Riboflavin and Its Effect on Dentin Bond Strength: Considerations for Clinical Applicability—An In Vitro Study

Bioactive collagen crosslinkers propose to render the dentin hybrid layer less perceptive to hydrolytic challenge. This study aims to evaluate whether bond strength of dental resin composite to dentin benefits from riboflavin (RB)-sensitized crosslinking when used in a clinically applicable protocol. A total of 300 human dentin specimens were prepared consistent with the requirements for a macro-shear bond test. RB was applied on dentin, either incorporated in the primer (RBp) of a two-step self-etch adhesive or as an aqueous solution (RBs) before applying the adhesive, and blue light from a commercial polymerization device was used for RB photoactivation. Bonding protocol executed according to the manufacturer’s information served as control. Groups (n = 20) were tested after 1 week, 1 month, 3 months, 6 months or 1 year immersion times (37 °C, distilled water). The different application methods of RB significantly influenced bond strength (p < 0.001) with a medium impact (η2p = 0.119). After 1 year immersion, post hoc analysis identified a significant advantage for RB groups compared to RBp (p = 0.018), which is attributed to a pH-/solvent-dependent efficiency of RB-sensitized crosslinking, stressing the importance of formulation adjustments. We developed an application protocol for RB-sensitized crosslinking with emphasis on clinical applicability to test its performance against a gold-standard adhesive, and are confident that, with a few adjustments to the application solution, RB-sensitized crosslinking can improve the longevity of adhesive restorations in clinics.

Effect of Thermocycling on the Bond Strength of Self-Adhesive Resin Cements Used for Luting CAD/CAM Ceramics to Human Dentin

The aim of the study was to evaluate the influence of thermocycling on the shear bond strength of self-adhesive, self-etching resin cements luted to human dentin and computer-aided design/computer-aided manufacturing (CAD/CAM) ceramics. Three modern self-adhesive dental cements (Maxcem Elite, RelyX U200, Panavia SA) were used to lute three CAD/CAM ceramics (IPS Empress CAD, IPS e.max CAD, IPS e.max ZirCAD) onto the dentin. One conventional cement (Panavia V5) served as a control. After preparation, the samples were subjected to thermocycling as a method of artificial aging of dental materials applied to simulate long-term use in oral conditions. Shear bond strength was evaluated according to PN-EN ISO 29022:2013-10 and failure modes were observed under a light microscope. Statistical analysis was performed. The study demonstrated that a combination of ceramics and cements directly impacts the bond strength. The highest bond strength was observed in Panavia V5, lower in Panavia SA and Maxcem Elite and the lowest–in RelyX U200. Adhesive failure between human dentin and cements was the most common failure mode. Moreover, thermocycling highly decreased bond strength of self-adhesive, self-etching cements.

Chemical Properties of Human Dentin Blocks and Vertical Augmentation by Ultrasonically Demineralized Dentin Matrix Blocks on Scratched Skull without Periosteum of Adult-Aged Rats

Vertical augmentation is one of the most challenging techniques in bone engineering. Several parameters, such mechano-chemical characteristics, are important to optimize vertical bone regeneration using biomaterials. The aims of this study were to chemically characterize human dentin blocks (calcified demineralized dentin matrix: CDM, partially demineralized dentin matrix: PDDM and completely demineralized dentin matrix: CDDM) (2 × 2 × 1 mm3) chemically and evaluate the behavior of PDDM blocks on non-scratched or scratched skulls without periosteum of adult rats (10–12 months old, female) as a vertical augmentation model. The dissolved efficiency of CDM showed 32.3% after ultrasonic demineralization in 1.0 L of 2% HNO3 for 30 min. The 30 min-demineralized dentin was named PDDM. The SEM images of PDDM showed the opening of dentinal tubes, nano-microcracks and the smooth surface. In the collagenase digestion test, the weight-decreasing rates of CDM, PDDM and CDDM were 9.2%, 25.5% and 78.3% at 12 weeks, respectively. CDM inhibited the collagenase digestion, compared with PDDM and CDDM. In the PDDM onlay graft on an ultrasonically scratched skull, the bone marrow-space opening from original bone was found in the bony bridge formation between the human PDDM block and dense skull of adult senior rats at 4 and 8 weeks. On the other hand, in the cases of the marrow-space closing in both non-scratched skulls and scratched skulls, the bony bridge was not formed. The results indicated that the ultrasonic scratching into the compact parietal bone might contribute greatly to the marrow-space opening from skull and the supply of marrow cells, and then bony bridge formation could occur in the vertical augmentation model without a periosteum.

