Effect of adhesive strategy on resin cement bonding to dentin.

OBJECTIVE: The cement bonding strategy and the polymerization mode can influence the prognosis of indirect restorations. The microtensile bond strength (µTBS) and dentin endogenous enzymatic activity of a dual-cure resin cement (PV5) used in combination with two dentin surface conditioners (accelerator-enhancer primer, TP or universal adhesive, UA) were evaluated. MATERIALS AND METHODS: PV5 was used to lute composite overlays after dentin treatment with TP or UA. The resin cement was self-cured, SC (1 h at 37 °C) or dual-cured, DC (20 s light-cure followed by 15 min self-cure at 37°C). The µTBS test, fractographic analysis, and the in situ zymography evaluations were performed after 24 h (T0 ) or 1 yr (T12 ) of artificial storage. Data were statistically analyzed (α = 0.05). RESULTS: TP/DC obtained the highest adhesive strengths (45 ± 9 and 36.6 ± 8), while UA/SC (17 ± 8 and 11 ± 4) the lowest, both at T0 and T12 , respectively. DC resulted in superior bonding values than the SC, independent of the dentin surface treatment (p < 0.05). The type of adhesive, curing mode and aging influenced the gelatinolytic activity (p < 0.05). CONCLUSIONS: The dual-cure resin cement used in combination with its accelerator-enhancer primer showed superior bonding performances with respect to universal adhesive. Dual-curing the resin cement was determinant to enhance bonding capability over time, independent of the adhesive strategy. CLINICAL RELEVANCE: Clinicians must be aware to faithfully follow manufacturer's recommendation regarding the adhesive strategy suggested with the resin cement used.

Selective adhesive luting: A novel technique for improving adhesion achieved by universal resin cements.

OBJECTIVE: We aimed to introduce the concept of "Selective adhesive luting-SAL" which is explained through clinical steps and supported by preliminary laboratory evidence. CLINICAL CONSIDERATIONS: Cementation with rubber dam is difficult to perform in case of short abutment teeth and/or subgingival crown margins. By means of universal resin cements/universal adhesive systems, which can be employed in self-adhesive as well as adhesive luting procedures, this paper presents a novel technique allowing clinicians to perform reliable cementation where rubber dam isolation is difficult. The SAL technique entails the application of a universal adhesive system only on easily accessible abutment surfaces, enabling simultaneous adhesive and self-adhesive luting in different portions of the abutment. The SAL clinical workflow is explained through prosthodontic rehabilitation of maxillary right central incisor affected by microdontia and restored with a lithium-disilicate crown. Furthermore, our laboratory microshear bond strength study supports the rationale behind SAL application demonstrating higher bond strength even when the adhesive resin is placed only on one portion of the cementation substrate. CLINICAL SIGNIFICANCE: This article advocates the application of SAL technique in clinical situations where effective adhesive luting is uncertain, since it can improve the adhesion between the tooth and universal resin cements.

Resin composite cements: Current status and a novel classification proposal.

OBJECTIVES: Currently, a classification of resin cements that includes relatively recently formulated ("universal") cements is lacking. Furthermore, the terminology used to define different resin cements in the scientific reports is inconsistent. Accordingly, this work aims to: (i) propose a novel classification of resin composite cements; (ii) disambiguate the term "universal cements" and (iii) present an overview of the properties of these cements. METHODS: An analysis of peer-reviewed literature (PubMed search), as well as market research on definitive resin composite cements were performed. RESULTS: A tendency toward simplified and versatile luting materials was observed both in the scientific literature and on the dental market with the advent of self-adhesive/one-step resin cements. However, additional priming procedures were necessary to improve their bonding performance in certain clinical situations. Hence, several cements that can be applied both in adhesive and self-adhesive mode were introduced. These cements are associated with a universal adhesive resin, that can be used as a tooth and/or restorative material primer, without the need for other priming systems, regardless of the substrate. These systems should be considered truly universal. Therefore, we hereby suggested a new classification of resin-based cements: (1) adhesive/multi-step; (2) self-adhesive/one-step; (3) universal cements (one- or multi-step). Despite promising in vitro results, clinical trials and long-track laboratory studies are necessary to confirm the reliability of the universal cements. CONCLUSIONS: This review presented the current advances in the field of resin-based cements, which are reflected in the proposed classification. The term "universal cement" was disambiguated, which will help standardize the terminology used in published research. CLINICAL SIGNIFICANCE: The classification of resin-based cements and a better understanding of the proper terminology will help standardize the terminology in published research, as well as improve the understanding of the clinical practitioners of the different indications and possible modalities of use of the available cements.

