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Bond Strength Durability of Different Adhesives to Dentin

After Aging in Two Different Solutions
Heba A. El-Deeba / Lamia E. Daifallab / Omar I. Badran0/ Enas H. Mobarakd

Purpose: To determine the influence of aging in artificial saliva compared to distilled water on the dentin microtensile bond strength (pTBS) of different adhesive systems.
Materials and Methods: Occlusal enamel and superficial dentin of 42 teeth were removed and roots were sec­
tioned to expose the pulp chamber for connecting the tooth segments to an intrapulpal pressure assembly. Accord­
ing to the tested adhesives, tooth segments were allocated to three groups (n = 14): an etch-and-rinse adhesive
(Adper Scotchbond Multi-Purpose, SBMP), a two-step self-etching adhesive (Clearfil SE Bond, CSE), and a singlestep self-etching adhesive (Clearfil S3 Bond, S3). Each adhesive system was applied to the dentin surface accord­
ing to its manufacturer’s instructions, while intrapulpal pressure was simulated. Resin composite (3M ESPE) was
built up in two increments of 2 mm each. Each bonded specimen was sectioned to obtain eight sticks
(0.8 ± 0.01 mm2*). Sticks of each group were divided equally (n = 56) according to the storage solution, either dis­
tilled water or artificial saliva. For each storage solution, half of the sticks of each subgroup (n = 28) was stored
for 24 h at 37°C and the other half was thermocycled for 10,000 cycles between 5°C and 55°C. Sticks were then
subjected to pTBS testing. Data were statistically analyzed using multifactor ANOVA with repeated measures and
Bonferroni’s post-hoc test (p < 0.05). Student’s t-test was used for pairwise comparison. Failure modes were de­
termined for all tested sticks using scanning electron microscopy.
Results: The decrease in bond strength of the three adhesives was significantly higher in distilled water than in ar­
tificial saliva. The predominant failure modes were adhesive and mixed.
Conclusion: The decrease in bond strength was more pronounced for specimens stored in distilled water than in

artificial saliva.
Keywords: distilled water, artificial saliva, adhesives, dentin bonding, bond strength, storage, microtensile bond

strength.
J Adhes Dent 2016; 18: 303-309.
doi: 10.3290/].jad.a36153

uring the last decades, adhesive dentistry has fo­
cused on enhancing bonding to dentin. Dental manu­
facturers achieved te c h n ic a l59 and chem ical ad­
vances15'61 to develop new dental adhesives with

D

3 Associate Professor, Restorative Dentistry Department, Faculty of Oral and
Dental Medicine, Cairo University, Egypt. Idea, performed the experiment,
wrote the manuscript.
b Associate Professor, Restorative Dentistry Department, Faculty of Oral and
Dental Medicine, Cairo University, Egypt. Contributed substantially to conduct­
ing the study, co-wrote manuscript.
c PhD Student, Restorative Dentistry Department, Faculty of Oral and Dental Medi­
cine, Cairo University, Egypt. Performed the experiment, co-wrote manuscript.
d Professor, Restorative Dentistry Department, Faculty o f Oral and Dental Medi­
cine, Cairo University, Egypt. Hypothesis, experimental design, constructed
setups, contributed substantially to discussion, proofread the manuscript.
Correspondence: Enas H. Mobarak, Cairo University, Faculty o f Dentistry,
709 El-Malak El Saleh, Cairo, 1 1 5 5 0 Egypt. Tel: + 0020-2-2206-6203;
e-mail: enasmobarak@hotmail.com

Vol 18, No 4, 2016

Submitted for publication: 22.01.15; accepted for publication: 26 .041 6

increased resin-dentin bond strength. Currently, several
adhesive systems are available, from the multi-step ver­
sions of etch-and-rinse and self-etching adhesives to the
more simplified single-step self-etching adhesives. Flowever, the loss of resin-dentin bond strength overtime still
affects the adhesive restorations and markedly reduces
their durability in vitro.8
To understand the causes behind bond degradation of
the adhesives being continually launched into the market,
both in vitro and in vivo experiments have been carried
out. The oral cavity is considered the ultimate testing en­
vironment to reveal the behavior of the adhesives, al­
though there diverse factors work simultaneously, making
it difficult to assess each factor’s contribution to bond
degradation separately. Flowever, in vitro models simulat­
ing certain in vivo conditions can provide less complex en­
vironments to assess the fundamental mechanisms of
bond degradation.55

