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Original Article

Influence of caries detection dye on bond strength of

sound and carious affected dentin: An in-vitro study
Udai Pratp Singh, AP Tikku1, Anil Chandra1, Kapil Loomba1, Lalit Chandra Boruah
Department of Conservative Dentistry and Endodontics, Kothiwal Dental College and Research Centre, Kanth Road, Moradabad,
1
Department of Conservative Dentistry and Endodontist Faculty of Dental Sciences, Lucknow, India

Abstract
Objectives: The objective of this study was to evaluate the influence of caries detection dye on the in-vitro tensile bond
strength of adhesive materials to sound and carious affected dentin.
Materials and Methods: Forty healthy and carious human molars were ground to expose superficial sound dentin and
carious affected dentin. Caries Detector dye was applied to sound and carious affected dentin and rinsed. Subsequently
the dentin was etched with 37% phosphoric acid and rinsed leaving a moist dentin surface. The adhesive (Single bond) was
applied in single layers and light cured. A posterior composite (Filtek Z 250) were used to prepare the bond strength specimens
with a 3 mm in diameter bonding area. Control and experimental groups were made with and without application of dye
respectively. Each group includes both sound and carious affected dentin. After 24 hour immersion in distilled water, tensile
bond strength (MPa) was measured using an Instron testing machine.
Results: Analysis of variance (ANOVA) was used to evaluate the data. The tensile bond strength were significantly less in
experimental subgroup than control subgroups.
Conclusion: The tensile bond strengths were higher in sound and carious affected dentin without application of caries
detection dyes.
Keywords: Caries detector dye; posterior composite; single bond; sound and carious affected dentin

INTRODUCTION Caries detector dyes have been developed to further help

the diagnosis and removal of dental caries, by differentiating
Tooth preparation is considered to be complete clinically between infected and affected dentin. The dye stains only
when the consistency of underlying dentin is hard to a the infected outer carious dentin.[2-4] Dye usage allows
sharp probe and free of caries. The carious process usually dentists to perform an ideal cavity preparation for adhesive
progresses as a series of exacerbations and remissions restorations.[5] To ensure that all carious dentin has been
that are characterized by periods of high production of removed, use of dye is indicated as the last step in tooth
acid that are responsible for the dissolution of the hard preparation.
tissues of the tooth. If allowed to proceed untreated, it
results in the progressive destruction of the tooth and The bonding mechanism for current adhesive agents is based
eventual infection of the dental pulp.[1] on the acid removal of the smear layer and demineralization
of the underlying dentin, which leaves an exposed collagen
Carious dentin has been identified by two layers of soft network. The application of hydrophilic primers followed by
dentin, the outer carious layer is infected unremineralizable the adhesive, which encapsulate this collagen network and
with irreversible deteriorated collagen fibers, with no form a resin impregnated layer or hybrid layer.[6]
odontoblastic processes, insensitive and therefore,
should be removed. The inner carious layer is uninfected, Few reports are available in literatures regarding the effect
remineralizable with reversibly denatured collagen fibers, of caries detection dye on the bond strength of sound and
alive with living odontoblastic processes, sensitive, and so carious affected dentin. The present study has been designed
should be preserved.[1,2] to evaluate the influence of caries detection dye on the in-
Address for correspondence: Access this article online
Dr. Udai Prtap Singh, Department of Conservative Dentistry Quick Response Code:
and Endodontics, Kothiwal Dental College and Research Centre, Website:
Kanth Road, Moradabad - 244 001, India. www.jcd.org.in
E-mail: drudaipratap@gmail.com
Date of submission: 01.10.2009 DOI:
Review completed: 14.12.2009 10.4103/0972-0707.80732
Date of acceptance: 03.07.2010

