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J Dent Res 85(4):349-353, 2006
© 2006 International and American Associations for Dental Research

RESEARCH REPORT
Biomaterials & Bioengineering

Development of a Self-etch Adhesive for Resin-modified Glass Ionomers

E. Coutinho1, K. Van Landuyt1, J. De Munck1, A. Poitevin1, Y. Yoshida2, S. Inoue3, M. Peumans1, K. Suzuki2, P. Lambrechts1, and B. Van Meerbeek1,*

1 Leuven BIOMAT Research Cluster, Department of Conservative Dentistry, School of Dentistry, Oral Pathology and Maxillo-Facial Surgery, Catholic University of Leuven, Kapucijnenvoer 7, B-3000 Leuven, Belgium;
2 Department of Biomaterials, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Science, Okayama, Japan; and
3 Division for General Dentistry, Hokkaido University Dental Hospital, Sapporo, Japan

* corresponding author, bart.vanmeerbeek{at}med.kuleuven.ac.be


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
The favorable self-adhesiveness of resin-modified glass ionomers (RMGIs) might be even further improved if the time-consuming and technically sensitive etch-and-rinse pre-treatment step with polyalkenoic acids could be avoided. We undertook this study to assess the effectiveness of an experimental self-etch adhesive for RMGIs that does not need to be rinsed off. Ultrastructural analysis and micro-tensile bond strength testing to enamel and dentin of a RMGI restorative material and a RMGI adhesive were performed after 4 different surface pre-treatments: no conditioning; 25% polyalkenoic acid; an experimental self-etch adhesive; and 37.5% phosphoric acid followed by the experimental self-etch adhesive. The use of an experimental self-etch adhesive increased the bond strength of RMGIs, especially after an additional conditioning step. Interfacial analysis showed the formation of a thin hydroxyapatite-containing hybrid layer. The self-etch technique enhances the user-friendliness of RMGIs and lowers their technique-sensitivity, while maintaining desirable characteristics of the conventional etch-and-rinse approach with polyalkenoic acids.

KEY WORDS: glass-ionomer cements • dental bonding • dental adhesives • self-etch


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Premature failure of dental restorations is still a major concern in routine clinical practice, and most restorations fail at their interface with tooth tissues, as a result of conditions such as secondary caries, marginal discolorations, and fractures (Mjör and Gordan, 2002). Nevertheless, recent reports have shown superior restoration retention rates of glass-ionomer-based materials when compared with other adhesive strategies (Peumans et al., 2005), despite their immediate lower bond strengths (Van Meerbeek et al., 2003). Chemical bonding to remaining hydroxyapatite crystals in enamel and dentin may be responsible for the enhanced durability of these bonds (Yoshida et al., 2000; Van Meerbeek et al., 2003).

In recent years, great attention has been given to the technique-sensitivity of resin-based adhesives, due to its influence on bond performance (Van Meerbeek et al., 2003). The self-etching approach has emerged as one of the main attempts to control the operator’s influence (Van Meerbeek et al., 2005), and, although generally associated with lower bond strengths, it eventually provided bond performances comparable with those achieved by the conventional 3-step technique in some cases (Shirai et al., 2005). Furthermore, it has also been suggested that bond performance of self-etch adhesives to enamel might be further improved by an additional prior conditioning step (Van Landuyt et al., 2005).

The main working hypotheses of this study were that (1) a trial self-etch adhesive enhances the adhesiveness of enamel and dentin to resin-modified glass-ionomers (RMGIs), and (2) a prior phosphoric-acid conditioning step even further increases the bond strength of tooth-RMGI interfaces.


   MATERIALS & METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Human third molars stored in 0.5% chloramine at 4°C were used within 1 mo after extraction following informed patient consent, as approved by the Commission for Medical Ethics of the Catholic University of Leuven. Mid-coronal dentin surfaces and mid-depth enamel surfaces were exposed by a low-speed diamond saw (Isomet 1000, Buehler, Lake Bluff, IL, USA) on which a bur-cut smear layer was produced (842KREF, Komet, Lemgo, Germany; MicroSpecimen Former, University of Iowa, Iowa City, IA, USA). An experimental self-etch adhesive was synthesized (exp-SE, GC, Tokyo, Japan) and 4 different surface pre-treatments performed according to the manufacturers’ instructions (TableGo): (1) no treatment; (2) Cavity Conditioner (GC, Tokyo, Japan); (3) exp-SE; and (4) Kerr Etchant (Kerr, Orange, CA, USA) followed by exp-SE. Afterward, the surfaces were restored with either a RMGI restorative material (Fuji II LC, GC, Tokyo, Japan) or a RMGI adhesive (FujiBOND LC, GC, Tokyo, Japan), followed by a resin composite (Gradia Direct, GC, Tokyo, Japan).


