Biomimetic strategy to stabilize the mechanical properties of caries-affected dentin matrix: A 12-month in vitro study.

PURPOSE: To evaluate the effect of dentin biomodification on the long-term strength of sound and caries-affected (CA) dentin as a strategy to stabilize the dentin matrix. The biomodification strategy utilized a naturally occurring proanthocyanidin-rich Vitis vinifera grape seeds (Vv), and compared with glutaraldehyde (GD). METHODS: Dentin from sound and carious human molars were sectioned from mid-coronal dentin. The temperature denaturation (Td) was assessed using differential scanning calorimetry in sealed pans. The inhibitory effect of the agents on the activity of recombinant MMP-2 and -9 were assessed using colorimetric assay. The ultimate tensile strength (UTS) of demineralized dentin were determined 24 hours after treatment and after 12 months storage in simulated body fluid. Data were statistically analyzed using ANOVA and post-hoc tests ( α= 0.05). RESULTS: There was no statistically significant difference in the Td between sound and CA dentin (P= 0.140); however, Vv and GD significantly increased the Td of both substrates (P< 0.001), indicating formation of collagen cross-linking. The activity of MMP-2 and MMP-9 were reduced by Vv and GD in a concentration dependent manner. The UTS of dentin matrix was significantly affected by treatments and storage times (P< 0.001). After a 12-month period, a significant decrease in UTS was observed for sound and CA, with complete solubilization of the CA dentin matrix. Vv and GD stabilized the UTS of both dentin substrates (P< 0.05). Sound and CA dentin matrix were susceptible to degradation after the 12-month period. Degradation of dentin matrix due to endogenous proteases activity was more pronounced in CA dentin. Dentin biomodification strategies increased the thermal stability and enhanced the long-term mechanical properties of both sound and CA dentin matrix. CLINICAL SIGNIFICANCE: Carious dentin matrix is more susceptible to breakdown over time than sound dentin; however, the degradation process can be impaired by dentin biomodification. This biomimetic strategy increases the long-term tensile strength of the dentin matrix. Reinforcement of caries-affected dentin may increase longevity of adhesive interfaces.

In vitro evaluation of enamel demineralization after Er:YAG and Nd:YAG laser irradiation on primary teeth.

OBJECTIVE: This in vitro study evaluated the influence of both Er:YAG and Nd:YAG laser irradiation on deciduous enamel demineralization. BACKGROUND DATA: Although there are still few studies on the use of the high-intensity laser for caries prevention in deciduous teeth, it is believed that its use on the dental structure can lead to a more acid-resistant surface. METHODS: Forty enamel samples obtained from 22 deciduous first molar teeth were ground and randomly divided into four groups (n = 10): group 1 (G1), no treatment (negative control); G2, fluoride (positive control); G3, Er:YAG laser (2 Hz, 60 mJ, 40.3 J/cm(2)); G4, Nd:YAG laser (80 mJ, 10 Hz, 0.8 W). After the surface treatment, the samples were submitted to an acid challenge that consisted of a 5-day immersion in demineralizing (3 h) and remineralizing solution (21 h). Next, a microhardness test was preformed. RESULTS: Analysis of variance (ANOVA) and Student Newman Keuls tests were performed (alpha = 5%). The percentage of lesion inhibition for each group was as follows: G2, 59.4%; G3, 35.7%; and G4, 40.4%. As regards the percentage loss of mineral volume, there was no statistical difference between groups G2 (444.37 +/- 146.42) and G3 (441.81 +/- 207.08) when compared with group G1 (281.03 +/- 134.57). All experimental groups presented a lower mineral loss compared with the non-irradiated samples (G4). CONCLUSION: The findings of the present study revealed that both Nd:YAG and Er:YAG lasers can be an alternative tool for enhancing deciduous enamel acid resistance.

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.

Effect of dentin biomodification using naturally derived collagen cross-linkers: one-year bond strength study.

