Effect of Dentin Moisture in Posterior Restorations Performed with Universal Adhesive: A Randomized Clinical Trial.

OBJECTIVES: This double-blind, randomized clinical trial evaluated the influence of dentin moisture on postoperative sensitivity (POS), as well as, on clinical performance in posterior bulk-fill composite restorations, using a universal adhesive, until 12 months after clinical service. METHODS AND MATERIALS: In accordance with a split-mouth design, 45 patients received posterior restorations, restored with a bulk-fill resin composite (Filtek Bulk Fill, 3M Oral Care) and a universal adhesive used in etch-and-rinse mode (SBU; Single Bond Universal Adhesive), which were applied on dry or moist dentin, with a cavity depth of at least 3 mm. Three operators placed 90 Class I/Class II restorations. Patients were evaluated for spontaneous and stimulated POS in the baseline, and after 48 hours, 7 days, and at 6 and 12 months. In addition, secondary parameters (marginal discoloration, marginal adaptation, fracture, and recurrent caries) were evaluated by World Dental Federation (FDI) criteria after 7 days and at 6 and 12 months. Statistical analyzes were performed using the Chi-square, Fisher exact, Friedman, Kruskall-Wallis, and Mann-Whitney tests (α=0.05). RESULTS: No significant spontaneous and stimulated POS was observed when SBU was applied in dry and moist dentin (p>0.05). A significant and higher risk of spontaneous POS (20.0%; 95%CI 10.9-33.82 for dry dentin and 22.22%; 95%CI 12.54-36.27 for moist dentin) occurred up to 48 hours after restoration placement for the dry and moist dentin groups (p<0.02). However, the POS intensity was mild up to 48 hours with no significant difference between dry and moist dentin groups (p>0.79). When secondary parameters were evaluated, no significant differences between the groups were observed. CONCLUSION: Dentin moisture did not influence POS in posterior bulk-fill composite restorations when associated with a universal adhesive applied in etch-and-rinse mode.

Evaluation of At-home Bleaching Times on Effectiveness and Sensitivity with 10% Hydrogen Peroxide: A Randomized Controlled Double-blind Clinical Trial.

OBJECTIVES: The aim of this randomized double-blind controlled clinical trial was to evaluate different protocols for at-home use of 10% hydrogen peroxide in whitening effectiveness and tooth sensitivity. METHODS: Seventy-two patients were selected according to the inclusion and exclusion criteria, with the upper central incisors having color A2 or darker according to the Vita Classical scale (VITA Zahnfabrik, Bad Säckingen, Germany) and randomized into two groups: 10% hydrogen peroxide applied once daily for 15 minutes (HP 15) or applied once daily for 30 minutes (HP 30). Bleaching was performed for 14 days in both groups. The color was evaluated before bleaching, during bleaching (1st and 2nd weeks), and 1 month after the bleaching treatment using the Vita Classical, Vita Bleachedguide 3D-MASTER, and Vita Easyshade spectrophotometer (VITA Zahnfabrik). Dental sensitivity was recorded by the patients using the numerical rating scale (0-4) and visual analogue scale (0-10 cm). Color data were evaluated by two-way analysis of variance (ANOVA) of repeated measures (group vs. treatment time). The Mann-Whitney test was performed to contrast the means (α=0.05). Tooth sensitivity was assessed by Fisher's exact test (p=1.00) and intensity of tooth sensitivity was evaluated by the Mann-Whitney test (α=0.05) for both scales. RESULTS: A significant whitening effect was observed after 2 weeks of bleaching for all color measurements (p=0.01), with no difference between HP 15 and HP 30 (p>0.05). Also, the absolute risk and intensity of tooth sensitivity were similar (47%; p>0.05). CONCLUSIONS: The effectiveness and tooth sensitivity of at-home bleaching carried out with 10% hydrogen peroxide applied for 15 minutes or 30 minutes are similar.

Arthritis in systemic lupus erythematosus is characterized by local IL-17A and IL-6 expression in synovial fluid.

