Facultad de Ciencias de la Salud

To evaluate the biomodification ability of lignin used as pre-treatment in human dentin before the application of an etch-and-rinse adhesive. Experimental hydroethanolic solutions with different cross-linking agents were used: 6.5% proanthocyanidins (PAC, from grape-seed extract); 2% cardanol (CARD, from cashew-nut shell liquid); lignin (LIG, from eucalyptus) at 1, 2 or 4% concentrations. The negative control (NC) was ethanol 50 v%. Extracted molars were prepared, and dentin microtensile bond strength ( TBS) was evaluated after 24 h water storage or 10,000 thermocycling aging. Further specimens were processed for SEM nanoleakage, micropermeability confocal microscopy evaluation and in situ degree of conversion (DC) through micro-Raman spectroscopy. Demineralized dentin sticks were submitted to a three-point bending test to evaluate the elastic modulus (E) before and after 1 min biomodification using the tested solutions. Moreover, it was also evaluated the mass changes and hydroxyproline (HYP) release after 4-weeks of water storage. Vibrational collagen crosslinking identification was evaluated through micro-Raman spectroscopy. The results were analyzed by analysis of variance (ANOVA) and Tukey’s test ( = 0.05). A significant reduction in TBS was observed in groups NC (p < 0.001) and CARD (p = 0.026). LIG-4% showed no significant reduction in TBS after aging (p = 0.022). Nanoleakage micrographs showed hybrid layer protection with all agents, but reduced micropermeability was attained only with lignin. Polymerization was negatively affected in the presence of all tested cross-linking agents, except LIG-1%. Lignin and cardanol increased the dentin E values, but only lignin reduced the mass loss in dentin specimens. Effective collagen crosslinking (1117 cm􀀀1 and 1235 cm􀀀1) was detected for all agents. HYP release was significantly lower with LIG-1% than NC (p < 0.001). Lignin was able to perform collagen cross-linking and prevent the degradation of unprotected dentin collagen, thereby improving the bonding performance of the composite restorations performed in this study.

Purpose: The aim of this study was to evaluate the chemomechanical properties, antibacterial activity, and cytotoxicity of an experimental adhesive resin containing halloysite nanotubes (HNT), doped with alkyl trimethyl ammonium bromide (ATAB). Materials and Methods: A filler of HNT doped with ATAB was obtained (ATAB:HNT) and incorporated (5 wt%) into a resin blend made of bisphenol A glycerolate dimethacrylate, 2-hydroxyethyl methacrylate and a photoinitiator/co-initiator system (GATAB:HNT). The same resin blend without ATAB:HNT was used as control (Ctrl). The ATAB:HNT filler was assessed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The two tested adhesives were evaluated for degree of conversion (DC) in vitro and in situ, softening in alcohol, dentin microtensile bond strength (μTBS), antibacterial activity, and cytotoxicity (n = 5). Results: SEM showed that the nanotubes had a characteristic tubular-needle morphology, while the TEM analysis confirmed the presence of ATAB inside the lumens of HNT. The incorporation of ATAB:HNT induced no reduction (p > 0.05) of the DC either in situ or in vitro. No difference was encountered after the softening challenge test (p > 0.05) and no difference was found in μTBS between the two adhesives, both at 24 h (p > 0.05) and after 6 months of storage in distilled water (p > 0.05). However, ATAB:HNT reduced Streptococcus mutans viability (p < 0.05) without a cytotoxic effect on pulp cells (p > 0.05). Conclusions: GATAB:HNT adhesive demonstrated appropriate polymerization without significant differences in softening after solvent immersion, while concomitantly maintaining reliable bond strength after 6 months of water aging. Moreover, the ATAB:HNT filler can provide antibacterial activity to the adhesive resin without affecting pulp cell viability.

Objective The purpose of this research was to investigate the effect per se of two shaping and cleaning techniques on the reduction of an in vitro multispecies biofilm. Materials and Methods A total of 39 freshly extracted monoradicular teeth for periodontal reason were decoronated. Roots were sectioned longitudinally. After autoclaving, a specific stressed biofilm was grown on the root halves that were subsequently reassembled in a silicone index. Two treatments (n = 9 each)—RaCe (Schottlander; Letchworth Garden City, United Kingdom) and ProTaper Gold (PTG; Dentsply Maillefer, Baillagues, Switzerland)—were tested; three noninstrumented samples served as a control group and three were rinsed with saline. Posttreatment samples were taken at three different levels of the root. Colony-forming units were counted after incubations. Additionally, three treatments (n = 5 each)—RaCe, PTG, and saline only—were evaluated under a confocal laser scanning microscope (CLSM). Statistical Analysis Statistical analysis was conducted using Tukey’s test and analysis of variance to evaluate the post-instrumentation bioburden. Results Both instrumentations were able to reduce the biofilm; however, differences were not present between them (p > 0.05). CLSM showed biofilm killing and disruption through mechanical shaping alone. Conclusions Intraradicular biofilm is reduced with mechanical shaping. There was no difference between RaCe and PTG systems in biofilm reduction despite differences in design, file sequence, and rotational speed.

