d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

Available online at www.sciencedirect.com

journal homepage: www.intl.elsevierhealth.com/journals/dema

The contribution of chemical bonding to the short- and long-term enamel bond strengths
Zhenliang Zhang a , Xiaomiao Wang a , Ling Zhang a , Bing Liang a , Tian Tang a , Baiping Fu a, , Matthias Hannig b
a

Hospital of Stomatology Afliated to Zhejiang University School of Medicine, Zhejiang University School of Stomatology, Yanan Road 395, Hangzhou 310006, China b Clinic of Operative Dentistry, Periodontology and Preventive Dentistry, Saarland University, Homburg/Saar, Germany

a r t i c l e
Article history:

i n f o

a b s t r a c t
Objectives. MDP (10-methacryloyloxydecyl dihydrogenphosphate) has been proven to possess chemical bonding ability to tooth hard tissues, but its contribution to the enamel bond strength has not been recognized. The aim of this study was to investigate the contribution of chemical bonding to the short- and long-term bovine enamel micro-tensile bond strengths (TBS). Methods. The acid-etched enamel surfaces were treated without any primer (control) or with one of three MDP-containing primers (containing different ratio of MDP/HEMA/Bis-

Received 25 July 2012 Received in revised form 15 December 2012 Accepted 25 April 2013

Keywords: Chemical bonding Micro-tensile bond strength Phosphoric acid esters Bovine enamel SEM

GMA, Kuraray Co.) for 5 s, water-sprayed and air-dried. Subsequently, the pretreated enamel surfaces were applied with an etch-and-rinse adhesive Durall Bond (Heraeus Kulzer) and placed with composite resin Durall VS (Heraeus Kulzer). The specimens were prepared for TBS tests after 24-h or 1-yr water storage. The etched enamel surfaces treated with or without MDP-containing primers were analyzed by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Results. The acid-etched enamel treated with the MDP-primers for a very short time could produce the greater enamel TBS than the control did (p < 0.05), and change enamel micromorphology. No signicant different TBS were found between 24-h and 1-yr water storage (p > 0.05). The chemical bonding of MDP on the enamel surfaces was re-conrmed by XPS. Signicance. The additional chemical bonding of MDP around the enamel crystallites of the etched enamel substrate could signicantly increase the short- and long-term enamel TBS, and their TBS surpass those of the etch-and-rinse adhesive alone. 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

1.

Introduction

Chemical bonding of carboxylic acids and phosphoric acid esters (PAEs) to enamel/hydroxyapatite (HAp) surfaces has

been studied by numerous researchers [19]. Chemical bonding at the resintooth hard tissue interfaces has been presumed to provide a strong and durable bond without any additional tooth preparations for macro-mechanical retention [911]. Whether carboxylic acids either decalcify or chemically

Corresponding author. Tel.: +86 0571 87217427; fax: +86 0571 87217427. E-mail addresses: fbp@zju.edu.cn, baiping fu@yahoo.com (B. Fu). 0109-5641/$ see front matter 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.dental.2013.04.009

e104

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

adsorb onto HAp depends on the dissolution rate of the respective calcium salts in their own acidic solutions, irrespective of concentration and pH [3]. Some PAEs used in contemporary self-etch dental adhesives could decalcify and chemically adhere to HAp simultaneously [7,8]. Hannig et al. demonstrated the inter- and intra-HAp crystallite hybridization by self-etch adhesives at the enamelresin interfaces without any nano-gaps [12]. Taken together, these ndings suggest a physisorption or chemisorption of self-etch primers onto enamel/HAp surfaces after the interaction of self-etch primers with the enamel crystallites [1317]. Chemical bonding at the enamel/HAp surfaces has been investigated using X-ray photon spectroscopy (XPS), X-ray diffraction (XRD), liquidand solid-state nuclear magnetic resonance (NMR), etc. in previous studies [19,18]. Ten-methacryloyloxydecyl dihydrogen phosphate (MDP) is one of the most promising PAEs used in self-etch adhesives such as Clearl SE Bond and Clearl S3 Bond [4,1923]. Recently, Yoshihara et al. detected a 4-nm layered structure around HAp powder after reaction with MDP [14]. Likewise, our previous research revealed that the self-etch adhesives containing either carboxylic acids or PAEs could decalcify and chemically adhere to HAp simultaneously [57,13]. However, the previously published data did not demonstrate that self-etch adhesives containing acidic functional monomers produce stronger bond strengths than etch-and-rinse adhesives do [2427]. Yoshihara et al. examined the chemical stability of new functional monomercalcium salts in combination with their previously determined bond strength results [28], and inferred that formation of stable monomer-calcium salts enhanced the immediate bond strength of self-etch adhesives [15]. Based on the hypothesis that similar surface micromorphology would result in similar bond strengths, Erickson et al. matched the enamel surface micromorphologies that resulted from treatments with self-etch adhesives or various concentrations of phosphoric acid, analyzed the corresponding enamel bond strengths, and deduced that chemical bonding of some self-etch adhesives cannot compete with the micromechanical interlocking produced by phosphoric acid etching [29]. All previously published data about chemical bonding at the enamel/HAp surfaces were not directly associated with the bond strengths obtained by micromechanical interlocking. We previously assumed that chemical bonding of self-etch adhesives on the residual HAp crystallites would make a minor contribution to the immediate mechanical bond strengths [13]. Up to now, it has not been claried how much chemical bonding at the enamel surfaces contributes to the shortand long-term mechanical bond strengths. The null hypothesis tested in this study was that chemical bonding on the etched enamel substrate would make a minor contribution to the short- and long-term enamel bond strengths obtained by micromechanical interlocking, and long-term water storage would deteriorate the resinenamel interfaces. The aim of this experimental study was to investigate (1) the synergistic contribution of chemical bonding on the etched enamel surface to short- and long-term micromechanical bond strengths; (2) the effect of MDP-containing experimental primers on the micromorphology of the etched enamel surfaces; and (3) the resinenamel interfaces after short- and long-term storage in water.