Arthritis is a common clinical feature of systemic lupus erythematosus (SLE) and is usually non-erosive, as opposed to rheumatoid arthritis (RA). While RA synovial pathology has been extensively studied, little is known about the pathophysiology of lupus arthritis. Here, we aimed to explore the cytokine and cellular compartments in synovial fluids of SLE patients with arthritic manifestations. Acellular synovial fluid and paired serum samples from SLE patients (n = 17) were analyzed with cytokine bead array for T helper-associated cytokines. From two SLE patients, synovial fluid mononuclear cells (SFMC) could also be captured and were analyzed by multiparameter flow cytometry to dissect T cell, B cell, monocyte and dendritic cell phenotypes. SLE-derived SFMC were further stimulated in vitro to measure their capacity for producing interferon (IFN)-γ and interleukin (IL)-17A. All patients fulfilled the ACR 1982 classification criteria for SLE. Clinical records were reviewed to exclude the presence of co-morbidities such as osteoarthritis or overlap with RA. IL-17A and IL-6 levels were high in SLE synovial fluid. A clear subset of the synovial CD4+ T cells expressed CCR6+ , a marker associated with T helper type 17 (Th17) cells. IL-17A-production was validated among CD4+ CCR6+ T cells following in-vitro stimulation. Furthermore, a strong IFN-γ production was observed in both CD4+ and CD8+ cells. Our study shows high IL-17A and IL-6 levels in synovial fluids of patients with lupus arthritis. The Th17 pathway has been implicated in several aspects of SLE disease pathogenesis and our data also point to Th17 involvement for lupus arthritis.

Tooth Sensitivity After Dental Bleaching With a Desensitizer-containing and a Desensitizer-free Bleaching Gel: A Systematic Review and Meta-analysis.

OBJECTIVES:: A systematic review and meta-analysis were performed to evaluate the risk and intensity of tooth sensitivity (TS) after dental bleaching with a desensitizer-containing and a desensitizer-free bleaching gel in adult patients. Color change and risk of gingival sensitivity was also evaluated. METHODS:: A comprehensive search was performed MEDLINE via PubMed, Scopus, Web of Science, Latin American and Caribbean Health Sciences Literature database (LILACS), Brazilian Library in Dentistry (BBO), EMBASE and Cochrane Library, and System for Information on Grey Literature in Europe (SIGLE) without restrictions to identify randomized clinical trials. Abstracts from the annual conference of the International Association for Dental Research (1990-2016), unpublished and ongoing trials registries, dissertations, and theses were also searched. The quality of the evidence was rated using the Grading of Recommendations: Assessment, Development and Evaluation (GRADE) approach. DATA:: After duplicates were removed, 1352 articles were identified. After title and abstract screening, only 47 studies remained for qualitative evaluation. Most of the studies had unclear risk of bias. No difference between groups were observed for the risk ratio of TS (risk ratio = 0.99; 95% confidence interval [CI] = 0.74-1.33); intensity of TS (standardized difference in means [SMD] = 0.04; 95% CI = 0.79-0.70); color change in shade guide units (SMD - 0.04; 95% CI = 0.50-0.42); color change in ΔE* (SMD = 0.41 (95% CI = 0.07-0.89); and risk ratio of gingival irritation (SMD = 1.05; 95% CI = 0.81-1.36). Except for the risk of TS, graded as moderate quality of evidence, all other outcomes were rated as low and very low quality. CONCLUSIONS:: Incorporating desensitizers in the bleaching gel did not reduce the risk of TS, and the quality of this evidence was considered moderate. On the other hand, the intensity of TS, color change, and risk of gingival irritation was similar between groups, but the quality of the evidence for these outcomes was graded as low or very low, thus reducing the level of confidence in these outcomes.

FoxP3+ CXCR5+ CD4+ T cell frequencies are increased in peripheral blood of patients with primary Sjögren's syndrome.

We recently explored the expression of CXCR5 on T and B cells from peripheral blood of patients with primary Sjögren's syndrome (SS). Here we investigated the frequency of CD25+ FoxP3+ CD4+ regulatory T cells (Tregs ) among CXCR5+ CD4+ follicular cells in the same cohort of patients. We confirm that the frequency of Tregs among follicular T cells is increased in SS patients and also provide novel data showing an increased frequency of PD-1 expressing cells among CXCR5+ FoxP3+ CD4+ T cells.

Effectiveness of and Dental Sensitivity to At-home Bleaching With 4% and 10% Hydrogen Peroxide: A Randomized, Triple-blind Clinical Trial.

