Photo-crosslinkable hydrogel incorporated with bone matrix particles for advancements in dentin tissue engineering.

The objective of this study was to create injectable photo-crosslinkable biomaterials, using gelatin methacryloyl (GelMA) hydrogel, combined with a decellularized bone matrix (BMdc) and a deproteinized (BMdp) bovine bone matrix. These were intended to serve as bioactive scaffolds for dentin regeneration. The parameters for GelMA hydrogel fabrication were initially selected, followed by the incorporation of BMdc and BMdp at a 1% (w/v) ratio. Nano-hydroxyapatite (nHA) was also included as a control. A physicochemical characterization was conducted, with FTIR analysis indicating that the mineral phase was complexed with GelMA, and BMdc was chemically bonded to the amide groups of gelatin. The porous structure was preserved post-BMdc incorporation, with bone particles incorporated alongside the pores. Conversely, the mineral phase was situated inside the pore opening, affecting the degree of porosity. The mineral phase did not modify the degradability of GelMA, even under conditions of type I collagenase-mediated enzymatic challenge, allowing hydrogel injection and increased mechanical strength. Subsequently, human dental pulp cells (HDPCs) were seeded onto the hydrogels. The cells remained viable and proliferative, irrespective of the GelMA composition. All mineral phases resulted in a significant increase in alkaline phosphatase activity and mineralized matrix deposition. However, GelMA-BMdc exhibited higher cell expression values, significantly surpassing those of all other formulations. In conclusion, our results showed that GelMA-BMdc produced a porous and stable hydrogel, capable of enhancing odontoblastic differentiation and mineral deposition when in contact with HDPCs, thereby showing potential for dentin regeneration.

Effects on dentin nanomechanical properties, cell viability and dentin wettability of a novel plant-derived biomodification monomer.

OBJECTIVES: To evaluate the effects of dentin biomodification agents (Proanthocyanidin (PAC), Cardol (CD) and Cardol-methacrylate (CDMA) on dentin hydrophilicity by contact angle measurement, viability of dental pulp stem cells (DPSCs) and nanomechanical properties of the hybrid layer (HL). METHODS: CDMA monomer was synthesized from cardol through methacrylic acid esterification. Human extracted third molars were used for all experiments. For nanomechanical tests, specimens were divided in four groups according to the primer solutions (CD, CDMA, PAC and control) were applied before adhesive and composite coating. Nanomechanical properties of the HL were analyzed by nanoindentation test using a Berkovich probe in a nanoindenter. Wettability test was performed on dentin surfaces after 1 min biomodification and measured by contact angle analysis. Cytotoxicity was assessed by a MTT assay with DPSCs after 48 and 72 h. Data were analyzed with Student's t test or Two-way ANOVA and Tukey HSD test (p < 0.05). RESULTS: CD and CDMA solutions achieved greater hydrophobicity and increased the water-surface contact angles when compared to PAC and control groups (p < 0.05). PAC group showed a greater reduction of elastic modulus in nanoindentation experiments when compared to CD and CDMA groups (p < 0.05) after 4 months of aging. CD inhibited cell proliferation compared to all further materials (p < 0.05), whilst CDMA and PAC indicated no cell cytotoxicity to human DPSCs. SIGNIFICANCE: Cardol-methacrylate provided significantly higher hydrophobicity to dentin and demonstrated remarkable potential as collagen crosslinking, attaining the lowest decrease of HL's mechanical properties. Furthermore, such monomer did not affect pulp cytotoxicity, thereby highlighting promising feasibility for clinical applications.

Biocompatibility, bioactivity, porosity, and sealer/dentin interface of bioceramic ready-to-use sealers using a dentin-tube model.

This study evaluated the biocompatibility, bioactivity, porosity, and sealer/dentin interface of Sealer Plus BC (SP), Bio-C Sealer (BIOC), TotalFill BC Sealer (TF), and AH Plus (AHP). Dentin tubes filled with the sealers and empty tubes (control group) were implanted in the subcutaneous tissue of rats for different periods (n = 6 per group/period). Number of inflammatory cells (ICs), capsule thickness, von Kossa reaction, interleukin-6 (IL-6) and osteocalcin (OCN) were evaluated. Porosity and voids in the interface dentin/sealers were assessed by micro-computed tomography. The data were submitted to ANOVA/Tukey's tests (α = 0.05). Greater capsule thickness, ICs and IL-6 immunolabeling cells were observed in AHP. No significant difference in thickness of capsule, ICs, and IL-6- immunolabeling cells was detected between SP and TF, in all periods, and after 30 and 60 days between all groups. At 60 days all groups had reduction in capsule thickness, ICs and IL-6 immunolabeling cells. Von Kossa-positive and birefringent structures were observed in the capsules around the sealers. BIOC, SP, and TF exhibited OCN-immunolabeling cells. All sealers had porosity values below 5%, besides low and similar interface voids. BIOC, SP and TF are biocompatible, bioactive, and have low porosity and voids. The dentin-tube model used is an alternative for evaluating bioceramic materials.

