In vitro antimicrobial activity of endodontic sealers, MTA-based cements and Portland cement.

The aim of this study was to evaluate the antimicrobial activity of different root-end filling materials - Sealer 26, Sealapex with zinc oxide, zinc oxide and eugenol, white and gray Portland cement, white and gray MTA-Angelus, and gray Pro Root MTA - against six different microorganism strains. The agar diffusion method was used. A base layer was made using Müller-Hinton agar (MH) and wells were formed by removing the agar. The materials were placed in the wells immediately after manipulation. The microorganisms used were: Micrococcus luteus (ATCC9341), Staphylococcus aureus (ATCC6538), Escherichia coli (ATCC10538), Pseudomonas aeruginosa (ATCC27853), Candida albicans (ATCC 10231), and Enterococcus faecalis (ATCC 10541). The plates were kept at room temperature for 2 h for prediffusion and then incubated at 37 degrees C for 24 h. Triphenyltetrazolium chloride 0.05% gel was added for optimization, and the zones of inhibition were measured. Data were subjected to the Kruskal-Wallis and Dunn tests at a 5% significance level. The results showed that all materials had antimicrobial activity against all the tested strains. Analysis of the efficacy of the materials against the microbial strains showed that Sealapex with zinc oxide, zinc oxide and eugenol and Sealer 26 created larger inhibition halos than the MTA-based and Portland cements (P < 0.05). On the basis of the methodology used, it may be concluded that all endodontic sealers, MTA-based and Portland cements evaluated in this study possess antimicrobial activity, particularly the endodontic sealers.

Influence of cavity preparation and curing method on the marginal seal of resin composite restorations: an in vitro evaluation.

OBJECTIVE: To evaluate the influence of cavity design and photocuring method on the marginal seal of resin composite restorations. METHOD AND MATERIALS: Seventy-two bovine teeth were divided into 2 groups: group 1 received box-type cavity preparations, and group 2 received plate-type preparations. Each group was divided into 3 subgroups. After etching and bonding, Z250 resin composite (3M Espe) was applied in 2 equal increments and cured with 1 of 3 techniques: (1) conventional curing for 30 seconds at 650 mW/cm2; (2) 2-step photocuring, in which the first step was performed 14 mm from the restoration for 10 seconds at 180 mW/cm2 and the second step was performed in direct contact for 20 seconds at 650 mW/cm2; or (3) progressive curing using Jetlite 4000 (J. Morita) for 8 seconds at 125 mW/cm2 and then 22 seconds at 125 mW/cm2 up to 500 mW/cm2. The specimens were thermocycled for 500 cycles and then submitted to dye penetration with a 50% silver nitrate solution. Microleakage was assessed using a stereomicroscope. Data were analyzed using analysis of variance and Tukey test (5% level of significance). RESULTS: A statistically significant difference was found between groups when a double interaction between photocuring and cavity preparation was considered (P = .029). CONCLUSIONS: No one type of cavity preparation or photocuring method prevented micro-leakage. The plate-type preparation showed the worst dye penetration when conventional and progressive photocuring methods were used. The best results were found using the 2-step photocuring with the plate-type preparation.

Pro- and anti-inflammatory cytokines produced by human monocytes challenged in vitro with Paracoccidioides brasiliensis

Monocytes and macrophages play a central role in innate and adaptive immune response against systemic fungal infections. Imbalances in suppressor or stimulatory cytokine secretion caused by these cells may influence disease development, microorganism death, and the nature of the adaptive immune response. This study analyzed the monocyte cytokine profiles of healthy individuals challenged with high and low virulent strains of P. brasiliensis and mRNA cytokine expression kinetics by reverse transcription polymerase chain reaction (RT-PCR). Peripheral blood monocytes from healthy volunteers were cultured in vitro with and without virulent (Pb18) or low virulence (Pb265) strains from P. brasiliensis viable yeast cells. Interleukin-1 beta (IL-1β), IL-6, IL-8, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor-beta (TGF-β1) were measured in culture supernatants by enzyme immunoassay (ELISA), and mRNA cytokine expression was determined by RT-PCR at 0, 4, 8, 12, 18 and 48 hr. Both P. brasiliensis strains induced monocyte production of IL-1β, IL-6, IL-10 and TNF-α. Pb18 induced higher levels of IL-1β, IL-6, and IL-10 than Pb265. IL-8 and TGF-β1 levels were not significantly different from those cultured without stimulus. The mRNA cytokine expression was similar to supernatant cytokines measured by ELISA. In vitro monocyte challenge with virulent P. brasiliensis strain induces earlier and higher levels of pro- and anti-inflammatory cytokines than low virulence strain.