OBJECTIVES: To evaluate the risk for and intensity of tooth sensitivity and color change of at-home dental bleaching with 4% and 10% hydrogen peroxide (HP). METHODS: For this study, 78 patients were selected according to the inclusion and exclusion criteria and randomized into two groups: HP 4 (White Class 4%, FGM) and HP 10 (White Class 10%, FGM). In both groups, the at-home bleaching was performed for a period of 30 minutes twice a day for two weeks. The color was assessed by Vita Classical, Vita Bleachedguide 3D-MASTER and spectrophotometer Vita Easyshade (Vita Zahnfabrik) at baseline, during bleaching (first and second weeks) and after bleaching (one month). Patients recorded their tooth sensitivity using a numeric rating scale (0-4) and visual analog scale (0-10). Data from color change (DeltaE data) was submitted to two-way analysis of variance. The color change data in Delta SGU from the two shade guide units were compared with the Mann Whitney test. The risk of tooth sensitivity was evaluated by χ2 test and the intensity of tooth sensitivity from both scales was evaluated by a Mann-Whitney test (α=0.05). RESULTS: The absolute risk and intensity of tooth sensitivity was higher in the group that used HP 10 than the one that used HP 4. Data from change in the number of shade guide units and color variation after one month of bleaching for both groups showed significant whitening, with no difference between groups. CONCLUSIONS: At-home bleaching is effective with 4% and 10% HP concentrations, but 10% HP increased the absolute risk and intensity of tooth sensitivity during at-home bleaching.

Diminished CXCR5 expression in peripheral blood of patients with Sjögren's syndrome may relate to both genotype and salivary gland homing.

Genetic investigations of Sjögren's syndrome (SS) have identified a susceptibility locus at p23.3 of chromosome 11, which contains the CXCR5 gene. C-X-C motif chemokine receptor 5 (CXCR5) is a chemokine receptor expressed on B and T cell subsets, and binds the chemotactic ligand C-X-C motif chemokine ligand 13 (CXCL13). In this study we aimed to link the genetic association with functional effects and explore the CXCR5/CXCL13 axis in SS. Expression quantitative trait loci analysis of the 11q23.3 locus was performed using B cell mRNA expression data from genotyped individuals. Lymphocyte surface markers were assessed by flow cytometry, and CXCL13 levels by a proximity extension assay. CXCR5+ and CXCL13+ cells in minor salivary glands were detected using immunohistochemistry. Our results demonstrated that SS-associated genetic polymorphisms affected the expression of CXCR5 (P < 0·01). Notably, a decreased percentage of CXCR5+ cells, with lower CXCR5 expression, was observed for most circulating B and T cell subsets in SS patients, reaching statistical significance in CD19+ CD27+ immunoglobulin (Ig)D+ marginal zone (P < 0·001), CD19+ CD27+ IgD- memory (P < 0·05) and CD27-IgD double-negative (P < 0·01) B cells and CD4+ CXCR3- CCR6+ Th17 cells (P < 0·05). CXCL13 levels were increased in patient plasma (P < 0·001), and immunohistochemical staining revealed expression of CXCL13 and higher numbers of CXCR5+ cells (P < 0·0001) within focal infiltrates and interstitially in salivary glands of SS patients. In conclusion, we link a genetic susceptibility allele for SS to a functional phenotype in terms of decreased CXCR5 expression. The decrease of CXCR5+ cells in circulation was also related to homing of B and T cells to the autoimmune target organ. Therapeutic drugs targeting the CXCR5/CXCL13 axis may be useful in SS.

Detection and follow-up of fibroblast growth factor receptor 3 expression on bone marrow and circulating plasma cells by flow cytometry in patients with t(4;14) multiple myeloma.

The t(4;14)(p16;q32) translocation, found in 15% of multiple myeloma (MM) cases, indicates a poor prognosis. Plasma cells (PC) with t(4;14) ectopically express the fibroblast growth factor receptor 3 (FGFR3) tyrosine kinase receptor, which has potential transforming activity and may represent a therapeutic target. To detect FGFR3 protein expression, bone marrow (BM) aspirate from 200 consecutive newly diagnosed (n = 116) or relapsing (n = 74) MM patients was studied by flow cytometry (FC) using anti-CD138 and anti-FGFR3 antibodies. FC data was compared to real time quantitative-polymerase chain reaction (RQ-PCR) of the IGH-MMSET and FGFR3 transcripts. An IGH-MMSET transcript was found in 24/200 patients (12%). In 20 of these, FC detected CD138(+)/FGFR3(+) cells. No expression of FGFR3 was detected in the 4 FGFR3(-) cases by RQ-PCR. FGFR3 was never expressed on PC without t(4;14). Circulating PC (CPC) were detected in patients with (11/11) and patients without (13/41) t(4;14). In 2/8 t(4;14) cases studied longitudinally, coexisting FGFR3(+) and FGFR3(-) CPC were observed. Fluorescent in situ hybridisation (FISH) analysis of the FGFR3(-) subclones showed deletion of the der(14) in one patient. In conclusion, as a supplemental method to RQ-PCR or FISH, FC analysis of FGFR3 expression is a reliable and routinely available method for the detection and management of new therapeutic approaches of t(4;14) MM.

