Adhesión a dentina parte I: características del sustrato y sistemas adhesivos / Dentin bonding part I: substrate characteristics and bonding systems

La Odontología Restauradora contemporánea no puede concebirse sin la adhesión de los materiales restauradores a las estructuras dentarias. En mu-chos procedimientos restauradores, no sólo el esmal-te sino también la dentina se encuentra involucrada, por lo tanto, la adhesión a esta última juega un rol cla-ve en el éxito clínico. No obstante, todavía supone un gran desafío lograr una adhesión estable y predeci-ble en el tiempo. El propósito del presente artículo es analizar las características principales de la dentina como sustrato adhesivo y describir los sistemas ad-hesivos actuales y sus mecanismos de acción (AU)

Contemporary Restorative Dentistry cannot be conceived without the adhesion of restorative materials to dental structures. In many restorative procedures not only the enamel, but also the dentin is involved, therefore dentin bonding plays a key role in their success. However, it is still a great challenge to achieve stable and predictable dentin bonding over time. The aim of this article is to analyze the main characteristics of dentin as an adhesive substrate and to describe current adhesive systems and their bonding mechanisms (AU)

Hypophosphatasia.

Hypophosphatasia (HPP) is a group of inherited disorders characterised by the impaired mineralisation of bones and/or teeth and low serum alkaline phosphatase (ALP) activity. It is caused by a mutation in the ALPL gene encoding the tissue-non-specific isoenzyme of ALP (TNSALP) resulting in a loss of function. The disease is highly heterogenous in its clinical expression ranging from stillbirth without mineralised bone to the mild form of late adult onset with symptoms and signs such as musculoskeletal pain, arthropathy, lower-extremity fractures, premature loss of teeth or an incidental finding of reduced serum ALP activity. A classification based on the age at diagnosis and the presence or absence of bone symptoms was historically used: perinatal, prenatal benign, infantile, childhood, adult and odontohypophosphatasia. These subtypes are known to have overlapping signs and complications. Three forms of HPP distinguishable by their genetic characteristics have been described: severe, moderate and mild. Severe forms of HPP (perinatal and infantile severe) are recessively inherited, whereas moderate HPP may be dominantly or recessively inherited. The biochemical hallmark of HPP is persistently low serum ALP for age and increase in natural substrates of TNSALP, pyridoxal 5'-phosphate and phosphoethanolamine supported by radiological findings. The diagnosis is confirmed by ALPL sequencing. A multidisciplinary team of experts is essential for the effective management. Calcium restriction is recommended in infants/children to manage hypercalcaemia. A targeted enzyme replacement therapy for HPP has become available and correct diagnosis is crucial to allow early treatment.

An in vitro study on dentin demineralization and remineralization: Collagen rearrangements and influence on the enucleated phase.

Dentin remineralization is of clinical relevance in the therapy of caries and dentin hypersensitivity. This study is aimed at gaining more insights on a molecular scale, through IR spectroscopy, into dentin demineralization and remineralization. The dentin demineralization by ethylenediaminetetraacetic acid, EDTA (17%, 2 h) significantly altered the secondary structure distribution of collagen, upon loss of interaction with calcium ions. To investigate dentin remineralization, previously demineralized human dentin slices were soaked in Dulbecco's Phosphate Buffered Saline (DPBS) or Hank's Balanced Salt Solution HBSS, in close contact with three commercial cements used as sustained releasing sources of Ca2+ and OH- ions (i.e. calcium hydroxide- and calcium silicate-based cements). IR spectroscopy showed the occurrence of remineralization under these conditions. Collagen did not lose its ability to chelate Ca2+, and these interactions allowed collagen to rearrange into a conformation similar to that of sound dentin. This process appeared slower in HBSS than DPBS, as also shown by the lower degree of maturation of the inorganic phase enucleated in the former medium (amorphous calcium phosphate versus B-type carbonated apatite). Collagen appeared to act as a spatial constraint to crystal deposition, affecting crystallinity and carbonate content of the enucleated phase. Remineralization was found to strongly depend on the calcium releasing ability of the cements. The fast formation of a rough apatite biocoating may represent a favorable clinical condition in the context of mineralized tissue regeneration.