2.
2.1.

Materials and methods

Specimen preparations

Thirty-two non-carious bovine mandibular incisors were extracted from 8 cows in a local slaughterhouse, stored in the respective bottle containing 0.1% thymol solution at 4 C, and were used for this study within 1 month after extraction. Four incisors extracted from the same cow were randomly divided into 4 groups according to the different surface treatments. Twenty-four incisors from 6 cows were randomly assigned into 4 groups as above-mentioned. All labial enamel surfaces were ground under copious running water serially with 300-, 600-, 1200- and 2500-grit SiC paper for 30 s, each ending with 4000grit SiC paper for 1 min. The enamel surfaces were etched with 37% phosphoric acid (ETCH-37, Bisco, Inc., Schaumburg, IL, USA) for 15 s, water-sprayed for 30 s, and thoroughly air-dried. Subsequently, they were treated with one of three experimental (MDP-containing) primers (Ex-1, Ex-2, Ex-3, Table 1) for 5 s or without any experimental primer (control). Afterwards, the enamel surfaces were thoroughly water-sprayed for 1 min and totally air-dried. The etch-and-rinse adhesive Durall Bond (non-acidic functional monomers, thus, no chemical reaction with enamel/hydroxyapatite) (Heraeus Kulzer, Hanau, Germany) was applied to the pre-treated enamel surfaces, gently air-dried, and light-cured for 20 s. Finally, two 2-mm increments of the composite resin Durall VS (Heraeus Kulzer, Hanau, Germany) were placed, and light-cured for 40 s, respectively. All the light-curing procedures were done using a light-curing unit (MACO, SLC-VIIIB, Hangzhou, China) with a light output of 800 mW/cm2 . All the materials used in the study are summarized in Table 1.

2.2.

Micro-tensile bond strengths ( TBS)

After water storage at room temperature for 24 h, 12 specimens were perpendicularly sectioned through the resinenamel interfaces using a low-speed saw (Isomet 1000, Buehler, Lake Bluff, IL, USA) under continuous water cooling. Subsequently, they were prepared into beams of about 0.9 mm 0.9 mm 8 mm. The TBS tests were performed with a Micro Tensile Tester (Bisco Co., USA) at a tensile speed of 1 mm/min until fracture. The dimension of the fractured surface was measured. The TBS was calculated in MPa. After water storage at room temperature for 1 yr (water change every week), another 12 specimens were prepared and subjected to TBS tests as mentioned above. After either 24-h or 1-yr water storage, the TBS tests were repeated three times using 4 bovine incisors per cow per time (24 incisors from 6 cows).

2.3.

Scanning electron microscopy (SEM)

The labial enamel surfaces of another 4 bovine incisors were treated with phosphoric acid and experimental primers as above-mentioned. These specimens were treated neither with adhesive nor composite resin. Furthermore, two debonded specimens were randomly selected from each group after TBS tests. All the specimens were dried with a series of

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

e105

Table 1 List of materials used in the study. Product and manufacturer (LOT No.)
Etch-37 Bisco, Inc., Schanmburg, IL, USA (0900005544) Durall bond Heraeus Kulzer, Hanau, Germany (232.08) Ex-1 Kuraray, Co., Japan (040218) Ex-2 Kuraray, Co., Japan (040218) Ex-3 Kuraray, Co., Japan (040218) Durall VS Heraeus Kulzer, Hanau, Germany (21206)