Dendritic cells recruitment and in vivo priming of CD8+ CTL induced by a single topical or transepithelial immunization via the buccal mucosa with measles virus nucleoprotein.

The buccal mucosa, a prototype of pluristratified mucosal epithelia, contains a network of directly accessible class II(+) epithelial dendritic cells (DC), similar to skin Langerhans cells. We showed that a single buccal immunization with measles virus nucleoprotein (NP), by either topical application onto or intradermal injection in the buccal mucosa, induced in vivo priming of protective class I-restricted specific CD8(+) CTL. Both routes of immunization with NP induced a rapid recruitment of DC into the mucosa, which peaked at 2 h and decreased by 24 h. Treatment of mice with Flt3 ligand resulted in an increased number of DC in the buccal mucosa and enhanced the frequency of IFN-gamma-producing NP-specific effectors and the NP-specific CTL response generated after buccal immunization with NP. Finally, NP-pulsed bone marrow-derived DC induced NP-specific IFN-gamma-producing cells upon adoptive transfer to naive mice. These data demonstrate that a viral protein delivered to DC of the buccal mucosa induces in vivo priming of protective anti-viral CD8(+) CTL.

Measles virus exploits dendritic cells to suppress CD4+ T-cell proliferation via expression of surface viral glycoproteins independently of T-cell trans-infection.

Dendritic cells (DC) have been proposed to play a pivotal role in transient immune suppression induced by measles virus (MV) infection. In the present study, we show that DC-induced suppression of T-cell proliferation was not mediated by IL-10 or IFNalpha/beta, which are released following infection of DC, but required cell contacts between MV-infected DC and T cells. Human sera containing neutralizing anti-MV antibodies, as well as anti-MV hemagglutinin (HA) or fusion protein (F) mAbs, were found (i) to reverse suppression and (ii) to restore DC allostimulatory capacity. Interestingly, DC-induced T-cell suppression was associated with both phenotypic and functional DC maturation, as demonstrated by IL-12 production and chemotaxis to MIP-3beta. These data suggest that MV infection turns on the maturation program of DC allowing migration to draining lymph nodes, where potent T-cell immune suppression might be achieved via cell surface expression of HA and F glycoproteins, independently of T cell trans-infection.

Combined natural killer cell and dendritic cell functional deficiency in KARAP/DAP12 loss-of-function mutant mice.

KARAP/DAP12 is a transmembrane polypeptide with an intracytoplasmic immunoreceptor tyrosine-based activation motif (ITAM). KARAP/DAP12 is associated with several activating cell surface receptors in hematopoietic cells. Here, we report that knockin mice bearing a nonfunctional KARAP/DAP12 ITAM present altered innate immune responses. Although in these mice NK cells are present and their repertoire of inhibitory MHC class I receptors is intact, the NK cell spectrum of natural cytotoxicity toward tumor cell targets is restricted. KARAP/DAP12 loss-of-function mutant mice also exhibit a dramatic accumulation of dendritic cells in muco-cutaneous epithelia, associated with an impaired hapten-specific contact sensitivity. Thus, despite its homology with CD3zeta and FcRgamma, KARAP/DAP12 plays a specific role in innate immunity, emphasizing the nonredundancy of these ITAM-bearing polypeptides in hematopoietic cells.

Antitumor effects of the mouse chemokine 6Ckine/SLC through angiostatic and immunological mechanisms.

