Thaiane Rodrigues Aguiar

Dentin biomodification: strategies, renewable resources and clinical applications.

OBJECTIVES The biomodification of dentin is a biomimetic approach, mediated by bioactive agents, to enhance and reinforce the dentin by locally altering the biochemistry and biomechanical properties. This review provides an overview of key dentin matrix components, targeting effects of biomodification strategies, the chemistry of renewable natural sources, and current research on their potential clinical applications. METHODS The PubMed database and collected literature were used as a resource for peer-reviewed articles to highlight the topics of dentin hierarchical structure, biomodification agents, and laboratorial investigations of their clinical applications. In addition, new data is presented on laboratorial methods for the standardization of proanthocyanidin-rich preparations as a renewable source of plant-derived biomodification agents. RESULTS Biomodification agents can be categorized as physical methods and chemical agents. Synthetic and naturally occurring chemical strategies present distinctive mechanism of interaction with the tissue. Initially thought to be driven only by inter- or intra-molecular collagen induced non-enzymatic cross-linking, multiple interactions with other dentin components are fundamental for the long-term biomechanics and biostability of the tissue. Oligomeric proanthocyanidins show promising bioactivity, and their chemical complexity requires systematic evaluation of the active compounds to produce a fully standardized intervention material from renewable resource, prior to their detailed clinical evaluation. SIGNIFICANCE Understanding the hierarchical structure of dentin and the targeting effect of the bioactive compounds will establish their use in both dentin-biomaterials interface and caries management.

Galloyl moieties enhance the dentin biomodification potential of plant-derived catechins

Proanthocyanidin-rich plant-derived agents have been shown to enhance dentin biomechanical properties and resistance to collagenase degradation. This study systematically investigated the interaction of chemically well-defined monomeric catechins with dentin extracellular matrix components by evaluating dentin mechanical properties as well as activities of matrix metalloproteinases (MMPs) and cysteine-cathepsins (CTs). Demineralized dentin beams (n=15) were incubated for 1h with 0.65% (+)-catechin (C), (-)-catechin gallate (CG), (-)-gallocatechin gallate (GCG), (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC) and (-)-epigallocatechin-3-gallate (EGCG). The modulus of elasticity (E) and the fold increase in E were determined by comparing specimens at baseline and after treatment. Biodegradation rates were assessed by differences in percentage of dry mass before and after incubation with bacterial collagenase. The inhibition of MMP-9 and CT-B by 0.65, 0.065 and 0.0065% of each catechin was determined using fluorimetric proteolytic assay kits. All monomeric catechins led to a significant increase in E. EGCG showed the highest fold increase in E, followed by ECG, CG and GCG. EGCG, ECG, GCG and CG significantly lowered biodegradation rates and inhibited both MMP-9 and CT-B at a concentration of 0.65%. Overall, the 3-O-galloylated monomeric catechins are clearly more potent than their non-galloylated analogues in improving dentin mechanical properties, stabilizing collagen against proteolytic degradation, and inhibiting the activity of MMPs and CTs. The results indicate that galloylation is a key pharmacophore in the monomeric and likely also in the oligomeric proanthocyanidins that exhibit high cross-linking potential for dentin extracellular matrix.

Mimicking the Hierarchical Functions of Dentin Collagen Cross-Links with Plant Derived Phenols and Phenolic Acids

Proanthocyanidins (PACs) are secondary plant metabolites that mediate nonenzymatic collagen cross-linking and enhance the properties of collagen based tissue, such as dentin. The extent and nature of cross-linking is influenced by the composition and specific chemical structure of the bioactive compounds present in certain PAC-rich extracts. This study investigated the effect of the molecular weight and stereochemistry of polyphenol compounds on two important properties of dentin, biomechanics, and biostability. For that, purified phenols, a phenolic acid, and some of its derivatives were selected: PAC dimers (A1, A2, B1, and B2) and a trimer (C1), gallic acid (Ga), its esters methyl-gallate (MGa) and propyl-gallate (PGa), and a pentagalloyl ester of glucose (PGG). Synergism was assessed by combining the most active PAC and gallic acid derivative. Mechanical properties of dentin organic matrix were determined by the modulus of elasticity obtained in a flexural test. Biostability was evaluated by the resistance to collagenase degradation. PACs significantly enhanced dentin mechanical properties and decreased collagen digestion. Among the gallic acid derivatives, only PGG had a significant enhancing effect. The lack of observed C1:PGG synergy indicates that both compounds have similar mechanisms of interaction with the dentin matrix. These findings reveal that the molecular weight of polyphenols have a determinant effect on their interaction with type I collagen and modulates the mechanism of cross-linking at the molecular, intermolecular, and inter-microfibrillar levels.