Principal ingredients
37% Phosphoric acid etchant, bonzalkonium chloride

pH

Steps of application
Applied and left untouched for 15 s, rinsed, totally dried

Methacrylate solvent, silicon dioxide, benzoi methyl ether, photopolymerization catalyst 30% (w/w) MDP/HEMA/Bis-GMA = 1:1:1 (w/w), solvent: distil water/ethanol=1:1 (v/v) 30% (w/w) MDP/HEMA/Bis-GMA = 2:1:1 (w/w), solvent: distil water/ethanol = 1:1 (v/v) 30% (w/w) MDP/Bis-GMA = 1:1 (w/w), solvent: distil water/ethanol = 1:1 (v/v) Urethanedimethacrylate, silicon dioxide (0.020.07m)

2 2 2

Applied and gently air-blown to a lm, light-cured for 20 s Applied for 5 s, water-rinsed for 1 min, totally dried Applied for 5 s, water-rinsed for 1 min, totally dried Applied for 5 s, water-rinsed for 1 min, totally dried Two 2 mm increments light-cured for 40 s, respectively

Bis-GMA, bisphenol A diglycidyl methacrylate; HEMA, 2-hydroxyethyl methacrylate; MDP, 10-methacryloyloxydecyl dihydrogenphosphate.

alcohol solutions, gold-sputtered, and analyzed with an SEM (ZEISS ULTRA 55, Germany). During specimen preparation for TBS tests, an additional 1 mm thick specimen was prepared from each group. After the cross-sectioned surfaces of these specimens were treated with 0.1 M HCl for 20 s to delineate the resinenamel interfaces, the specimens were prepared as above-mentioned for the SEM observations.

2.4.

X-ray photoelectron spectroscopy (XPS)

The labial enamel surfaces of another 4 bovine incisors were treated in the same way as those prepared for SEM analysis. The specimens were dried with a series of alcohol solutions, and subsequently prepared in high vacuum for XPS (AXIS ULTRADLD , Kratos, England). XPS was performed using a monochromatic Al K X-ray source under the emission current of 8 mA, and accelerating voltage of 15 kV.

2.5.

Statistics

Statistical analysis was performed with the SPSS software (version 17.0, SPSS Inc., Chicago, IL, USA). Factorial design ANOVA was used to statistically analyze all TBS data. Post hoc LSD multiple comparisons were applied to evaluate the statistical differences in TBS among the four groups after 24-h or 1-yr water storage.

3.
3.1.

Results
Micro-tensile bond strength

Fig. 1 Mean enamel micro-tensile bond strengths measured in this study (Means SD (n), MPa). Whiskers indicate 95% condence intervals. (n: indicates the numbers of the beams for the TBS. Ex-1: aqueous-ethanol (1:1) solution of 30% MDP/HEMA/Bis-GMA = 1:1:1 (w/w); Ex-2: aqueous-ethanol (1:1) solution of 30% MDP/HEMA/Bis-GMA = 2:1:1 (w/w); Ex-3: aqueous-ethanol (1:1) solution of 30% MDP/Bis-GMA = 1:1 (w/w)).

All the TBS data are schematically presented in Fig. 1. No pretesting failure occurred in this study. The TBS of the control group were signicantly lower than those of three experimental groups (p < 0.05), regardless of 24-h and 1-yr water storage. No signicantly different TBS were found either among the three tests within each group, or among the three

experimental groups, or between 24-h and 1-yr water stored specimens (p > 0.05).

3.2.

SEM observations

The polished enamel surface etched with phosphoric acid revealed a distinct etching pattern with exposure of enamel

e106

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

Fig. 2 Micromorphology of enamel surfaces. (a) The highly polished enamel surface etched with phosphoric acid revealing a distinct etching pattern with exposure of enamel prism rods due to the preferential interprismatic apatite dissolution. The acid-etched enamel surfaces were further treated with the experimental MDP-containing primers (Ex-1 (b), Ex-2 (c), Ex-3 (d)), revealing enamel HAp crystallites covering the etched enamel surfaces with many micro-porosities (bd). (magnications = 20,000 fold, bar = 1 m).

prism rods due to the preferential interprismatic tissue dissolution (Fig. 2a). The acid-etched enamel surfaces treated with three MDP-containing experimental primers for 5 s exhibited a variable network-like layer of enamel HAp crystallites on the etched enamel surfaces (Fig. 2bd). The resinenamel crosssections revealed intimate contacts with a few porosities after 24-h (Fig. 3a, c, e, and g) or 1-yr water storage (Fig. 3b, d, f, h). Fracture failure of the debonded specimens was mainly a mixed failure in the hybrid layer (Fig. 4). Enamel surfaces of control specimens and 24-h water stored experimental specimens were covered partly with some adhesives residues (Fig. 4ac). Enamel hydroxyapaptite crystallites were clearly visible in control specimens, but not clearly visible in 24h water stored experimental specimens (Fig. 4b). Fractures of debonded experimental specimens after 1-yr water storage took place mainly at the adhesivecomposite interfaces, revealing microlled materials (Fig. 4d).

