Bonding performance and ultramorphology of the resin-dentine interface of contemporary universal adhesives.

OBJECTIVES: This study aimed at evaluating the microtensile bond strength (µTBS) and the resin-dentine ultramorphology (24 h and 10 months ageing) of contemporary universal adhesives applied in self-etch (SE) or etch-and-rinse (ER) mode. MATERIALS AND METHODS: Sixty-four sound human molars were collected and randomly allocated in 4 main experimental groups (n = 16) according to the adhesive system employed and subsequently divided into two subgroups depending on their application mode SE or ER (n = 8): ZipBond X (ZBX-SE; ZBX-ER), Prime and Bond Active (PBA-SE; PBA-ER), Clearfil Universal Bond Quick (CBQ-SE; CBQ-ER) or Scotchbond Universal (SCH-SE; SCH-ER). The specimens were cut into sticks with a cross-sectional area of approximately 0.9 mm2 and subjected to µTBS testing at 24 h or after 10 months of ageing in artificial saliva (AS). Five representative fractured specimens from each group were analysed using field-emission scanning electron microscopy (FE-SEM). Resin-dentine slabs (Ø 0.9mm2) from each experimental group were immersed in Rhodamine B and subsequently analysed using confocal microscopy analysis (CLSM). The µTBS results were analysed using a two-way ANOVA and Newman-Keuls multiple-comparison test (α = 0.05). RESULTS: ZBX, PBA and SCH exhibited greater µTBS values than CQB at 24 h in both SE and ER modes (p < 0.05). CQB showed a significant decrease in µTBS values after ageing both when used in SE and ER mode (p < 0.05). ZBX-ER exhibited no significant differences in the µTBS test after ageing (p > 0.05), while a significant drop in µTBS was seen in SCH-ER and APB-ER after 10-month ageing (p < 0.05). Clear signs of degradation were evident in the resin-dentine interface created with CQB regardless of the application mode or the ageing time. In APB-ER and SCH-ER groups, such signs of degradation were evident after ageing in AS. ZBX showed slight dye infiltration both when used in ER and SE mode. CONCLUSIONS: The long-term bonding performance of modern universal adhesives is usually influenced by the adhesive strategy employed; self-etching application should be prioritised during dentine bonding. Moreover, the use of shortened bonding protocols may compromise the quality of the resin-dentine interface and the bonding performance of most modern universal adhesives. CLINICAL RELEVANCE: The use of etch-and-rinse bonding procedures, as well as "shortened" application protocols should be eluded when using modern universal adhesives in dentine. However, new generation universal adhesives based on innovative chemical formulations may probably allow clinicians to achieve long-term bonding performance with such simplified system also when employed in ER mode.

Modifications in Glass Ionomer Cements: Nano-Sized Fillers and Bioactive Nanoceramics.

Glass ionomer cements (GICs) are being used for a wide range of applications in dentistry. In order to overcome the poor mechanical properties of glass ionomers, several modifications have been introduced to the conventional GICs. Nanotechnology involves the use of systems, modifications or materials the size of which is in the range of 1-100 nm. Nano-modification of conventional GICs and resin modified GICs (RMGICs) can be achieved by incorporation of nano-sized fillers to RMGICs, reducing the size of the glass particles, and introducing nano-sized bioceramics to the glass powder. Studies suggest that the commercially available nano-filled RMGIC does not hold any significant advantage over conventional RMGICs as far as the mechanical and bonding properties are concerned. Conversely, incorporation of nano-sized apatite crystals not only increases the mechanical properties of conventional GICs, but also can enhance fluoride release and bioactivity. By increasing the crystallinity of the set matrix, apatites can make the set cement chemically more stable, insoluble, and improve the bond strength with tooth structure. Increased fluoride release can also reduce and arrest secondary caries. However, due to a lack of long-term clinical studies, the use of nano-modified glass ionomers is still limited in daily clinical dentistry. In addition to the in vitro and in vivo studies, more randomized clinical trials are required to justify the use of these promising materials. The aim of this paper is to review the modification performed in GIC-based materials to improve their physicochemical properties.

Biocompatibility and osteogenic assessment of experimental fluoride-doped calcium-phosphate cements on human dental pulp stem cells.

OBJECTIVES: This study investigated the impact of some specific experimental calcium phosphate cements doped with different fluoride salts (FDCPCs) concentrations on the basal functions of human Dental Pulp Stem Cells (hDPSCs). Furthermore, this study also examined the migration, as well as the mineralisation through osteogenic differentiation. METHODS: Experimental FDCPCs were formulated using different concentrations of calcium/sodium fluoride salts [(5 wt%: VS5F), (10 wt%: VS10F), (20 wt%: VS20F)]. A fluoride-free calcium phosphate (VS0F) was used as a control. The hDPSCs were assessed to evaluate their self-renewal and migration activity in the presence of eluates of the different FDCPCs. A viability assay in osteogenic conditions was carried out, along with the differentiation potential through Alkaline Phosphatase Activity (ALP), and Alizarin Red Staining (ARS). Moreover, the gene expression of specific markers (RUNX2, ALP, COL1α1, OCN, OPN, DSPP, MEPE, and DMP-1) was also evaluated. RESULTS: All the tested FDCPD had no influence on cell migrations, but they caused a decrease in cell viability in osteogenic conditions when not diluted. Conversely, the eluants of VS20F showed a positive effect on stem cell differentiation. This result was corroborated through ALP activity, ARS assay. Moreover, upregulation of specific gene markers such as RUNX2, DMP-1, and DSPP was observed in hDPSCs, especially when treated with VS20F. SIGNIFICANCE: The experimental FDCPC tested in this study exhibits a dose-dependent capacity to promote mineralisation in osteogenic environment. The FDCPC-VS20F seems to be the most promising experimental material suitable for developing of pulp-capping materials with osteogenic and bioactive properties.

