obtain two beams from each tooth and they were
subjected to microtensile bond testing and the data was recorded. Statistical analysis was
made to evaluate any difference between the three etching groups. Once the microtensile
bond strength test was complete, the type of failure mode in each sample was analyzed
with the help of scanning electron microscope (SEM).
3.2 Research Question

Based on the previous in vitro studies (Hashimoto et al. 2002, 99-105) which
found that excessive pretreatment of dentin with phosphoric acid would create a deep
demineralized poor resin infiltrated zone decreasing dentin bond strength, and
considering the lack of in vivo systematic observations on bond strength of over-etched
dentin, the following question is posed: Does increase of acid etching on vital dentin has
an impact on microtensile bond strength?
3.3 Hypotheses
It is hypothesized that due to the vital and fresh condition of dentin in vivo after
cavity preparation, the variation on phosphoric acid time application will translate into
similar microtensile bond strength.
The specific aims of this research are:
A. Investigate the influence of different phosphoric acid application on vital dentin
microtensile bond strength.

23
B. Investigate any association between different acid etching time and fracture mode.
Null Hypothesis
There is no difference in microtensile bond strengths with different etch times on
vital dentin.
Alternate Hypothesis
With different etch times on vital dentin, there is a difference in the mean
microtensile bond strengths, specifically as the etching times increases, a decrease in
microtensile bond strength will be observed.
3.4 Outcome of interest

Based on the hypothesis that excessive etching of dentin will significantly reduce
the microtensile bond strength of resin dentin bond, the 80 seconds group was expected
to see lower bond strengths than 5 and 20 seconds. On the other hand, 5 seconds may
represent enough time to achieve bond strength similar to 20 seconds.
3.5 Operational Definition
Specimen or Beam: A rectangular 2x2 mm sample of the resin-dentin interface
obtained from the extracted tooth.
3.6 Variables
Etching time is the independent variable and microtensile bond strength measured
in Megapascals (MPa) is the dependent variable.
3.7 IRB approval
This study involved three phases in order to be completed. The first phase was the
placement of the restorations in vivo; the second was the mechanical testing evaluation of
resin-dentin interfaces created in vital dentin after different acid etching times, and the
third was the processing of the samples for scanning electron microscope evaluation of
debonded interfaces.
24
The in vivo phase was conducted at Guarulhos University (Brazil) where teeth
were restored and extracted. The human subject’s office at Guarulhos University (UNG
CEPPE) reviewed and approved the project.
The second and third phases, namely mechanical testing and fracture mode
evaluations, were conducted in the University of Iowa, College of Dentistry after the
approval by Iowa Human Subjects Office. The IRB approval number for this project is
200901773.
3.8 Teeth Sample

The aim of the research was to evaluate the influence of different acid etching
times on microtensile bond strength of a simplified two-step etch-and-rinse adhesive
applied on vital dentin. All in vivo procedures were conducted at Guarulhos University,
Sao Paul, Brazil. A total of 26 subjects in the age group of 18-25 years were selected to
participate in this study. The inclusion criteria used for subject selection were:
a. Having premolars scheduled to be extracted for orthodontic reasons.
b. No caries or hypoplasia in these premolars selected.
c. Absence of history of spontaneous pain.
d. Absence of apical pathology.
e. Willingness to participate in the study.
All premolars (at least one tooth per subject) selected in this study were diagnosed
for extraction as part of the orthodontic treatment plan proposed by the department of
Orthodontics in the same university.
After receiving the informed consent signed from the patient or parent, a standard
class I occlusal cavity preparation was performed on each tooth preceded by local
anesthesia and rubber dam placement for complete isolation. Class I cavities were
designed with an occlusal depth of ±1mm below the dentin-enamel junction (DEJ). The
occlusal preparation was centered at the middle of the occlusal surface with a smooth
25
even pulpal floor in the dentin using a pear shaped Brassler™ diamond bur. The
buccolingual dimension was between 2.5-3mm and the total depth of about 2.5-3 mm.
The premolars were randomly divided based on the acid etching times of 5 (n=8),
20 (n=9) and 80 seconds (n=9).
A 34% Ortho Phosphoric acid (CAULK® 34% Tooth conditioner gel,
Dentsply/Caulk, Table 1) was used to etch the enamel and dentin. The gel was applied
first on the enamel cavosurface and then extended to the dentin pulpal floor. Once in
dentin, a dental assistant with a digital watch precisely checked the time.
The acid was water rinsed for 15 seconds and the excess of water removed with
high suction from the surrounding areas of the tooth followed by blot dry of the cavity.
In the end, dentin was considered ready to receive the adhesive if superficial moisture
was present.
A generous amount of simplified two-step etch-and-rinse adhesive (Prime Bond
NT™, Dentsply, Caulk, Table 1) was applied to the cavity preparation with a disposable
microbrush. The acid etched dentin was kept fully wet by the adhesive for 20 seconds.
Excess of solvent could be removed by applying a gently dry, clean air form the syringe
for at least 5 seconds. A uniform glossy appearance indicated the removal of the solvent.
The adhesive was light cured for 20 seconds.
A thin layer of microhybrid resin-based composite (Esthet-X™, Dentsply, Caulk,
Table 1) shade A1 was first placed in the pulpal floor of the cavity. This increment was
then light cured for 40 seconds as per manufacturer’s recommendations using a light-
curing unit.
To obtain consistency and to minimize inter operator discrepancies in the
procedure; a single well-trained clinician performed all restorative procedures. Shortly
after the completion of the restoration, the tooth was extracted following conventional
techniques. Patient and/or parents were given post-operatory recommendations as per
Oral Surgery Department of the same university. Existing soft tissues were removed and
26
the teeth were stored in 100% humidity wrapped in water moist gauze and stored in
refrigerator at 32º F until shipping to the Unviversity of Iowa, College of Dentistry.