MDP-containing experimental primers (Ex-1, Ex-2, Ex-3) were similar (Fig. 5). Compared with the peak intensity (C 1s) of the etched enamel surface (Fig. 5a), the peak intensity of the etched enamel surfaces treated with the MDP-containing primers (Fig. 5bd) signicantly increased. The narrow-scan spectrum of the C 1s region of the etched enamel revealed a backbone (C C, C H, C C) peak at 284.6 eV, C O peak at 286.1 eV, and C O peak at 288.1 eV (Fig. 6a). The narrow-scan spectra of the C 1s region of enamel treated with the three MDP-containing primers were the same, revealing a backbone (C C, C H, C C) peak at 284.6 eV, C O peak around 286.1 eV, and COO peak at 288.6 eV (Fig. 6bd).

4.

Discussion

3.3.

XPS

The wide-scan spectra of the acid-etched enamel surfaces and the acid-etched enamel surfaces treated with the

The risk of random errors and the heterogeneity of teeth origin in this study could be reduced to a minimum, because highly polished labial enamel samples from the same cow were used for the TBS test, and TBS tests were repeated three times within each group, using 4 bovine incisors per cow per time. Phosphoric acid etching of the labial bovine enamel surface in this study produced the typical etching pattern with

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

e107

Fig. 3 SEM micromorphology of enamelresin interfaces The micromorphology revealed intimate contacts with a few porosities at the enamelresin interfaces (a, control; c, Ex-1; e, Ex-2; g, Ex-3) and even after 1-yr water storage (b, control; d, Ex-1; f, Ex-2; h, Ex-3) (magnications = 5000 fold, bar = 2 m).

e108

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

Fig. 4 SEM micromorphology of fractured surfaces. The micromorphology of the enamel site of debonded specimens mainly revealed a mixed failure mode in the hybrid layer (ad). Enamel hydroxyapatite crystallites are clearly visible in the control group after either 24-h (a) or 1-yr water storage (c) and are faintly visible in experimental groups after 24-h water storage (b). The debonded specimens of experimental groups after 1-yr water storage mainly revealed a mixed failure at adhesivecomposite interfaces (d). A, adhesive; E, enamel hydroxyapatite crystallites (magnications = 1000 fold, bar = 50 m; high magnications = 20,000 fold, bar = 2 m).

preferential interprismatic tissue dissolution (Fig. 2a). Extension of etching time will not improve the enamel bond strengths due to the excessive loss of the enamel substrate [30]. Therefore, a relatively short etching time (15 s) was applied in this study. Moreover, the etched enamel surfaces treated with the different MDP-containing primers for a short time revealed that the etched enamel surfaces were covered by a layer of variable network-like/bril-like HAp crystallites (Fig. 2bd). These SEM observations suggest that the nonacidic functional monomers (HEMA, Bis-GMA) could inuence the enamel etching patterns. The different enamel etching patterns might be attributed to the specic viscosity and hydrophilicity of the primers that resulted from the different ratios of the monomers (HEMA, Bis-GMA) in solutions of the MDP-containing primers. These ndings could be explained by the fact that partial decalcication of HAp crystallites resulted from the interaction of MDP-containing primers with HAp and re-deposited onto the enamel surfaces due to the hardly soluble calcium salts of MDP [4,8]. Moreover, the fractography of debonded specimens in control groups revealed clearly visible enamel hydroxyapatite crystallites partly and adhesive mixed

with fractured hydroxyapatite crystallites partly (Fig. 4a and c), indicating solely micro-mechanical interlocking at the interface. In contrast, the fractography of debonded specimens in experimental groups revealed faintly visible enamel hydroxyapatite crystallites partly and adhesive mixed with fractured hydroxyapatite crystallites partly (Fig. 4b), indicating some chemical interaction between the hydroxyapatite crystallites and MDP. The evidence of chemisorption between phosphoric acid esters (including MDP) and HAp was proved in previous studies [4,7,8,1417]. HEMA and Bis-GMA of the three MDP-containing primers should be completely washed away, after the etched enamel surfaces treated with MDP-containing primers had been thoroughly water-sprayed and dried with a series of alcohol solutions prior to XPS analysis. However, the wide-scan spectra revealed that the peak intensity (C 1s) signicantly increased after the etched enamel surfaces were treated with the primers, indicating that some ingredients of the primers (carbon) were left on the enamel surfaces. Furthermore, the narrow-scan spectra (C 1s) of the etched enamel surfaces treated with the MDP-containing primers in this