Dentin Biomodification Potential Depends on Polyphenol Source

Although proanthocyanidins (PACs) modify dentin, the effectiveness of different PAC sources and the correlation with their specific chemical composition are still unknown. This study describes the chemical profiling of natural PAC-rich extracts from 7 plants using ultra high pressure/performance liquid chromatography (UHPLC) to determine the overall composition of these extracts and, in parallel, comprehensively evaluate their effect on dentin properties. The total polyphenol content of the extracts was determined (as gallic acid equivalents) using Folin-Ciocalteau assays. Dentin biomodification was assessed by the modulus of elasticity, mass change, and resistance to enzymatic biodegradation. Extracts with a high polyphenol and PAC content from Vitis vinifera, Theobroma cacao, Camellia sinensis, and Pinus massoniana induced a significant increase in modulus of elasticity and mass. The UHPLC analysis showed the presence of multiple types of polyphenols, ranging from simple phenolic acids to oligomeric PACs and highly condensed tannins. Protective effect against enzymatic degradation was observed for all experimental groups; however, statistically significant differences were observed between plant extracts. The findings provide clear evidence that the dentin bioactivities of PACs are source dependent, resulting from a combination of concentration and specific chemical constitution of the complex PAC mixtures.

A galloylated dimeric proanthocyanidin from grape seed exhibits dentin biomodification potential

Grape seeds are a rich source of polyphenols, especially proanthocyanidins (PACs), and are also known for the presence of galloylated oligomeric PACs (OPACs). The present study focuses on the phytochemical methodology for grape seed (O)PACs and their potential role as dentin biomodifiers to be used in restorative and reparative dentistry. A new method using centrifugal partition chromatography (CPC) was developed for the preparative separation of the grape seed (O)PACs. Orthogonal phytochemical profiling of the resulting CPC fractions was performed using C18 and diol HPLC, normal phase HPTLC, and IT-TOF MS analysis. A galloylated procyanidin dimer (1) was isolated from a CPC fraction in order to evaluate its potential to enhance dentin bio-mechanical properties. Moreover, it helped to evaluate the impact of the galloyl moiety on the observed bioactivity. Structure elucidation was performed using ESI-MS, 1D and 2D NMR analyses. For the first time, (1)H iterative full spin analysis (HiFSA) was performed on this type of molecule, enabling a detailed proton chemical shift and coupling constant assignment. The CPC fractions as well as 1 showed promising results in the dentin stiffness bioassay and indicate that they may be used as dental intervention biomaterial.

Effect of different adhesion strategies on fiber post cementation: Push-out test and scanning electron microscopy analysis

Aim: The aim of this study was to investigate the effect of phosphoric acid etching and the dentin pre-treatment with sodium hypochlorite (NaOCl) on the push-out bond strength between fiber post and root canal dentin. Materials and Methods: Root canals of 48 human incisors were selected, post spaces were prepared and assigned to four groups: G1-37% phosphoric acid (15 s); G2-5.25% NaOCl (2 min) +37% phosphoric acid (15 s); G3-37% phosphoric acid (60 s); and G4-5.25% NaOCl (2 min) +37% phosphoric acid (60 s). Fiber post cementation was performed with two-step etch-and-rinse adhesive system/dual-cured resin cement according to the manufacturers recommendation. After 24 h, each root was sectioned transversally into three slices (cervical, middle and apical) and the bond strength of each section was determined using a push-out bond strength test. Morphology analysis of the bonded interface was evaluated using a scanning electron microscopy. Push-out strength data (MPa) were analyzed by Analysis of Variance and Tukey-Kramer (α = 0.05). Results: Considering the NaOCl pre-treatment, no statistically significant differences were observed among groups; however, when the phosphoric acid was applied during 60 s in the apical portion without NaOCl pre-treatment, the bond strength was statistically significant increased. Conclusion: The NaOCl pre-treatment did not improve the bond strength of adhesive luting cement to root canal dentin. The findings suggest that the use of 37% phosphoric acid for 60 s may have a beneficial effect on bond strength in the apical root third.