27
Table 1 - Materials, composition and manufacturer
MATERIAL COMPOSITION MANUFACTURER

Phosphoric acid 34%
Phosphoric Acid
Acid etchant Highly dispensed silicon dioxide
Colorant
Water
Prime & Bond NTTM
Di- and Trimethacrylate resins
PENTA (dipentaerythritol penta
acrylate monophosphate) DENTSPLY Caulk
simplified two-
Nanofillers-Amorphous Silicon
step etch-and-
Dioxide
rinse adhesive
Photoinitiators
Stabilizers
Cetylamine hydrofluoride
Acetone
Resin-Based Esthet- XT M
Composite Microhybrid resin based composite
28
Figure 1 - Procedure of tooth preparation
Sequence of images depicting initial prophylaxis, rubber dam placement, cavity

preparation and measurement of pulpal depth of the cavity preparation with a periodontal
probe.
29
Figure 2 - Procedure of restoration
Sequence of images showing application of the phosphoric acid into the cavity
preparation, application of the simplified two-step etch-and-rinse
rinse adhesive and insertion
of the resin-based
based composite layers
30
Upon receiving the teeth, a well-trained operator (AG) proceeded with the
microtensile sample preparation and mechanical testing.
Specimen sample preparation for microtensile bond strength analysis
Teeth were flattened horizontally at the occlusal surface by slightly grinding the tip
of the cusps. The roots were cut flat to about 2 mm below the CEJ using a saw machine
and a diamond disc (Isomet 1000™, Buehler). This was done to facilitate mounting of the
teeth on 2×2 inch Plexy glass support (Figure 3). The Isomet trimmer uses rotary
diamond blades at varying speed to allow sectioning of the tooth to any desired thickness
by varying the distance between the cutting discs.
Using a sticky wax, each tooth was attached to the center of the Plexy-glass square
support followed by sectioning in a saw machine. The goal of the sectioning was to
obtain two beams out of each tooth, one from the mesial and one from the distal surface.
In order to achieve this, the mesial beam was distinguished from the distal by marking it
with a blue highlight marker. Each beam was further trimmed to an dumbbell shape at the
interface of resin and dentin to about 0.8 mm diameter using the CNC specimen former
(University of Iowa, IA, USA), which is a computerized system used to obtain specimens
for microtensile bond strength (Figure 3).

31
Figure 3 - Tooth sectioning
Sequence of images showing a grinded premolar prior to its stabilization with a stick wax
over the Plexy-glass
glass square support; sectioned premolar with two beam identified with
different water resistant shades; beam attached to attached to a Plexy by aid of a sticky
stick
wax; beam attached to a special device prior trimming of the resin
resin-dentin
dentin interface; final
dumbbell sample obtained and ready to be mechanically tested
32
Out of the 26 teeth, 52 beams were obtained an

andd trimmed to a dumbbell shape.
During mechanical testing of the samples 8 failures were observed before testing and
were considered pre-test

test failures. These failures were from the following groups: two
failures in the 5 seconds group, one failure in 20 sec

seconds
onds group and five failures in the 80
seconds group.
Before mechanical testing, eeach

ach dumbbell was examined under a
stereomicroscope for any preparation defects and their diameter measured using a digital
caliper. Tensile testing was performed at a crosshe