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

e109

Fig. 5 Wide-scan Spectra of XPS. The wide-scan spectra of the acid-etched enamel surface (a) and of the etched enamel surface treated with three MDP-containing experimental primers (b, Ex-1; c, Ex-2; d, Ex-3). The etched enamel surfaces treated with the three MDP-containing primers (bd) show an increased C 1s peak intensity signicantly, when compared with the etched enamel surface (a).

study are exactly the same or very similar to those published by Yoshida et al. [4], suggesting a chemical bonding layer on the enamel. Moreover, the etch-and-rinse adhesive (Durall Bond) cannot chemically react with HAp, because it does not contain any acidic monomers [19]. Supposedly, the residual individual HAp crystallites within the enamel substrate were intimately enclosed by MDP [8,9], forming nano-layering structures [14,16], which were subsequently covered by and co-polymerized with the adhesive. Up to date, it has not been clearly elucidated how chemical bonding contributes to the mechanical bond strengths at the resinenamel interfaces. Moszner et al. assumed that chemical bonding makes only minor contributions to the immediate mechanical bond strengths, as we previously presumed [13], but contributes to the long-term bond strengths, which are associated with the chemical stability of chemical bonding and the adhesives themselves [31]. Moreover, previous studies did not directly demonstrate that chemical bonding was associated with an increase in bond strengths [15,29]. Furthermore, it has been inferred that chemical bonding plays an important role in the bond strengths by analyzing formation of stable monomer-calcium salts, and matching enamel surface morphology between both treatments with various concentrations of phosphoric acid and self-etch adhesives in combination with the previously determined bond strengths [15,29]. The ndings of the present study directly demonstrated that a signicant contribution of chemical bonding was made to the etched enamel surface with regard to the short- and

Fig. 6 Narrow-scan Spectra of XPS. Narrow-scan spectrum (a) of the C 1s region of the etched enamel surface reveal a peak at 284.6 eV representing backbone (C C, C H, and C C) bindings, a peak at 286.1 eV representing C O binding, and a peak at 288.1 eV representing C O binding. Narrow-scan spectra of the C 1s region of enamel surfaces treated with MDP-containing primers (b, Ex-1; c, Ex-2; d, Ex-3) are the same, revealing a peak at 284.6 eV representing backbone (C C, C H, and C C) bindings, a peak around 286.1 eV representing C O binding, and a peak at 288.6 eV representing COO binding.

long-term mechanical enamel bond strengths. Nevertheless, it is very difcult to completely exclude additional micromechanical inter-locking by MDP through nano-roughing of the enamel surface [18]. However, in the present study MDPcontaining primers were applied for a very short time (5 s), while phosphoric acid treatment was 15 s. In addition, MDP with a pH of around 2 reveal a lower etching ability than phosphoric acid with a pH of less than 1. Furthermore, in our pre-study experiments, we proved that an extension of the phosphoric acid etching time from 15 s to 20 s did not improve the enamel bond strength (our unpublished data). It is in accordance with previous reports on the inuence of etching time on enamel bond strength [30]. Thus, the bond strength increase observed in the present study might be mainly attributed to chemical bonding phenomenon, whereas

