Effect of silver diamine fluoride on the longevity of the bonding properties to caries-affected dentine.

OBJECTIVES: This study aimed to evaluate the adhesive properties in dentine after the application of silver diamine fluoride (SDF) on carious dentine lesions immediately and after 2 years of water storage. METHODS: 96 human molars used were subjected to artificial dentine caries production, and then randomly divided into 12 experimental groups according to 1. application of an SDF solution (carious dentine lesion without SDF treatment [control], with 12 % silver diamine fluoride [SDF 12 %] or 38 % silver diamine fluoride [SDF 38 %]); 2. Universal adhesives (Clearfil Universal Bond Quick [CUQ] and Single Bond Universal [SBU]); 3. adhesive strategy (etch-and-rinse [ER] and self-etch [SE]). After restoration, the specimens were sectioned and submitted to the microtensile bond strength test (µTBS) and energy-dispersive X-ray spectrometry analysis (SEM/EDX). All tests were performed immediately and after 2 years of water storage. Data from the µTBS were analyzed using four-way ANOVA and Tukey's test (α = 0.05). RESULTS: Only the interaction of factors 'SDF' vs 'time' was significant (p = 0.03). After 2 years of storage, the groups where SDF was applied showed higher µTBS values compared to the control group. No significant decrease in µTBS values was observed for SBU when comparing immediate and 2-year results, but a significant reduction in µTBS values was observed after 2 years for CUQ. CONCLUSION: Independent of the adhesive strategy, the use of SDF may be a promising alternative to maintain the bonding of universal adhesives to carious dentinal lesions. CLINICAL SIGNIFICANCE: This study may clarify and support clinicians regarding the longevity of resin-based restoration in caries-affected dentine treated with silver diamine fluoride.

Effect of the Absence of HEMA on the Bonding Properties of Universal Adhesive Systems Containing 10-MDP: An In Vitro Study.

OBJECTIVES: To evaluate the absence of 2-hydroxyethyl methacrylate (HEMA) on the adhesive properties with enamel and dentin of universal adhesive systems containing 10-methacryloyloxydecyl dihydrogen phosphate (MDP). METHODS AND MATERIALS: One hundred and twelve caries-free third molars were used to test adhesion to dentin (n=64) and enamel (n=48). For each substrate, teeth were divided into eight experimental groups: four different adhesives each using two adhesive strategies. The adhesives used were: (1) Scotchbond Universal (SBU, 3M Oral Care, St Paul, MN, USA) as a HEMA-containing universal adhesive; (2) Gluma Bond Universal (GBU, Kulzer, Hanau, Germany); (3) Solare Universal Bond (SUB, GC, Tokyo, Japan); and (4) Zipbond Universal (ZIP, SDI, Victoria, Australia) as HEMA-free universal adhesives. The adhesive strategies used were etch-and-rinse (ER) and self-etch (SE). For dentin tests, the occlusal third of the crown of all teeth was removed and an adhesive protocol was applied. After completing the restoration, specimens were sectioned into bonded sticks (0.8 mm 2) and tested for microtensile bond strength (µTBS), in situ degree of conversion (DC), and nanoleakage (NL) by scanning electron microscopy. For enamel tests, teeth were sectioned into four parts (buccal, lingual, and proximal), and an adhesive protocol was applied. After completing the restoration, the specimens were tested for their microshear bond strength (µSBS). For in situ degree of conversion (DC) and nanoleakage (NL) evaluation of enamel, the specimens were sectioned in slices to be evaluated. The data for each substrate were subjected to two-way ANOVA and Tukey's test (α=0.05) for each property evaluated. RESULTS: The SBU and ZIP adhesives showed the highest µSBS, and DC (dentin and enamel) and lower NL (dentin) values compared to GBU and SUB (p=0.001). However, SBU showed better results in terms of µTBS and µSBS (SE strategy), and DC (dentin and enamel) than ZIP. Strategy ER presented higher values of µTBS and µSBS when compared to strategy SE (p=0.001), except for SBU. CONCLUSION: The effect of the absence of HEMA in commercial universal adhesive systems on enamel and dentin adhesive properties appears to be material-dependent.

Are universal adhesives in etch-and-rinse mode better than old 2-step etch-and-rinse adhesives? One-year evaluation of bonding properties to dentin.

OBJECTIVES: This study compared the bonding properties of dentin of three 2-step etch-and-rinse adhesives (2-ERAs) to those of three universal adhesives (UAs) applied with an etch-and-rinse strategy (ER), immediately and after 1 year of water storage. METHODS: Sixty caries-free molars were divided into 6 groups according to the adhesive systems used (n = 10). The 2-ERA systems included were: 1) Adper Single Bond 2 (SB), 2) Tetric N-Bond (TB), and 3) Ambar (AM); and the UAs systems were: 4) Single Bond Universal (SBU) 5) Tetric N-Bond Universal (TBU), and 6) Ambar Universal (AMU). The occlusal third of each tooth was removed and the adhesives were applied. After the composite build-up, specimens were sectioned and tested for microtensile bond strength (µTBS) and nanoleakage (NL) immediately and after 1 year of water storage. In situ degree of conversion (DC) was only evaluated in the immediate time. For water sorption (WS), solubility (SO), and mass change (MC) tests, 48 disk-shaped specimens were prepared (n = 8) and assessed according to ISO 4049:2009. RESULTS: UAs showed higher µTBS and lower NL values than 2-ERAs did after 1 year of water storage (p = 0.001). Regarding DC, 2-ERAs showed higher DC values than UAs (p = 0.001). Regarding WS, 2-ERAs showed higher WS values than those of UAs (p = 0.00001), except for AM and AMU. Lower WS was observed for AM than for other 2-ERAs (p = 0.00001). CONCLUSION: The use of UAs applied with the ER strategy seems to be a more effective technique for maintaining adhesion to dentin substrate than 2-ERAS. CLINICAL SIGNIFICANCE: This study may support clinicians in selecting the most adequate adhesive system to be used in ER strategy in dentin, demonstrating that UAs were more effective, considering the longevity of the resin restorations.

