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 collagen 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.

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.

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.

The hydrolytic degradation of resin-dentine interfaces depend upon the type of the in vitro ageing strategy employed in the experimental design. Direct water exposure remains the quickest method to age the resin-dentine bonds. However, the use of SPP may better simulate the in vivo scenario. However, the application of a separate hydrophobic solvent-free adhesive layer may reduce the hydrolytic degradation and increase the longevity of resin-dentine interfaces created with simplified adhesives.

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 phytochemical methodology for grape seed (O)PACs and their potential role as dentin bio-modifiers 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, 1H 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 it may be used as dental intervention material.

Proanthocyanidins (PACs) find wide applications for human use including food, cosmetics, dietary supplements, and pharmaceuticals. The chemical complexity associated with PACs has triggered the development of various chromatographic techniques, with countercurrent separation (CCS) gaining in popularity. This study applied the recently developed DESIGNER (Depletion and Enrichment of Select Ingredients Generating Normalized Extract Resources) approach for the selective enrichment of trimeric and tetrameric PACs using centrifugal partition chromatography (CPC). This CPC method aims at developing PAC based biomaterials, particularly for their application in restoring and repairing dental hard tissue. A general separation scheme beginning with the depletion of polymeric PACs, followed by the removal of monomeric flavan-3-ols and a final enrichment step produced PAC trimer and tetramer enriched fractions. A successful application of this separation scheme is demonstrated for four polyphenol rich plant sources: grape seeds, pine bark, cinnamon bark, and cocoa seeds. Minor modifications to the generic DESIGNER CCS method were sufficient to accommodate the varying chemical complexities of the individual source materials. The step-wise enrichment of PAC trimers and tetramers was monitored using normal phase TLC and Diol-HPLC-UV analyses. CPC proved to be a reliable tool for the selective enrichment of medium size oligomeric PACs (OPACs). This method plays a key role in the development of dental biomaterials considering its reliability and reproducibility, as well as its scale-up capabilities for possible larger-scale manufacturing.

The aim was to assess the nanohardness (H) and the reduced modulus of elasticity (Er ) of 2.5% titanium tetrafluoride (TiF4 ) modified dentin, before and after an erosive challenge with 0.3% citric acid (CA). Exposed dentin surfaces were divided into two groups (n = 5): (1) Control-no dentin pretreatment with TiF4 prior to etching with CA, and (2) Experimental-dentin pretreatment with TiF4  + CA. The H and the Er of intertubular dentin were measured using a triboindenter at different time points: baseline for both groups, after using 2.5% TiF4 for the experimental group, and after using CA for both the experimental and the control groups. Scanning electron microscope and energy dispersive X-ray spectroscopy (EDS) analysis of the dentin surfaces were undertaken at the same time points for both groups. Two-way ANOVA for randomized block design was applied. There was significant interaction between the application of the TiF4 solution and different time points (p = 0.001 for H and p < 0.001 for Er ), identified by Tukey's test. Erosive challenge provided a significant decrease in H and Er mean values. The TiF4 solution caused a significant increase in H and Er values, but no significant differences were found between post-TiF4 and post-CA application. TiF4 application produced a precipitate surface layer on intertubular and intratubular dentin. EDS analysis indicated the presence of titanium. The H and Er of the dentin surface were greatly increased after application of 2.5% TiF4 . TiF4 may modify the micromorphology of the dentin surface and produces an erosive resistance surface.

A self-adhesive resin cement can provide a significantly stronger bond than a three-step etch-and-rinse adhesive and resin cement when used to lute fiber posts. SUMMARYThe bond strength of fiber posts luted with resin cements was evaluated after two storage times in different regions of a post space. A total of 40 single-rooted human teeth were endodontically treated and prepared for cementation of fiber posts (White Post DC). In groups 1 and 3 (G1 and G3, respectively), posts were luted with RelyX ARC, whereas the posts in groups 2 and 4 (G2 and G4, respectively) were luted with RelyX Unicem. After one month of storage at 100% humidity, G1 and G2 were transversally sectioned in 1.7-mm slices of the cervical (C), middle (M), and apical (A) thirds of the post space and submitted to push-out testing at 1 mm/min. After nine months of storage, the roots of G3 and G4 underwent the same process. Mean values were analyzed using the Mann-Whitney and Kruskal-Wallis tests (a¼0.05). The bond strengths in G2 (C¼4.2662.29; M¼4.6763.54; A¼7.2764.30) were statistically higher than in G1 (C¼3.8161.07; M¼1.5761.62; A¼1.9961.60) in the middle and apical thirds (p¼0.001). Bond strengths in G4 (C¼3.3661.39; M¼4.4962.17; A¼3 . 8 361 . 9 2 ) w e r e h i g h e r t h a n i n G 3 (C¼2.1360.47; M¼0.9461.05; A¼0.9561.02) in all evaluated regions ( p¼0.02, p,0.001, and p,0.001, respectively). When comparing the root regions for each group, G1 had higher , 2011, 36-6, 643-648 values in the cervical third than the middle third (p¼0.02). The self-adhesive resin cement showed better results than the conventional resin cement at both storage times. For both materials a similar performance among the three root regions was found. Storage time did not influence the shear bond strength.