Chemometric analysis of Hymenoptera toxins and defensins: A model for predicting the biological activity of novel peptides from venoms and hemolymph

When searching for prospective novel peptides, it is difficult to determine the biological activity of a peptide based only on its sequence. The trial and error approach is generally laborious, expensive and time consuming due to the large number of different experimental setups required to cover a reasonable number of biological assays. To simulate a virtual model for Hymenoptera insects, 166 peptides were selected from the venoms and hemolymphs of wasps, bees and ants and applied to a mathematical model of multivariate analysis, with nine different chemometric components: GRAVY, aliphaticity index, number of disulfide bonds, total residues, net charge, pI value, Boman index, percentage of alpha helix, and flexibility prediction. Principal component analysis (PCA) with non-linear iterative projections by alternating least-squares (NIPALS) algorithm was performed, without including any information about the biological activity of the peptides. This analysis permitted the grouping of peptides in a way that strongly correlated to the biological function of the peptides. Six different groupings were observed, which seemed to correspond to the following groups: chemotactic peptides, mastoparans, tachykinins, kinins, antibiotic peptides, and a group of long peptides with one or two disulfide bonds and with biological activities that are not yet clearly defined. The partial overlap between the mastoparans group and the chemotactic peptides, tachykinins, kinins and antibiotic peptides in the PCA score plot may be used to explain the frequent reports in the literature about the multifunctionality of some of these peptides. The mathematical model used in the present investigation can be used to predict the biological activities of novel peptides in this system, and it may also be easily applied to other biological systems. © 2011 Elsevier Inc.

A comparative study of the physical properties of conventional and Grander-modified adhesive systems

Purpose: The objective of this study was to evaluate the influence of the Grander technology in reducing the surface tension and contact angle of a self-etch adhesive system. Methods: Distilled water and Xeno III self-etch adhesive system (Dentsply) were modified by physical contact with the Flexible unit Grander system to revitalize water, for 48 hours, resulting in four groups: Group 1 - Xeno III under normal conditions; Group 2 - Xeno III modified by Grander; Group 3 - distilled water under normal conditions; Group 4 - distilled water system modified by the Grander system;. Surface tension and contact angle (dentin substrate and titanium plate) of the adhesive system and water in normal and Grander-modified conditions was measured with a goniometer. Data were analyzed by ANOVA and Tukey test (5%). Results: Grander technology reduced the surface tension of water but not of the adhesive system Xeno III. The contact angle property of the adhesive ystem was not affected. Conclusion: Grander technology did not interfere with the surface tension and contact angle physical properties of the Xeno III self-etch adhesive system. Copyright: © 2011 Gonçalves et al.;.

Analysis of hybrid layer thickness, resin tag length and their correlation with microtensile bond strength using a total etch adhesive to intact dentin.

The objective of this study was to evaluate the use of a two-step total etch and rinse adhesive, the correlation between the hyybrid layer thickness (HL) and bond strength (BS), and between resin tag length (RT) and bond strength in the same teeth, and also to evaluate the fracture patterns of the tested specimens. Ten human molars were used for the restorative procedure and then sectioned in two halves (mesio-distally). The materials used were Adper Single Bond 2, 3M ESPE, Ultra etch gel, Ultradent and Filtek Z250, 3M ESPE. One half were utilized to measure the HL thickness and RT length through light microscopy analysis (400x), and the other half was subject to a microtensile test to measure the BS. The fractured surfaces were analyzed by scanning electron microscopy and fracture patterns classified. The Pearson correlation test was applied (p = 0.05). The results of the analyses of each specimen then were correlated: mean HL thickness = 4.39 (0.48) microm, mean length of RT = 9.94 (1.69) microm, mean BS = 23.98 (10.24) MPa. A statistically significant correlation between HL thickness and bond strength was found (r = 0.93). The two step etch and rinse adhesive system, showed a strong correlation between HL thickness and bond strength. The most common fractures were adhesive, followed by cohesive in resin.

Influence of cutting instruments and adhesive systems on hybrid layer formation.