Fatigue and failure mode analyses of glass infiltrated 5Y-PSZ bonded onto dentin analogues.

The purpose of this study was to evaluate the fatigue survival of 5Y-PSZ zirconia infiltrated with an experimental glass and bonded onto dentin analogues. Disc-shaped specimens of a 5Y-PSZ (Katana UTML Kuraray Noritake) were cemented onto dentin analogs (NEMA G10) and divided into four groups (n = 15): Zctrl Group (control, without infiltration); Zglz Group (Glaze, compression surface); Zinf-comp Group (Experimental Glass, compression surface); Zinf-tens Group (Experimental Glass, tension surface). Surface treatments were varied. Cyclic fatigue loading, oblique transillumination, stereomicroscope examination, and scanning electron microscopy were performed. Fatigue data were analyzed (failure load and number of cycles) using survival analysis (Kaplan-Meier and Log-Rank Mantel-Cox). There was no statistically significant difference in fatigue survival between the Zglz, Zctrl, and Zinf-comp groups. The Zinf-tens group presented a significantly higher failure load when compared to the other groups and exhibited a different failure mode. The experimental glass effectively infiltrated the zirconia, enhancing structural reliability, altering the failure mode, and improving load-bearing capacity over more cycles, particularly in the group where the glass was infiltrated into the tensile surface of the zirconia. Glass infiltration into 5Y-PSZ zirconia significantly enhanced structural reliability and the ability to withstand loads over an increased number of cycles. This approach has the potential to increase the durability of zirconia restorations, reducing the need for replacements and save time and resources, promoting efficiency in clinical practice.

The effects of re-irradiation on the chemical and morphological properties of permanent teeth.

This study aimed to assess the in vitro effects of re-irradiation on enamel and dentin properties, simulating head and neck cancer radiotherapy retreatment. Forty-five human permanent molars were classified into five groups: non-irradiated; irradiated 60 Gy, and re-irradiated with doses of 30, 40, and 50 Gy. Raman spectroscopy, scanning electron microscopy (SEM), and energy dispersive x-ray spectroscopy (EDS) were employed for analysis. Raman spectroscopy assessed intensity, spectral area, and specific peaks comparatively. Statistical analysis involved Kolmogorov-Smirnov and One-Way ANOVA tests, with Tukey's post-test (significance level set at 5%). Significant changes in irradiated, non-irradiated, and re-irradiated enamel peaks were observed, including phosphate (438 nm), hydroxyapatite (582 nm), phosphate (960 nm), and carbonate (1070 nm) (p < 0.05). Re-irradiation affected the entire tooth (p > 0.05), leading to interprismatic region degradation, enamel prism destruction, and hydroxyapatite crystal damage. Dentin exhibited tubule obliteration, crack formation, and progressive collagen fiber fragmentation. EDX revealed increased oxygen percentage and decreased phosphorus and calcium post-reirradiation. It is concluded that chemical and morphological changes in irradiated permanent teeth were dose-dependent, exacerbated by re-irradiation, causing substantial damage in enamel and dentin.

Effect of Nisin-based pretreatment solution on dentin bond strength, antibacterial property, and MMP activity of the adhesive interface.