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  		Table. Materials, Composition, and Application Directions Followed in This Study
 
Micro-tensile Bond Strength (µTBS) Testing and Fractographic Analysis
After one-week storage in distilled water at 37°C, 4 central rectangular beams (2 x 2 x 8 mm) were sectioned and trimmed into an hourglass shape with a cross-sectional area of approximately 1 mm2 (5835KREF, Komet, Lemgo, Germany; MicroSpecimen Former, University of Iowa, Iowa City, IA, USA). Subsequent to fixation to a Ciucchi’s jig, specimens were tested in tension at a crosshead speed of 1 mm/min by means of a universal testing device (LRX, Lloyd, Hampshire, UK). We derived the µTBS by dividing the imposed force at time of fracture by the initial bond area, and assigned 0 MPa values to all pre-testing failures. Statistical differences were examined by the Kruskal-Wallis non-parametric statistical analysis ({alpha} = 0.05). After µTBS testing, all samples were processed for scanning electron microscopy (SEM; Philips XL30, Eindhoven, The Netherlands), by common specimen processing procedures (Perdigão et al., 1995), so that the mode of failure could be determined.

Transmission Electron Microscopy (TEM) Interfacial Characterization
Additional dentin surfaces were prepared and pre-treated as described above. Subsequently, they were covered by a 1-mm layer of RMGI (either Fuji II LC or FujiBOND LC, GC, Tokyo, Japan) and sealed with an unfilled resin (Clearfil Protect Liner F, Kuraray, Osaka, Japan). After surfaces were stored for 1 wk in distilled water at 37°C, we obtained TEM samples by sectioning central rectangular beams and further processing for TEM analysis (Van Meerbeek et al., 1998). Non-demineralized, unstained, 70- to 90-nm thin sections were cut by means of a diamond knife (Diatome, Bienne, Switzerland) in an ultra-microtome (Ultracut UCT, Leica, Vienna, Austria) and examined under TEM (Philips CM10, Eindhoven, The Netherlands).


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Main findings in the µTBS test are graphically displayed in Fig. 1Go. In general, higher bond strength values were associated with the RMGI restorative material when compared with the RMGI adhesive. Lower bond strengths were also linked to the non-conditioned groups, in both enamel and dentin. Non-statistically-different bond strength values were found when either the polyalkenoic acid or the exp-SE was used, but significantly higher values have been found for the phosphoric-acid-etched samples.


Figure 1
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  		Figure 1. µTBS results for the RMGI adhesive and restorative applied to enamel and dentin by the experimental surface treatments. Numbers above columns indicate mean values and respective standard deviation (between brackets); n stands for total number of specimens and ptf for number of pre-testing failures (to which 0 MPa values were attributed during the statistical analysis). Groups identified by different letters below their respective columns are statistically different (p < 0.05).

 
Fractographic analysis showed a higher number of pre-testing failures associated with non-conditioned groups, mainly by adhesive debonding. Specimens with higher bond strengths exhibited a greater proportion of cohesive failures in enamel/dentin or RMGI. All fractures involving RMGIs disclosed a typical porous surface, whether manipulation was manual or automatic. However, manual mixing of RMGIs did increase the porous aspect of the fractured surface when compared with automatic mixing. In some cases, it was even possible to correlate some of these pores to crack initiation sites and determine the crack propagation path.

Different specific structural features have been found for both the RMGI adhesive and restorative material, when bonded according to the 4 investigated surface treatments (Figs. 2Go, 3Go). A clearly distinct hybrid layer was present in all situations, but with different structural characteristics. For the non-conditioned groups and those treated with polyalkenoic acid and exp-SE, it showed a common aspect, with a thickness of 0.5 µm, and contained heterogeneously dispersed hydroxyapatite crystals. Nevertheless, in specimens treated with phosphoric acid followed by exp-SE, a 10x thicker hybrid layer was found (5 µm), but it was completely depleted of hydroxyapatite crystals. Remnants of the smear layer were also particularly identified in the non-conditioned groups, but not in the others. The gel phase was observed only in specimens pre-treated with polyalkenoic acid, immediately on the top of the hybrid layer. Non-particulate absorption layers were not obviously identified in this work and were clearly absent in the groups treated with exp-SE.