Duplicated or Hybridized Peptide Functional Domains Promote Oral Homeostasis.

Proteins that have existed for millions of years frequently contain repeats of functional domains within their primary structure, thereby improving their functional capacity. In the evolutionary young statherin protein contained within the in vivo-acquired enamel pellicle (AEP), we identified a single functional domain (DR9) located within the protein N-terminal portion that exhibits a higher affinity for hydroxyapatite and more efficient protection against enamel demineralization compared to other native statherin peptides. Thus, we tested the hypothesis that multiplication of functional domains of naturally occurring pellicle peptides amplifies protection against enamel demineralization. In addition, a specific amino acid sequence from histatin 3 (RR-14) was introduced to the hybrid peptides for further testing. Enamel specimens were sectioned to 150-µm thickness and randomly grouped as follows: DR9, DR9-DR9, DR9-RR14, statherin, histatin 1, or distilled water (control). After submersion for 2 h at 37°C, the specimens were placed in 2 mL demineralization solution for 12 d at 37°C. Upon sample removal, the remaining solution was subjected to colorimetric assays to determine the amount of calcium and phosphate released from each specimen. DR9-DR9 amplified protection against enamel demineralization when compared to single DR9 or statherin. Notably, the hybrid peptide DR9-RR14 demonstrated relatively strong protection when the antimicrobial property of these peptides was tested against Candida albicans and Streptococcus mutans. DR9-RR14 was able to maintain 50% of the antifungal activity compared with RR14 for C. albicans and similar values of S. mutans killing activity. This study has pioneered the functional exploration of the natural peptide constituents of the AEP and their evolution-inspired engineered peptides. The knowledge obtained here may provide a basis for the development of stable (proteinase-resistant) synthetic peptides for therapeutic use against dental caries, dental erosion, and/or oral candidiasis.

Demineralization-remineralization dynamics in teeth and bone.

Biomineralization is a dynamic, complex, lifelong process by which living organisms control precipitations of inorganic nanocrystals within organic matrices to form unique hybrid biological tissues, for example, enamel, dentin, cementum, and bone. Understanding the process of mineral deposition is important for the development of treatments for mineralization-related diseases and also for the innovation and development of scaffolds. This review provides a thorough overview of the up-to-date information on the theories describing the possible mechanisms and the factors implicated as agonists and antagonists of mineralization. Then, the role of calcium and phosphate ions in the maintenance of teeth and bone health is described. Throughout the life, teeth and bone are at risk of demineralization, with particular emphasis on teeth, due to their anatomical arrangement and location. Teeth are exposed to food, drink, and the microbiota of the mouth; therefore, they have developed a high resistance to localized demineralization that is unmatched by bone. The mechanisms by which demineralization-remineralization process occurs in both teeth and bone and the new therapies/technologies that reverse demineralization or boost remineralization are also scrupulously discussed. Technologies discussed include composites with nano- and micron-sized inorganic minerals that can mimic mechanical properties of the tooth and bone in addition to promoting more natural repair of surrounding tissues. Turning these new technologies to products and practices would improve health care worldwide.

Relationship between analysis of laser speckle image and Knoop hardness on softening enamel.

BACKGROUND: In this study is presented the correlation between laser speckle images and enamel hardness loss. METHODS: In order to shift the enamel hardness, a dental demineralization model was applied to 32 samples of vestibular bovine teeth. After they were cleaned, cut and polished, the samples were divided into 4 groups and immersed in 30ml of a cola-based soft drink for 10, 20, 30 and 40min twice a day for 7 consecutive days with half the surface protected by two layers of nail polish. Each sample was analyzed by Knoop hardness and laser speckle imaging. RESULTS: Pearson's correlation analysis demonstrated that the laser speckle image technique presents a strong correlation with the hardness loss of the enamel (r=0.7085, p<0.0001). This finding is corroborated by Blend & Altman analysis, in which the data presented a constant behavior throughout the whole interval. For both analyses, more than 95% of the data is within the confidence interval, as expected. CONCLUSION: This work demonstrates, for the first time to our knowledge, an empirical model for correlating laser speckle images with the loss of tooth enamel hardness.