OBJECTIVE: To evaluate the effect of a Nisin-based dentin pretreatment solution on microtensile bond strength, antibacterial activity, and matrix metalloproteinase (MMP) activity of the adhesive interface. MATERIALS AND METHODS: 100 human molars were sectioned to expose dentin. The teeth were assigned to five groups (n = 20), according to the dentin pretreatment: 0.5%, 1.0%, or 1.5% Nisin; 0.12% chlorhexidine (positive control), and no solution (negative control), and divided into 2 subgroups: no aging, and thermomechanical aging. Specimens were etched with 37% H3PO4 for 15 s and submitted to the dentin pretreatment. Then, they were bonded with an adhesive (Adper Single Bond 2) and a resin composite for microtensile bond strength (µTBS) evaluation. Antibacterial activity against Streptococcus mutans was qualitatively examined using an agar diffusion test. Anti-MMP activity within hybrid layers was examined using in-situ zymography. Data were analyzed with two-factor ANOVA and post-hoc Tukey's test (α = 0.050). RESULTS: For µTBS, significant differences were identified for the factors "solutions" (p = 0.002), "aging" (p = 0.017), and interaction of the two factors (p = 0.002). In the absence of aging, higher µTBS was observed for the group 0.5% Nisin. In the presence of aging, all groups showed similar µTBS values. All Nisin concentrations were effective in inhibiting the growth of S. mutans. Endogenous MMP activity was more significantly inhibited using 0.5% and 1.0% Nisin (p < 0.050). CONCLUSION: 0.5% and 1.0% Nisin solutions do not adversely affect resin-dentin bond strength and exhibit a potential bactericidal effect against S. mutans. Both concentrations effectively reduce endogenous gelatinolytic activity within the hybrid layer. CLINICAL RELEVANCE: The use of 0.5% and 1.0% Nisin solutions for dentin pretreatment potentially contributes to preserving the adhesive interface, increasing the longevity of composite restorations.

Microfabrication-based engineering of biomimetic dentin-like constructs to simulate dental aging.

Human dentin is a highly organized dental tissue displaying a complex microarchitecture consisting of micrometer-sized tubules encased in a mineralized type-I collagen matrix. As such, it serves as an important substrate for the adhesion of microbial colonizers and oral biofilm formation in the context of dental caries disease, including root caries in the elderly. Despite this issue, there remains a current lack of effective biomimetic in vitro dentin models that facilitate the study of oral microbial adhesion by considering the surface architecture at the micro- and nanoscales. Therefore, the aim of this study was to develop a novel in vitro microfabricated biomimetic dentin surface that simulates the complex surface microarchitecture of exposed dentin. For this, a combination of soft lithography microfabrication and biomaterial science approaches were employed to construct a micropitted PDMS substrate functionalized with mineralized type-I collagen. These dentin analogs were subsequently glycated with methylglyoxal (MGO) to simulate dentin matrix aging in vitro and analyzed utilizing an interdisciplinary array of techniques including atomic force microscopy (AFM), elemental analysis, and electron microscopy. AFM force-mapping demonstrated that the nanomechanical properties of the biomimetic constructs were within the expected biological parameters, and that mineralization was mostly predominated by hydroxyapatite deposition. Finally, dual-species biofilms of Streptococcus mutans and Candida albicans were grown and characterized on the biofunctionalized PDMS microchips, demonstrating biofilm-specific morphologic characteristics and confirming the suitability of this model for the study of early biofilm formation under controlled conditions. Overall, we expect that this novel biomimetic dentin model could serve as an in vitro platform to study oral biofilm formation or dentin-biomaterial bonding in the laboratory without the need for animal or human tooth samples in the future.

Optical properties of dental zirconia, bovine dentin, and enamel-dentin structures.

OBJECTIVE: To evaluate the optical properties and the relative translucency parameter of Ceramill ZI White (3Y-TZP) and Ceramill Zolid FX White (5Y-PSZ) zirconia ceramic systems and compare them with those of the bovine dentin and enamel/dentin structures. MATERIALS AND METHODS: 3Y-TZP and 5Y-PSZ zirconia ceramic systems were evaluated. A 0.5-mm-thick 3Y-TZP (3Y-NC.5), 0.5-mm-thick (5Y-NC.5), and 1.4-mm-thick (5Y-C.14) were used. A 0.5-mm-thick dentin specimens and 1.4-mm-thick enamel/dentin specimens (n = 5) were obtained from anterior bovine maxillary teeth. Scattering, absorption, transmittance, and albedo coefficient were calculated using Kubelka-Munk's model. Data were statistically analyzed using Kruskal-Wallis and Mann-Whitney tests (p < 0.001), and goodness-of-fit coefficient (GFC). Relative translucency parameter differences were evaluated using translucency thresholds. RESULTS: Reflectance, scattering, absorption, and transmittance properties were wavelength dependent. Good matches (GFC ≥ 0.999) in spectral reflectance were observed between 0.5-mm-thick dentin and 1.4-mm-thick enamel/dentin, and 3Y-NC.5 and 5Y-NC.5. Scattering was the main optical extinction process during light interaction with zirconia and dental structures, as indicated by albedo coefficient. Translucency differences were acceptable only for 3Y-NC.5 and the dentin structure, and 5Y-C.14 and the enamel/dentin structure. CONCLUSIONS: Optical properties of 3Y-TZP and 5Y-PSZ dental zirconia differed from each other and from bovine dental structures. Nevertheless, 3Y-TZP showed similar relative translucency parameter to bovine dentin. CLINICAL SIGNIFICANCE: To achieve the best esthetic results in restorative dentistry, it is crucial for clinicians to know about the optical properties of 3Y-TZP and 5Y-PSZ and to be able to compare these properties with those of dental structures.