Influence of cutting instruments and The aim of this study was to analyze the hybrid layer in noncarious dentin prepared by different cutting instruments and restored with composite resin. The cavities were randomly prepared in 40 specimens using a high-speed diamond bur (KG Sorensen 1013) and an ultrasonic tip (CVDentus C22). The cavities were restored with composite resin by varying the adhesive system between the Adper™ Single Bond (2 x 1 system, primer+adhesive) and the Prompt L-Pop™ (3 x 1 system, self-etching). The restorations were hemisected longitudinally and analyzed in the SEM (Scanning electron microscopy) in order to evaluate the hybrid layer and resinous tags characteristics, using scores ranging from 1 to 6. The Pearson test revealed a high correlation coefficient and good significance levels for both intra- and inter-raters values (r=0.90). The data were statistically analyzed using the Mann-Whitney test (P≤0.05). A larger proportion of regular hybrid layers with numerous tags were observed in the dentin prepared using the high-speed diamond burs and restored with a 2 × 1 adhesive system. Alternatively, the 3 × 1 adhesive system promoted the generation of a thin hybrid layer with few tags. After preparation using an ultrasonic tip revealed few or no tags after the preparation and 2 × 1 or 3 × 1 adhesive system application. The high-speed diamond burs produced a dentin surface that was more favorable to restorative material adhesion than the ultrasonic tips, regardless of the adhesive system used.

One-year clinical evaluation of composite restorations in posterior teeth: Effect of adhesive systems

This clinical study assessed the performance of posterior composite resins applied with the Adper™ Single Bond Plus (SB) and Adper ™ Scotchbond SE (SE) adhesive systems and Filtek ™ Supreme Plus composite resin, using modified US Public Health Service criteria. A total of 97 restorations were placed in posterior teeth by two calibrated operators. Application of the materials followed manufacturers' instructions. The restorations were evaluated by two examiners at baseline and after one year. Statistical analyses were conducted using the proportion test at a significance level of 5% (p<0.05). All the restorations evaluated (ie, 100%) received an alpha rating for the criteria of marginal discoloration and marginal integrity at baseline. At one year, for marginal discoloration, 64.6% of SB and 61.2% of SE received an alpha rating. For marginal integrity, 72.9% of SB and 77.6% of SE received an alpha rating. The other restorations received bravo ratings for both criteria. None of the teeth that received the restorative systems presented caries lesions around the restorations. A total of eight teeth presented postoperative sensitivity one week after baseline, five with SB and three with SE; the symptom had disappeared one year later. One year later, composite resin restorations using either adhesive system showed satisfactory clinical performance.

Correlation between hybrid layer thickness, resin tag length and microtensile bond strength of a self-etching adhesive system

The purpose of this study was to evaluate the correlation between the hybrid layer thickness, resin tag length and resin bond strength of a self-etching adhesive system to sound dentin tissue in vivo. After performing restorative procedures and tooth extractions, ten specimens were sectioned in a mesiodistal direction. One dental section was used for light microscope analysis, in which both the resin tag length and hybrid layer thickness were measured, while the other section was analyzed using a microtensile test (0.5 mm/min). The fractured surface of the latter section was characterized using a stereoscopic magnifying glass (40x magnifcation). The results were subject to statistical analysis using the Pearson Correlation Test (a = 0.05). The hybrid layer thickness, resin tag length and resin bond strength mean values were 2.19 microm (0.34), 4.34 microm (0.28) and 9.73 MPa (5,55), respectively. In addition, correlation tests between the resin tag length and the resin bond strength (r=0.014) and also between the hybrid layer thickness and bond strength (r=0.43), showed no statistically significant correlation. The microtensile bond strength of Adper Prompt L Pop self-etching adhesive system does not depend on hybrid layer thickness or resin tag length.

Bond strength of adhesive systems with different solvents to dry and wet dentin

Aim: This study evaluates bond strength between dentin and composite using adhesives with different solvents to dry and wet dentin. Materials and methods: Ninety bovine incisors were used; the vestibular surfaces were worn by the exposure of an area with a diameter of 4 mm of dentin. The specimens were divided into 6 groups, according to the type of adhesive used and hydratation stals: Group SB-wet: Single Bond 2 in wet dentin, Group SBdry: Single Bond 2 in dry dentin, Group SL-wet: Solobond M in wet dentin, Group SL-dry: Solobond M in dentin dry. Group XPwet: XP Bond in wet dentin, Group XP-dry: XP Bond in dentin dry. They were cut to obtain specimens in the shape of stick with 1 × 1 mm and subjected to microtensile test in universal testing machine with a cross speed of 1mm/min. The data were analyzed with ANOVA and Tukey's tests (5%). Results: ANOVA showed significant differences for surface treatment and interaction, but no difference was found for adhesive factor. The Tukey's test showed that the samples with wet dentin shown higher values of bond strength. Conclusion: The adhesive did not influence in the bond strength. The groups with wet dentin showed higher values of bond strength than groups with dry dentin.