Impact of three endocrine disruptors, Bisphenol A, Genistein and Vinclozolin on female rat enamel.

Concerns about the potential adverse effectsof endocrine disruptors (EDs) have been increasingover the last three decades. BisphenolA (BPA), genistein (G) and vinclozolin (V) arethree widely used EDs sharing similar effects.Since populations are exposed to many diverseEDs simultaneously, we demonstratedrecently their impact alone or combined onmale rat tooth enamel. The purpose of thisstudy was therefore to assess their effects onfemale rat tooth enamel in order to understandwhy they are differentially sensitive. Ratswere exposed daily in utero and after birth tolow doses of EDs during the critical fetal andsuckling periods when amelogenesis takesplace. Enamel of rats exposed to EDs presentedopaque areas of hypomineralization. Theproportion of affected rats was the highestin the groups of rats treated with BPA aloneand higher in males than in females (in all thegroups). Comparison of enamel key gene expressionlevels showed modulations of Klk4and Enamelin in males but no significant variationsin females. These findings show thatfemale rats are less affected than males bythe three EDs chosen in this study and suggestthat enamel hypomineralization may differbetween males and females.

In Vitro Comparative Study of Two Different Bleaching Agents on Micro-hardness Dental Enamel.

OBJECTIVE: To evaluate the effect of home-use bleaching agent containing 16% Carbamide Peroxide (CP) and in-office bleaching agent containing 38% Hydrogen Peroxide (HP) on enamel micro-hardness. STUDY DESIGN: An in vitroexperimental study. PLACE AND DURATION OF STUDY: Department of Operative Dentistry and Science of Dental Materials at Dr. Ishrat-ul-Ebad Khan Institute of Oral Health Sciences, Dow University of Health Sciences and Material Engineering Department of NED University of Engineering and Technology, Karachi, from July to December 2014. METHODOLOGY: Atotal of 90 enamel slabs from 45 sound human 3rd molar were randomly divided into 3 groups. Each group contained 30 specimens (n=30). Group 1 was kept in artificial saliva at 37°C in incubator during the whole experiment. However, Groups 2 and 3 were treated with power whitening gel and tooth whitening pen respectively. After bleaching session, specimens were thoroughly rinsed with deionized water again for 10 seconds and then stored in artificial saliva at 37°C in incubator. Artificial saliva was changed after every 2 days. The Vickers hardness tester (Wolpert 402 MVD, Germany) was adjusted to a load of 0.1 kg (100 gm) and dwell time of 5 seconds. Three Vickers were performed on each specimen using a hardness tester according to the ISO 6507-3:1998 specification. Micro-hardness measurements were performed before and after bleaching at day 1, 7 and 14. RESULTS: In the control group, the baseline micro-hardness was 181.1 ±9.3 which was reduced after the storage on day 1, 7 and 14 (p = 0.104). In Group 2, baseline micro-hardness was 180.4 ±10.1 which was reduced to 179.79 ±10.0 units after day 1. Whereas, on day 7 and 14, the values of micro-hardness were 179.8 ±10 and 179.7 ±10.29, respectively (p=0.091). Furthermore, the baseline micro-hardness in Group 3 was 174.0 ±22.9 units which was reduced to 173 ±23 on day 1, 170 ±30 on day 7 and 173 ±23 on day 14 (p = 0.256). The statistically insignificant difference was found among micro-hardness values of different bleaching agents (p = 0.118). CONCLUSION: Bleaching with 38% Hydrogen Peroxide (HP) and 16% Carbamide Peroxide (CP) resulted in insignificant effect on surface micro-hardness of enamel.