Radiotherapy in the head and neck region influences the chemical and mechanical properties of intraradicular dentin.

OBJECTIVES: To investigate the chemical and mechanical properties of intraradicular dentin submitted to radiotherapy. MATERIALS AND METHODS: Sixteen mandibular incisors were divided into two groups (n = 8): non-irradiated and irradiated. The irradiated teeth were obtained from head and neck radiotherapy patients, with a total dose ranging from 70.2 to 72 Gy divided into 1.8 Gy daily. After sample preparation, intraradicular dentin slices of each root third were evaluated by Raman spectroscopy, energy dispersive spectroscopy and Knoop microhardness test. Data were analyzed by Two-way ANOVA and Tukey's test (α = 0.05). RESULTS: In Raman spectroscopy, carbonate and amide III showed a significant difference for irradiation and third (carbonate p = 0.021 and p < 0.001; amide III p < 0.001 and p = 0.001, respectively). For amide I, there was a significant difference for third (p < 0.001). For carbonate/mineral ratio, there was a significant difference for irradiation (p = 0.0016) and third (p < 0.001), with the irradiated middle third showing the lowest values. For amide I/amide III ratio, there was a significant difference for irradiation (p = 0.005) in the cervical third. In energy dispersive spectroscopy, carbon (p = 0.004; p = 0.020), phosphorus (p < 0.001; p = 0.009) and calcium (p = 0.008; p = 0.007) showed differences for irradiation and third, with the irradiated groups presenting lower values in cervical and middle thirds. For calcium/phosphorus ratio, there was a significant difference for irradiation (p < 0.001) in cervical and middle thirds. Regarding microhardness, there was a significant difference for irradiation (p < 0.001), with all irradiated groups showing lower microhardness values. CONCLUSIONS: The radiotherapy altered the chemical and mechanical properties of intraradicular dentin, mainly in the cervical and middle root thirds.

Glass ionomer cement with calcium-releasing particles: Effect on dentin mineral content and mechanical properties.

OBJECTIVE: to evaluate the effect a glass ionomer cement (GIC) containing hydroxyapatite (HAp) or calcium silicate (CaSi) particles on mineral content and mechanical properties of demineralized dentin. Ion release and compressive strength (CS) of the cements were also evaluated. METHODS: GIC (Fuji 9 Gold Label, GC), GIC+ 5%HAp and GIC+ 5%CaSi (by mass) were evaluated. Ion release was determined by induced coupled plasma optical emission spectroscopy (Ca2+/Sr2+) or ion-specific electrode (F-) (n = 3). A composite (Filtek Z250, 3 M ESPE) was used as control in remineralization tests. Demineralized dentin discs were kept in contact with materials in simulated body fluid (SBF) at 37 °C for eight weeks. Mineral:matrix ratio (MMR) was determined by ATR-FTIR spectroscopy (n = 5). Dentin hardness (H) and elastic modulus (E) were determined by nanoindentation (n = 10). CS was tested after 24 h and 7d in deionized water (n = 12). Data were analyzed by ANOVA/Tukey test (α = 0.05). RESULTS: Ca2+ and Sr2+ release was higher for the modified materials (p < 0.05). Only GIC+ 5%HAp showed higher F- release than the control (p < 0.05). All groups showed statistically significant increases in MMR, with no differences among them after 8 weeks (p > 0.05). No differences in dentin H or E were observed among groups (p > 0.05). HAp-modified GIC showed increased initial CS, while adding CaSi had the opposite effect (p < 0.05). After 7 days, GIC+ 5%CaSi presented lower CS in relation to control and GIC+ 5%HAp (p < 0.05). SIGNIFICANCE: GIC modification with HAp or CaSi affected CS and increased ion release; however, none of the groups showed evidence of dentin remineralization in comparison to the negative control.

Novel resin-based material containing ß-tricalcium phosphate nanoparticles for the reduction of dentin permeability.