e110

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

the additional nano- and micro-retentive effect caused by 5-s treatment with MDP-containing primers might not be signicantly relevant. The SEM ndings in this study did not reveal substantial micro-morphological changes of the resinenamel interfaces over time (Fig. 3). Thus, the hypothesis of this study that chemical bonding to the etched enamel substrate would make only minor contributions to the short- and long-term mechanical enamel bond strengths, and that long-term water storage would deteriorate the resinenamel interfaces was totally rejected. Confusingly, why self-etch adhesives which possess an additional chemical capacity for bonding to enamel/HAp do not yield stronger bond strengths than the etch-and-rinse adhesives do [29,3235]? The bonding instability of selfetch adhesives has been conrmed by microtensile bond strength tests and marginal analysis of restorations [3638]. We presumed that the soluble or slightly soluble calcium salts resulting from the reaction of self-etch adhesives with the enamel surface would weaken enamel bond strength due to the interference of soluble or slightly soluble calcium salts within the hybrid layer during the bonding procedure. Likewise, the etched enamel surfaces treated with MDP-containing primers in this study were thoroughly water-sprayed and air-dried, therefore, soluble calcium salts resulting from the reaction of MDP with the enamel were totally washed away. Subsequently, molecular layers of the monomer (MDP) that chemisorbed onto the HAp crystallites of the etched enamel surfaces were co-polymerized with the adhesive (Durall Bond). This might help to explain why the chemical bonding of MDP around the HAp crystallites of the etched enamel substrate has a synergistic effect on the enamel bond strength obtained by purely micromechanical interlocking. The enamel treatment procedure chosen in the present study is markedly different from the selective enamel etching procedure reported in the literature [22,39,40]. Compared with the application of self-etch adhesives alone, the use of mild/moderate self-etch adhesives subsequently to enamel pre-etching by phosphoric acid could greatly improve the enamel bonding performance over time [29,32,41], since enamel pre-etching will provide much more retentive potential for micromechanical interlocking [22,39,40]. Also, the enamel bond strengths of mild/moderate self-etch adhesives could be increased by enamel pre-etching to similar values as obtained with three-step etch-and-rinse adhesives [33,34]. However, it should be kept in mind that the selective enamel etching is performed on the enamel surface followed by application of mild/moderate self-etch adhesives without any water rinsing. In this situation, the soluble or slightly soluble calcium salts resulting from the interaction of the self-etch adhesives with enamel/HAp will remain within the hybrid layer. This fact might explain why the enamel bond strengths of mild/moderate self-etch adhesives could be greatly improved by the pre-etching with phosphoric acid, but did not surpass those of etch-and-rinse adhesives [29,3235]. Resinenamel bonding of the etch-and-rinse adhesive in the present study was stable at least over 1-yr water storage (Figs. 1 and 3), which is consistent with results of previous in vitro and in vivo studies on etch-and-rinse adhesives [22,24,26,4248]. Usually, etch-and-rinse adhesives reveal better bonding performance than self-etch adhesives [9,29,3235].

Additional chemisorption effects of MDP and the hydrolytic stability of its calcium salts contribute to the long-term durability of enamel bonding (Figs. 13) [4,49]. In addition, the nano-layer formed by MDP-Calcium salts could protect the hybrid layer against hydrolyzation [17,50]. The SEM ndings in the present study also reveal an intimate contact at the enamelresin interfaces after 24-h or 1-yr water storage (Fig. 3). These micromorphological ndings are in accordance with the results of the present TBS tests. Chemical bonding to tooth hard tissues by specic functional monomers, and the hydrolytic stability of the corresponding monomer-calcium salts should be studied for improvement of the bonding performance in the future.

5.

Conclusion

The additional chemical bonding of MDP around HAp crystallites of the etched enamel substrate could signicantly increase the enamel bond strength and maintain it at least over 1-yr water storage.

Acknowledgements
This research was nancially supported by the Natural Scientic Research Fund of Zhejiang, China (Y2080045) and the National Natural Science Foundation of China (20973152). The authors were grateful to Mr. Kemin Jiang (Ningbo Institute of Materials Technology & Engineering, Chinese Academy of Sciences) for extensive technical assistance in X-ray photon spectroscopy and associate professor Xiuyang Li (Department of Epidemiology & Health Statistic, Zhejiang University) for statistical assistance. We also thank Kuraray Co., for providing the functional monomers.

references

[1] Fukuda R, Yoshida Y, Nakayama Y, Okazaki M, Inoue S, Sano H, et al. Bonding efcacy of polyalkenoic acids to hydroxyapatite, enamel and dentin. Biomaterials 2003;24:18617. [2] Yoshida Y, Van Meerbeek B, Nakayama Y, Snauwaert J, Hellemans L, Lambrechts P, et al. Evidence of chemical bonding at biomaterialhard tissue interfaces. Journal of Dental Research 2000;79:70914. [3] Yoshida Y, Van Meerbeek B, Nakayama Y, Yoshioka M, Snauwaert J, Abe Y, et al. Adhesion to and decalcication of hydroxyapatite by carboxylic acids. Journal of Dental Research 2001;80:15659. [4] Yoshida Y, Nagakane K, Fukuda R, Nakayama Y, Okazaki M, Shintani H, et al. Comparative study on adhesive performance of functional monomers. Journal of Dental Research 2004;83:4548. [5] Fu B, Yuan J, Qian W, Shen Q, Sun X, Hannig M. Evidence of chemisorption of maleic acid to enamel and hydroxyapatite. European Journal of Oral Sciences 2004;112:3627. [6] Fu B, Shen Q, Qian W, Sun X, Hannig M. Interfacial interaction of tartaric acid with hydroxyapatite and enamel. Journal of Materials Science Materials in Medicine 2005;16:82731.