303

The null hypotheses were: 1) there is no difference between dentin microtensile bond strengths when specimens are stored for 24 h in distilled water vs artificial saliva. 3) there is no difference be­ tween dentin microtensile bond strengths when thermocycled either in distilled water or in artificial saliva. the current study was performed to examine the influence of artificial saliva. Therefore.32 Then. Each tooth was attached upside down with its occlusal surface placed on the center of a circle drawn on a glass 304 slab to facilitate centrally embedding it in a chemically cur­ ing polyester resin (Polyester # 2 1 2 1 . and a sin­ gle-step self-etching adhesive. 8 inch.000 thermocycles corre­ spond to approximately one year of in vivo functioning. Nevertheless. thermocycling is used as a model at­ tempting to test bond degradation. Adper Scotchbond Multi-Purpose (SBMP. Clearfil SE Bond (CSE. 3M ESPE) shade A3 was applied in two increments of 2 mm each. A second cut was made parallel to the occlusal surface. cleaning of the pulp chamber (b). Selected teeth were stored in phosphate buffer solution containing 0. Each adhesive system was applied according to the manufacturer’s instructions (Table 1) while intrapulpal pressure was simulated.or even long-term periods to assess bond durability. IL. Teflon plate fixation (c). USA). Solingen. 3M ESPE. 2 mm gingival to the cementoenamel junction. Buehler) for 10 s to produce a standard­ ized smear layer and surface roughness. each of which was polymerized for 20 s using Bluephase C5 (Ivoclar Vivadent. Schaan.48 Dentin surfaces were then wet polished using 600-grit silicon carbide paper (MicroCut. It allows bonded speci­ mens to be subjected to extreme temperatures. Previous articles10’16 suggested that 1 0. Japan). USA) under a stream of water to remove occlusal enamel and superficial dentin to obtain a flat dentin surface (midcoronal portion). whether the interaction between artificial sa­ liva as a storage solution. Kuraray Noritake). Clearfil S3 Bond (S3. butterfly needle insertion and embedding of the specimen (d).02% sodium azide at 4°C until being used. Eternal Chemical. Remnants of pulp tissue in the pulp cham ber were removed using an excavator (Carl M artin. many studies have used distilled water as a storage solution for short. Hsein. leaving the crown and 1 to 2 mm of hard tooth structure gingival to the cementoenamel junction ex­ posed using the same protocol described previously by Mobarak et al.33 This was done to facilitate trimming the tooth perpendicular to its longitudinal axis using a dia­ mond disk (Isomet. in laboratory settings. Germany) w ithout touching the walls of the pulp chamber (Fig lb ) . and adhesive restorations would differ from that of distilled water storage solution has yet to be validated.11'14’17’22’35’36'39-40'54’60 Others preferred to mimic the clinical situation and used artificial saliva as an immersion solution. Liechtenstein) with an intensity of > 500 m W /cm 2. MATERIALS AND METHODS Specimen Preparation A total of 42 sound human third molars were selected for the study. Buehler. For instance. with or without thermocy­ cling. Lake Bluff. MN. St Paul.30 Restorative Procedures Prepared specimens were divided into three groups (n = 14) according to the tested adhesives: an etch-and-rinse adhe­ sive. Resin composite (Filtek Z250. on the bond strength dura­ bility of adhesives with different bonding strategies to dentin. thereby ex­ posing the pulp chamber (Fig la ). Light intensity was checked The Journal of Adhesive Dentistry . Tokyo. Taiwan).El Deeb et al Fig 1 Specimen preparation for intrapupal pressure simulation: ob­ taining the coronal section (a). the spec­ im ens were connected to th e intra pu lpa l pressure assembly during bonding (Fig 1 c-e) following the same procedures described by Mobarak.6'8’23’24’43 Furthermore. compared to distilled water. used alone or in combination with the thermocycling process. a two-step self-etching adhesive. 2) there is no difference between dentin microtensile bond strengths with or without thermocycling. bonded speci­ mens in the specially constructed incubator while connected to intrapulpal pressure assembly (e). Kuraray Noritake.