Influence of the activation mode on long-term bond strength and endogenous enzymatic activity of dual-cure resin cements.

OBJECTIVE: To investigate the long-term microtensile bond strength (µTBS), interfacial nanoleakage expression (NL), and adhesive stability of dual-cure resin cements with/out light activation to dentin. MATERIALS AND METHODS: Composite overlays (N = 20) were luted to deep dentin surfaces with RelyX Ultimate (RXU, 3M) or Variolink EstheticDC (VAR, Ivoclar-Vivadent). A universal adhesive was used for bonding procedures (iBond universal, Heraeus Kulzer). The resin cements were either self-cured (SC; 1 h at 37 °C) or dual-cured (DC; 20s light-cure followed by 15 min self-cure at 37 °C). Specimens were submitted to µTBS immediately (T0) or after 1 year of laboratory storage (T12). The fracture pattern was evaluated using scanning electron microscopy (SEM). Data were statistically analyzed with two-way ANOVA/Tukey test. Further, the NL was quantified and analyzed (chi-square test) and in situ zymography was performed to evaluate the endogenous enzymatic activity within the hybrid layer (HL) at T0 and T12 (Mann-Whitney test). The significance level for all statistical tests was set at p = 0.05. RESULTS: DC resulted in higher bond strength and decreased fluorescence at the adhesive interface, irrespective of the material and the storage period (p < 0.05). Significantly lower bonding performances (p < 0.05) and higher endogenous enzymatic activity (p < 0.05) were observed within the HL at T12 compared to T0 in all tested groups. CONCLUSIONS: Light-curing the dual-cure resin cements, more than the cement materials, accounted for good bonding performances and higher HL stability over time when used with a universal adhesive. Clinical significance The curing condition influences the bonding performances of dual-cure resin cements to dentin when used with a universal adhesive.

The Influence of Different Bleaching Protocols on Dentinal Enzymatic Activity: An In Vitro Study.

This study aimed to investigate matrix metalloproteinase (MMP) activity in human dentin using in-situ and gelatin zymography, after at-home and in-office bleaching, related to their clinical exposure times. Dentin specimens (n = 5) were treated with 35% hydrogen peroxide (50 min per session/4 sessions), 10% carbamide peroxide (180 min/21 sessions), or no treatment. All were subjected to in-situ zymography. Dentin slices were, subsequently, obtained, covered with fluorescein-conjugated gelatin, and examined with confocal laser-scanning microscopy. The fluorescence intensity was quantified and statistically analyzed using one-way ANOVA and Bonferroni tests (α = 0.05). Furthermore, gelatin zymography was performed on protein extracts obtained from dentin powder (N = 8 teeth), treated with hydrogen peroxide or carbamide peroxide, with different exposure times (10/50 min for hydrogen peroxide; 252/1260 min for carbamide peroxide). The results of the in-situ zymography showed no statistical differences between the bleached specimens and the control group, with a medium level of gelatinolytic activity expressed in the dentin tubules. The results of gelatin zymography showed an increased expression of pro-MMP-9 in carbamide peroxide groups. The expression of pro-MMP-2 decreased in all the experimental groups. The bleaching treatments performed on the enamel of sound teeth do not influence dentinal enzymatic activity. However, when unprotected dentin tissue is bleached, matrix metalloproteinases are more expressed, particularly when carbamide peroxide is used, proportional to the exposure time.

Biochemical and immunohistochemical analysis of tissue inhibitor of metalloproteinases-1 in human sound dentin.