Molecular diagnosis of hypophosphatasia and differential diagnosis by targeted Next Generation Sequencing.

Hypophosphatasia (HPP) is a rare inherited skeletal dysplasia due to loss of function mutations in the ALPL gene. The disease is subject to an extremely high clinical heterogeneity ranging from a perinatal lethal form to odontohypophosphatasia affecting only teeth. Up to now genetic diagnosis of HPP is performed by sequencing the ALPL gene by Sanger methodology. Osteogenesis imperfecta (OI) and campomelic dysplasia (CD) are the main differential diagnoses of severe HPP, so that in case of negative result for ALPL mutations, OI and CD genes had often to be analyzed, lengthening the time before diagnosis. We report here our 18-month experience in testing 46 patients for HPP and differential diagnosis by targeted NGS and show that this strategy is efficient and useful. We used an array including ALPL gene, genes of differential diagnosis COL1A1 and COL1A2 that represent 90% of OI cases, SOX9, responsible for CD, and 8 potentially modifier genes of HPP. Seventeen patients were found to carry a mutation in one of these genes. Among them, only 10 out of 15 cases referred for HPP carried a mutation in ALPL and 5 carried a mutation in COL1A1 or COL1A2. Interestingly, three of these patients were adults with fractures and/or low BMD. Our results indicate that HPP and OI may be easily misdiagnosed in the prenatal stage but also in adults with mild symptoms for these diseases.

Effect of Solvents on Dentin Collagen Cross-linking Potential of Carbodiimide.

PURPOSE: This study compared the dentin collagen cross-linking potential of carbodiimide (EDC) prepared in three most commonly used solvents in dental adhesive systems: water, ethanol, and acetone. MATERIALS AND METHODS: Thirty-eight extracted caries-free human permanent molars were used in this study. Demineralized dentin beams were prepared and cross linked by 0.3 M EDC in water, acetone, or ethanol. The modulus of elasticity of demineralized dentin, the resistance of dentin matrix to enzymatic degradation, the swelling ratio, and the mass change of demineralized dentin matrix were examined to compare the cross-linking efficacy of EDC in their respective solvents. RESULTS: The modulus of elasticity of demineralized dentin cross linked by EDC in acetone was significantly higher (p < 0.05) than demineralized dentin cross linked by EDC in ethanol and EDC in water. Furthermore, the ultimate tensile strength of demineralized dentin cross linked by EDC in water and ethanol dropped significantly following enzymatic degradation, while the ultimate tensile strength of demineralized dentin cross linked by EDC in acetone was preserved. The swelling ratio of demineralized dentin cross linked by EDC in acetone and ethanol was significantly lower (p < 0.05) than that of demineralized dentin cross linked by EDC in water. Conversely, the mass change of demineralized dentin cross linked by EDC in acetone was significantly higher (p < 0.05) than demineralized dentin cross linked by EDC in water and EDC in ethanol. CONCLUSION: The dentin collagen cross-linking potential of EDC could be enhanced by using acetone as a solvent.

Study on the influence of leucine-rich amelogenin peptide (LRAP) on the remineralization of enamel defects via micro-focus x-ray computed tomography and nanoindentation.