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Singh, et al.: Influence of caries detection dye on bond strength of dentin

vitro tensile bond strength of adhesive materials to sound Table 1: Materials and their composition used in the
and carious affected dentin. study
Name of material Composition
MATERIALS AND METHODS Caries detector(Kurary, 1% acid red 52 solution in propylene glycol.
Osaka, Japan)
Materials along with their composition used in the present Single Bond (3M ESPE) Water, ethanol, HEMA (2-Hydorxy-
study are summarized in Table 1. ethylmethacrylate), BIS-GMA (bisphenol
A diglycidyl ether dimethacrylate), a
photoinitiator system and a methacrylate
Experimental groups functional copolymer of polyacrylic and
Forty freshly extracted (both carious and non-carious) polyitaconic acids.
FiltekTM Z 250 (3M ESPE) Filler -Zirconia/Silica. Filler loading 61% by
human mandibular molar teeth were selected for this (Posterior composite) volume (without silane treatment) particle
study. Teeth were rinsed thoroughly under running tape size range of 0.01 to 3.5 µm, composite
water to remove blood and saliva. Teeth were rendered resin composed of BIS-GMA (bisphenol
free of debris and calculus using scaler. The samples were A diglycidyl ether dimethacrylate),UDMA
(urethane dimethacrylate) and BIS-EMA
stored in normal saline at room temperature until they (bisphenolA polyethylene glycol diether
were subjected to the experimental procedure. dimethacrylate).

For carious affected dentin, twenty samples with coronal

caries extending approximately halfway through the 40 samples
dentin were used in this study. The buccal carious surface
was ground parallel to the long axis of the tooth to expose GROUP A (Control) n=20 GROUP B (Experimental) n=20
a flat surface of normal dentin surrounding the carious Without application of with application of
caries detection dye caries detection dye
lesion. The buccal enamel was grinded with the help of
carborundum disc, made smoothened by the sand paper
(silicon carbide, 220-600 grit) and washed copiously with Subgroup A1 Subgroup A2 Subgroup B1 Subgroup B2
(sound dentin) (affected dentin) (sound dentin) (affected dentin)
distilled water. n=10 n=10 n=10 n=10

Figure 1: Flow chart showing distribution of samples into

To obtain carious affected dentin, grinding was performed
groups
using combined criteria of visual examination and staining
with caries detector dye (Kurary, Japan) as described[7] that dentin surface as per the manufacturer’s instruction and
is the dentin was hard to an explorer and no longer stained
photo cured for 10 sec using QTH light source (3M Curing
bright red with caries detector dye [Figure 1].
Light 2500).
The samples in Group-A (n=20, control group) were without A plastic cylindrical mould with a internal diameter of 3
application of caries detection dye on both sound and mm and length 4 mm was placed atop the bonded surface.
carious affected dentin surfaces. The samples in Group-B
A flexible orthodontic wire to be used during testing
(n=20, experimental group) were with application of
procedure was inserted into first layer of composite resin
caries detection dye on sound and carious affected dentin
inside the mould before photo curing .The composite resin
surfaces. The control and experimental group were further
divided into two subgroups A1 and A2 and B1 and B2 with 10 was build up in increments and each layer was cured for
samples in each as shown in the following flowchart. 20 sec. After complete curing, the plastic mold was easily
removed with the help of tweezers.
All the samples were embedded in the acrylic resin blocks
Debonding procedure was performed in tension on Instron
of uniform size. Buccal surface of each sample was exposed
from acrylic resin block. universal testing machine at a crosshead speed 0.5 mm/
min. Tensile bond strength of each group was calculated
In control subgroups, dentin surfaces were etched with in Mpa. After testing, the fracture mode of each specimen
37% phosphoric acid gel for 15 sec while in experimental was determined visually under 5X magnification.
subgroups all samples were etched after application of
caries detection dye. RESULTS

After etching, it was rinsed with spray water and dried The bond strength results are presented in Table 2. Tensile
leaving a moist dentin surface for application of the strength is significantly higher in sound dentine from
bonding agent. The adhesive resin (single bond) was carious affected dentine in control subgroups as well as
applied in a single layer on the sound and carious affected experimental subgroups [Table 3].