In vitro evaluation of surface roughness of 4 resin composites after the toothbrushing process and methods to recover superficial smoothness.

OBJECTIVE: This study evaluated the efficiency of repolishing, sealing with surface sealant, and the joining of both in decreasing the surface roughness of resin-based composites after a toothbrushing process. METHOD AND MATERIALS: Ten specimens of each composite (Alert, Z100, Definite, and Prodigy Condensable), measuring 2 mm in thickness and 4 mm in diameter, were made and submitted to finishing and polishing processes on both sides of the specimens using the Sof-Lex system. The specimens were then subjected to toothbrushing (30,000 cycles), and surface roughness (Ra) was analyzed with a Surfcorder SE 1700 profilometer. The upper surface of each composite was etched with 37% phosphoric acid, and the surface-penetrating sealant Protect-it was applied on 1 surface. The roughness of these surfaces was again measured. On the other side, the surface of the specimen was repolished, and the efficiency of this procedure was measured using the profilometer. The surface roughness resulting from the joining of the 2 methods was verified by applying, in the final stage, the surface-penetrating sealant on the repolished surface. Data were analyzed with analysis of variance and Tukey test (P <.05). RESULTS: Results showed that the lowest surface roughness values were obtained for Definite, Z100, and Prodigy Condensable after the repolishing process and after the repolishing plus sealing. For Alert, the joining of repolishing plus sealing promoted the lowest values of surface roughness. CONCLUSION: Of the resin-based composites, Alert demonstrated the highest values of surface roughness in all the techniques tested.

In vitro evaluation of marginal leakage in bonded restorations, with mechanical or chemical-mechanical (Carisolv) removal of carious tissue

This study carried out an in vitro evaluation and comparison of the occurrence of marginal leakage in bonded restorations using mechanical or chemical-mechanical (Carisolv) removal of carious tissue. For that purpose, 40 extracted decayed human molars were divided into 4 groups: GI (burs + Prime & Bond NT + TPH), GII (Carisolv + Prime & Bond NT + TPH), GIII (burs + SBMP + Z100) and GIV (Carisolv + SBMP + Z100). After accomplishment of the restorations and thermal cycling, the teeth were exposed to dye, sectioned and qualitatively evaluated. The results demonstrated that the system of removal of carious tissue did not influence the results of microleakage at any of the cavity margins. At dentinal margins, use of the Prime & Bond NT + TPH restorative system allowed the occurrence of less microleakage than the SBMP + Z100 system.

In vitro antimicrobial efficiency of a mouthwash containing triclosan/gantrez and sodium bicarbonate

Several antiseptic substances have been used as adjuncts to routine mechanical procedures of oral hygiene, based on their antimicrobial effects. The objective of this study was to assess in vitro the antimicrobial efficiency of 2 mouthwash containing Triclosan/Gantrez and sodium bicarbonate in comparison to both positive and negative controls. Standard strain samples of Escherichia coli, Pseudomonas aeruginosa, Actinomyces viscosus and Bacillus subtilis were used. Samples of Streptococcus mutans and Gram-negative bacilli were collected from 20 volunteers (10 with a clinically healthy periodontium and 10 presenting biofilm-associated gingivitis). Evaluation of the antimicrobial activity was performed by determining the minimal inhibitory concentration (MIC). The results indicated that the test solution inhibited the growth of both Gram-negative and Gram-positive microorganisms from the volunteers' saliva as well as that of the standard strains at the MIC dilution of 1:20, whereas the MIC dilution of 0.12% chlorhexidine against the same bacteria was 1:80. Thus, even though the tested mouthrinse solution presented an in-vitro antimicrobial activity superior to that of a placebo, it was inferior to that of chlorhexidine.

In vitro evaluation of the effect of natural orange juices on dentin morphology

The patient's diet has been considered an important etiological factor of dentin hypersensitivity. The frequent ingestion of acidic substances can promote the loss of dental structure or remove the smear layer. The purpose of this study was to evaluate the degree of smear layer removal and dentinal tubules exposure by different natural orange juices. Extracted human teeth were submitted to manual scaling in order to develop the smear layer. Seventy dentin samples were obtained and distributed into the following groups: Control, lime orange, lime, valência orange, navel orange, mandarin, and tangerine. Each group included 2 methods of application: Topical and topical + friction. After preparation for SEM analysis, photomicrographs were assessed by a blind calibrated examiner using an index system. The Kruskal-Wallis test indicated a significant influence of the orange juices on smear layer removal. Significant difference was observed between navel orange, valência orange, mandarin and the control group (p < 0.05). These orange juices resulted in greater removal of the smear layer and greater opening of dentinal tubules. The comparison between the application methods for each group using the Mann-Whitney test showed that friction increased smear layer removal significantly only for lime orange and lime. The data suggest that certain natural orange juices are more effective in terms of smear layer removal and dentinal tubules exposure than others.