Regeneration of severely damaged enamel (e.g. deep demineralized lesions) is currently not possible, because the structural units of enamel crystal construction are removed after its maturation. The aim of this in vitro study was to evaluate the effect of surface impregnation by leucine-rich amelogenin peptide (LRAP) on the remineralization of eroded enamel using micro-focus x-ray computed tomography (µCT). Fifteen bovine enamel blocks were embedded in resin and three zones (sound, demineralization, and remineralization) were defined on each specimen. Lesions were prepared by immersing the samples in demineralization solution for 7 d. The samples were soaked in distilled water or 60 or 120 µg mL(-1) solution of LRAP in water for 30 min. After the surface treatment, specimens were incubated in artificial saliva for either 5 or 10 d at 37 °C. The amount of mineral gain (dΔZ%) and the relative changes in the lesion depth (dLD%), obtained from µCT, were used to evaluate the effect of LRAP on the remineralization of lesions. The effects of LRAP on cross-sectional integrated hardness ΔINH were studied after 10 d using nanoindentation. ANOVA test was used to determine the effect of time and/or LRAP concentration on dΔZ%, dLD% and ΔINH mean values. Tukey's analysis was used for multiple comparison testing (α = 0.05). Analysis of µCT data showed significant effect of time and LRAP concentration on the dΔZ% (p = 0.013, p = 0.003) and the dLD% (p < 0.001, p = 0.002) mean values. The nanoindentation hardness was significantly improved by 120 µg mL(-1) LRAP (p = 0.02). Also, the peptide treatment affected the mineral distribution throughout the lesion by inhibiting of superficial deposition. This study showed that the treatment of eroded lesions in enamel by LRAP can improve and regulate the pattern of remineralization in vitro.

An in vitro comparison of quantitative light-induced fluorescence-digital and spectrophotometer on monitoring artificial white spot lesions.

PURPOSE: The aim of this study was to evaluate the efficacy of quantitative light-induced fluorescence-digital (QLF-D) compared to a spectrophotometer in monitoring progression of enamel lesions. METHODS: To generate artificial caries with various severities of lesion depths, twenty bovine specimens were immersed in demineralizing solution for 40 days. During the production of the lesions, repeat measurements of fluorescence loss (ΔF) and color change (ΔE) were performed in six distinct stages after the demineralization of the specimens: after 3, 5, 10, 20, 30, and 40 days of exposure to the demineralizing solution. Changes in the ΔF values in the lesions were analyzed using the QLF-D, and changes in the ΔE values in lesions were analyzed using a spectrophotometer. The repeated measures ANOVA of ΔF and ΔE values were used to determine whether there are significant differences at different exposure times in the demineralizing solution. Spearman's rank correlation coefficient was analyzed between ΔF and ΔE. RESULTS AND CONCLUSION: The ΔF values significantly decreased based on the demineralizing period (p<0.001). Relatively large changes in ΔF values were observed during the first 10 days. There were significant changes in L(*), a(*), b(*), and ΔE values in lesions with increasing demineralizing duration (p<0.001). A strong correlation was observed between ΔF and ΔE with p=-0.853 (p<0.001). The results support the efficacy of QLF-D in monitoring color changes. Our findings demonstrate that QLF-D are a more efficient and stable tool for early caries detection.

Enamel and dentin bond strength, interfacial ultramorphology and fluoride ion release of self-etching adhesives during a pH-cycling regime.

PURPOSE: This study evaluated the effects of pH cycling on fluoride release and bond strength of two self-etching adhesive systems to both enamel and dentin. The ultramorphology of the interfaces produced by the adhesive systems were also analyzed. MATERIALS AND METHODS: The buccal surfaces of bovine incisors were flattened to expose enamel and dentin, which were bonded with either Clearfil Protect Bond (CPB) or One-Up Bond F Plus (OBP). The bonded samples were prepared for microtensile bond strength (µTBS) testing, fluoride ion release, and transmission electron microscopy. pH cycling comprised demineralization (8 h/day) and remineralization (16 h/day) cycles for 8 days. The µTBS data were analyzed by two-way ANOVA, while fluoride release was analyzed using the Friedman and Wilcoxon tests. RESULTS: The adhesives presented similar bond strengths to enamel. However, the dentin bond strength of CPB was higher than that of OBP. pH cycling did not influence enamel or dentin µTBS. The amount of fluoride released from the bonded enamel and dentin was low and varied among the groups. The morphological evaluation showed that the thickness of the dentin hybrid layers was similar for both adhesives. CONCLUSION: The pH-cycling regime did not affect enamel or dentin bond strengths. In enamel, both the self-etching adhesives tested presented similar bond strengths, but in dentin, Clearfil Protect Bond showed higher dentin bonding than One-Up Bond F Plus.