OBJECTIVES: Matrix metalloproteases (MMPs) are a family of enzymes that operate a proteolytic activity at the level of the extracellular matrix. MMPs are regulated by tissue inhibitors of metalloproteinases (TIMPs) that can ubiquitously bind different enzyme forms. The study aims to identify a morfo-functional association between TIMP-1 and MMP-2 and -9 in human dentin. MATERIALS AND METHODS: Proteins were extracted from demineralized human sound dentin powder and centrifuged to separate two aliquots with different molecular weights of proteins, higher and lower than 30 kDa. In each aliquot, the evaluation of the presence of TIMP-1/MMP-2 and TIMP-1/MMP-9 was performed using co-immunoprecipitation/immunoblotting analysis. The distribution of TIMP-1, in association with MMP-2 and -9, was investigated using a double immunohistochemical technique. Furthermore, the activity of TIMP-1 was measured by reverse zymography, where acrylamide gel was copolymerized with gelatin and recombinant MMP-2. RESULTS: Co-immunoprecipitation/immunoblotting analysis showed the association TIMP-1/MMP-2 and TIMP-1/MMP-9 in human sound dentin. Electron microscopy evaluation revealed a diffuse presence of TIMP-1 tightly associated with MMP-2 and -9. Reverse zymography analysis confirmed that TIMP-1 present in human dentin is active and can bind different MMPs isoforms. CONCLUSIONS: The strict association of TIMP-1 with MMP-2 and -9 in situ appeared a constant finding in the human sound dentin. CLINICAL RELEVANCE: Considering the role of TIMP-1, MMP-2, and MMP-9 within the connective tissues, clinically applicable protocols could be developed in the future to increase or decrease the level of TIMPs in human dentin to regulate the activity of MMPs, contributing to reduce caries progression and collagen degradation.

Endogenous Enzymatic Activity in Dentin Treated with a Chitosan Primer.

The aim of this study was to evaluate the effect of different concentrations of chitosan polymer on dentinal enzymatic activity by means of gelatin and in situ zymography. Human dentin was frozen and ground in a miller. Dentin powder aliquots were demineralized with phosphoric acid and treated with three different concentrations of lyophilized chitosan polymer (1, 0.5 and 0.1 wt%) dissolved in distilled water. Dentin proteins were extracted from each experimental group and electrophoresed under non-reducing conditions in 10% SDS-PAGE containing fluorescein-labeled gelatin. After 48 h in the incubation buffer at 37 °C, proteolytic activity was registered under long-wave UV light scanner and quantified by using Image J software. Furthermore, additional teeth (n = 4) were prepared for the in situ zymographic analysis in unrestored as well as restored dentin pretreated with the same chitosan primers. The registered enzymatic activity was directly proportional to the chitosan concentration and higher in the restored dentin groups (p < 0.05), except for the 0.1% chitosan primer. Chitosan 0.1% only showed faint expression of enzymatic activity compared to 1% and 0.5% concentrations. Chitosan 0.1% dissolved in water can produce significant reduction in MMPs activity and could possibly contribute to bond strength preservation over time.

Color match of single-shade restorations after professional dental bleaching: An in vitro study.

Background: Restorative dentistry aims to reproduce natural tooth shades through resin composites that must be layered to obtain colors, opacities, and translucencies, and therefore, clinical success is operator skill dependent. Aims: The purpose of this study is to evaluate the color shift of single-shade composite restorations before and after dental bleaching. Materials and Methods: Eighty human extracted posterior teeth were restored with four single-shade composites (Omnichroma OM; Clearfil Majesty ES-2 Universal CL; Essentia Universal ES; Venus Diamond One VE) (n = 20 each). Standardized V class cavities were prepared on buccal side. VITA Easyshade V spectrophotometer was used to register VITA color and color coordinates 24 h before (T0), 24 h after (T1), and 1 week after (T2) dental bleaching (Opalescence Boost PF 40%). Color differences (ΔEab) and (ΔWID) were calculated and subjected to statistical analysis. Statistical Analysis: Categorical variables were analyzed using Pearson Chi-square, and data from color coordinates were analyzed using one-way analysis of variance and Tukey's multiple comparison test with Bonferroni correction. Paired t-tests were performed to compare continuous measures between groups and treatment time. Results: Instrumental evaluations revealed statistically significant differences between materials (P < 0.05) with lower values for ES and VE samples followed by CL and OM at T0 and T1. At T2, OM and CL ΔEab values decrease getting closer to ES and VE. Conclusion: Single-shade composites seem to match with the surrounding bleached tooth.

The influence of finishing line and luting material selection on the seating accuracy of CAD/CAM indirect composite restorations.