Introduction: This study compared bacterial percolation and sealer penetration of a novel obturation technique with the ones of warm vertical condensation technique. Methods and Materials: A bacterial percolation test was carried out with 80 single rooted human teeth divided into 5 groups; A (n=20): warm vertical condensation and AH-Plus, B (n=20): CPoint with AH-Plus, C (n= 20): CPoint with EndoSequence BC, +ve Control (n=10): teeth with no canal obturation, -ve Control (n=10): teeth with no access cavity. The samples were inoculated with a multispecies bacterial incoulum. Bacterial percolation was evaluated by turbidity. Confocal laser scanning microscopy (CLSM) was used to observe the presence of gaps and voids. Further 48 extracted human mandibular single-canal premolars were used to determine the sealer penetration. Slices of the samples were observed by CLSM to evaluate tubules penetration of the sealer. Kaplan Meyer survival, ANOVA one way and Tuckey HSD test and a Wilcoxon signed-rank test were utilised. Results: A Kaplan-Meier test showed no significant difference overall (P>0.05) between groups A, B and C. At 43 days, the group B showed a significantly inferior ability to prevent bacterial passage (P<0.05). The group C showed a deeper sealer penetration than group A and B with statistically significant differences (P<0.05) for the total penetration (ANOVA one way and Tukey HSD). A Wilcoxon signed-rank test showed statistically significant differences for the penetration in the middle-and apical third of the 3 groups. Conclusion: Based on this in vitro study, the single polymer-cone obturation technique with a resin based- and bioceramic based-sealer behaved similarly to the warm vertical obturation technique in preventing bacterial passage. The bioceramic sealer showed the deepest penetration but did not fully prevent bacterial leakage.

This study aimed at evaluating the e cacy of a novel silver-citrate root canal irrigation solution (BioAKT) on smear layer removal, sealer penetration after root canal instrumentation and antibacterial activity. Single-root teeth were endodontically treated, sealed with an epoxi-amine resin sealer and irrigated using: Group I: 5.25% sodium hypochlorite (NaOCl); Group II: silver-citrate solution (BioAKT); Group III: phosphate bu er solution (PBS); Group IV: 17% ethylenediaminetetraacetic acid (EDTA). Smear layer removal and silver deposition at the coronal, middle and apical portion of each canal were analyzed using scanning electron microscopy (SEM) and energy-dispersive x-ray spectroscopy (EDS). Sealer penetration into dentinal tubules at coronal, middle and apical portion was assessed through dye-assisted confocal microscopy (CSM). Both SEM and CSM micrographs were evaluated by two examiners ( = 0.86), who were blind to the irrigation regimens; scores were given according to the degree of penetration of the sealer. Data analysis included Pearson’s x2 and Sidak’s multiple comparisons. Dentin discs were polished and sterilized. Enterococcus faecalis biofilms were grown using a continuous-flow bioreactor under anaerobic conditions for 72 h. Specimens were irrigated with the tested solutions, and bacterial viability was assessed using a tetrazolium salt assay (MTT). Statistical analysis included one-way ANOVA and Student’s post-hoc t-test (p < 0.05). BioAKT and EDTA were the most e cient solutions both in removing the smear layer and allowing sealer penetration. However, at the apical portion BioAKT performed significantly better compared to EDTA both in smear layer removal and sealer penetration (p < 0.05). BioAKT and NaOCl showed comparable antibacterial e ect (p = 0.53). In conclusion, BioAKT represents a suitable smear layer removal agent, which allows for reliable sealer penetration at the apical portion of the root canal system and o ers significant antibacterial properties.

This study aimed at evaluating the microtensile bond strength (MTBS) and fractographic features of dentine-bonded specimens created using universal adhesives applied in etch-and-rinse (ER) or self-etching (SE) mode in combination with modern ion-releasing resin-modified glass-ionomer cement (RMGIC)-basedmaterials after load cycling and artificial saliva aging. Two universal adhesives (FTB: Futurabond M+, VOCO, Germany; SCU: Scotchbond Universal, 3M Oral Care, USA) were used. Composite build-ups were made with conventional nano-filled composite (AURA, SDI, Australia), conventional resin-modified glass ionomer cement (Ionolux VOCO, Germany), or a (RMGIC)-based composite (ACTIVA, Pulpdent, USA). The specimens were divided in three groups and immersed in deionized water for 24 h, load-cycled (350,000 cycles; 3 Hz; 70 N), or load-cycled and cut into matchsticks and finally immersed for 8 months in artificial saliva (AS). The specimens were cut into matchsticks and tested for microtensile bond strength. The results were analyzed statistically using three-way ANOVA and Fisher’s LSD post hoc test (p < 0.05). Fractographic analysis was performed through stereomicroscope and FE-SEM. FTB showed no significant drop in bond strength after aging. Unlike the conventional composite, the two RMGIC-based materials caused no bond strength reduction in SCU after load-cycle aging and after prolonged aging (8 months). The SEM fractographic analysis showed severe degradation, especially with composite applied on dentine bonded with SCU in ER mode; such degradation was less evident with the two GIC-based materials. The dentine-bond longevity may be influenced by the composition rather than the mode of application (ER vs. SE) of the universal adhesives. Moreover, the choice of the restorative material may play an important role on the longevity of the finalrestoration. Indeed, bioactive GIC-based materials may contribute to maintain the bonding performance of simplified universal adhesives over time, especially when these bonding systems are applied in ER mode.