The effect of mouthwashes containing biguanides on the progression of erosion in dentin.

BACKGROUND: Dental erosion is caused by frequent exposure to acids without the involvement of microorganism. This study analyzed the effect of biguanides (polyhexamethylene biguanide - PHMB and chlorhexidine - CHX) on dentin erosion due to their possible influence on the enzymatic degradation of the demineralized organic matrix. METHOD: Sixty bovine dentin specimens were prepared. On both sides of their surface, nail varnish was applied to maintain the reference surfaces for the determination of dentin loss. Samples were cyclically de- and remineralized for 6 days. Demineralization was performed with a 0.87 M citric acid solution (6×5 min daily). Thereafter, samples were treated with distilled water (negative control), 0.12% CHX (positive control), 0.07% PHMB, Sanifill Perio Premium™ (0.07% PHMB plus 0.05% NaF), or F solution (0.05% NaF) for 1 min and then subjected to enzymatic challenge for 10 min using a bacterial collagenase (Clostridium hystoliticum, 100 µg/ml). Dentin loss was assessed using profilometry (µm) daily. Data were analyzed using 2-way repeated measures-ANOVA and Bonferroni's test (p < 0.05). RESULTS: Dentin loss progressed significantly for all groups during the 6 days. After the 3rd day, Sanifill Premium™, CHX, and PHMB significantly reduced dentin erosion compared to control. On the 6th day, the lowest mean (±SD) dentin loss was observed for Sanifill Perio Premium™ (94.4 ± 3.9 µm). PHMB and CHX led to intermediate dentin loss (129.9 ± 41.2 and 135.3 ± 33.5 µm, respectively) that was significantly lower than those found for negative control (168.2 ± 6.2 µm). F (157.4 ± 6.1 µm) did not significantly differ from negative control. CONCLUSIONS: Sanifill Perio Premium™ mouthwash has a good potential to reduce dentin loss, which might be associated with the presence of PHMB.

Effect of cariogenic challenge on bond strength of adhesive systems to sound and demineralized primary and permanent enamel.

PURPOSE: To evaluate the effect of cariogenic challenge on the bond strength of adhesive systems to sound and artificially demineralized enamel of primary and permanent teeth. MATERIALS AND METHODS: Eighty molars (40 primary, 40 permanent) were randomly assigned to 16 groups (n = 5) according to the type of tooth (primary [PRIM] or permanent [PERM]), enamel condition (sound [S] or demineralized [DEM]), treatment after the restorative procedure (control [C] or cariogenic challenge [pH]), and adhesive system (Adper Single Bond [SB] or Clearfil SE Bond [SE]). Teeth from the DEM group were subjected to cariogenic challenge by pH cycling prior to restorative procedures and pH group specimens were subjected to cariogenic challenge before the microshear test. One of two adhesive systems was applied to the flat enamel surfaces and composite cylinders (0.45 mm2) were built. The microshear bond test was performed. The data (MPa) were subjected to ANOVA and Tukey's test (α = 0.05). RESULTS: No statistically significant differences were detected between the adhesive systems. The S groups exhibited higher bond strength values than the DEM groups, as did C groups compared to pH groups. PERM groups also had higher bond strength values than PRIM groups, excluding PRIM S and PERM S, which had similar values. CONCLUSION: The bond strength to demineralized enamel of primary teeth was lower than to the demineralized enamel of permanent teeth. Cariogenic challenge negatively influenced enamel bond strength, regardless of the type of tooth and adhesive system.

Effect of incorporation of remineralizing agents into bleaching gels on the microhardness of bovine enamel in situ.