Mouse 6Ckine/SLC (secondary lymphoid tissue chemokine) is a chemotactic factor for dendritic cells, T cells, and NK cells in vitro. In addition, mouse 6Ckine/SLC interacts with the chemokine receptor CXCR3, as do several chemokines with antiangiogenic properties. These dual properties of mouse 6Ckine/SLC were tested for the induction of an antitumor response by transducing the C26 colon carcinoma tumor cell line with a cDNA encoding mouse 6Ckine/SLC. The C26-6CK-transduced cells showed reduced tumorigenicity in immunocompetent or in nude mice. Part of this effect was likely due to angiostatic mechanisms as shown by immunohistochemistry and Matrigel assay. C26-6CK tumors were also heavily infiltrated with leukocytes, including granulocytes, dendritic cells, and CD8+ T cells. In vivo, anti-CD8 treatment increased the tumorigenicity of the C26-6CK tumor cells, and tumor-infiltrating CD8+ T cells had the phenotype of memory effector cells, suggesting the induction of cytotoxic tumor-specific T lymphocytes. On the other hand, anti-asialo-GM1 depletion also increased the tumorigenicity of C26-6CK cells, supporting the participation of NK cells. Finally, tumor-infiltrating dendritic cells had the phenotype and functional features of immature dendritic cells. Overall, these results suggest that mouse 6Ckine/SLC has strong antitumor effects by inducing both angiostatic, CD8+ T cell-mediated, and possibly NK-mediated tumor resistance mechanisms.

Dendritic cell biology and regulation of dendritic cell trafficking by chemokines.

DC (dendritic cells) represent an heterogeneous family of cells which function as sentinels of the immune system. They traffic from the blood to the tissues where, while immature, they capture antigens. Then, following inflammatory stimuli, they leave the tissues and move to the draining lymphoid organs where, converted into mature DC, they prime naive T cells. The key role of DC migration in their sentinel function led to the investigation of the chemokine responsiveness of DC populations during their development and maturation. These studies have shown that immature DC respond to many CC and CXC chemokines (MIP-1 alpha, MIP-1 beta, MIP-3 alpha, MIP-5, MCP-3, MCP-4, RANTES, TECK and SDF-1) which are inducible upon inflammatory stimuli. Importantly, each immature DC population displays a unique spectrum of chemokine responsiveness. For examples, Langerhans cells migrate selectively to MIP-3 alpha (via CCR6), blood CD11c+ DC to MCP chemokines (via CCR2), monocytes derived-DC respond to MIP-1 alpha/beta (via CCR1 and CCR5), while blood CD11c- DC precursors do not respond to any of these chemokines. All these chemokines are inducible upon inflammatory stimuli, in particular MIP-3 alpha, which is only detected within inflamed epithelium, a site of antigen entry known to be infiltrated by immature DC. In contrast to immature DC, mature DC lose their responsiveness to most of these inflammatory chemokines through receptor down-regulation or desensitization, but acquire responsiveness to ELC/MIP-3 beta and SLC/6Ckine as a consequence of CCR7 up-regulation. ELC/MIP-3 beta and SLC/6Ckine are specifically expressed in the T-cell-rich areas where mature DC home to become interdigitating DC. Altogether, these observations suggest that the inflammatory chemokines secreted at the site of pathogen invasion will determine the DC subset recruited and will influence the class of the immune response initiated. In contrast, MIP-3 beta/6Ckine have a determinant role in the accumulation of antigenloaded mature DC in T cell-rich areas of the draining lymph node, as illustrated by recent observations in mice deficient for CCR7 or SLC/6Ckine. A better understanding of the regulation of DC trafficking might offer new opportunities of therapeutic interventions to suppress, stimulate or deviate the immune response.

Effects of characterised Pseudomonas aeruginosa exopolysaccharides on adherence to human tracheal cells.

This study evaluated, in vitro, the role of different Pseudomonas aeruginosa exopolysaccharides (EPS) in mediating adherence to human respiratory epithelial cells. Two mucoid and non-mucoid isogenic pairs of P aeruginosa strains isolated from patients with cystic fibrosis (CF) and bronchiectasis were used. Adherence was tested with human tracheal epithelial cell lines from CF and normal fetuses. The CF cells bound significantly more bacteria than the normal cells. The strain from the bronchiectasis patient was significantly more adherent than that from the CF patient and this difference was consistently most marked with the non-mucoid variant and with normal epithelial cells. The differing behaviour of mucoid CF and non-mucoid bronchiectasis strains reflected the chemical composition of their EPS: mainly alginate in the former and neutral polysaccharides in the latter. Additive inhibition experiments with chemically characterised EPS indicated that neutral polysaccharides associated with alginate may act as ligands for the adherence of P. aeruginosa to CF epithelial cells.