Dental adhesives and strategies for displacement of water/solvents from collagen fibrils

OBJECTIVES To evaluate the influence of temperature of evaporation in adhesive systems with different solvents on the apparent modulus of elasticity and mass change of macro-hybrid layers modified by proanthocyanidins (PACs). METHODS Adhesive resin beams (A) from Single Bond Plus (SB), Excite (EX) and One Step Plus (OS) were prepared after solvent evaporation at 23°C or 40°C (n=12). Macro-hybrid layers (M) (n=12) were prepared using demineralized dentin beams sectioned from extracted human third molars. The demineralized dentin specimens were infiltrated with each one of the three adhesive systems at 23°C or 40°C; with or without prior dentin treatment with PACs for 10min. The apparent modulus of elasticity (E) and mass change (Wmc, %) of adhesives beams and resin-infiltrated specimens were assessed in dry and wet conditions after immersion in water (24h, 1, 3 and 6 months). The E was statistically analyzed by Tukey-Kramer test and the Wmc, % by Kruskal Wallis, and Dunn (α=0.05). RESULTS Solvent evaporation at 40°C resulted in higher E values for adhesive resin beams at all storage conditions, regardless of the adhesive system (p<0.05). Increased mass loss (3 months: -0.01%; 6 months: -0.05%) was observed in One Step resin beams (p≤0.05). In the macro-hybrid layer models the pretreatment with PACs along with solvent evaporation at 40°C increased E and decreased the Wmc, % (3 months: -2.5; 6 months: 2.75%) for adhesives evaluated over time (p<0.05). No significant differences in ratio (resin/dentin) were found for the macro-hybrid layers (p>0.05). SIGNIFICANCE Improved solvent evaporation at higher temperature, and increased collagen cross-linking induced by PACs, enhanced the mechanical properties resulting in highly stable macro-hybrid layers over 6 months storage.

Changes in Water Sorption and Solubility of Dental Adhesive Systems after Cigarette Smoke

Aim. To evaluate the effect of cigarette smoke on water sorption and solubility of four adhesive systems. Materials and Methods. Sixteen disks of each adhesive system were prepared (Adper Scotchbond Multipurpose Adhesive (SA); Adper Scotchbond Multipurpose Adhesive System (Adhesive + Primer) (SAP); Adper Single Bond Plus (SB); Adper Easy One (EO)). Specimens were desiccated until a constant mass was obtained and divided into two groups (n = 8). One-half of the specimens were immersed in deionized water, while the other half were also immersed, but with daily exposure to tobacco smoke. After 21 days, disks were measured again and stored in desiccators until constant mass was achieved. Data were calculated according to ISO specifications and statistically analyzed. Results. The tobacco smoke only significantly affected the water sorption and solubility of EO. There were significant differences in both analyses among materials tested. The SB exhibited the highest water sorption, followed by EO, which demonstrated significantly higher solubility values than SB. The SA and SAP showed low water sorption and solubility, and there were no significant differences between the two. Conclusion. Regardless of smoke exposure, both simplified adhesive systems presented an inferior performance that could be related to the complex mixture of components in such versions.

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.

Effect of agitation and storage temperature on water sorption and solubility of adhesive systems.

The purpose of this study was to evaluate the influence of storage temperature and flask agitation on the water sorption (WS) and solubility (SL) of simplified adhesive systems. Seventy-two disc-shaped specimens were prepared according to the adhesive system (water/ethanol-based: Adper Single Bond 2; and water-based: One Coat Bond SL) and experimental conditions tested (mechanical agitation and storage temperature). Statistical analysis (3-way ANOVA, alpha=5%) found significantly greater WS and SL means for the water/ethanol-based system when compared to the water-based. Irrespective of factors studied, significant differences in WS and SL were noted between cold and room temperatures, with greater values been obtained at 1°C, and lower ones at 20°C. Agitation provided increased WS for both materials at all temperatures, but did not affect their SL. The mechanical agitation of the flask may negatively affect the dynamics of diffusion of simplified adhesive systems, even at extremely cold or warm temperatures.