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Singh, et al.: Influence of caries detection dye on bond strength of dentin

Table 2: The mean value of tensile strength within control group and experimental group
Control group-A Experimental group-B
(Without application of caries detection dye) (With application of caries detection dye)
Subgroup A1 Subgroup A2 Subgroup B1 Subgroup B2
(sound dentin) (caries affected dentin) (sound dentin) (caries affected dentin)
No. of sample 10 10 10 10
Mean (in MPa) 22.4280 13.1940 18.5530 9.4500
S.D. 1.3765 1.2460 0.8353 1.1043
S.E. 0.4353 0.3940 0.2641 0.3323

Table 3: ANOVA table for tensile strength in subgroups

Source of variation D. F. Sum of square Mean sum of square Variance ratio
Between subgroups 3 985.780 328.593 F =245.018
Error 36 48.280 1.341 P=0
Total 39 1034.059

Table 4: Comparison of tensile strength between The mechanism by which caries disclosing agents selectively
control (without application of caries detection dye) stain only carious, irreversibly demineralized dentin
and experimental subgroup (with application caries has been determined. It was originally thought that the
detection dye) solutions were staining bacteria directly. It is now known
Comparison ‘t’ value ‘P’ value that the stain is the result of bacterial demineralization.
A1 vs. A2 15.7274 P<.001* Both basic fuchsin and acid red stain the collagen fibers
B1 vs. B2 20.7897 P<.001*
exposed by the bacteria caused dentin demineralization
process.[1]
Since 'F' is highly significant hence there are significant
difference in tensile strength of with and without The results of this study show that the mean value of
application of dye in experimental and control group. tensile bond strength of single bond was higher in the
control subgroup than experimental subgroups which can
There are significant differences between the control be explained as that caries-affected dentin contain some
subgroups (A1 vs A2) and experimental subgroups (B1 vs substances that interfere with free radical generation
B2) in tensile strength values. (P<.001) [Table 4]. or propagation, leading to improper polymerization of
resins in such dentin. The peritubular dentin matrix of
caries-affected dentin, which take up much more toluidine
DISCUSSION
blue stain and exhibit more intense metachromasia than
Caries- detector dyes have proven to be useful in the normal peritubular dentin, suggests the presence of
identification and removal of carious dentin. These agents mucopolysaccharides or glycoprotiens. These molecules
made from basic fuschin in a propylene glycol base reliably may interfere with resin wetting of fine porosities within
stain only the dentin that is infected with bacteria and both intertubular and peritubular dentin and/or may
irreversibly demineralized without staining the affected interfere with conversion of adhesive monomers to
dentin. Therefore, the presence of stain reliably determines polymeric network.[9] Fusayama et al, observed affected
the part of dentin to be removed.[8,9] dentin to have turbid, transparent and subtransparent
zones. There is limited information on the structure of
Early formulations of the caries detector included a 5% basic these zones as well as conflicting evidences about their
fuchsin solution in propylene glycol as a solvent; however, properties. General perception is that transparent dentin
it was replaced with 1% acid red 52 solution in the same is sclerotic and hypermineralized due to tubular occlusion
solvent as a substitute dye because fuchsin is believed to which might act as a barrier to penetration of primers and
be carcinogenic. The acid red disclosant can accurately bonding agents.
determine dentin status.[4] Studies demonstrated that
dentin containing less than 10000 CFU/mg was normally Although the adhesive resin may have followed the primer,
not disclosed by the fuschin dye, whereas counts of greater it may not have copolymerized well with the primer. Thus
than 550,000 CFU /mg dentin were readily stained. This the adhesion of resins to caries-affected dentin may be
suggests that the dye can be used to approximate the inferior to that of normal dentin, due to weaker collagen
bacterial load of the dentinal surface because a certain and/or weaker resin even though most of the tubules
bacterial mass apparently needs to be present before the in such dentin are filled with mineral deposits. These
dye is absorbed by the dentin.[3] intratubular crystals are not well-packed and are softer