Influence of CVD diamond tips and Er:YAG laser irradiation on bonding of different adhesive systems to dentin

Aim: The aim of this study was to compare the microtensile bond strength of three adhesive systems, using different methods of dentin preparation. Materials and methods: A hundred and eight bovine teeth were used. The dentin from buccal face was exposed and prepared with three different methods, divided in 3 groups: Group 1 (DT)- diamond tip on a high-speed handpiece; Group 2 (CVD)-CVD tip on a ultrasonic handpiece; Group 3 (LA)-Er: YAG laser. The teeth were divided into 3 subgroups, according adhesive systems used: Subgroup 1-Adper Single Bond Plus/3M ESPE (SB) total-etch adhesive; Subgroup 2-Adper Scotchbond SE/3M ESPE (AS) selfetching adhesive; Subgroup 3-Clearfil SE Bond/Kuraray (CS) selfetching adhesive. Blocks of composite (Filtek Z250-3M ESPE) 4 mm high were built up and specimens were stored in deionized water for 24 hours at 37°C. Serial mesiodistal and buccolingual cuts were made and stick-like specimens were obtained, with transversal section of 1.0 mm2. The samples were submitted to microtensile test at 1 mm/min and load of 10 kg in a universal testing machine. Data (MPa) were subjected to ANOVA and Tukey's tests (p < 0.05). Results and conclusion: Surface treatment with Diamond or CVD tips associated with Clearfil SE Bond adhesive produced significantly lower bond strength values compared to other groups. Surface treatment with Er: YAG laser associated with Single Bond Plus or Clearfil SE Bond adhesives and surface treatment with CVD tip associated with Adper Scotchbond SE adhesive produced significantly lower bond strength values compared to surface treatment with diamond or CVD tips associated with Single Bond Plus or Adper Scotchbond SE adhesives. Clinical significance: Interactions between laser and the CVD tip technologies and the different adhesive systems can produce a satisfactory bonding strength result, so that these associations may be beneficial and enhance the clinical outcomes.

Effects of dentin dry/moist condition on the immediate adhesive performance of different solvent-based etch-and-rinse adhesivesystems

Purpose: The purpose of this research was to analyze and measure, under optical microscopy, the hybrid layer thickness and resin tags length, as well as the microtensile bond strength of two conventional adhesive systems when applied to dry and moist dentinal substrate. Methods: Thirty-two extracted human molars were randomly distributed into four groups according to the adhesive systems (XP Bond and Prime&Bond 2.1) and moisture condition (dry and moist). In Groups I and II, XP adhesive system was applied on dry and moist dentin, respectively; while Groups III and IV received PB adhesive system, in the same way as was done in Groups I and II, respectively. After adhesive and restorative procedures, all specimens were sectioned along their long axes; one hemi-tooth sample was subjected to the microtensile bond strength test while the other was decalcified and serially sectioned into six micron thick slices and sequentially mounted on glass slides. These sections were stained by the Brown and Brenn method for posterior analysis and measurement of the hybrid layer and resin tags under a light microscope with a micrometric ocular 40/075. Results: Data were analyzed using two-way ANOVA and Tukey's test (α=0.05). For the variable hybrid layer thickness, XP showed no significant differences between dry and moist dentin (5.2 μm and 5.5 μm, respectively), but for PB, hybrid layer was significantly thicker for moist (4.0 μm) than for dry dentin (3.0 μm). For the variable resin tags length XP showed 17.9 μm length for dry dentin and 20.8 μm for moist dentin; PB 11.7 μm for dry and 12.69 μm for moist dentin;there was no significant differences between them, independent of the moisture condition. For the variable microtensile bond strength, XP showed 38.0 MPa for dry dentin and 44.5 MPa for moist dentin; and PB showed 22.7 MPa for dry dentin and 20.8 MPa for dry dentin no significant difference was observed between moist and dry dentin for XP (p=0.2) and PB (p=0.7), but XP was presented significantly higher bond strength values than PB in both moisture conditions (p=0.003 for dry and p=0.002 for moist). Conclusion: The two-step butanol-based etch-and-rinse adhesive XP Bond presented a superior behavior with regard to the hybrid layer thickness, length of resin tags and bond strength to dry and moist dentin substrates when compared with two-step acetone-based adhesive system Prime&Bond2.1. © 2013 Elsevier Ltd.