AIM: This study evaluated the effect of adding calcium or fluoride to 35% hydrogen peroxide (HP) bleaching gel and the effect of human saliva on the microhardness of sound and demineralized enamel, using an in situ model. MATERIALS AND METHODS: Cylindrical bovine enamel specimens (3 × 2 mm) were divided into two groups (n = 30): sound enamel (SE) and demineralized enamel (DE). Each group was divided into three subgroups, according to the bleaching gel: 35% HP; 35% HP + calcium; 35% HP + fluoride. After bleaching therapy, the specimens were fixed to intraoral devices worn by 10 volunteers for 7 days. Surface enamel microhardness (SMH) was measured before and after bleaching procedures, and after 1 and 7 days of saliva exposure. Data were analyzed by Repeated Measures ANOVA (5%). RESULTS: The variable time resulted in significant differences for SE and DE groups (p = 0.001). For SE, significantly lower SMH was detected for control at post-bleaching period in comparison to the baseline and after 7 days. For DE, the lowest mean values were obtained before bleaching, and the addition of calcium to the peroxide significantly increased enamel SMH. The exposure to human saliva resulted in increased SMH. CONCLUSION: The addition of potential remineralizing agents into bleaching gels might play an important role in maintaining the microhardness of sound enamel and in inducing remineralization of artificially demineralized enamel right after bleaching, and the remineralizing action of human saliva might minimize the deleterious effects of bleaching gels on enamel. Clinical significance: The incorporation of calcium into HP bleaching gel might be beneficial for the initial phases of the bleaching procedure.

Effect of flavonoids on the mechanical properties of demineralised dentine.

OBJECTIVES: This study compared the effect of three flavonoids: proanthocyanidin, naringin and quercetin on the modulus of elasticity (MOE) and ultimate tensile strength (UTS) of demineralised dentine. METHODS: Thirty teeth were sectioned into 0.5mm×1.7mm×7mm beams for MOE measurement. Another 30 non-carious molars were sectioned into 0.5mm×0.5mm thick dentine beams for UTS testing. Demineralised specimens were divided into three groups according to treatments: 6.5% proanthocyanidin, 6.5% quercetin and 6.5% naringin. Specimens were kept in their respective solutions and tested at baseline, 10min, 30min, 1h and 4h. The MOE of each specimen was determined using a three-point bending test at a crosshead speed of 0.5mm/min. For UTS evaluation, each specimen was tested in tension until failure using a crosshead speed of 1mm/min. Means and standard deviation were calculated. Two-way ANOVA and Tukey test were used to evaluate the effect of flavonoid treatment and treatment duration on MOE and UTS. RESULTS: Both MOE and UTS were significantly affected by flavonoid treatment (p<0.001) and treatment duration (p<0.001). Interaction of the two factors was significant for MOE (p<0.001), but not for UTS (p>0.05). Flavonoid treatment improved the mechanical properties of demineralised dentine in the order: proanthocyanidin>quercetin>naringin. It took a longer time for the flavonoids to produce a significant change in UTS, when compared to MOE. CONCLUSION: Proanthocyanidin was more effective than quercetin and naringin in improving biomechanical properties of dentine matrix, thereby enhancing preventive and reparative dental therapies. CLINICAL SIGNIFICANCE: Despite its larger molecular size, proanthocyanidin was more effective than quercetin and naringin, in enhancing the biomechanical properties of demineralised dentine.

Anti-proteolytic capacity and bonding durability of proanthocyanidin-biomodified demineralized dentin matrix.

Our previous studies showed that biomodification of demineralized dentin collagen with proanthocyanidin (PA) for a clinically practical duration improves the mechanical properties of the dentin matrix and the immediate resin-dentin bond strength. The present study sought to evaluate the ability of PA biomodification to reduce collagenase-induced biodegradation of demineralized dentin matrix and dentin/adhesive interfaces in a clinically relevant manner. The effects of collagenolytic and gelatinolytic activity on PA-biomodified demineralized dentin matrix were analysed by hydroxyproline assay and gelatin zymography. Then, resin-/dentin-bonded specimens were prepared and challenged with bacterial collagenases. Dentin treated with 2% chlorhexidine and untreated dentin were used as a positive and negative control, respectively. Collagen biodegradation, the microtensile bond strengths of bonded specimens and the micromorphologies of the fractured interfaces were assessed. The results revealed that both collagenolytic and gelatinolytic activity on demineralized dentin were notably inhibited in the PA-biomodified groups, irrespective of PA concentration and biomodification duration. When challenged with exogenous collagenases, PA-biomodified bonded specimens exhibited significantly less biodegradation and maintained higher bond strengths than the untreated control. These results suggest that PA biomodification was effective at inhibiting proteolytic activity on demineralized dentin matrix and at stabilizing the adhesive/dentin interface against enzymatic degradation, is a new concept that has the potential to improve bonding durability.