A dhesive: Apply. Thermocycling was done for 10. The Netherlands) to determine the mode of failure.56 The artificial saliva was pre­ pared as CaCi2 (0.05 was consid­ ered statistically significant. tw o-com po nen t tw o-step self-e tc h in g ad hesive syste m (Kuraray N orita ke.25’57 The aver­ Vol 18.01 mm2. HEMA. Fareham. leave u n d istu rb e d fo r 2 0 s. water. CA. 2016 age of the pTBS values for the sticks originating from the same tooth (two per tooth) was calculated considering the tooth as a statistical unit .50 Microtensile Bond Strength Measurment Each stick was then fixed to the m icrotensile strength jjgi3. KH2 P0 4 (4.Ndiethano l-p-toluidin e. USA) and stressed in ten­ sion using a universal testing machine (Lloyd Instruments. HEMA. arti­ ficial saliva for 24 h. MDP bis-GMA. Milano. Orange. and artificial saliva with thermocy­ cling). From each specimen. The failure mode was classified as: type 1: adhesive failure at the dentin side.8 ± 0. compositions. hydrophobic dim e thacrylate.29 Wjth a cyanoacrylate adhesive (Rocket heavy. Eind­ hoven. yielding 28 sticks per group.12’42 Statistical Analysis Statistical analysis was performed using multifactor ANOVA (followed by Bonferroni’s post-hoc test).Ndiethanol-p-toluidine. in which pTBS was the dependent variable while the adhesive system type. version 15 for MS Windows. polyalkenoic acid A dhesive: HEMA. and application procedures M a te r ia l (m a n u fa c tu r e r ) C o m p o s itio n A p p lic a tio n p r o c e d u r e s E tchant: Apply fo r 1 5 s. rinse w ith w a te r spray fo r 1 5 s. storage solution. Sticks that failed before testing were assigned a bond strength of zero MPa and were included in the statistical analysis . as well as the pTBS values of each adhesive system with both storage solutions under each storage con­ dition (baseline and thermocycled). HEMA. p < 0. hydrophobic d im e th a c ry la te . S t Paul. MN. No 4. eth ano l. Philips Electronic Instruments. Adper S cotchb ond M ulti-Purpose.0 mmol/l).000 cycles of 5°C to 55°C.and y-axes to obtain multiple sticks with a cross-sectional area of 0. UK) at a crosshead speed of 0. CA. 8 central sticks were selected. USA) S cotchbond U niversal E tchant: 32% by w eigh t phospho ric acid etch in g gel (pH < 1) P rim er: Ethanol.L-cam phorquinone. USA). leave un d istu rb e d fo r 5 s. th e n dry w ith a m ild oil-free a ir s tre a m fo r 5 s. leave u n d istu rb e d fo r 1 5 s.5 m m /m in until failure. hydrophilic d im e th a c ry la te . N.37 Every day. lig h t cure fo r 1 0 s. and HEPES buffer (20 m m o l/l ) ]. D . type 3: mixed failure (adhesive failure at the dentin side/cohesive failure in the adhesive layer). P rim er: Apply o n to th e visib ly m o is t prepared to o th surface. SPSS. The