crosshead
ad speed of 1 mm/min in a calibrated
materials testing machine (Zwick, GmbH & Co, Germany, Figure 4) and subjected to
tensile strength. Data were collected and transferred to an appropriate sheet.
Figure 4 - Dumbbell sample on Dirks device
Image of a dumbbell sample inserted on Dirks device and set in the testing machine prior
microtensile bond strength testing
33
3.9 Scanning electron microscope (SEM)
After submitting the dumbbell to the testing microtensile bond strength, the
fractured specimens were mounted on round aluminum stubs using cyanoacrylate glue
(Zapit, Dental Ventures of America, CA). A gold sputter coater (SCD-040) was used to
sputter a layer of about 15µm thickness on the samples. The failure mode was observed
under SEM (Amray 1820-D, Boston, MA, USA). The fracture surface of each specimen
was recorded as one of the four failure types: cohesive failure in the dentin, cohesive
failure in the resin composite, joint failure at the adhesive interface and mixed failure
involving both cohesive failure in dentin or resin composite and adhesive interface.
3.10 Statistical Methods
Descriptive statistics were computed using the tooth as a statistical unit. One-way
ANOVA with post-hoc Tukey-Kramer’s test was used to determine whether there was
significant difference in microtensile bond strength between the three etching times.
Possible association between failure type and group of etching times was assessed using
Fisher’s exact test. In addition, the Shapiro-Wilks test was conducted to test normality.
All tests employed a 0.05 level of statistical significance. SAS for Windows (v9.1, SAS
Institute Inc, Cary, NC, USA) was used for the data analysis.
34
CHAPTER 4: RESULTS
A total of 52 beams were obtained from 26 teeth samples among the 3 groups.
There were 8 beams which failed before testing for the microtensile bond strength. These
beams were considered as pre test failures. There were two failures in the 5 seconds
group, one failure in 20 seconds group and five failures in the 80 seconds group. The
remaining intact 44 beams were used for testing the microtensile bond strength and
fracture mode analysis.
4.1 Microtensile bond strength evaluation

Descriptive statistics were computed from the recorded data using one-way
ANOVA with post-hoc Tukey-Kramer’s test to determine any significant differences in
microtensile bond strength among the three etching times.
Statistical analyses were applied to test the hypothesis in two types of data
collected. All data collected are included in Table 2. There were 8 specimen samples
which failed before the tensile loading and were treated as left-censored data and
assigned a bond strength of 1MPa based on half the preload value of 1 Newton applied in
material testing machine i.e. 0.5newton/0.5mm2 = 1MPa (Vachiramon et al. 2008, 178-
185)
The pretest failure data were included the data as 1MPa and used in the statistical
analysis. The descriptive statistics are summaries in Tables 3 and 4 by the surface and the
etching time, including the mean and standard deviation values, and by the two
conditions with and without considering the failure mode as 1 MPa.
Table 5 includes mean bond strengths for all groups with and without the pretest
failure. For 5 seconds group the bond strengths were 32.49 +/- 9.27 MPa with the pretest
failure data included at 1MPa and 38.35 +/- 11.63 without including the pretest failure.
Mean bond strengths for 20 seconds groups was 36.41 +/- 11.17 MPa with the pretest
failure considered at 1MPa and 38.09 +/- 8.57 without including the pretest failure. Mean
bond strengths for 80 seconds groups was 19.08 +/- 10.33 MPa with the pretest failure
35
included at 1MPa and 28.82 +/- 7.35 without including the pretest failure. Figure 5 and 6
shows the comparison in the mean tensile bond strength with and without including the
pretest failure.
36
Table 2: Data for microtensile bond strength in MPa for each group
Tooth Group TBS_M TBS_D AVG_TBS
1 5 27.91 24.34 26.125
2 5 55.1 25.79 40.445
3 5 24.51 23.76 24.135
4 5 41.27 43.85 42.56
5 5 39.81 44.78 42.295
6 5 34.32 1 17.66
7 5 33.91 37.01 35.46
8 5 1 61.43 31.215
9 20 52.79 49.95 51.37
10 20 25.95 29.46 27.705
11 20 48.98 44.92 46.95
12 20 58.39 29.94 44.165
13 20 1 31.16 16.08
14 20 29.24 34.31 31.775
15 20 28.19 35.57 31.88
16 20 36.9 51.87 44.385
17 20 40.19 26.59 33.39
18 80 21.55 34.24 27.895
19 80 30.41 1 15.705
20 80 1 13.1 7.05
21 80 1 19.97 10.485
22 80 1 21.77 11.385
23 80 34.1 38.21 38.21
24 80 24.07 1 12.535
25 80 40.43 1 20.715
26 80 26.62 28.95 27.785

37
Table 2 continued
TBS_M = mesial microtensile bond strength in MPa of mesial beam
TBS_D = distal microtensile bond strength in MPa of distal beam
Avg_TBS = Average tensile bond Strength of mesial and distal beams
Groups =Based on etching times of 5, 20and 80 seconds
Note: Pre test failures are recorded as 1 MPa