OBJECTIVES: This study aimed to assess the seating accuracy of resin composite CAD/CAM overlay restorations, employing various preparation designs and luting materials (pre-heated composite (HC) or resin cement (RC)). METHODS: A human molar's STL file was utilized to create 100 3D-printed resin tooth replicas, randomly distributed into 5 groups (n=20) based on finishing line preparation designs: 1) Rounded shoulder above the dental equator - DE (SA); 2) Chamfer above the DE (CA); 3) Butt joint above the DE (BJ); 4) Rounded shoulder below the DE (SB); 5) Chamfer below the DE (CB). Digital impressions were acquired for all replicas, and restorations milled using Tetric CAD (Ivoclar Vivadent). The restorations were luted with HC (Tetric Prime, Ivoclar Vivadent) or RC (RelyX Universal, 3M). Seating accuracy was evaluated through digital scans during try-in without any luting agent and post-cementation using a 3D analysis software (Geomagic wrap, 3D Systems). Data were statistically analyzed using Two-Way ANOVA (p<0.05). RESULTS: The type of luting material (RC vs HC), preparation design, and their interactions significantly impacted 3D seating of the restorations (p<0.001). HC exhibited higer volumetric increase than RC. BJ and CA designs consistently demonstrated superior seating accuracy, irrespective of the luting material used. CONCLUSIONS: The utilization of pre-heated composite resin could negatively influence the seating of overlay restorations, probably due to its higher viscosity when compared to the resin cement. However, when HC is selected as luting agent, preparation designs lacking internal angles are recommended for enhancing the precision of overlays seating.

Comparisons of ammonia- and water-based silver-containing solutions on dentin bonding and enzymatic activity: 1-yr evaluation.

OBJECTIVE: To evaluate the effects of an ammonia-based and a water-based silver-containing solutions on bonding performance and matrix-metalloproteinases (MMPs) activity of a universal adhesive to dentin after 1 year of artificial aging. METHODS: Mid-coronal dentin surfaces of 60 sound human molars were exposed and the following groups were formed according to the surface pre-treatment and etching mode of the universal adhesive (Zipbond Universal, SDI) (n = 10): G1) Zipbond in the self-etch mode (ZSE); G2) Riva Star (SDI) applied before ZSE; G3) Riva Star Aqua (SDI) applied before ZSE; G4) Zipbond in the etch-and-rinse mode (ZER); G5) Riva Star applied before ZER; G6) Riva Star Aqua applied before ZER. The specimens were sectioned and subjected to microtensile bond strength (µTBS) test at baseline (T0) and after 12 months (T12) of artificial storage. Scanning electron microscope (SEM) and energy dispersive spectroscopy analysis (EDS) were also conducted. Three additional molars per group were processed for the in situ zymography analysis at T0 and T12. Data were statistically analyzed (p < 0.05). RESULTS: Dentin pre-treatments and aging decreased bonding values, regardless of the etching mode (p < 0.05). No differences in µTBS were observed between the two silver-containing solutions, both at T0 and T12. Riva Star Aqua and etching significantly increased the MMPs activity, independent of the storage period (p < 0.05). SIGNIFICANCE: Dentin surface pre-treatment with silver-containing solutions negatively affects the bonding performances of resin composite restorations placed with a universal adhesive. However, the ammonia-based product Riva Star might show better stability in the long term, due to lower activation of MMPs.

Polishing matters / Unfolding peri-implantitis.

The goal of this section ist o bring our readers eye-catching and high quality images related to novel techniques, clinical challenges and emerging trends relevant to esthetic dentistry. The section portrays the clinical image, the photographer, the camera and a short caption of the story behind the photograph.

Glutaraldehyde-based desensitizers' influence on bonding performances and dentin enzymatic activity of universal adhesives.