Effects of surface treatment, hydration and application method on the bond strength of a silorane adhesive and resin system to dentine

Objectives: To evaluate the effects of surface treatment, surface hydration (SH) and application method (AM) on the tensile bond strength of the Silorane Adhesive System (SAS) to dentine. Methods: Ninety bovine teeth were used. For the control group (n = 10), each dentine surface was treated according to the manufacturer's instructions of the SAS. The remaining teeth were randomly distributed into two groups (n = 40), according to the type of dentine surface treatment (ST) - 37% phosphoric acid or Er:YAG Laser prior to the application of the SAS. Each group was further divided into 2 subgroups (n = 20), according to the SH status: dry (D) or wet (W). Each subgroup was further divided into 2 subgroups (n = 10), according to the application method [AM: Active (AC) mode or Passive (PA) mode]. A coat of resin composite (Filtek P90) was applied on the surface. Artificial ageing was performed with a thermo-mechanical cycling machine. The specimens were sectioned into 1 mm × 1 mm × 10 mm sticks and stressed to failure using a universal testing machine. The remaining teeth in each group were used for Scanning Electron Microscopy to examine the fractured area. Data were subjected to a three-way ANOVA, Tukey's test and Dunnet's test (α = 0.05). Results: The ANOVA showed significant differences for SH and AM, but not for ST. For SH, the results of Tukey's test were (in MPa): D-14.9(±3.8)a, W-17.1(±4.3)b; and for AM: PA-14. 9(±4.2)a, AC-17.1(±3.9) b. Conclusions: Acid etching, when combined with a moist dentine surface and the use of primer agitation, improves the bond strength of the SAS to dentine. Clinical Significance: According to the results of the present in vitro study, modification of the application protocols for the silorane-based adhesive system may improve its clinical performance. © 2012 Published by Elsevier Ltd.

Durability of Adhesion between Feldspathic Ceramic and Resin Cements: Effect of Adhesive Resin, Polymerization Mode of Resin Cement, and Aging

Purpose: Adhesive cementation is an important step for restorations made of feldspathic ceramic as it increases the strength of such materials. Incorrect selection of the adhesive resin and the resin cement to adhere to the ceramic surface and their durability against aging can affect the adhesion between these materials and the clinical performance. This study evaluated the effect of adhesive resins with different pHs, resin cements with different polymerization modes, and aging on the bond strength to feldspathic ceramic. Materials and Methods: One surface of feldspathic ceramic blocks (VM7) (N = 90) (6.4 × 6.4 × 4.8 mm3) was conditioned with 10% hydrofluoric acid for 20 seconds, washed/dried, and silanized. Three adhesive resins (Scotchbond Multi-Purpose Plus [SBMP], pH: 5.6; Single Bond [SB], pH: 3.4; and Prime&Bond NT [NT], pH: 1.7) were applied on the ceramic surfaces (n = 30 per adhesive). For each adhesive group, three resin cements with different polymerization modes were applied (n = 10 per cement): photo-polymerized (Variolink II base), dual polymerized (Variolink II base + catalyst), and chemically polymerized (C&B). The bonded ceramic blocks were stored in water (37°C) for 24 hours and sectioned to produce beam specimens (cross-sectional bonded area: 1 ± 0.1 mm2). The beams of each block were randomly divided into two conditions: Dry, microtensile test immediately after cutting; TC, test was performed after thermocycling (12,000×, 5°C to 55°C) and water storage at 37°C for 150 days. Considering the three factors of the study (adhesive [3 levels], resin cement [3 levels], aging [2 levels]), 18 groups were studied. The microtensile bond strength data were analyzed using 3-way ANOVA and Tukey's post hoc test (α= 0.05). Results: Adhesive resin type (p < 0.001) and the resin cement affected the mean bond strength (p= 0.0003) (3-way ANOVA). The NT adhesive associated with the chemically polymerized resin cement in both dry (8.8 ± 6.8 MPa) and aged conditions (6.9 ± 5.9 MPa) presented statistically lower bond strength results, while the SBMP adhesive resin, regardless of the resin cement type, presented the highest results (15.4 to 18.5 and 14.3 to 18.9 MPa) in both dry and aged conditions, respectively (Tukey's test). Conclusion: Application of a low-pH adhesive resin onto a hydrofluoric acid etched and silanized feldspathic ceramic surface in combination with chemically polymerized resin cement did not deliver favorable results. The use of adhesive resin with high pH could be clinically advised for the photo-, dual-, and chemically polymerized resin cements tested. © 2012 by the American College of Prosthodontists.

