The stabilization of dentin collagen with biocompatible crosslinking agents may be of clinical importance to improve dentin bond strength. The present study aimed to evaluate the effect of three collagen crosslinking agents on the ultimate tensile strength (UTS) of undemineralized and demineralized dentin. Ten freshly extracted sound molars were sectioned into 0.5 ؋ 0.5 mm 2 thick beams. The beams were either demineralized or kept undemineralized. Then, specimens were subdivided into four groups according to treatments-PBS solution (control), 5% glutaraldehyde (GD), 0.5% proanthocyanidin PBS solution (PA), and 0.625% genipin PBS solution (GE). Specimens were kept in their respective solutions for either 4 or 40 h. To assess UTS, specimens were subjected to tensile forces at a crosshead speed of 1 mm/min. Statistical analysis was performed using two-way ANOVA and Fisher's PLSD test (p < 0.05). Statistically significant increases in UTS were observed for demineralized dentin after PA and GE dentin treatment, when compared with those of the control group. Dentin treated with GD showed no statistically significant differences in UTS when compared with that the control. Undemineralized dentin revealed no significant differences as compared to that of the control, regardless of the collagen crosslinkers. The application of two naturally occurring crosslinkers, i.e., PA and GE, to dentin collagen significantly improves UTS, indicating its potential value in restorative dentistry.

It is thought that increasing the strength of the dentin matrix using crosslinking agents may improve both the strength and the durability of resin-dentin bonds. The purpose of this study was to evaluate the effect of two collagen crosslinking agents (glutaraldehyde, GD and grape seed extract, GSE) on the modulus of elasticity of demineralized dentin. Sound molar fragments were fully demineralized and divided into five groups according to the type and concentration of crosslinking agents: 2.5% GD; 5% GD, 25% GD; 0.65% GSE; 6.5% GSE. Specimens were immersed in their respective solution and tested at baseline, 10 min, 30 min, 1 h, 2 h, 4 h. The elastic modulus of dentin was significantly affected by the treatment (p < 0.01) and exposure time (p < 0.01). There was a statistically significant interaction between the two factors evaluated (treatment vs. time p < 0.01). Mean baselines values varied between 4.8 and 6.2 MPa in water; after 4 h of treatment the values increased between 34.9 and 242.5 MPa, that were treatment time and agent dependent. The use of these collagen crosslinkers to increase the stiffness of demineralized dentin, was both concentration and time dependent.

Type I collagen is a major component of the hybrid layer, and improvement of its mechanical properties may be advantageous during bonding procedures.Objective-To investigate the effect of three different cross-linking agents (Glutaraldehyde [GD], Grape seed extract [GSE], and Genipin [GE]) on the tensile bond strength (TBS) of resindentin bonds.Materials and Methods-Sixty-four sound human molars were collected and their occlusal surfaces were ground flat to expose dentin. Dentin surfaces were etched using a phosphoric acid and then teeth were randomly divided according to the dentin treatment: Control group (no treatment), 5% GD, 6.5% GSE or 0.5% GE. Teeth were restored either with One Step Plus or Adper Single Bond Plus adhesive systems and resin composite. After 24 hours, teeth were sectioned to produce a cross-sectional surface area of 1.0 mm 2 and tested for tensile bond strength. Data were statistically analyzed using ANOVA and Fisher's PLSD tests (p< 0.05). There was a statistically significant interaction between factors (treatment and adhesive p<0.001). Treatment affected TBS (p< 0.0001), while no differences were observed between the adhesive systems (p = 0.6961).Conclusion-Chemical modification to the dentin matrix promoted by GD and GSE, but not GE, resulted in increased bond strength. The application of selective collagen cross-linkers during adhesive restorative procedures may be a new approach to improve dentin bond strength properties.