OBJECTIVES: To evaluate the influence of two glutaraldehyde-based desensitizers (L: GLUMA Desensitizer, Heraeus Kulzer and G: GLUMA Desensitizer PowerGel) prior to the adhesive procedures on microtensile bond strength (µTBS) to dentin and endogenous enzymatic activity. METHODS: Noncarious human third molars (N = 48) were cut to expose middle coronal dentin. Six experimental groups were formed according to the dentin pre-treatment (L or G) and the universal adhesives (IBU - iBond universal, Kulzer or AU - Adhese Universal, Ivoclar Vivadent) used in the self-etch mode (n = 8): 1) L/IBU; 2) G/IBU; 3) IBU; 4) L/AU; 5) G/AU; 6) AU. Specimens were cut into sticks and stressed until failure after 24 h (T0) or 1 yr of aging (T12). Additional 4 teeth were used for in situ zymography evaluation and data were statistically analyzed (α = 0.05). RESULTS: Dentin pre-treatment, adhesive and aging statistically influenced bond strength and enzymatic activity (P<0.001). AU demonstrated higher bond strength values than IBU (P<0.001). The L resulted in higher bond strength compared to the G and control groups (P<0.001). aging statistically influenced bonding performance, especially when no dentin pre-treatment was performed (P<0.001). In situ zymography revealed that at baseline the control groups exhibited lower interfacial fluorescence compared to the experimental groups, irrespective of the adhesive used (P<0,001). However, after 1 yr of artificial storage, no differences were found among the groups (P>0.05). CONCLUSIONS: Glutharldeadeyde-based products increased bond strength and determined a stabilization of the adhesive interface over time apparently not related to the MMPs inhibition. CLINICAL SIGNIFICANCE: The results of this in vitro study suggest that the application of glutaraldehyde-based desensitizers prior to the adhesive procedures when associated with universal adhesives could result in increased bond strength and stabilization of the adhesive interface over time.

Clinical longevity of direct and indirect posterior resin composite restorations: An updated systematic review and meta-analysis.

OBJECTIVES: To answer the PICO(S) question: Is there a difference in clinical longevity between direct and indirect resin composite restorations placed on permanent posterior teeth? DATA: Randomized controlled clinical trials (RCTs) investigating direct and indirect resin composite restorations in posterior permanent teeth were considered. SOURCES: Several electronic databases were searched, with no language or date restrictions. The revised Cochrane Collaboration's tool for assessing risk of bias (RoB-2) was used to analyze the studies; meta-analyses were run and the certainty of evidence was assessed by the GRADE tool. A subgroup meta-analysis was performed for resin composite restorations placed on posterior worn dentition. STUDY SELECTION: Twenty-three articles were included in qualitative synthesis, while 8 studies were used for meta-analyses. According to the RoB-2 tool, 5 studies were ranked as "low risk", 7 had "some concerns", while 11 papers were rated as "high risk" of bias. There were no statistically significant differences in short-term (p = 0.27; RR=1.54, 95% CI [0.72, 3.33]), medium-term (p = 0.27; RR=1.87, 95% CI [0.61, 5.72]) and long-term longevity (p = 0.86; RR=0.95, 95% CI [0.57, 1.59]). The choice of restorative technique had no influence on short-term survival of resin composite restorations placed on worn dentition (p = 0.13; RR=0.46, 95% CI [0.17, 1.25]). The certainty of evidence was rated as "very low". CONCLUSIONS: Direct and indirect resin composite restorations may show similar clinical longevity in posterior region, regardless of the observation period or substrate (wear-affected and non-affected dentition). The very low quality of evidence suggests that more long-term RCTs are needed to confirm our results.

An Insight into Enamel Resin Infiltrants with Experimental Compositions.

Resin infiltration is a conservative treatment of initial enamel carious lesions. Only one infiltrant material is available on the market (Icon, DMG), and research is now investigating new chemical compositions so as to further exploit the benefits of the resin infiltration technique. A literature search of the articles testing the effects of different formulations on mechanical properties, resin penetration ability, remineralizing, and antibacterial activities was conducted. Of 238 articles, 29 resulted in being eligible for the literature review. The formulations investigated were all different and consisted in the inclusion of hydrophobic monomers (i.e., BisEMA, UDMA), solvents (ethanol, HEMA), alternative etchants (PAM) or molecules with antibacterial or bioactivity features (i.e., AgNP, YbF3, MTZ, chitosan, DMAMM, HAp, MC-IL, NACP, PUA, CHX) and microfilled resins. Information on the long-term performances of the tested experimental materials were scarce. The combination of TEGDMA with hydrophobic monomers and the inclusion of a solvent alternative to ethanol reinforced mechanical properties of the materials. Hybrid-glass materials demonstrated an enhanced remineralization capacity. Techniques such as tunnelization increased the penetration depth and preserved the recourse to less-conservative treatments. Combining the min-invasive infiltrant approach with remineralizing and bacteriostatic properties would be beneficial for therapeutic and economical aspects, according to the principles of minimally invasive dentistry.