Figure 2
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  		Figure 2. Representative TEM photomicrographs and SEM fractographs of the RMGI adhesive applied to dentin by the experimental surface treatments. Important microstructural features have been identified: unaffected dentin (Ud); dentin tubule (T); hybrid layer (Hy); gel-phase (G-P); matrix in RMGI (M); glass particle in RMGI (Gp). (A1,A2) Non-demineralized and unstained specimen of the untreated group revealing a thin, 0.5-µm, hybrid layer and absence of the gel-phase. (A3) Fractured dentin side after µTBS testing, showing a typical adhesive failure pattern characterized by the furrows from a bur-cut smear layer. (B1,B2) Non-demineralized and unstained specimen of the polyalkenoic acid group, revealing an underlying thin, 0.5-µm, hybrid layer covered by the gel-phase, and heterogeneously distributed hydroxyapatite crystals within the hybrid layer. (B3) Fractured dentin side after µTBS testing, showing a typical mixed failure pattern characterized by adhesive debonding on the right side and cohesive failure of the RMGI on the left side. Notice the typical porous appearance of fractured RMGIs. (C1,C2) Non-demineralized and unstained specimen of the exp-SE group, revealing a thin, 0.5-µm, hybrid layer, absence of the gel-phase, and heterogeneously distributed hydroxyapatite crystals within the hybrid layer. (C3) Fractured dentin side after µTBS testing, showing a typical mixed failure pattern. (D1,D2) Non-demineralized and unstained specimen of the phosphoric acid group, unveiling a 5-µm-thick hybrid layer and absence of the gel-phase or hydroxyapatite crystals within the hybrid layer. (D3) Fractured dentin side after µTBS testing, showing a typical cohesive failure in RMGI.

 

Figure 3
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  		Figure 3. Representative TEM photomicrographs and SEM fractographs of the RMGI restorative applied to dentin. (A1,A2) Non-demineralized and unstained specimens of the exp-SE group revealing a thin, 0.5-µm, hybrid layer, absence of the gel-phase, and heterogeneously distributed hydroxyapatite crystals within the hybrid layer. (A3) Fractured dentin side after µTBS testing, showing cohesive failure in RMGI. (B1,B2) Non-demineralized and unstained specimens of the phosphoric acid group, unveiling a 5-µm-thick hybrid layer and absence of the gel-phase or hydroxyapatite crystals within the hybrid layer. (B3) Fractured dentin side after µTBS testing, showing cohesive failure in RMGI. Unaffected dentin (Ud); dentin tubule (T); hybrid layer (Hy); matrix in RMGI (M); glass particle in RMGI (Gp).

 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
Although favorable clinical retention rates of glass-ionomer-based materials have already been comprehensively verified (Peumans et al., 2005), very little is known about their bonding mechanisms (Mjör and Davidson, 1999). This gap in knowledge is particularly reflected in the conflicting terminology used by different research groups to characterize the interface with dental hard tissues, hence the urgent need for consistency (Tyas, 2003). The ionomer-tooth interface itself was first described as an ion-exchange layer (Wilson et al., 1983), and was later referred to as: a zone of interaction (Ngo et al., 1997); an interdiffusion zone or hybrid-like layer (Ferrari and Davidson, 1997); an indistinct zone (Pereira et al., 1997); a "structureless" or absorption layer (Sidhu and Watson, 1998); and an interphase (Sennou et al., 1999). Only recently were further developments made possible through high-resolution microscopy, which allowed for the identification of different zones or layers within the interface, leading to the realization that many of the features described before were equivalent (Yoshida et al., 1999; Tay et al., 2001; Van Meerbeek et al., 2003).

The actual bonding mechanism of RMGIs to tooth tissues has been recently determined to be two-fold, by micromechanical interlocking (Tay et al., 2001; Yip et al., 2001) and by chemical interaction (Yoshida et al., 2000; Fukuda et al., 2003). Micromechanical interlocking is achieved by impregnation of a partially demineralized layer on the top of the dentin substrate with a high-molecular-weight polycarboxyl-based polymer (Van Meerbeek et al., 2003), akin to the hybrid layer in resin-based interfaces. This layer was clearly identified in all groups investigated, but was particularly thick on phosphoric-acid-etched samples impregnated by exp-SE (Fig. 2Go). The combination of a thicker hybrid layer with a low-molecular-weight self-etching polymer may explain the higher initial bond strengths obtained for these groups (Fig. 1Go).