Purpose. This study investigated the long-term resin-dentin bond strength of dentin biomodified by proanthocyanidin-rich (PA) agents. Materials and Methods. Forty molars had their coronal dentin exposed, etched, and treated for 10 minutes with 6.5% grape seed extract (GSE), 6.5% cocoa seed extract ethanol-water (CSE-ET), 6.5% cocoa seed extract acetone-water (CSE-AC), and distilled water (CO). Samples were restored either with One-Step Plus (OS) or Adper Single-Bond Plus (SB). Bond strength test was performed immediately or after 3, 6, and 12 months. Results. Higher µ TBS were observed for GSE immediately (SB- 62.9 MPa; OS- 51.9 MPa) when compared to CSE-ET (SB- 56.95 MPa; OS- 60.28 MPa), CSE-AC (SB- 49.97 MPa; OS- 54.44 MPa), and CO (SB- 52.0 MPa; OS- 44.0 MPa) (P < 0.05). CSE outcomes were adhesive system and solvent dependant. After 12 months storage SB results showed no difference among treatment types (GSE- 57.15 MPa; CSE/ET- 54.04 MPa; CSE/AC- 48.22 MPa; CO- 51.68 MPa; P = 0.347),while OS results where treatment dependent (GSE- 42.62 MPa; CSE/ET- 44.06 MPa; CSE/AC- 41.30 MPa; CO- 36.85 MPa; P = 0.036). Conclusions. GSE and CSE-ET agents provided enhanced immediate adhesion and stabilization to demineralized dentin after long-term storage, depending on adhesive system.

Characterization of biomodified dentin matrices for potential preventive and reparative therapies.

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 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 interaction, biodegradation rates, proteoglycan 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 decreased remarkably following biomodification of dentin matrices after 24h collagenase digestion. A significant decrease in the proteoglycan 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 the control. The 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 months water storage. Biomodification of dentin matrices using chemical agents not only affects the collagen biochemistry, but also involves interaction with proteoglycans. Tissue biomodifiers interact differently with dentin matrices and may provide the tissue with enhanced preventive and restorative/reparative abilities.

Long-term effect of carbodiimide on dentin matrix and resin-dentin bonds.

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 h were performed using microtensile bond strength (microTBS) test. All data (MPa) were individually analyzed using ANOVA and Tukey HSD tests (alpha = 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 muTBS, 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.

One-year stability of resin-dentin bonds created with a hydrophobic ethanol-wet bonding technique.

UNLABELLED: Dentin bonding performed with hydrophobic resins using ethanol-wet bonding should be less susceptible to degradation but this hypothesis has never been validated. OBJECTIVES: This in vitro study evaluated stability of resin-dentin bonds created with an experimental three-step BisGMA/TEGDMA hydrophobic adhesive or a three-step hydrophilic adhesive after one year of accelerated aging in artificial saliva. METHODS: Flat surfaces in mid-coronal dentin were obtained from 45 sound human molars and randomly divided into three groups (n=15): an experimental three-step BisGMA/TEGDMA hydrophobic adhesive applied to ethanol (ethanol-wet bonding-GI) or water-saturated dentin (water-wet bonding-GII) and Adper Scotchbond Multi-Purpose [MP-GIII] applied, according to manufacturer instructions, to water-saturated dentin. Resin composite crowns were incrementally formed and light-cured to approximately 5mm in height. Bonded specimens were stored in artificial saliva at 37 degrees C for 24h and sectioned into sticks. They were subjected to microtensile bond test and TEM analysis immediately and after one year. Data were analyzed with two-way ANOVA and Tukey tests. RESULTS: MP exhibited significant reduction in microtensile bond strength after aging (24h: 40.6+/-2.5(a); one year: 27.5+/-3.3(b); in MPa). Hybrid layer degradation was evident in all specimens examined by TEM. The hydrophobic adhesive with ethanol-wet bonding preserved bond strength (24h: 43.7+/-7.4(a); one year: 39.8+/-2.7(a)) and hybrid layer integrity, with the latter demonstrating intact collagen fibrils and wide interfibrillar spaces. SIGNIFICANCE: Coaxing hydrophobic resins into acid-etched dentin using ethanol-wet bonding preserves resin-dentin bond integrity without the adjunctive use of MMPs inhibitors and warrants further biocompatibility and patient safety's studies and clinical testing.