The influence of selective enamel etch and self-etch mode of universal adhesives' application on clinical behavior of composite restorations placed on non-carious cervical lesions: A systematic review and meta-analysis.

BACKGROUND: To answer the PICOS question: "Is the risk of retention loss equal for SEE and SE approach when universal adhesives and composite restorations are indicated for restoring NCCLs?" METHODS: Web of Science, PubMed, Cochrane Central Register of Controlled Trials, Scopus, Scientific Electronic Library Online, LILACS, Google Scholar™ and OpenGrey were searched. Only randomized controlled clinical trials in which NCCLs were restored with composites and universal adhesives applied in SEE or SE mode were included. The articles were assessed for the risk of bias, after which meta-analyses were run (fixed-effects model was applied; heterogeneity was explored using Cochran Q test and I2 statistics; α = 0.05) and the certainty of evidence was assessed by the GRADE tool. RESULTS: Fifteen articles were included in qualitative, while 7 articles were included in quantitative analysis. Seven studies were judged as "low" risk of bias, while 8 were considered as "unclear" risk of bias. Statistically significant difference for retention were observed at 6- and 18/24 months (p = 0.05; OR=0.42, 95% CI [0.18, 0.99]; very low certainty of evidence and p = 0.007; OR=0.31, 95% CI [0.13, 0.72]; low certainty of evidence, respectively), favoring SEE approach. No other significant differences in clinical outcomes were observed between SEE and SE approach (very low certainty of evidence). SIGNIFICANCE: When restoring NCCLs, clinicians might consider applying universal adhesives in SEE mode since it could lead to more predictable retention compared to SE approach up to 2 years of follow-up.

The influence of finishing lines and margin location on enamel and dentin removal for indirect partial restorations: A micro-CT quantitative evaluation.

OBJECTIVES: This in vitro research aimed to quantitatively evaluate the enamel and dentin tissue removal and the residual adhesion surface area (RAS) after different margin designs and locations for indirect partial restorations (IPR). METHODS: A human molar was scanned using a Micro-CT and the STL file obtained was used to 3D-print 50 resin-tooth replicas. IPR standardized preparations were performed. The specimens were randomly assigned to 5 groups (n = 10), according to preparation and margin location to the dental equator (DE): 1) Rounded shoulder above the DE (SA); 2) Hollow chamfer above the DE (CA); 3) Butt joint above the DE (BJ); 4) Rounded shoulder below the DE (SB); 5) Chamfer below the DE (CB). Then, the tooth replicas were scanned and each STL file was aligned and superimposed to the original STL model file. Data of enamel and dentin volume removal and RAS were assessed and statistically analyzed (one-way ANOVA and Kruskal-Wallis tests for the two dental substrates respectively). Significance was set at p<0.05. RESULTS: Differences in dental tissue reductions were related to the margin location. Above the equator, SA, CA, and BJ performed comparably (p>0.05). Below the equator, CB was significantly more conservative in enamel reduction than SB (p<0.05) and showed the highest enamel adhesive surface exposure among the tested preparations (p<0.05). CONCLUSIONS: When the preparation margin is placed above DE, BJ determines a greater exposure of dentin, reducing the adhesive surface in enamel. Below DE, SB seems to be more aggressive in terms of tissue removal compared to CB. CLINICAL SIGNIFICANCE: The results of this in vitro study suggest that in teeth requiring partial restoration with the margin below the dental equator, a chamfer preparation would be more conservative than a shoulder preparation. When above the equator, preparations with flat designs would expose more dentine providing a worse substrate for adhesion.

Bonding to dentin using an experimental zirconium oxynitrate etchant.