38
Table3 - Mean microtensile bond strength by surfaces and etching times without
including the failure mode.
Group Mean_TBS Mean_TBS/M Mean_TBS/D Std_Dev Std_Dev/M Std_Dev/D
5 sec 38.35 36.69 37.28 11.63 10.07 13.93
20 sec 38.09 40.08 37.09 8.57 12.22 9.42
80 sec 28.82 29.53 28.63 7.35 6.96 7.83
Analysis variable: Microtensilebond strength
Groups = Based on etch times of 5, 20 and 80 seconds
Mean_TBS = Average mean microtensile bond strength in MPa
Mean_TBS/M = Mean microtensile bond strength in MPa of mesial beams
Mean_TBS/D = Mean microtensile bond strength in MPa of distal beams
Std_Dev = Standard deviation
Std_Dev/M= Standard deviation of mesial beams
Std_Dev/D= Standard deviation of distal beams

39
Table 4 - Mean microtensile bond strength by surfaces and etching times with the
pre test failure as 1MPa
Group Mean_TBS Mean_TBS/M Mean_TBS/D Std_Dev Std_Dev/M Std_Dev/D
5 sec 32.49 32.23 32.75 9.27 15.69 18.19
20 sec 36.41 35.74 37.09 11.17 17.33 9.42
80 sec 19.08 30.02 17.69 10.33 15.29 14.58
Analysis Variable: Microtensile bond strength
Mean_TBS = Average Mean microtensile bond strength in MPa
Mean_TBS/M = Mean microtensile bond strength in MPa of mesial beams
Mean_TBS/D = Mean microtensile bond strength in MPa of distal beams
Std_Dev = Standard deviation

Std_Dev/M Standard deviation of mesial beams
Std_Dev/D Standard deviation of distal beams

40
Table5 – Comparison of mean microtensile bond strength (MPa) with and without
pretest failures
Groups Mean_TBS PTF [1MPa] Mean_TBS without PTF

A A
5 sec 32.49 38.35
A A
20 sec 36.40 38.09
B A
80 sec 19.08 28.82
Mean_TBS PTF[1MPa] = Mean microtensile bond strength in MPa
including pretest failure as 1 MPa in data
Mean_TBS without PTF = Mean microtensile bond strength in MPa without
including pre test failure
Letters A and B = Represents group comparison of the mean tensile
bond strength. Means with same letters represent

no significant statistical difference.
41
Figure 5 - Graph showing data comparison of mean microtensile bond strength with
and without including the pretest failure
45
40
35
30
25 5 seconds
20 20 seconds
15 80 seconds
10
0
PTF WITHOUT PTF
PTF: Pre test failure
Y axis: Mean microtensile bond strength in MPa
X axis: 5, 20 and 80 seconds groups with and without pretest failure

42
Figure 6 - Graph showing comparison of mean microtensile bond strength
PTF: Pre test failure
Y axis: Mean microtensile bond strength in MPa
X axis: Groups based on etching time of 5, 20 and 80 seconds

43
4.2 Statistical analysis
4.2.1 Statistical results without the pretest failures data
Comparisons of microtensile bond strength for mesial surfaces
The results from one-way ANOVA procedure revealed that there was no
statistically significant effect for the etching time on the micro-tensile bond strength for
mesial surfaces, F (2, 18)=1.84, p=0.1875.That is, the data showed that there was no
significant difference in microtensile bond strength among three etching times.
Comparisons of microtensile bond strength for distal surfaces
statistically significant effect for the etching time on the micro-tensile bond strength for
distal surfaces, F (2, 18) =1.18, p=0.3298. That is, the data showed that there was no
significant difference in micro-tensile bond strength among three etching times.
Comparisons of microtensile bond strength at tooth level
statistically significant effect for the etching time on the micro-tensile bond strength at
tooth level, F(2, 22)=2.76, p=0.0851. That is, the data showed that there was no
significant difference in micro-tensile bond strength among three etching times (Table 5).
If there was a missing value at either distal or mesial surface, the average value was equal
to the one value left.
4.2.2 Statistical results with the pretest failure data
Comparisons of microtensile bond strength at mesial surface
mesial surface, F (2, 23) =2.33, p=0.1195. That is, the data showed that there was no
44
significant difference in micro-tensile bond strength among three etching times at mesial
surface.
Comparisons of micro-tensile bond strength at distal surface
tooth level, F(2, 22)=2.30, p=0.0972. That is, the data showed that there was no
significant difference in micro-tensile bond strength among three etching times on distal
surfaces
Comparisons of microtensile bond strength at tooth level
The results from one-way ANOVA procedure revealed that there was statistically
significant effect for the etching time on the micro-tensile bond strength at tooth level, F
(2, 23) =6.91, p=0.0045. The post-hoc Tukey-Kramer’s test indicated that the mean
micro-tensile bond strengths at distal surface observed in 20 seconds and 5 seconds
groups were significantly greater than that observed in 80 seconds group, while there was
no significant difference between 20 and 5 etching times. Table 5 shows the results from
Tukey-Kramer’s tests.
4.3 Results from scanning electronic microscopy