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

e111

[7] Fu B, Sun X, Qian W, Shen Y, Chen R, Hannig M. Evidence of chemical bonding to hydroxyapatite by phosphoric acid esters. Biomaterials 2005;26:510410. [8] Fukegawa D, Hayakawa S, Yoshida Y, Suzuki K, Osaka A, Van Meerbeek B. Chemical interaction of phosphoric acid ester with hydroxyapatite. Journal of Dental Research 2006;85:9414. [9] Van Meerbeek B, Peumans M, Poitivin A, Mine A, Van Ende A, Neves A, et al. Relationship between bond-strength tests and clinical outcomes. Dental Materials 2010;26:e10021. [10] Ali SA, Williams DF. The characteristics and performance of dentine bonding agents. Clinical Materials 1993;14:24354. [11] Laswell HR, Welk DA. Rationale for designing cavity preparations. Dental Clinics of North America 1985;29:2419. [12] Hannig M, Bock H, Bott B, Hoth-Hannig W. Inter-crystallite nanoretention of self-etching adhesives at enamel imaged by transmission electron microscopy. European Journal of Oral Sciences 2002;110:46470. [13] Fu B, Bi L, Hannig M. Micro-mechanical interlocking and chemical bonding of self-etching adhesives at the resintooth hard tissue interfaces. International Journal of Clinical Dentistry 2008;1:13956. [14] Yoshihara K, Yoshida Y, Nagaoka N, Fukegawa D, Hayakawa S, Mine A, et al. Nano-controlled molecular interaction at adhesive interfaces for hard tissue reconstruction. Acta Biomaterials 2010;6:357382. [15] Yoshihara K, Yoshida Y, Hayakawa S, Nagaoka N, Torii Y, Osaka A, et al. Self-etch monomer-calcium salt deposition on dentin. Journal of Dental Research 2011;90:6026. [16] Yoshihara K, Yoshida Y, Hayakawa S, Nagaoka N, Irie M, Ogawa T, et al. Nanolayering of phosphoric acid ester monomer on enamel and dentin. Acta Biomaterialia 2011;7:318795. [17] Yoshida Y, Yoshihara K, Nagaoka N, Hayakawa S, Torii Y, Ogawa T, et al. Self-assembled nano-layering at the adhesive interface. Journal of Dental Research 2012;91:37681. [18] Iwai H, Nishiyama N. Effect of calcium salt of functional monomer on bonding performance. Journal of Dental Research 2012;91:10438. [19] Van Landuyt KL, Snauwaert J, De Munck J, Peumans M, Yoshida Y, Poitevin A, et al. Systematic review of the chemical composition of contemporary dental adhesives. Biomaterials 2007;28:375785. [20] Yu X, Liang B, Jin X, Fu B, Hannig M. Comparative in vivo study on the desensitizing efcacy of dentin desensitizers and one-bottle self-etching adhesives. Operative Dentistry 2010;35:27986. [21] Jiang Q, Pan H, Liang B, Fu B, Hannig M. Effect of saliva contamination and decontamination on bovine enamel bond strength of four self-etching adhesives. Operative Dentistry 2010;35:194202. [22] Peumans M, De Munck J, Van Landuyt K, Poitevin A, Lambrechts P, Van Meerbeek B. Eight-year clinical evaluation of a 2-step self-etch adhesive with and without selective enamel etching. Dental Materials 2010;26:117684. [23] Van Meerbeek B, Yoshihara K, Yoshida Y, Mine A, De Munck J, Van Landuyt KL. State of the art of self-etch adhesives. Dental Materials 2011;27:1728. [24] De Munck J, Van Landuyt K, Peumans M, Poitevin A, Lambrechts P, Braem M, et al. A critical review of the durability of adhesion to tooth tissue: methods and results. Journal of Dental Research 2005;84:11832. [25] Ikeda T, De Munck J, Shirai K, Hikita K, Inoue S, Sano H, et al. Effect of fracture strength of primeradhesive mixture on bonding effectiveness. Dental Materials 2005;21: 41320. [26] Peumans M, De Munck J, Van Landuyt K, Lambrechts P, Van Meerbeek B. Three-year clinical effectiveness of a self-etch

[27]

[28]

[29]

[30]

[31]

[32]

[33]

[34]

[35]

[36]

[37]

[38]

[39]

[40]

[41]

[42]

[43]