Inactivation of human immunodeficiency virus type 1 in blood samples stored as high-salt lysates.

Blood samples to be tested for the presence of parasite DNA by using specific DNA probes are routinely stored in our laboratory as high-salt lysates (HSL). To safeguard against the risk of accidental infection with etiological agents such as the human immunodeficiency virus type 1 (HIV-1) while manipulating large numbers of blood samples in preparation for DNA probing, we determined the residual infectivity of HIV-1 after exposure to HSL components. Both high-titer virus stocks or provirus-carrying cells, suspended either in tissue culture medium or freshly drawn blood, were completely inactivated upon contact with the HSL components. This was verified by the absence of any detectable HIV-1-specific antigen in the supernatants of long-term cultures and the absence of virus-specific DNA fragments after amplification by polymerase chain reaction with DNA from such cultures as target DNA. These results support the conclusion that the virus is in fact completely inactivated by contact with the HSL components, rendering blood specimens stored as HSL noninfectious in regard to HIV-1.

High salt lysates: a simple method to store blood samples without refrigeration for subsequent use with DNA probes.

Blood specimens to be tested for the presence of Plasmodium falciparum using specific DNA probes can be stored as high salt lysates (HSL) without refrigeration. The lysates are prepared from 100 microliter blood samples by a simple 3-step procedure using 2 volumes of H2O to lyse the erythrocytes (step I), 1 volume of a detergent/EDTA mix to lyse the parasites (step II), followed by the addition of 1 volume cesium trifluoroacetate (CsTFA) (step III). The parasite DNA was found to be undegraded, as shown by the unaltered pattern of repetitive sequences obtained after storage of up to 1 month at 37 degrees C, due to the inhibition of DNA degrading enzymes by the cesium salt. The bulk of protein can be removed from the samples by a 1-step precipitation. The addition of 0.3 volumes of a mixture of ethanol: chloroform: isoamyl alcohol (2.5:1:0.04 v/v) precipitates greater than 90% of the proteins from the lysates, leaving greater than 86% of the parasite DNA in the supernatant. The reduced protein content of the samples, when applied to solid supports, results in an increased signal: background ratio on autoradiograms.

Kinetic and molecular properties of the dihydrofolate reductase from pyrimethamine-sensitive and pyrimethamine-resistant clones of the human malaria parasite Plasmodium falciparum.

Dihydrofolate reductase (DHFR) (5,6,7,8-tetrahydrofolate: NADPH+-oxidoreductase; EC 1.5.1.3) was partially purified by affinity chromatography from three clones of the human malaria parasite Plasmodium falciparum. The three clones were representative of pyrimethamine-sensitive (clone 3D7) and pyrimethamine-resistant (clone HB3 and clone 7G8) parasites with ID50 values of 0.53 nM (3D7), 210 nM (HB3), and 540 nM (7G8), when tested in vitro against the drug. The specific activities of the partially purified DHFR differed by less than a factor of 2 between the sensitive clone 3D7 (442 +/- 39 nmol min-1 mg-1 protein) and the resistant clones HB3 (634 +/- 25 nmol min-1 mg-1 protein) and 7G8 (565 +/- 85 nmol min-1 mg-1 protein). The number of catalytic sites in partially purified DHFR from the three clones was similar and ranged from 151 to 194 pmol mg-1 protein. The Km value for NADPH was similar in all three clones (4.5-11.6 microM). The Km value for dihydrofolate was altered 13-fold comparing the sensitive clone 3D7 (3.2 +/- 0.6 microM) with the resistant clone HB3 (42.6 +/- 1.6 microM), with the Km for the resistant clone 7G8 falling in between (11.9 +/- 1.2 microM). The inhibition constants for pyrimethamine increased from 0.19 +/- 0.08 nM (3D7) to 2.0 +/- 0.3 nM (HB3) to 8.9 +/- 0.8 nM (7G8). The inhibition by pyrimethamine of the sensitive clone 3D7 was noncompetitive and competitive for the two other clones. The titration of partially purified DHFR with pyrimethamine revealed a 500-fold increase in the concentration of the drug needed to inhibit the DHFR activity by 50%, when the sensitive clone 3D7 (0.18 +/- 0.02 nM) was compared to the resistant clone 7G8 (95 +/- 16 nM). From the comparison of the specific activities and the catalytic center activities with the Km values for the substrate and the inhibition constants for pyrimethamine, both of which are altered in the resistant clones, we conclude that the molecular mechanism for pyrimethamine resistance in the three clones studied is not based on an overproduction of the DHFR but is due to a decreased affinity to antifolates by a structurally altered enzyme.