Immediately over the hybrid layer, a gray intermediate layer or gel-phase was also identified in the groups treated with polyalkenoic acid (Yoshida et al., 1999; Tay et al., 2001), which also encompassed globular structures referred as "multilocular phases" (Tay et al., 2004) (Fig. 2Go). The gel-phase is formed by the interaction of carboxyl groups from the polyalkenoic acid and calcium from the partially demineralized dentin (Yoshida et al., 1999; Tay et al., 2001). Such chemical interaction between polyalkenoic acids and calcium was demonstrated not only on hydroxyapatite blocks (Yoshida et al., 2000), but also on enamel and dentin (Fukuda et al., 2003). According to the adhesion/decalcification concept (AD-concept), the nature of this resulting calcium salt is especially important, since its dissolution rate controls the adherence to or decalcification of hydroxyapatite (Yoshida et al., 2001; Yoshioka et al., 2002). Therefore, the physical presence of the gel-phase attached to the tooth surface suggests that the resulting calcium polycarboxylate salt is stable in this particular system (Fig. 2Go). In addition, the presence of a hydroxyapatite-coated collagen fibril network on the exp-SE groups may also offer the possibility of chemical interactions (Figs. 2Go, 3Go), since low-molecular-weight polymers contained in self-etch adhesives have also demonstrated the ability to bond to hydroxyapatite (Yoshida et al., 2004). Additional chemical interactions may well explain the good clinical retention rates of glass-ionomer-based materials (Peumans et al., 2005), since the ability to adhere chemically to a substrate has been associated with bond durability (Venables, 1984). Regrettably, the same rationale also applies to the phosphoric-acid-etched groups, on which the lack of a reactive substrate, such as hydroxypapatite crystals, may jeopardize their bond effectiveness over time (De Munck et al., 2004).

Although non-particulate thick absorption layers have been previously identified immediately below the RMGI substrate on TEM photomicrographs (Tay et al., 2004; Yiu et al., 2004) and by other techniques (Pereira et al., 1997; Sidhu and Watson, 1998), they were not clearly identified in this work. One possible explanation is their own mechanism of formation, which restricts the existence of an absorption layer to the surroundings of dentin tubules (Mjör and Davidson, 1999). Accordingly, earlier works with high-resolution microscopy techniques have not been able to disclose, unambiguously, absorption layers on TEM photomicrographs (Tay et al., 2001). Nonetheless, this remains an interesting topic for future investigations, since the presence of HEMA in the composition of exp-SE might influence the water uptake from the underlying vital dentin (Tay et al., 2002). The presence of a permeable membrane on ionomer-tooth interfaces may bring some positive effects, such as compensation for polymerization shrinkage (Sidhu et al., 2002) and a reparative mechanism for crack propagation. Therefore, it was not possible to establish a precise relationship between the type of conditioning and the absorption layer characteristics.

Higher bond strengths were found for the restorative RMGI, which is probably due to its higher cohesive strength (Tyas, 2003), since compositions of both materials are qualitatively similar (TableGo). Another factor contributing to higher bond strengths may be attributed to fewer pores in the restorative material, due to automatic mixing (Miguel et al., 2001).

In conclusion, the self-etch technique is a very promising approach to ionomer-tooth interfaces, since it enhances the user-friendliness of RMGIs and lowers their technique-sensitivity. Such outcome is achieved while maintaining desirable characteristics of the conventional etch-and-rinse approach with polyalkenoic acids, namely, no significant decrease in immediate bond strength, the possibility of chemical interaction with the substrate, and consequent enhanced bond durability.


   ACKNOWLEDGMENTS
 
We thank Dominique Crombez and Bernadette Vanderschueren (Catholic University of Leuven) for extensive technical assistance. All commercial and experimental materials were generously donated by the manufacturers. This study was supported by the Toshio Nakao Chair for Adhesive Dentistry and the CAPES Foundation.

Received August 24, 2005; Last revision November 7, 2005; Accepted December 2, 2005


   REFERENCES
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 ABSTRACT
 INTRODUCTION
 MATERIALS & METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
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