OBJECTIVE: To investigate, by means of microtensile bond strength test (µTBS), nanoleakage expression analysis (NL), gelatin zymography and in situ zymography, the effects of an experimental metal salt-based zirconium oxynitrate etchant [ZrO(NO3)2,] - ZON with two simplified adhesives on long-term bond strength and endogenous enzymatic activities. METHODS: Middle/deep coronal dentin surfaces (N = 32) were conditioned either with a traditional 37 % H3PO4 etchant (TE) or with ZON. Further, a single-component etch-and-rinse adhesive (EF) or a universal adhesive (AU) were applied and µTBS and NL tests were performed. Additional freshly extracted teeth were processed for gelatin zymography and in situ zymography evaluation. The tests were performed at baseline and (T0) and after 1-year-aging (T12). Bond strength and in situ zymography results were analyzed using analysis of variance (ANOVA) (three-way and one-way, respectively), while Chi-squared test was used for the NL results. Statistical significance was preset at α = 0.05. RESULTS: All the investigated factors (adhesive system, dentin conditioner and aging) significantly influenced µTBS, with the AU and ZON performing better compared to EF and TE, respectively, and with lower bond strength values after aging (p < 0.05). Incremented silver nitrate deposits were observed at the adhesive interfaces after aging, especially for the TE groups (p < 0.05). Further, the experimental groups treated with ZON had significantly lower levels of enzymatic activity compared to TE, as shown by gelatin and in situ zymography (p < 0.05). CONCLUSIONS: The experimental etchant demonstrated promising results in hybrid-layer preservation over time when used with simplified bonding systems, and could therefore be recommended in the clinical practice.

Effect of an ethanol cross-linker on universal adhesive.

OBJECTIVE: To evaluate the effects of N,N'-dicyclohexylcarbodiimide (DCC), an ethanol-based dentin cross-linker, on the immediate and long-term microtensile bond strength (µTBS) and nanoleakage expression of a universal adhesive employed in self-etch mode (SE) or etch-and-rinse mode (ER). The effect of DCC on the dentinal MMP activity was also investigated by means of in-situ zymography. METHODS: Eighty freshly extracted human molars were sectioned to expose mid-coronal dentin surfaces. The teeth were assigned to one of the following groups, according to the dentin surface priming/adhesive approach: (G1): DCC pre-treatment and Scotchbond Universal (SBU) in ER mode; (G2): SBU in ER mode; (G3): DCC pretreatment and SBU in SE mode; (G4): SBU in SE mode. µTBS test was performed immediately (T0) or after 1-year aging (T12) in artificial saliva. Ten additional teeth per group were prepared for nanoleakage evaluation (N = 5) and for in-situ zymography (N = 5). RESULTS: Three-factor analysis of variance revealed significant difference for the variables DCC pretreatment, application mode and aging (p < 0.05) for both microtensile bond strength testing and in-situ zymography. Nanoleakage analysis revealed reduced marginal infiltration of DCC experimental groups both at T0 and T12. SIGNIFICANCE: The use of an ethanol-based primer containing DCC appears to be promising in preserving the stability of the adhesive interface of a universal adhesive, especially in the SE mode.

Chlorhexidine preserves the hybrid layer in vitro after 10-years aging.

OBJECTIVE: The present study investigated the ability of a chlorhexidine (CHX)-containing primer (0.2% aqueous solution) to inhibit dentinal enzymes, preserve the hybrid layer (HL) and remain within the HL, after 10 years of aging in artificial saliva at 37°C. METHODS: Non-carious extracted molars were assigned to two groups, cut into slabs exposing middle/deep dentin, etched and bonded with Adper Scotchbond 1XT (SB1XT) with or without 0.2% CHX aqueous solution pretreatment. Composite build-ups were made, and the specimens were cut in 1-mm thick bonded sticks. In situ zymography was performed on freshly prepared specimens (T0) and specimens aged for 10 years (T10-yr) at 37°C in artificial saliva, to investigate endogenous gelatinolytic activity within the HL. At T10-yr, specimens were also decalcified and embedded in epoxy resin for TEM analysis. Micro-Raman spectroscopy was performed at T0 and T10-yr to evaluate the chemical profiles in intertubular dentin and the HL. RESULTS: In situ zymography showed less pronounced enzymatic activity in the CHX-pretreated group (p<0.05) regardless of aging, maintaining a similar level of fluorescence at T0 and T10-yr (p>0.05). TEM results showed that 98% of the HL had been degraded in the control group, while 95% of the HL was intact in the experimental group. Moreover, all the Raman spectra peaks assigned to CHX could be identified only in the CHX-pretreated group (T0 and T10-yr). SIGNIFICANCE: In vitro, CHX remains in the HL after 10 years with its inhibitory effect preserved. This may be the underlying factor for HL preservation after this long aging period.