Selective modulation by PARP-1 of HIF-1α-recruitment to chromatin during hypoxia is required for tumor adaptation to hypoxic conditions.

BACKGROUND: The adaptation to hypoxia is mainly controlled by the HIF transcription factors. Increased expression/activity of HIF-1α correlates with poor prognosis in cancer patients. PARP-1 inhibitors are used in the clinic to treat BRCAness breast/ovarian cancer and have been shown to regulate the hypoxic response; therefore, their use could be expanded. METHODS: In this work by integrating molecular/cell biology approaches, genome-wide ChIP-seq, and patient samples, we elucidate the extent to which PARP-1 exerts control over HIF-1-regulated genes. RESULTS: In human melanoma, PARP-1 and HIF-1α expression are strongly associated. In response to a hypoxic challenge poly(ADP-ribose) (PAR) is synthesized, HIF-1α is post-transcriptionally modified (PTM) and stabilized by PARylation at specific K/R residues located at its C-terminus. Using an unbiased ChIP-seq approach we demonstrate that PARP-1 dictates hypoxia-dependent HIF-recruitment to chromatin in a range of HIF-regulated genes while analysis of HIF-binding motifs (RCGTG) reveals a restriction on the recognition of hypoxia responsive elements in the absence of PARP-1. Consequently, the cells are poorly adapted to hypoxia, showing a reduced fitness during hypoxic induction. CONCLUSIONS: These data characterize the fine-tuning regulation by PARP-1/PARylation of HIF activation and suggest that PARP inhibitors might have therapeutic potential against cancer types displaying HIF-1α over-activation.

The Multifactorial Role of PARP-1 in Tumor Microenvironment.

Poly(ADP-ribose) polymerases (PARPs), represent a family of 17 proteins implicated in a variety of cell functions; some of them possess the enzymatic ability to synthesize and attach poly (ADP-ribose) (also known as PAR) to different protein substrates by a post-translational modification; PARPs are key components in the cellular response to stress with consequences for different physiological and pathological events, especially during neoplasia. In recent years, using PARP inhibitors as antitumor agents has raised new challenges in understanding their role in tumor biology. Notably, the function of PARPs and PAR in the dynamic of tumor microenvironment is only starting to be understood. In this review, we summarized the conclusions arising from recent studies on the interaction between PARPs, PAR and key features of tumor microenvironment such as hypoxia, autophagy, tumor initiating cells, angiogenesis and cancer-associated immune response.

Effects of Polyacrylic Acid Pre-Treatment on Bonded-Dentine Interfaces Created with a Modern Bioactive Resin-Modified Glass Ionomer Cement and Subjected to Cycling Mechanical Stress.

OBJECTIVES: Resin-modified glass ionomer cements (RMGIC) are considered excellent restorative materials with unique therapeutic and anti-cariogenic activity. However, concerns exist regarding the use of polyacrylic acid as a dentine conditioner as it may influence the bonding performance of RMGIC. The aim of this study was to evaluate the effect of different protocols for cycling mechanical stress on the bond durability and interfacial ultramorphology of a modern RMGIC applied to dentine pre-treated with/without polyacrylic acid conditioner (PAA). METHODS: The RMGIC was applied onto human dentine specimens prepared with silicon-carbide (SiC) abrasive paper with or without the use of a PAA conditioner. The specimens were immersed in deionised water for 24 h then divided in 3 groups. The first group was cut into matchsticks (cross-sectional area of 0.9 mm2) and tested immediately for microtensile bond strength (MTBS). The second was first subjected to load cycling (250,000 cycles; 3 Hz; 70 N) and then cut into matchsticks and tested for MTBS. The third group was subjected to load cycling (250,000 cycles; 3 Hz; 70 N), cut into matchsticks, and then immersed for 8 months storage in artificial saliva (AS); these were finally tested for MTBS. The results were analysed statistically using two-way ANOVA and the Student⁻Newman⁻Keuls test (α = 0.05). Fractographic analysis was performed using FE-SEM, while further RMCGIC-bonded dentine specimens were aged as previously described and used for interfacial ultramorphology characterisation (dye nanoleakage) using confocal microscopy. RESULTS: The RMGIC applied onto dentine that received no pre-treatment (10% PAA gel) showed no significant reduction in MTBS after load cycling followed by 8 months of storage in AS (p > 0.05). The RMGIC⁻dentine interface created in PAA-conditioned SiC-abraded dentine specimens showed no sign of degradation, but with porosities within the bonding interface both after load cycling and after 8 months of storage in AS. Conversely, the RMGIC⁻dentine interface of the specimens with no PAA pre-treatment showed no sign of porosity within the interface after any of the aging protocols, although some bonded-dentine interfaces presented cohesive cracks within the cement after prolonged AS storage. However, the specimens of this group showed no significant reduction in bond strength (p < 0.05) after 8 months of storage in AS or load cycling (p > 0.05). After prolonged AS storage, the bond strength value attained in RMGIC⁻dentine specimens created in PAA pre-treated dentine were significantly higher than those observed in the specimens created with no PAA pre-treatment in dentine. CONCLUSIONS: PAA conditioning of dentine prior to application of RMGIC induces no substantial effect on the bond strength after short-term storage, but its use may increase the risk of collagen degradation at the bonding interface after prolonged aging. Modern RMGIC applied without PAA dentine pre-treatment may have greater therapeutic synergy with saliva during cycle occlusal load, thereby enhancing the remineralisation and protection of the bonding interface.