In vitro antimicrobial activity of acroseal, polifl and epiphany against Enterococcus faecalis

Using the agar diffusion method, this study evaluated the in vitro antimicrobial activity of the commercial endodontic sealers Acroseal and Epiphany, a castor-oil based experimental sealer, Polifl, and a primer agent (Epiphany self-etching primer), against Enterococcus faecalis. Zinc oxide and eugenol cement (ZOE) served as control. Five wells per dish were made at equidistant points and immediately flled with the test and control materials. After incubation of the dishes at 37°C for 24 h and 48 h, the diameter of the zones of microbial growth inhibition produced around the wells was measured (in mm) with a millimeter rule. After 48 h, the diameters of the zones of microbial growth inhibition were the same as those observed at 24 h, only the substances continued to diffuse. Epiphany and Polifl did not show antibacterial activity (no formation of zones of microbial growth inhibition). The primer produced the largest zones of inhibition (17.62 mm) followed by Acroseal (7.25 mm) and ZOE (7.12 mm). E. faecalis was resistant to Epiphany and Polifl, while the primer and Acroseal sealer were effective against this microorganism under the tested conditions.

Characterization and in vitro cytocompatibility of an acid-etched titanium surface

The aims of this study were to characterize the microstructure of a commercially pure titanium (cpTi) surface etched with HCl/H 2SO 4 (AE-cpTi) and to investigate its in vitro cytocompatibility compared to turned cpTi (T-cpTi). T-cpTi showed a grooved surface and AE-cpTi revealed a surface characterized by the presence of micropits. Surface parameters indicated that the AE-cpTi surface is more isotropic and present a greater area compared to T-cpTi. The oxide film thickness was similar between both surfaces; however, AE-cpTi presented more Ti and O and less C. Osteoblastic cell proliferation, alkaline phosphatase activity, and bone-like nodule formation were greater on T-cpTi than on AE-cpTi. These results show that acid etching treatment produced a surface with different topographical and chemical features compared to the turned one, and such surface modification affected negatively the in vitro cytocompatibility of cpTi as demonstrated by decreasing culture growth and expression of osteoblastic phenotype.

In vitro evaluation of the effect of dietary acids and toothbrushing on human root dentin permeability

OBJECTIVES: The purpose of this in vitro study was to quantify the alterations in human root dentin permeability after exposure to dietary acids and to evaluate the effect of toothbrushing after acid application. METHOD AND MATERIALS: Extracted human third molars had their crowns sectioned above the CEJ, pulp tissue removed, and cervical root dentin exposed using a high-speed bur (approximately 1 mm in depth of substance loss). From each root fragment, one specimen was prepared. A total of 25 specimens were used and distributed randomly into five groups. The specimens were attached to a hydraulic pressure apparatus to evaluate the alterations of root dentin permeability after exposure to different acids. Dentin permeability was measured after the following sequential steps: (1) treatment with EDTA for 3 minutes to obtain the maximum permeability; (2) root planing to create a smear layer; (3) exposure to different acidic substances for 5 minutes (vinegar, cola drink, lemon juice, white wine, and orange juice); and (4) brushing for 3 minutes. RESULTS: All acidic substances increased dentin permeability after root planing. Lemon juice produced higher values for permeability when compared to the other substances (P = .009); moreover, orange juice showed similar results (P < .02) except when compared to vinegar (P = .12). Brushing right after acid exposure significantly reduced dentin permeability except in the vinegar group (P = .07). CONCLUSION: Under the experimental conditions, dietary acids increased root dentin permeability, and immediate brushing reduced permeability levels.