Once the bond strength was evaluated in each sample, the fracture mode was
observed under SEM (Amray 1820-D) to evaluate the predominant type of failure. The
list of fracture mode is represented in Table 6. All fractures are grouped in any of the 4
categories which include cohesive failure in dentin, cohesive failure in resin composite,
adhesive failure or joint failure and mixed failure if the fracture was in the adhesive layer
and dentin and/or resin composite.
The failure mode was examined at 90 times magnification and analyzed using
Fisher’s exact test. The results revealed no association between groups of etching times
and failure association. Table 6 shows the failure mode in each group based on etching
time. 5 seconds (Figure 7) and 20 seconds group (Figure 8) showed majority of their
45
failures as cohesive failures in resin composite and dentin followed by mixed failures and
a few failures as joint failure.
80 seconds group (Figure 9) showed about 40% of the failure as mixed failures
with about 40 % of joint failures with only about 20% of failures as cohesive failure in
dentin with no cohesive failures in resin composite.

46
Table 6 - Failure mode results
Group N Type 1 Type 2 Type 3 Type 4
20 14 4 2 2 6
40 17 6 2 3 6
80 13 3 0 5 5
Groups = Based on etch times of 5, 20and 80 seconds
N = Sample size
Type 1 = Cohesive failure at dentin
Type 2 = Cohesive failure at resin based composite
Type 3 = Adhesive failure
Type 4 = Mixed (adhesive failure + dentin/ resin based composite or both)

47
Figure 7 - Graph showing the fracture modes in 5 seconds group
5 Secs
Cohesive/Dentin Cohesive/RBC
Mixed= Adhesive+Dentin/RBC Adhesive
14%
29%
43% 14%
A chart representing a 5 seconds group and their percentage of failure mode

48
Figure 8:: Graph showing the fracture modes in 20 seconds group
20 Secs
Mixed=Adhesive+Dentin/RBC Adhesive
18%
35%
35%
12%
A chart representing a 20 seconds group and their percentage of failure mode

49
Figure 9:: Graph showing the failure modes in 80 seconds group
80 Secs
Mixed=Adhesive+Dentin/RBC Adhesive
23%
38%
0%
39%
A chart representing an 80 second group and their percentage of failure mode

50
Figure 10 - Graph showing a comparison of the fracture modes from each group.
6
5 Cohesive/Dentin
4
Cohesive/RBC
3
Mixed-Adhesive
Adhesive +
2
Dentin/RBC
1 Adhesive
0
5 Secs 20 Secs 80 Secs
A graph comparing the failure mode among the 3 etching groups

51
Figure 11 – Cohesive failure in dentin
Picture 1
Picture 2
Cohesive fractures in dentin

entin in 5 seconds (picture1) and 20 seconds (picture
picture 2) groups
52
Figure 12 - Cohesive failure in composite resin
Picture 3
Cohesive fracture in resin based composite

omposite from 20 seconds group
53
Figure 13 - Joint failures
Picture 4
Picture 5
Picture 4 and 5 showing jjoint
oint failure in the adhesive
54
Figure 14: Mixed failures
Picture 6
Picture 7
Picture 6 and picture 7 showing mixed failure