significance of the effect of each inde­ pendent variable and its interactions was determined. manufacturers. Corona. C learfil SE Bond. Dental Ventures of America. Japan) C learfil S3 Bond. type 2 : cohesive failure in the adhesive layer. using a Demetron LED radiom eter (Kerr. th re e -ste p etch-and-rinse ad hesive system (3M ESPE. [\laN3 (0. th e n lig h t cure fo r 1 0 s.3 mmol/l).El Deeb et al Table 1 Material specifications. MDP: methacryloyloxydecyl dihydrogen phosphate. th e n dry w ith g e n tle a ir flo w fo r 5 s. KOI (30 m mol/l). HEMA: 2-hydroxyethyl methacrylate. water with thermocycling.28 Bonding and resin composite buildup were done while the tooth segments were subjected to 15 mm Hg simulated intrapulpal pressure.7 m m ol/l). s ilana ted colloidal silica Bond: Apply one c o a t and a g e n tle oil-free a ir stre am fo r 5 s. th e n ge ntly th in w ith a m ild oil-free a ir stre am fo r 5 s a t a d ista n ce o f 2 cm fro m d e n tin surface. Stu­ dent’s t-test was used to compare the pTBS values of each adhesive system with or without thermocyling in each stor­ age solution. Lloyd Instruments). s ila n a te d co llo id a l silica Apply 2 0 s. USA).2 m m ol/l). Failure Mode Analysis Both fractured sections of each stick (dentin side and resin composite side) were mounted on an aluminum stub. ph oto-initiator. The sticks col­ lected from each tooth were evenly distributed over the four study groups (water for 24 h. Data were analyzed using SPSS for Windows (Statistical Package for Social Sciences. The tensile force at failure was recorded and con­ verted to tensile stress in MPa units using computer soft­ ware (Nexygen-MT. water. with a dwell time of 30 s and a transfer tim e 10 s . IL. lig h t cure fo r 1 0 s. gold sputter coated. N. and storage condition were the indepen­ dent variables. The specimens were then immersed in artificial saliva 37 for 24 h at 37°C in a spe­ cially constructed large incubator to accommodate the in­ trapulpal pressure assembly. MgCI2 -6H 20 (0. Bond: MDP bis-GMA. specimens were sectioned along the x. or type 4 : mixed failure (adhesive failure at the dentin side/cohesive failure in the adhesive layer/cohesive failure in resin com­ posite). and observed with a scanning electron mi­ croscope (SEM 515. dl-cam phorquinone.31 305 . bis-GMA P rim er: Apply. Tokyo. The frequency of each mode was expressed in per­ cent for each group . Bis-GMA: bis-phenol-A giycidyl methacrylate. one-com pon ent sin g le -ste p s e lf-e tc h in g ad hesive syste m (Kuraray N oritake) P rim er: 10-M DP HEMA. After 24 h. the pH of all solutions used in the study was checked using a digital pH m eter (Hanna. Chicago. Italy) and the solutions were changed . a ir dry fo r 5 s w ith high-pressure a ir s tre a m .