[44]

adhesive in cervical lesions. European Journal of Oral Sciences 2005;113:5128. Breschi L, Mazzoni A, Ruggeri A, Cadenaro M, Di Lenarda R, De Stefano Dorigo E. Dental adhesion review: aging and stability of the bonded interface. Dental Materials 2008;24:90101. Van Landuyt KL, Yoshida Y, Hirata I, Snauwaert J, De Munck J, Okazaki M, et al. Inuence of the chemical structure of functional monomers on their adhesive performance. Journal of Dental Research 2008;87:75761. Erickson RL, Barkmeier WW, Latta MA. The role of etching in bonding to enamel: a comparison of self-etching and etch-and-rinse adhesive systems. Dental Materials 2009;25:145967. Legler LR, Retief DH, Bradley EL. Effects of phosphoric acid concentration and etch duration on enamel depth of etch: an in vitro study. American Journal of Orthodontics and Dentofacial Orthopedics 1990;98:15460. Moszner N, Salz U, Zimmermann J. Chemical aspects of self-etching enameldentin adhesives: a systematic review. Dental Materials 2005;21:895910. Van Landuyt KL, Kanumilli P, De Munck J, Peumans M, Lambrechts P, Van Meerbeek B. Bond strength of a mild self-etch adhesive with and without prior acid-etching. Journal of Dentistry 2006;34:7785. Frankenberger R, Lohbauer U, Roggendorf MJ, Naumann M, Taschner M. Selective enamel etching reconsidered: better than etch-and-rinse and self-etch? Journal of Adhesive Dentistry 2008;10:33944. Erickson RL, Barkmeier WW, Kimmes NS. Bond strength of self-etch adhesives to pre-etched enamel. Dental Materials 2009;25:118794. Carvalho AP, Turbino ML. Can previous acid etching increase the bond strength of a self-etching primer adhesive to enamel? Brazilian Oral Research 2009;23: 16974. Hashimoto M, Ohno H, Sano H, Kaga M, Oguchi H. In vitro degradation of resindentin bonds analyzed by microtensile bond test, scanning and transmission electron microscopy. Biomaterials 2003;24:3795803. Yiu CK, King NM, Pashley DH, Suh BI, Carvalho RM, Carrilho MR, et al. Effect of resin hydrophilicity and water storage on resin strength. Biomaterials 2004;25:578996. Abdalla AI, Feilzer AJ. Four-year water degradation of a total-etch and two self-etching adhesives bonded to dentin. Journal of Dentistry 2008;36:6117. Gamborgi GP, Loguercio AD, Reis A. Inuence of enamel border and regional variability on durability of resindentin bonds. Journal of Dentistry 2007;35:3716. Reis AF, Giannini M, Pereira PN. Effects of a peripheral enamel bond on the long-term effectiveness of dentin bonding agents exposed to water in vitro. Journal of Biomedical Materials Research Part B: Applied Biomaterials 2008;85:107. Ibrahim IM, Elkassas DW, Yousry MM. Effect of EDTA and phosphoric acid pretreatment on the bonding effectiveness of self-etch adhesives to ground enamel. European Journal of Dentistry 2010;4:41828. Frankenberger R, Krmer N, Petschelt A. Long-term effect of dentin primers on enamel bond strength and marginal adaptation. Operative Dentistry 2000;25:119. Loguercio AD, Bittencourt DD, Baratieri LN, Reis A. A 36-month evaluation of self-etch and etch-and-rinse adhesives in noncarious cervical lesions. Journal of the American Dental Association 2007;138:50714. Pashley DH, Tay FR, Breschi L, Tjderhane L, Carvalho RM, Carrilho M, et al. State of the art etch-and-rinse adhesives. Dental Materials 2011;27:116.

e112

d e n t a l m a t e r i a l s 2 9 ( 2 0 1 3 ) e103e112

[45] Carvalho RM, Manso AP, Geraldeli S, Tay FR, Pashley DH. Durability of bonds and clinical success of adhesive restorations. Dental Materials 2012;28:7286. [46] Loguercio AD, Moura SK, Pellizzaro A, Dal-Bianco K, Patzlaff RT, Grande RH, et al. Durability of enamel bonding using two-step self-etch systems on ground and unground enamel. Operative Dentistry 2008;33:7988. [47] Ansari ZJ, Sadr A, Moezizadeh M, Aminian R, Ghasemi A, Shimada Y, et al. Effects of one-year storage in water on bond strength of self-etching adhesives to enamel and dentin. Dental Materials Journal 2008;27:26672.

[48] Peumans M, De Munck J, Van Landuyt KL, Kanumilli P, Yoshida Y, Inoue S, et al. Restoring cervical lesions with exible composites. Dental Materials 2007;23:74954. [49] Inoue S, Koshiro K, Yoshida Y, De Munck J, Nagakane K, Suzuki K, et al. Hydrolytic stability of self-etch adhesives bonded to dentin. Journal of Dental Research 2005;84: 11604. [50] Yoshida Y, Yoshihara K, Hayakawa S, Nagaoka N, Okihara T, Matsumoto T, et al. HEMA inhibits interfacial nano-layering of the functional monomer MDP. Journal of Dental Research 2012;91:10605.