55
Figure 15: Adhesive failure in the hybrid layer
Picture 8
Picture 8 Higher magnification
View of dentin side of specimen of 80 second group in low magnification
showing failure in the hybrid layer (Picture 8). Higher magnification of picture 8 of
specimen from 80 seconds groups showing failure in the bottom of the hybrid layer.
56
CHAPTER 5: DISCUSSION
When performing an adhesive restoration, clinicians are daily challenged with
several steps involved that might be difficult to control, of which conditioning of enamel
and dentin with acid to a specific time is one of the variables among the many factors that
influence this adhesion process. For instance, there is a thought that phosphoric acid
should be on enamel and dentin within periods of 30 and 15 seconds, respectively as
recommended by various manufactures. However some cavity preparations for restoring
direct or indirect restorations may involve large areas of exposed dentin to which need to
be etched with acid uniformly. Conceivably, the dentin located on some surfaces of those
preparations may undergo uneven or longer periods of acid conditioning than anticipated
depending on the manner it is applied. So in this study the effect of extended etching time
on vital dentin was evaluated in comparison to some in vitro studies that have been
carried out to evaluate the same effect on the dentin (Abu-Hanna and Gordan 2004, 105-
110;Abu-Hanna, Gordan, and Mjor 2004, 28-33;Hashimoto et al. 2002, 99-105;Jacques
and Hebling 2005, 103-109) and primary dentin (Bolanos-Carmona et al. 2006a, 1121-
1129;Sardella et al. 2005, 355-362) by measuring bond strengths with either tensile bond
strength or shear bond strength tests.
Consistent with previous in vitro studies, the results of this study showed a
reduction in the bond strengths with increase in etching time on vital dentin and the null
hypothesis was rejected within the limits of the study and when pretest failure were
included in the data. In all these studies the results have been consistent that excessive
etching of dentin has an inverse effect on the bond strengths. Most of these tests were
microtensile bond testing which is a suitable approach in evaluating the joint interface.
57
The groups with excessive etching time of dentin showed a drop in the bond strength
irrespective of some variation with the type of adhesive system, or methods and these
results were comparable to the results in this study.
Major limitation of this study was the sample size. When a sample and power
analysis was applied to the existing data, the total sample size needed to obtain a
detectable difference between three groups with different etching times including the
pretest failure were 375 premolars at 80% power , 0.252 effect size and standard
deviation as 11. The total sample size increased to 492 premolars when the power was
increased to 90% to detect any contrast with the same effect size and standard deviation.
It is not uncommon to have some failure of samples during trimming or sectioning of
beams before testing them for the microtensile bond strength testing and in these cases
the bond strength of these samples is considered as 1 MPa and included in the data as left
censored data for statistical analysis. On the contrary, when the pretest failures were
excluded from the data, the total sample size needed to obtain a detectable difference
between those groups were 54 at 80% power, 0.197 effect size and 10 as standard
deviations. The total sample size increased to 69 when the power was increased to 90% to
detect any contract with same effect size and standard deviation. There was a limitation
to obtain these many subjects to fulfill this statistical requirement.
Bond strengths with altered etching time produced a distinguished difference in
the microtensile bond strength in the overetched group which were comparable to other
in vitro studies, justifying that a deeper depth of demineralization and incomplete
penetration of monomers of the adhesive to the base of the demineralized dentin. It is
reported that monomer diffusion and polymerization is competitive in the deminerialized
dentin in that the polymerization process will increase in the molecular weight which
reduces the diffusion rate in this dentin and demonstrated non resin impregnated dentin in
the groups with prolonged etching time (Nakabayashi, Watanabe, and Arao 1998, 379-
58
385). On the other hand 5 seconds group showed no significant difference in bond
strength from the 20 seconds group. This could be due to the fact that the sample sizes in
each of these groups were small for analyzing any statistical significant difference. At the
same time when sample size was estimated using sample and power analysis to detect
any possible difference between the groups showed that at 80% or 90% power, the
sample size needed where very high yet there was a difference between the 80 second
group and the other two groups even with a limitation of samples when the pretest
failures were included. This raises a question and further research areas of lower etching
times on vital dentin and its affect on the durability and bond strengths at the interface of
the joint. Previous in vitro study (Abu-Hanna and Gordan 2004, 105-110;Abu-Hanna and
Gordan 2004, 105-110) comparing the shear bond strength to different etching times had
no significant adverse effect on bond integrity with etching times less than 15 seconds.
This relates to optimum depth of demineralization and complete infiltration of the
adhesive monomer to the base of the hybrid layer leaving no unprotected mineral
depleted collagen.(Eick et al. 1997, 306-335)
A comparison was made in this study with the different etch times with one group
having excessive etching time of 80 seconds on vital dentin. Putting this into clinical
perspective, there may be various clinical situations while etching a large area of tooth
structure where extensive dentin is being exposed. In spite of being aware of the
manufacturer’s recommendations of etching time with the particular etching agents,
controlling the time on dentin to the specific time could be a challenge combined with the
coordination of etching of enamel to be optimally prepared with longer etch times with
up to 60 seconds (Gardner and Hobson 2001, 64-67) compared to dentin for optimum
bonding surface and dentin not being over etched at the same time. Control of etching
time is important; particularly over etching can be detrimental to adhesive strength,
durability and among the many variables which affect the bond integrity, this is a variable
which is completely determined by the operator.