1 (4.78). However.6 (4. Meanwhile. SBMP specimens mostly showed failure types 2 (cohesive failure in adhesive layer) and 3 (mixed failure: adhesive failure at the dentin side/cohesive failure in the adhesive layer).3 (5. type 4 was the predominant mode after 24 h storage in either solution. t-test).6) bB p tf/tn t = 1 /2 8 p tf/tn t = pre-test faiIure/totaI number of tested sticks. no signifi­ cant e ffe ct was found for the adhesive system type (p = 0 .8) aB ptf/tn t = 3 /2 8 22.001). whereas type 3 was frequently recorded after thermocycling in either solution. Figure 2 shows the percentages of failure modes of all tested groups. failure type 4 (mixed failure: adhe­ sive failure at dentin side/cohesive failure in the adhesive layer/cohesive failure in resin composite) was the most common mode for SBMP fractured specimens. mean pTBS values for all tested adhesives significantly decreased (p < 0.1 (6.1) aA ptf/tn t = 0 /2 8 29.8) bB ptf/tn t = 1 /2 8 15. After thermocycling in both solutions. the storage condition (p < 0. standard de­ viations) are presented in Table 2.9 (4.3 (3. as well as their interaction (p < 0. Following either 24 h storage or thermocycling in distilled water. The interaction among the three variables was not significant (p = 0.7) aB ptf/tn t = 3 /2 8 23. CL 0) CD CO O ) x: CI CSE CD Distilled water i Artificial saliva | Distilled water Type 1: Adhesive failure at the dentin side Artificial saliva Type 2: Cohesive failure in the adhesive layer CN Distilled water CO CO Artificial saliva CL o >> o E a3 i— CO CO CSE •CuO Type 3: Mixed failure (adhesive failure at the dentin side + cohesive failure in the adhesive layer) Distilled water Artificial saliva Type 4: Mixed failure (adhesive failure at the dentin side + cohesive failure in the adhesive layer + cohesive failure in resin composite) Distilled water Artificial saliva Distilled water CO CO Artificial saliva 20% 40% 60% 80% 100% RESULTS Multifactor ANOVA revealed a significant effect for the stor­ age solution (p < 0.1) aA ptf/tn t = 0 /2 8 Thermocycling 16.2 (4. SBMP fractured specimens predominantly showed failure type 1 (adhesive failure at the dentin side). means with different superscript small letters are statistically significantly different (p < 0. The descriptive statistics (means.7) bB ptf/tn t = 2 /2 8 14. S3: Clearfil S3 Bond.001).01). The pTBS values of each adhesive system after 24 h stor­ age either in distilled water or artificial saliva were not signifi­ cantly different. within columns for each adhesive system with each storage solution. the difference between the pTBS val­ ues for all tested adhesives thermocycled in distilled water and artificial saliva were significantly different (p > 0.3 (6.01).8 3 ) or its interaction with the storage solution (p = 0.05. while when stored in artificial saliva for 24 h.1) aA p tf/tn t = 0 /2 8 32. means with different superscript capital letters are statistically significantly different (p < 0.5)aA p tf/tn t = 1 /2 8 33.5 (7.81) and storage condition (p = 0. The Journal of Adhesive Dentistry . Within rows for each adhesive system. After thermocy­ cling in artificial saliva. CSE: C le a rfil SE Bond.El Deeb et al Table 2 Mean (SD) microtensile bond strength pTBS in MPa of tested groups Adhesive system Adper Scotchbond Multi-Purpose Clearfil SE Bond Clearfil S3 Bond D istilled w ater A rtificial saliva D istilled w ater A rtificial saliva D istilled w ater A rtificial saliva 24 h 34.95) variables. The CSE fractured specimens exhibited mostly type 4 failure when stored for 24 h or thermocycled in either solution. For S3 fractured specimens. 306 Fig 2 The percentages of the recorded modes of fail­ ure.3) aB ptf/tn t = 2 /2 8 24. SBMP: Adper Scotchbond Multi-Purpose.9 (4.7 (5. t-test).3) aA p tf/tn t = 0 /2 8 30.4) aA ptf/tn t = 0 /2 8 34.01).7 (6.05.