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Singh, et al.: Influence of caries detection dye on bond strength of dentin

than well-packed apatite even through they are more acid- solution Quintessence Int 1988;19:397-401.
2. List G, Lommel TJ, Tilk MA, Murdoch HG. Use of a dye in caries
resistant.[10-15] identification. Quintessence Int 1987;18:343-5.
3. Anderson MH, Loesche WJ, Charbeneav GT. Bacteriologic study of a
basic fuschin caries-disclosing dye. J Prosthet Dent 1985;54:51-5.
In relation to the influence of dyes on the adhesion of 4. Kidd EA, Joyston-Bechal S, Smith MM, Allan R, Howe L, Smith SR.
FiltekTM Z250, the decrease in the bond strength may be The use of a caries detector dye in cavity preparation. Br Dent J
due to dye solution remaining in sound and affected dentin 1989;167:132-4.
5. Fusayama T. Ideal cavity preparation for adhesive composites. Asian J
as mentioned in other studies.[16] Single bond is an adhesive Aesthet Dent 1993;1:55-62.
that needs to have close contact with the dentin substrate to 6. Nakabayashi N. Current development in adhesive materials. Esthet Dent
Update 1994;5:42-5.
produce the desired bond strength. Dye remaining trapped 7. Nakajima M, Sano H, Urabe I, Tagami J, Pashley DH. Bond strength
in dentin may adversely affect the wetting of dentin by of single-bottle dentin adhesive to caries affected dentin. Oper Dent
materials, thereby decreasing micromechanical retention of 2000;25:2-10.
8. Shimizu C, Yamashita Y, Ichijo T, Fusayama T. Carious change of dentin
these materials.[17] Resin materials are sensitive to previous observed on long span ultra thin sections. J Dent Res 1981;60:1826-1931.
dentin contamination. During our study, it was observed 9. Fusayama T. Two layers of carious dentin, diagnosis and treatment.
Oper Dent 1979;4:63-70.
that despite the application on sound dentin and carious 10. Ogawa K, Yamashita Y, Ichijo T, Fusayama T. The ultra structure and
affected dentin, caries-detecting dyes, even after being hardness of the transparent layer of human carious dentin. J Dent Res
1983;62:7-10.
rinsed and acid etched, were not completely removed, as 11. Kanca S. Effect of resin primer solvents and surface wetness on resin
evidenced by some samples with sound tissue remaining composite bond strength to dentin. Am J Dent 1992;5:213-5.
lightly colored which might have influenced the results. 12. Tay FR, Gwinnett AI, Pong KU, Wei SH. The overwet phenomenon an
optical micromorphological study of surface moisture in the acid etched,
resin-dentin interface. Am J Dent 1995;9:43-8.
CONCLUSION 13. Xie J, Powers JM, McGuckin RS. In-vitro bond strength of two adhesives
to enamel and dentin under normal and contaminated condition. Dent
Mater 1993;9:295-9.
Within the limitation of the present study, it is concluded 14. Pashley DH, Ciucchi B, Sano H, Horner JA. Permabilty of dentino
adhesive agent. Quintessence Int 1993;24:618-31.
that the tensile bond strength was higher in sound and 15. Powers JM, Finger WJ, Xie J. Bonding of composite resin to
carious affected dentin in both control and experimental contaminated human enamel and dentin. J Prosthodont 1995;4:28-32.
group without application of caries detection dyes than 16. Demacro FF, Matos AB, Matson E, Powers JM. Dyes for caries detection
influence sound dentin bond strength. Oper Dent 1998;23:294-8.
that with the application of caries detection dyes. 17. Shimizu C, Inokoshi S, Bushita M, Hosoda H, Fusayama T. Caries
detector for pulpless teeth. Oper Dent 1983;8:94-8.

REFERENCES
1. Fusayama T. Clinical guide for removing caries using a caries detecting Source of Support: Nil, Conflict of Interest: None declared.

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