59
Besides etching time, there are many other variables which determine the integrity
of the joint and durability of the restoration in the oral environment. Some variables like
the choice of adhesive systems, their application methods or the type of restorative
material can be controlled by the operator but some variables like age of the patient, the
type or location of dentin as a substrate can still have significant effect on the joint
integrity. Vital dentin is a diverse structure for bonding. The dentin pulp complex
demonstrates permeability that is characteristic of dentin which induces diffusion of the
substrates responsible for the intrinsic wetness of exposed dentin (Pashley and Carvalho
1997, 355-372) and most monomers in the adhesive systems being hydrophobic can
result in incomplete infiltration due to outward movement of fluid from the dentinal
tubules and thus interfere with the adhesive mechanism. Extrinsic wetness of the dentin
with moisture or saliva contamination can also affect the bond strength (Barghi, Knight,
and Berry 1991, 130-135); however, in this study the restorations were performed under
optimal moisture control with the use of rubber dam.
The subjects included in this study were in the age group of 18 - 25 years and at
this age the dentin is considered as young dentin. This type of dentin can behave
differently as a substrate when compared to dentin with increasing age as there is clinical
evidence that with increasing age, there is dentinal sclerosis which can affect bonding and
related restoration loss rate. (Van Meerbeek et al. 1993, 195-202; Van Meerbeek et al.
1993, 195-202). However some studies show no difference in bond strengths with young
and aged dentin (Brackett et al. 2008, 282-286)
An etch-and-rinse system with acetone based solvent in which primer and
adhesive are combined in one bottle was implemented. The solvent in this adhesive
drives the water away and facilitate a better affinity of the adhesive to penetrate into the
demineralized collagen rich dentin (Hashimoto et al. 2002, 99-105;Hashimoto et al. 2000,
406-411) and many in vitro studies have shown good bonding results with this system
however, due the technique sensitivity of this adhesive the in vivo results are lower in
60
long term (Van Meerbeek et al. 2003a, 215-235) due to intrinsic wetness of the dentin.
Acetone acts as an excellent monomer solvent but at the same time it evaporates quickly
after being applied and leaves a thick viscous adhesive fluid that does not penetrate
completely in the exposed collagen. The amount and concentration of acetone as solvent
in a important and when applied in thin multiple layers it resulted in increased monomer
conversion (Holmes et al. 2007, 1506-1512). This makes the use of acetone based
adhesive system more challenging due to technique sensitivity. Adding to this when
dentin is over etched, it produces deeper areas of demineralized collagen and makes
viscous adhesive fluid harder to penetrate to the complete depth as acetone evaporates
quickly.
Every specimen sample tested for the microtensile bond strength was analyzed for
failure modes using a SEM and the percentage failures were calculated. Majority of the
failures were in the mixed failures in all the groups with the large percentage of mixed
failures in the adhesive interface and dentin. With the etching time, there was an increase
in adhesive failures and the failures occurred at the base of the adhesive hybrid layer in
the 80 second groups. This is suggestive that there could have been discrepancies
between depth of demineralization and adhesive penetration and when stress occurred in
these areas, the failures occurred in these weaker areas. On the contrary, the 5 and 20
seconds groups showed more cohesive failures than the adhesive failures. However there
was no statistical significant association with the type of failure mode and the etching
times.
The study was performed on short term duration which would generate various
questions about long term durability in vital dentin treated with extended etching time.
This could bring the better insight and fate of the unprotected collagen especially when
left for a long time period in association with acetone as solvent. The limitations of such
long term in vivo studies could be the recruiting of subjects for the desired time frame
since most of the subjects who are commonly recruited are subjects who are undergoing
61
orthodontic treatment and need their teeth extracted for orthodontic reasons and waiting
long term in these cases who interfere with the orthodontic treatment.
Further studies are suggested in comparing vital dentin with different types of
adhesives systems’ response to different etching times. Nonoleakege analysis can also
play a vital role in analyzing vital dentin treated with the extended etching times to
evaluate the fate of the unprotected collagen and its degradation over a time due to
incomplete monomer infiltration. It would be valuable to assess the areas of deeper dentin
that has lesser intertubular dentin than superficial dentin to which the adhesive monomers
penetrates.
Further studies should be continued on vital dentin to evaluate any differences by
increasing the sample size and power of the statistical procedures as this was a limitation
of this research and the area of great consideration should be etching under 15 seconds
which did not produce any statistically significant difference which could provide
valuable information if any.

62
CHAPTER 6: CONCLUSIONS
Etch-and-rinse adhesive system requires etching with phosphoric acid as a
separate step when bonding RBC to enamel and dentin. However, differences on etching
time, particularly on dentin has an influence on the resin-dentin interface. This study
evaluated the effect of three different etching times (5, 20 and 80 seconds) on
microtensile bond strength of RBC bonded to vital dentin with 2 steps etch and rinse
adhesive system. The results inferred that the group with overextended etch time of 80
seconds, inversely effected the microtensile bond strength by lowering it and thus
rejecting the null hypothesis. However there was no difference with the other two groups
of 5 and 20 seconds. These results were achieved when pretest failures as 1MPa were
included in the data but when they were excluded from data, there was no statistically
significant difference among the groups. On the other hand, the 5 seconds group and the
20 seconds groups had no statistical difference with the available sample size. The failure
mode analysis provided evidence that the majority of the fractures occurred in the
overextended etch group where the joint failures and mixed failures representing
weakened areas when compared to cohesive failures in dentin or RBC but these failures
had no statistical significance with the duration of etching time.
In conclusion, within the limitations of this study, over extended etching times on
vital dentin associated with 2-step total etch bonding system showed lower bond
strengths There was no association with the percentage of failure and the etching time in
spite of the majority of the failures in the over etched groups being adhesive failures.
Further research is needed to evaluate the impact of shorter acid etching time on
longevity of in vitro and in vivo resin-dentin interfaces.