OBJECTIVES: To synthesize and characterize a novel dentin adhesive containing Beta-Tricalcium Phosphate (ß-TCP) nanoparticles and test its ability to reduce dentin permeability (dP). METHODS: Experimental adhesives were prepared by mixing Bis-GMA, TEGDMA, HEMA (50/25/25 wt.%), photo-initiators, and inhibitors. The following groups were tested: Experimental adhesives without ß-TCP (Exp.); with 10 wt.% ß-TCP (Exp.10 wt.% ß-TCP); with 15 wt.% ß-TCP (Exp.15 wt.% ß-TCP), Scotchbond Multi-Purpose (SBMP) and Clearfil SE Protect Bond (CFPB). Degree of conversion (DC%, 10 and 20 s); Flexural Strength (FS), Knoop Hardness (KHN), and Cell Viability (OD%) tests were performed. dP was evaluated by hydraulic conductance, using human dentin disks (n=12), at three-time intervals: post-EDTA (T0); post-treatment (T1); and post-erosion/abrasion cycling (T2). Data were statistically analyzed (α=0.05). RESULTS: For all groups, exposure time for 20 s presented a higher DC% than for 10 s. For FS, filled adhesives did not differ from unfilled and from CFPB. Experimental adhesives did not differ among them and showed lower KHN than the commercial products. Cell viability did not differ among adhesives, except Exp. 15 wt.%, which showed lower OD% than Exp., Exp. 10% and, CFPB. For dP, only Exp.10 and 15 wt.% ß-TCP did not present difference between the times T1 and T2. After cycling, Exp.10 wt.% ß-TCP presented lower permeability than Exp. and CFPB. CONCLUSIONS: The incorporation of 10 wt.% ß-TCP nanoparticles into the resin-based dental material did not affect its mechanical properties and biocompatibility, and promoted the greatest reduction in dentin permeability, sustaining this effect under erosive/abrasive challenges. CLINICAL SIGNIFICANCE: A novel resin-based dental material containing ß-TCP nanoparticles was able to reduce dentin permeability, maintaining its efficacy after erosive/abrasive challenges. The synthesized material did not affect dental pulp cell viability and might be promising for other conditions that require dental remineralization, such as tooth wear and dental caries.

Fracture resistance and bonding performance after antioxidants pre-treatment in non-vital and bleached teeth.

This study aimed to evaluate the effect of antioxidant solutions on fracture strength and bonding performance in non-vital and bleached (38% hydrogen peroxide) teeth. One hundred and eighty dentin specimens were obtained, 60 for each test: fracture strength, hybrid layer thickness, and bond strength. The groups (n=10) were randomly composed according to post-bleaching protocol: REST - restoration, without bleaching; BL - bleaching + restoration; SA - bleaching, 10% sodium ascorbate solution, and restoration; AT - bleaching, 10% α-tocopherol solution, and restoration; CRAN - bleaching, 5% cranberry solution, and restoration; CAP - bleaching, 0.0025% capsaicin solution, and restoration. Data were analyzed with ANOVA, Kruskal-Wallis, Dunn, and Qui-Square tests (α=0.05). The highest fracture strength values were observed in REST (1508.96 ±148.15 N), without significant difference for the bleached groups (p>0.05), regardless of the antioxidant use. The hybrid layer thickness in the group that was not subjected to bleaching (REST) was significantly higher than in any other group. The bond strength in the bleached and antioxidants-treated groups (SA, AT, CRAN, CAP) has no differences with the bleached group without antioxidants (BL). Adhesive failures were predominant in the groups that did not receive the antioxidant application. In conclusion, the evaluated antioxidants did not show an effect on the fracture strength, hybrid layer thickness, or bond strength of dentin bleached after endodontic treatment. The application of 10% sodium ascorbate, 10% alpha-tocopherol, 5% cranberry, or 0.0025% capsaicin solutions is not an effective step and should not be considered for the restorative protocols after non-vital bleaching.

Demineralized dentin matrix technique - a comparison of different demineralizing solutions.

This study aimed to evaluate the microstructure formed after the chemical treatment of teeth, for the development of autogenous grafts from the demineralized dentin matrix (DDM) technique, in order to identify the most efficient demineralizing solution. The specimens, originating from the root and coronal portion, were submitted to ultrasonic cleaning and drying in an oven for 1h at 100 ºC. Then, the density was determined by Archimedes' principle for each specimen, using distilled water as immersion liquid. The samples were separated into five groups: Control group: negative control, Distilled water;EDTA group: positive control, trisodium EDTA; NaOCl group: 2.5% sodium hypochlorite; HCl-0.6M group: 0.6M hydrochloric acid; and H2O2/H2SO4 group: hydrogen peroxide and sulfuric acid. Each specimen was immersed for 1h in the corresponding group descaling solution at 60 ºC. Subsequently, the mass loss and density of the treated specimens were determined by Archimedes' principle. Ultimately, the specimens of each group were characterized by microtomography, Scanning Electron Microscopy, and Energy Dispersive Spectrometry X-ray (SEM-EDS). The results demonstrated that the H2O2/H2SO4 solution allowed the formation of interconnected micropores, suggesting better pore structures for application in scaffolds, when compared to the other studied solutions.