Cytotoxicity of adhesive systems of different hydrophilicities on cultured odontoblast-like cells

This study evaluated the cytotoxicity of experimental adhesive systems (EASs) on odontoblast-like cells. Paper discs (n=132) were impregnated with 10 μL of each EAS-R1, R2, R3, R4, and R5 (in an ascending order of hydrophilicity), followed by photoactivation. R1 and R2 are nonsolvated hydrophobic blends, R3 represents a simplified etch-and-rinse adhesive system, and R4 and R5 represent simplified self-etch adhesive systems. Discs were immersed in Dulbecco's modified Eagle's medium for 24 h to obtain eluates applied on MDPC-23 cell cultures. No material was applied on discs used as control (R0). Cell viability [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], total protein (TP) production, alkaline phosphatase (ALP) activity, type of cell death, and degree of monomer conversion Fourier transform infrared (%DC-FTIR) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (α=0.05). Considering R0 (control) as having 100% of cell viability, R1, R2, R3, R4, and R5 reduced the metabolic activity of cells by 36.4, 3.1, 0.2, 21.5, and 65.7%, respectively, but only R1 and R5 differed from R0. Comparing with R0, lower TP production was observed for R1, R4, and R5, while ALP activity decreased for R1 and R5. Necrotic cell death was predominant for all EASs, but only R1, R4, and R5 differed from R0. Only R5 presented a different apoptotic cell death ratio from R0. R1 presented the lowest %DC (ca. 37%), whereas R4 and R5 presented the highest (ca. 56%). In conclusion, R2 and R3 were not toxic to the MDPC-23 cells, suggesting that the degree of hydrophilicity or %DC of the EASs alone were not responsible for their cytopathic effects. © 2013 Wiley Periodicals, Inc.

Transdentinal cytotoxicity of experimental adhesive systems of different hydrophilicity applied to ethanol-saturated dentin

The aim of this study was to evaluate the transdentinal cytotoxicity of experimental adhesive systems (EASs) with different hydrophilicity and dentin saturation solutions on odontoblast-like cells. One hundred 0.4-mm-thick dentin discs were mounted in in vitro pulp chambers and assigned to 10 groups. MDPC-23 cells were seeded onto the pulpal side of the discs, incubated for 48 h. The EASs with increasing hydrophilicity (R1, R2, R3 and R4) were applied to the occlusal side after etching and saturation of etched dentin with water or ethanol. R0 (no adhesive) served as controls. R1 is a non-solvated hydrophobic blend, R2 is similar to a simplified etch-and-rinse adhesive system and R3 and R4 are similar to self-etching adhesives. After 24 h, cell metabolism was evaluated by MTT assay (n = 8 discs) and cell morphology was examined by SEM (n = 2 discs). Type of cell death was identified by flow cytometry and the degree of monomer conversion (%DC) was determined by infrared spectroscopy (FTIR) after 10 s or 20 s of photoactivation. Data were analyzed by the Kruskal-Wallis and Mann-Whitney tests (α = 0.05). Dentin saturation with ethanol resulted in higher necrotic cell death ratios for R2, R3 and R4 compared with water saturation, although R2 and R3 induced higher SDH production. Photoactivation for 20 s significantly improved the %DC of all EASs compared with 10 s. A significant positive correlation was observed between the degree of hydrophilicity and %DC. In conclusion, except for R1, dentin saturation with ethanol increased the cytotoxicity of EASs, as expressed by the induction of necrotic cell death. © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Avaliação da eficiência de adesão de um cimento resinoso autocondicionante (self-adhesive) sobre esmalte e dentina: efeito termociclagem e condicionamento dental : estudo in vitro

Pós-graduação em Odontologia Restauradora - ICT