Objectives To characterize the interaction of 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide Hydrochloride (EDC) with dentin matrix and its effect on the resin-dentin bond. Methods Changes to the stiffness of demineralized dentin fragments treated with EDC/N-hydroxysuccinimide (NHS) in different solutions were evaluated at different time points. The resistance against enzymatic degradation was indirectly evaluated by ultimate tensile strength (UTS) test of demineralized dentin treated or not with EDC/NHS and subjected to collagenase digestion. Short- and long-term evaluations of the strength of resin-dentin interfaces treated with EDC/NHS for 1 hour were performed using microtensile bond strength (µTBS) test. All data (MPa) were individually analyzed using ANOVA and Tukey HSD tests (α=0.05). Results The different exposure times significantly increased the stiffness of dentin (p<0.0001, control - 5.15 and EDC/NHS - 29.50), while no differences were observed among the different solutions of EDC/NHS (p=0.063). Collagenase challenge did not affect the UTS values of EDC/NHS group (6.08) (p>0.05), while complete degradation was observed for the control group (p=0.0008, control - 20.84 and EDC/NHS - 43.15). EDC/NHS treatment did not significantly increase resin-dentin µTBS, but the values remained stable after 12 months water storage (p<0.05). Conclusions Biomimetic use of EDC/NHS to induce exogenous collagen cross-links resulted in increased mechanical properties and stability of dentin matrix and dentin-resin interfaces.

The ability of certain oligomeric proanthocyanidins (OPACs) to enhance the biomechanical properties of dentin involves collagen cross-linking of the 1.3–4.5 nm wide space via protein–polyphenol interactions. A systematic interdisciplinary search for the bioactive principles of pine bark has yielded the trimeric PAC, ent-epicatechin-(4β→8)-epicatechin-(2β→O→7,4β→8)-catechin (3), representing the hitherto most potent single chemical entity capable of enhancing dentin stiffness. Building the case from two congeneric PAC dimers, a detailed structural analysis decoded the stereochemistry, spatial arrangement, and chemical properties of three dentin biomodifiers. Quantum-mechanics-driven 1H iterative full spin analysis (QM-HiFSA) of NMR spectra distinguished previously unrecognized details such as higher order J coupling and provided valuable information about 3D structure. Detection and quantification of H/D-exchange effects by QM-HiFSA identified C-8 and C-6 as (re)active sites, explain preferences in biosynthetic linkage, and suggest their involvement in dentin cross-linking activity. Mapping of these molecular properties underscored the significance of high δ precision in both 1H and 13C NMR spectroscopy. Occurring at low- to subppb levels, these newly characterized chemical shift differences in ppb are small but diagnostic measures of dynamic processes inherent to the OPAC pharmacophores and can help augment our understanding of nanometer-scale intermolecular interactions in biomodified dentin macromolecules.

Biomodification of existing hard tissue structures, specifically tooth dentin, is an innovative approach proposed to improve the biomechanical and biochemical properties of tissue for potential preventive or reparative/regenerative therapies. The objectives of the study were to systematically characterize dentin matrices biomodified by proanthocyanidin-rich grape seed extract (GSE) and glutaraldehyde (GD). Changes to the biochemistry and biomechanical properties were assessed by several assays to investigate the degree of interactions, biodegradation rates, proteoglycans interaction, and effect of collagen fibril orientation and environmental conditions on the tensile properties. The highest degree of agent-dentin interaction was observed with GSE which exhibited the highest denaturation temperature, regardless of the agent concentration. Biodegradation rates remarkably decreased following biomodification of dentin matrices after 24hs collagenase digestion. A significant decreased in the proteoglycans content of GSE treated samples was observed using a micro-assay for glycosaminoglycans and histological electron microscopy, while no changes were observed for GD and control. Tensile strength properties of GD biomodified dentin matrices were affected by dentin tubule orientation, most likely due to the orientation of the collagen fibrils. Higher and/or increased stability of the tensile properties of GD and GSE-treated samples were observed following exposure to collagenase and 8 month water storage. Biomodification of dentin matrices using chemical agents not only affects the collagen biochemistry; it also involves interaction with proteoglycans. Tissue biomodifiers interact differently with dentin matrices and may provide the tissue with enhanced preventive and restorative/reparative abilities.

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Proanthocyanidins (PACs) are secondary plant metabolites that mediate non-enzymatic 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: PACs 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 combination of 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 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 modulate the mechanism of cross-linking at the molecular, inter-molecular, and inter-micro-fibrillar levels.