Final irrigation protocols can be used to promote stable long-term bond strength of AH Plus to dentin.

Irrigation solutions might affect dentin surface characteristics and, consequently, endodontic sealers adhesion. This study analyzed the effect of different final irrigation protocols on push-out bond strength (BS) of AH Plus to dentin seven days and 20 months after obturation. Scanning electron micrographs were obtained from the dentin surface of one sample/group after final irrigation. Canals of bovine incisors were instrumented and received final irrigation with (n=21): G1 - 2.5% sodium hypochlorite (NaOCl) + distilled water; G2 - 2.5% NaOCl + 17% EDTA; G3 - 2.5% NaOCl + 17% EDTA + 2.5% NaOCl; G4 - 2.5% NaOCl + 17% EDTA + 2% chlorhexidine (CHX); G5 - mixture 5% NaOCl + 18% etidronate (HEDP); and G6 - mixture 5% NaOCl + 10% tetrasodium EDTA (Na4EDTA). After irrigation, one root/group was split and images were obtained by scanning electron microscopy (SEM). The other 20 roots/group were filled with only AH Plus sealer. Three slices/root were used for push-out assessment seven days and 20 months after obturation. One-way analysis of variance and Tukey (α<0.05) were used to compare the results among experimental groups, and unpaired t-test (α<0.05) was used to compare the results of the same group over time. The photomicrographs showed that, excepting G1, all groups completely removed the smear layer from the samples. In G2 and G4, the opening of the dentin tubules enlarged. In G3, erosion was observed in the peritubular and intertubular dentin. Values of the BS in the seven days were G2=G3=G4=G5>G6=G1 and in the 20 months were G3=G5>G6=G4>G1=G2. G3, G5, and G6 presented values of BS in 20 months similar to the values of seven days (P>0.05). The final irrigation protocols tested produced dentin surfaces with different characteristics. Only G3 and G5 presented high BS values that were stable over time.

Impact of radiotherapy on the morphological and compositional structure of intra-radicular dentin.

Considering the side effects in the oral cavity and dental structures of radiotherapy (RDT) for head and neck cancer, this study aimed to evaluate the effects of RDT on the root dentin concerning the obliteration of dentinal tubules, the inorganic composition of intra-radicular dentin, and the integrity of collagen fibers. Thirty human canines were selected from a biobank and randomly divided into two groups (n=15). The samples were sectioned buccolingually, and a hemisection was used for structural analysis by scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometer (EDS). Low-vacuum SEM images were obtained at 2000-x magnification to observe the obliteration of the dentinal tubules. Moreover, compositional evaluation was performed using EDS. After RDT, the SEM and EDS analyses were repeated using the same methodology. RDT was applied fractionally at 2 Gy per day, 5 days per week, for 7 weeks, resulting in a total dose of 70 Gy. The collagen integrity of the irradiated and non-irradiated samples was analyzed using Masson's trichrome and picrosirius red staining polarization microscopy. Samples subjected to RDT exhibited dentinal tubule obliteration (p < 0.001); low integrity of type I and III collagen fibers (p < 0.05); compositional reduction of calcium (p = 0.012), phosphorus (p = 0.001), and magnesium (p < 0.001); an increased Ca/P ratio (p < 0.001). RDT affects the structure of dentinal tubules, the inorganic composition of intra-radicular dentin, and the collagen fiber integrity in the root dentin, which may interfere with the effectiveness and durability of dental procedures.

Enamel and Dentin Etching with Glycolic, Ferulic, and Phosphoric Acids: Demineralization Pattern, Surface Microhardness, and Bond Strength Stability.