Secondary Dentin Formation Mechanism: The Effect of Attrition

Human dentin consists of a primary layer produced during tooth formation in early childhood and a second layer which first forms upon tooth eruption and continues throughout life, termed secondary dentin (SD). The effect of attrition on SD formation was considered to be confined to the area subjacent to attrition facets. However, due to a lack of three-dimensional methodologies to demonstrate the structure of the SD, this association could not be determined. Therefore, in the current study, we aimed to explore the thickening pattern of the SD in relation to the amount of occlusal and interproximal attrition. A total of 30 premolars (50–60 years of age) with varying attrition rates were evaluated using micro-computerized tomography. The results revealed thickening of the SD below the cementoenamel junction (CEJ), mostly in the mesial and distal aspects of the root (p < 0.05). The pattern of thickening under the tooth cervix, rather than in proximity to attrition facets, was consistent regardless of the attrition level. The amount of SD thickening mildly correlated with occlusal attrition (r = 0.577, p < 0.05) and not with interproximal attrition. The thickening of the SD below the CEJ coincided with previous finite element models, suggesting that this area is mostly subjected to stress due to occlusal loadings. Therefore, we suggest that the SD formation might serve as a compensatory mechanism aimed to strengthen tooth structure against deflection caused by mechanical loading. Our study suggests that occlusal forces may play a significant role in SD formation.

Effect of Cement Layer Thickness on the Immediate and Long-Term Bond Strength and Residual Stress between Lithium Disilicate Glass-Ceramic and Human Dentin

This study tested whether three different cement layer thicknesses (60, 120 and 180 μm) would provide the same bonding capacity between adhesively luted lithium disilicate and human dentin. Ceramic blocks were cut to 20 blocks with a low-speed diamond saw under cooling water and were then cemented to human flat dentin with an adhesive protocol. The assembly was sectioned into 1 mm2 cross-section beams composed of ceramic/cement/dentin. Cement layer thickness was measured, and three groups were formed. Half of the samples were immediately tested to evaluate the short-term bond strength and the other half were submitted to an aging simulation. The microtensile test was performed in a universal testing machine, and the bond strength (MPa) was calculated. The fractured specimens were examined under stereomicroscopy. Applying the finite element method, the residual stress of polymerization shrinkage according to cement layer thickness was also calculated using first principal stress as analysis criteria. Kruskal–Wallis tests showed that the ‘‘cement layer thickness’’ factor significantly influenced the bond strength results for the aged samples (p = 0.028); however, no statistically significant difference was found between the immediately tested groups (p = 0.569). The higher the cement layer thickness, the higher the residual stress generated at the adhesive interface due to cement polymerization shrinkage. In conclusion, the cement layer thickness does not affect the immediate bond strength in lithium disilicate restorations; however, thinner cement layers are most stable in the short term, showing constant bond strength and lower residual stress.

Nanomechanical and molecular characterization of aging in dentinal collagen

Methylglyoxal (MGO) is an important molecule derived from glucose metabolism with the capacity of attaching to collagen and generating advanced glycation end-products (AGEs), which accumulate in tissues over time and are associated to aging and diseases. However, the accumulation of MGO-derived AGEs in dentin and their effect on the nanomechanical properties of dentinal collagen remain unknown. Thus, the aim of the present study was to quantify MGO-based AGEs in the organic matrix of human dentin as a function of age and associate these changes with alterations in the nanomechanical and ultrastructural properties of dentinal collagen. For this, twelve healthy teeth from <26 year-old and >50 year-old patients were collected and prepared to obtain crown and root dentin discs. Following demineralization, MGO-derived AGEs were quantified with a competitive ELISA assay. In addition, atomic force microscopy (AFM) nano-indentation was utilized to measure changes in elastic modulus in both peritubular (PT) and intertubular (IT) collagen fibrils. Finally, principal component analysis (PCA) was carried out to determine aging profiles for both crown and root dentin. Results showed increased presence of MGO AGEs in the organic matrix of dentin in the >50 year-old compared to <26 year-old samples in both crown and root. Furthermore, an overall increase in PT and IT collagen elasticity was observed in the >50 year-old group associated to ultrastructural changes in the organic matrix determined by AFM analysis. Furthermore, PCA loading plots suggested different 'aging profiles' in both crown and root dentin, which could potentially have important therapeutic implications in restorative and adhesive dentistry approaches. Overall, these results demonstrate that the organic matrix of human dentin undergoes aging-related changes due to MGO-derived AGEs with important changes in nanomechanical behavior of collagen that may impact diagnostic and restorative procedures in the elderly.