Physical chemical effects of zinc on in vitro enamel demineralization.

OBJECTIVES: Zinc salts are formulated into oral health products as antibacterial agents, yet their interaction with enamel is not clearly understood. The aim was to investigate the effect of zinc concentration [Zn(2+)] on the in vitro demineralization of enamel during exposure to caries-simulating conditions. Furthermore, the possible mechanism of zinc's action for reducing demineralization was determined. METHODS: Enamel blocks and synthetic hydroxyapatite (HAp) were demineralized in a range of zinc-containing acidic solutions (0-3565ppm [Zn(2+)]) at pH 4.0 and 37°C. Inductively coupled-plasma optical emission spectroscopy (ICP-OES) was used to measure ion release into solution. Enamel blocks were analysed by Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy (ATR-FTIR), and HAp by X-ray diffraction (XRD) and neutron diffraction (ND). RESULTS: ICP-OES analysis of the acidic solutions showed a decrease in [Ca(2+)] and [PO4(3-)] release with increasing [Zn(2+)]. FTIR revealed a α-hopeite (α-Zn3(PO4)2.4H2O)-like phase on the enamel surfaces at >107ppm [Zn(2+)]. XRD and ND analysis confirmed a zinc-phosphate phase present alongside the HAp. CONCLUSIONS: This study confirms that zinc reduces enamel demineralization. Under the conditions studied, zinc acts predominantly on enamel surfaces at PO4(3-) sites in the HAp lattice to possibly form an α-hopeite-like phase. CLINICAL SIGNIFICANCE: These results have a significant implication on the understanding of the fundamental chemistry of zinc in toothpastes and demonstrate its therapeutic potential in preventing tooth mineral loss.

The effect of saliva substitutes on enamel erosion in vitro.

OBJECTIVES: To investigate the effect of saliva substitutes on enamel erosion in vitro. METHODS: A total of 204 bovine enamel samples were embedded in acrylic resin and allocated to 17 groups (n=12). The specimens were eroded in an artificial mouth (3 days; 6×30 s/days, flow rate: 2 ml/min) using citric acid (pH: 2.5). Immediately after the erosive attacks, saliva substitutes (12 sprays, 3 gels) were applied. Between the erosive cycles the specimens were rinsed with artificial saliva (flowrate: 0.5 ml/min). A SnCl2/AmF/NaF-containing mouthrinse was used as positive control, water spray served as negative control. Enamel loss was measured profilometrically and the data were analyzed using one-way ANOVA followed by Scheffé's post hoc tests (p<0.05). RESULTS: Four saliva substitutes increased enamel erosion, probably due to the low pH or the content of citric acid. Several saliva substitutes were able to reduce enamel erosion significantly by 60-90% (in the range of the positive control). The protective potential of these products was in the range of the positive control (reduction of enamel loss to 30% of negative control). The erosion-protective potential of these high-viscous products is probably related to their film-forming properties, leading to a mechanical protection of the surface. CONCLUSION: Saliva substitutes containing a very low pH exhibit a distinct erosive potential, while most high-viscous products present an erosion-protective effect. It can be recommended that patients suffering from xerostomia and at high risk for dental erosion should use high-viscous saliva substitutes, but should avoid saliva substitutes with low pH or containing citric acid. CLINICAL SIGNIFICANCE: It can be recommended that patients suffering from xerostomia and at high risk for dental erosion should use high-viscous saliva substitutes, but should avoid saliva substitutes with low pH or containing citric acid.