This study evaluated the etching pattern, surface microhardness, and bond strength for enamel and dentin submitted to treatment with phosphoric, glycolic, and ferulic acids. Enamel and dentin blocks were treated with phosphoric, glycolic, and ferulic acid to evaluate the surface and adhesive interface by scanning electron microscopy (2000×). Surface microhardness (Knoop) was evaluated before and after etching, and microtensile bond strength was evaluated after application of a two-step adhesive system (Adper Single Bond 2, 3M ESPE) at 24 hours and 12 months storage time points. Analysis of variance (ANOVA) and Tukey's test showed a decrease in the microhardness values for both substrates after application of each acid (p<0.0001). The reduction percentage was significantly higher for enamel treated with phosphoric acid (59.9%) and glycolic acid (65.1%) than for ferulic acid (16.5%) (p<0.0001), and higher for dentin that received phosphoric acid (38.3%) versus glycolic acid (27.8%) and ferulic acid (21.9%) (p<0.0001). Phosphoric and glycolic acids led to homogeneous enamel demineralization, and promoted the opening of dentinal tubules, whereas ferulic acid led to enamel surface demineralization and partially removed the smear layer. The adhesive-enamel interface showed micromechanical embedding of the adhesive in the interprismatic spaces when phosphoric and glycolic acids were applied. Ferulic acid showed no tag formation. Microtensile bond strength at both time points, and for both substrates, was lower with ferulic acid (p=0.0003/E; p=0.0011/D; Kruskal Wallis and Dunn). The bond strength for enamel and dentin decreased when using phosphoric and glycolic acids at the 12-month time evaluation (p<0.05). Glycolic acid showed an etching pattern and microhardness similar to that of phosphoric acid. Ferulic acid was not effective in etching the enamel or dentin, and it did not provide satisfactory bond strength to dental substrates.

Dentine biomodification by sulphonamides pre-treatment: bond strength, proteolytic inhibition, and antimicrobial activity.

We evaluated the effects of dentine biomodification after pre-treatment with two sulphonamide carbonic anhydrase inhibitors (CAIs) of the N-[4-sulphamoylphenethylcarbamoyl]benzenesulphonamide type, investigating matrix metalloproteases activity, resin-dentine micro tensile bond strength, dentine surface wettability, and antimicrobial activities. Ninety-five sound-extracted human molars were selected for the study. Inhibitory effects were evaluated by gelatinase and collagenase activity tests and collagen degradation FT-IR spectroscopic analysis. Pre-treatment with the two CAIs kept the micro tensile values after 12 months of storage (32.23 ± 5.95) and cariogenic challenge (34.13 ± 2.71) similar to the initial, pre-treatment values (33.56 ± 4.34). A decreased Streptococcus mutans biofilm formation on dentine surfaces and antibacterial activity against planktonic bacteria were observed after CAI treatment. Dentine pre-treatment with sulphonamide CAIs maintained adhesion strength stability, allowed better dentine wettability, maintained matrix collagen, and showed anti-S. mutans activity.

Which self-etch acidic composition may result in higher dental bonds at the long-term? A network meta-analysis review of in vitro studies.

OBJECTIVES: This review evaluated the effects of the acidic composition of self-etch (SE) adhesives at the long-term bond strengths to dentin and enamel. DATA: The review followed the PRISMA Extension Statement for network meta-analysis. Studies were identified by a systematic search in PubMed, Web of Science, and Scopus databases. STUDY SELECTION: The inclusion criteria were in vitro studies that evaluated bond strength data of samples analyzed at both immediate and long-term (after aging simulation) periods and that were bonded to sound dentin/enamel using SE adhesives, with at least one group of adhesives being based on 10-MDP (10-methacryloyloxy-decyl-dihydrogen-phosphate; control) and the other group being comprised of alternative acidic monomers. Statistical analyses were conducted using two methods: standard pairwise meta-analysis (SPMA) and Bayesian network meta-analysis (NMA). Heterogeneity was assessed by using the Cochran Q test and I2 statistics. RESULTS: From 5220 studies identified, 87 met the eligibility criteria and 83 were meta-analyzed. Seventeen adhesives were based on 10-MDP and 44 systems were based on alternative acids. The resin-dentin/enamel bonds were predominantly reduced after aging (∼84% of cases). From the SPMA findings, the following acidic compositions showed lower bond strength values (effect size: mean difference [MD] with 95% confidence interval [95% CI]) than 10-MDP: 4-META (MD -4.99, 95% CI: -7.21, -2.78; p<0.001); sulfonic acids (MD -9.59, 95% CI -12.19, -6.98; p<0.001); unspecified phosphate esters (MD -8.89, 95% CI -17.50, -0.28; p = 0.04); or mixed acids (MD -11.0, 95% CI -13.62, -8.38; p<0.001). The dental bonds were benefited from the presence of 10-MDP upon longer aging (>6 months). From the NMA probabilistic findings, adhesives based on 10-MDP and phosphonic acids ranked as having the best and the worst bonding potential to dentin, respectively. More than one composition (phosphonic acids and mixed acids) ranked similarly to 10-MDP in enamel. The studies scored as having moderate risk of bias (58.6%), followed by low (39.1%) and high (2.3%) risk of bias. CONCLUSION: 10-MDP is an outstanding acidic monomer that contributes to higher bonds to dentin at the long-term. In enamel, there is no evidence that one acidic composition prevails over the other. CLINICAL SIGNIFICANCE: The acidic composition of SE adhesives affects the resistance of dental bonds after simulated aging, with 10-MDP playing a significant role in the adhesion to dentin but not to the enamel. REGISTRATION NUMBER: This report is registered at the Open Science Framework (osf.io/urtdf).