This leads to higher diffusion of water mole­ cules through the dentin/adhesive interface when stored in water than in artificial saliva. However. the higher is its flux (ie.El Deeb et al DISCUSSION The present study findings revealed that the use of different storage solutions for 24 h had no influence on the microtensile bond strength of the tested adhesives. prone to water sorption over time. as there was a significant difference between the microtensile bond strengths of the thermocycled adhesives in distilled water and those in artificial saliva. The concentration of water molecules is higher in distilled water than in saliva. the baseline results did not predict the future behavior of the tested adhesive systems. whether the MMP types activated and their activation rates would differ based on the storage me­ dium used (distilled water and artificial saliva) still requires investigation. Additionally. A study comparing the leachability of resin composite components when stored in dis­ tilled water or artificial saliva revealed a significant differ­ ence between the leached components in the two storage solutions. it cannot be considered as the appropriate means of testing the real performance of the adhesive res­ toration during clinical service. no published data on the effect of these storage media (artificial saliva vs distilled water) on dentin adhesive bond strength is available. which the literature1-19-41 .6-45-49 The degradation of incompletely infiltrated zones within the hybridized dentin may continue even in the absence of bacterial enzymes. which results in reduction of their mechanical proper­ ties. Hence. According to Fick’s first law of diffusion. the first null hypothesis failed to be rejected. it seems that the use of distilled water as a storage solution de­ creases the similarity of in vitro studies to the clinical find­ ings. uncut specimen. water remains entrapped at the resin/dentin interface. a dramatic decrease in the microtensile bond strength of the dentin adhesives was associated with therm ocycling in both media. The reduction in bond strength of S3 following thermocy­ cling could be due to the behavior of bonded interfaces as semi-permeable membranes. Diffu­ sion-dependent hydrolysis caused by hot water may acceler­ Vol 18. because the pTBS values of all tested adhesives decreased signifi­ cantly after thermocyling.including a recent clinical study38 . as hydrolysis of the colla­ gen peptides may occur and weaken the adhesive joint over tim e . thermocy­ cling causes repetitive contraction/expansion stress.18. As presented. It must also be borne in mind that application of simulated intrapulpal pressure during bonding increases convective fluid movement and leads to the formation of water channels through the polymerized adhesives. it has been reported that the glass-ionomer restorative materials possessed higher surface hardness when stored in human saliva compared to distilled water. which could jeopardize its polymerization efficacy and in turn lower its mechanical properties.18-27 Moreover. it has been shown that MMP acti­ vation is induced by dentin adhesives. Because the degree of hydrolysis depends on the amount of reactive ions and their degree of diffusion. No 4. On the other hand. which may eventually result in crack propagation along the resin/dentin interface. In addition to diffusion-dependent hydrolysis. 2-3'1147. To date. The commonly observed reduction in bond strength could be partially attributed to the chemical degradation of both collagen fib rils and resinous polymers by hydroly­ sis.46-51 the difference between the water results (48% to 50% de­ crease in bond strength) and the artificial saliva results (26% to 30% decrease in bond strength) could be explained on the basis of their differences in hydrolytic degradation potential. thus.47 Although distilled water was more aggressive as an aging medium. accelerating the degradation of the adhesive inter­ face. irrespective of the strategy employed (ie. rather than being exposed as a whole.40-45 Being highly hydrophilic.9 especially as the four sides of the cut sticks in the present study were exposed to the storage solution.20 Moreover.5 the higher the concentration difference of a certain particle is. CSE contains hydrophilic monomers. This is despite the fact that CSE contains 10-MDP.49 The bond strength reduction with the two-step self-etch­ ing adhesive (CSE) might be explained by both the hydro­ lytic degradation of polymers occurring after water sorption and the permeability of the adhesive layer. it is well known that duplicating the exact properties of human saliva 307 . the high thermal contraction/ expansion coefficient of the adhesives compared to tooth tissue generates repeated interfacial contraction/expansion stresses that could be similar to those in the oral cavity. which could contribute to the observed reduction in bond strength.37 The significant reduction of bond strength o f the SBMP adhesive system after 3 months’ storage in distilled water has been reported by others.26 These endogenous gelatinolytic/collagenolytic enzymes can degrade the collagen structure of the bonded interface over tim e.44-52 Recently. the second null hypothesis should be rejected.34 Based on the previous and the current findings. causing swelling and plasticization of the poly­ mers. 2016 ate the hydrolysis of unprotected collagen and extract poorly polymerized resin oligom ers. its permeation through the dentin/adhesive interface in this case).23 Nevertheless. the three adhesive systems contain HEMA.has reported to provide bond stability.58 The significant reduction of bond strength of the etchand-rinse adhesive system (SBMP) following thermocycling could be related to the discrepancies between the extent of demineralization and the depth of infiltration of the adhe­ sive resin. The third null hypothesis m ust also be re­ jected. etch-and-rinse or self-etching). as well as interaction between storage solution and resin composite constituents.51 Bauer and llie4 found that aging in artificial saliva rather than distilled water im­ proved the micromechanical properties of resin composites. HEMA is a hydro­ philic monomer which imbibes a large amount of water within the adhesive and the hybrid layer53 during thermocy­ cling. Such a mismatch has been claimed to affect the stability of the resin-dentin bond. it is very difficult to remove water from the adhesive resin of one-step self-etching adhesives.

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