63
APPENDIX
Failure mode 5 seconds group
Composite side Dentin side Composite side Dentin side

1 1
2 2
3 3
4 4
5 5
6 7
7 8
64

1 1
2 2
3 3
4 4
5 6
6 7
7 8
8 8
9
65

1 2
3 4
5 6
6 7
7 8
8 9
9
66
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Impact of Different Acid Etching Time On Microtensile Bond Streng

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University of Iowa

Iowa Research Online

Theses and Dissertations

Impact of different acid etching time on microtensile bond

Follow this and additional works at: https://ir.uiowa.edu/etd

Part of the Other Dentistry Commons

Copyright © 2009 Aadarsh Gopalakrishna

This thesis is available at Iowa Research Online: https://ir.uiowa.edu/etd/291

Follow this and additional works at: https://ir.uiowa.edu/etd

A thesis submitted in partial fulfillment

Thesis Supervisor: Assistant Professor Saulo Geraldeli

This is to certify that the Master’s thesis of

has been approved by the Examining Committee

Thesis Committee: ____________________________________

I would like to express my gratitude to Dr. Saulo Geraldeli, my thesis advisor,

thank him for all the extended support in literature development.

It is my great honor to have Dr. Deborah Cobb as my graduate program director

my thesis project. She made me feel home away from home.

statistics of this thesis and her valuable suggestions.

for the in vivo aspect of this research.

help to make me a better clinician and for his motivation to teach.

LIST OF TABLES -------------------------------------------------------------------------------- vi

LIST OF FIGURE -------------------------------------------------------------------------------- vii

2.1 Statement of problem----------------------------------------------------------4

Table 1: Materials, compositions and manufacturer------------------------------------------ 27

including the failure mode-------------------------------------------------------------- 36

the pretest failure as 1 MPa-------------------------------------------------------------37

Table 6: Failure mode results--------------------------------------------------------------------- 46

Figure 1: Procedure of tooth preparation------------------------------------------------------- 28

Figure 2: Procedure of restoration--------------------------------------------------------------- 29

Figure 3: Tooth sectioning------------------------------------------------------------------------ 31

Figure 4: Dumbell sample on Dirks device----------------------------------------------------- 32

Figure 5: Graph showing data comparison of mean microtensile bond strength

Figure 6: Graph showing data comparison of mean microtensile bond strength---------- 42

Figure 8: Graph showing the fracture modes in 20 seconds group-------------------------- 48

Figure 9: Graph showing the failure modes in 80 seconds group--------------------------- 49

Figure11: Cohesive failure in dentin------------------------------------------------------------ 51

Figure 12: Cohesive failure in composite resin------------------------------------------------ 52

Figure 13: Joint failures--------------------------------------------------------------------------- 51

Figure 14: Mixed failure-------------------------------------------------------------------------- 54

Figure 15: Adhesive failure in the hybrid layer------------------------------------------------ 55

dental adhesives. These adhesives, decrease the microleakage in the tooth-restoration

esthetically restoring the teeth with minimal or no tooth preparation.

RBC adheres to enamel and dentin by adhesives. However a major shortcoming

substrate, especially when compared to enamel bonding.

Whenever dentin is cut for placement of restorations, considerable quantities of

before its application in order to increase material retention.

Resin adhesives classified as etch-and-rinse adhesives require the removal of the

dissolution of mineral content, which are predominately hydroxyapatite crystals. This

demineralization versus monomer penetration of the adhesive, which is greatly

Gordan, and Mjor 2004, 28-33; Bolanos-Carmona et al. 2006, 1121-1129;Hashimoto et

Based on previous studies that excessive etching creates a deep demineralized

information as to the clinical importance of avoiding extended phosphoric acid

valuable source of information in terms of improving the clinical longevity of the

restoration and better success of treatment with tooth colored restorations.

CHAPTER 2: LITERATURE REVIEW

2.1 Statement of problem

2.2 Background of tooth structures and adhesion

Understanding different tooth structures is important for a successful adhesive

restorative dentistry as different tooth structures as substrate behave differently to dental

adhesive bonding systems. It would be important to understand the adhesion principal as

Tooth enamel is the hardest substance in the human body. It is of ectodermal

and 12% water by volume. Enamel is formed by ameloblasts. There is formation of