Comparison of collagen features of distinct types of caries-affected dentin.

OBJECTIVES: To compare the biodegradability, mechanical behavior, and physicochemical features of the collagen-rich extracellular matrix (ECM) of artificial caries-affected dentin (ACAD), natural caries-affected dentin (NCAD) and sound dentin (SD). METHODS: Dentin specimens from human molars were prepared and assigned into groups according to the type of dentin: ACAD, NCAD, or SD. ACAD was produced by incubation of demineralized SD with Streptococcus mutans in a chemically defined medium (CDM) with 1% sucrose for 7 days at 37 °C under anaerobic conditions. Specimens were assessed to determine collagen birefringence, biodegradability, mechanical behavior, and chemical composition. Data were individually processed and analyzed by ANOVA and post-hoc tests (α = 0.05). RESULTS: CDM-based biofilm challenge reduced loss, storage, and complex moduli in ACAD (p < 0.001), while the damping capacity remained unaffected (p = 0.066). Higher red and lower green birefringence were found in ACAD and NCAD when compared with SD (p < 0.001). Differently to ACAD, SD and NCAD presented higher biodegradability to exogenous proteases (p = 0.02). Chemical analysis of the integrated areas of characteristic bands that assess mineral quality (carbonate/phosphate and crystallinity index), mineral to matrix (phosphate/amide I) and post-translational modifications (amide III/CH2, pentosidine/CH2, and pentosidine/amide III) (p<0.05) showed that NCAD was significantly different from SD while ACAD exhibited intermediate values. CONCLUSIONS: CDM-based biofilm challenge produced a dentin ECM with decreased mechanical properties and increased collagen maturity. The compositional and structural conformation of the ACAD suggested that CDM-based biofilm challenge showed potential to produce artificial lesions by revealing a transitional condition towards mimicking critical features of NCAD. CLINICAL SIGNIFICANCE: This study highlights the importance of developing a tissue that mimics the features of natural caries-affected dentin ECM for in vitro studies. Our findings suggested the potential of a modified biofilm challenge protocol to produce and simulate a relevant substrate, such as caries-affected dentin.

Phosphoric acid containing proanthocyanidin enhances bond stability of resin/dentin interface.

Proanthocyanidin (PA) is a promising dentin biomodifier due to its ability to stabilize collagen fibrils against degradation by matrix metalloproteinases (MMPs); however, the most effective protocol to incorporate PA into bonding procedures is still unclear. This study evaluated the effect of dentin biomodification with a PA acid etchant on MMP activity, adhesive interface morphology and resin-dentin microtensile bond strength. Sound extracted human molars were flattened to expose dentin and acid-etched for 15 s according to the groups: EXP - experimental phosphoric acid; EXP+PA - experimental phosphoric acid 10% PA; TE - total-etching system; SE - self-etching system. Samples were restored with composite resin and stored in distilled water (37ºC). MMP activity and interface morphology were analyzed after 24 h by in situ zymography (n=6) and scanning electron microscopy (n=3), respectively. The resin-dentin microtensile bond strength (µTBS) was evaluated after 24 h and 6 months storage (n=6). Significantly higher MMP activity was detected in etched dentin compared with untreated dentin (p<0.05), but no difference among acid groups was found. Resin tags and microtags, indicative of proper adhesive system penetration in dentinal tubules and microtubules, were observed along the hybrid layer in all groups. There was no difference in µTBS between 24 h and 6 months for EXP+PA; moreover, it showed higher long-term µTBS compared with TE and EXP (p<0.05). The results suggest that 15 s of biomodification was not sufficient to significantly reduce MMP activity; nonetheless, EXP+PA was still able to improve resin-dentin bond stability compared with total- and self-etching commercial systems.