Effect of an acid diet allied to sonic toothbrushing on root dentin permeability: An in vitro study

This study quantified alterations in root dentin permeability after exposure to different acid beverages. Twenty-five third molars were sectioned below the cementoenamel junction, the root segment was collected, and the pulp tissue was removed. The root segments were connected to a hydraulic pressure apparatus to measure the permeability of root dentin after the following sequential steps, with 5 specimens in each: 1) phosphoric acid etching for 30 s (maximum permeability), 2) root planning to create new smear layer, 3) exposure to different acid substances for 5 min (orange, cola drink, vinegar, white wine, lemon juice), 4) toothbrushing with sonic toothbrush for 3 min, 5) toothbrushing with sonic toothbrush plus dentifrice for 3 min. Considering step I as 100%, the data were converted into percentage and each specimen was its own control. Data were analyzed statistically by Kruskal-Wallis and Dunn's post test at 5% significance level. All acidic substances increased dentin permeability significantly after scraping (p<0.05). Toothbrushing after exposure to acid substances decreased dentin permeability and the association with dentifrice accentuated the decrease (p<0.05), except for the specimens treated with cola drink. Thus, it may be concluded that all tested acid fruit juices increased dentin permeability, and toothbrushing with or without dentifrice can decrease root dentin permeability after dentin exposure to acid diet.

The in vitro antimicrobial activity of natural infant fluoride-free toothpastes on oral micro-organisms

Purpose: The objective of this study was to evaluate the antimicrobial activity of six toothpastes for infants: 3 fluoride-free experimental toothpastes - cashew-based, mango-based and without plant extract and fluoride compared with 2 commercially fluoride-free toothpastes and 1 fluoridated toothpastes. Methods: Six toothpastes for infants were evaluated in this study: (1) experimental cashew-based toothpaste; (2) experimental mango-based toothpaste; (3) experimental toothpaste without plant extract and fluoride (negative control); (4) First Teeth brand toothpaste; (5) Weleda brand toothpaste; and (6) Tandy brand toothpaste (positive control). The antimicrobial activity was recorded against Streptococcus mutans, Streptococcus sobrinus, Lactobacillus acidophilus, and Candida albicans using the agar plate diffusion test. Results: First Teeth, Weleda, mango-based toothpaste, and toothpaste without plant extract presented no antimicrobial effect against any of the tested micro-organisms. Cashew toothpaste had antimicrobial activity against S mutans, S sobrinus, and L acidophilus, but it showed no antimicrobial activity against C albicans. There was no statistical difference between the inhibition halo of cashew and Tandy toothpastes against S mutans and L acidophilus. Conclusions: Cashew fluoride-free toothpaste had inhibitory activity against Streptococcus mutans and Lactobacillus acidophilus, and these results were similar to those obtained for fluoridated toothpaste.

Blood cells attachment after root conditioning and PRP application: An in vitro study

Aim: Root conditioning is aimed at smear layer removal and at dental matrix collagen exposure, which may promote periodontal regeneration. This in vitro study assessed smear layer removal, collagen fiber exposure and the influence of PRP (platelet-rich plasma) application on adhesion of blood cells to the root surface using scanning electron microscopy (SEM). Materials and methods: Scaled root samples (n = 160) were set in five groups and conditioned with: group I - control group (saline solution); group II (EDTA 24%); group III (citric acid 25%); group IV (tetracycline hydrochloride 50 mg/ml); group V (sodium citrate 30%). Eighty samples were assessed using the root surface modification index (RSMI). The other eighty samples were set in two groups. The first group (n = 40) received PRP gel application with a soft brush and the second group (n = 40) received PRP application and then a blood drop. The fibrin clot formation was assessed in the first group and the blood cells adhesion was assessed in the second group using the BEAI (blood elements adhesion index). A previously trained, calibrated, and blind examiner evaluated photomicrographs. Statistical analysis was performed using the Kruskal-Wallis's and Dunn's tests. Results: Group III attained the best results for RSMI and BEAI. Moreover, it was the only group showing fibrin clot formation. Conclusion: Citric acid was the most efficient conditioner for smear layer removal, collagen fiber exposure and blood cell adhesion. Moreover, it was the only group showing fibrin clot formation after PRP application. Clinical significance: This study demonstrated that root conditioning followed by PRP application may favor blood cell adhesion on root surface which may optimize periodontal healing.

In vitro susceptibility of oral Candida albicans strains to different pH levels and calcium hydroxide saturated aqueous solution

Candida albicans is present in the oral cavity and in the whole digestive tract of humans and other animals, being frequently related to endodontic treatment failure. The present study determined the incidence of C. albicans in the oral cavity and the susceptibility of isolates to different pH values and saturated calcium hydroxide aqueous solution at pH 12.5. Sixty-five patients attending the Endodontic Clinic at the Sagrado Coração University participated in the study. The collected samples were cultivated in selective media for C. albicans and the isolates were tested in terms of resistance to both alkaline pH and saturated aqueous solution of calcium hydroxide. In relation to time variables, yeast viability was assessed by the Sabouraud's agar culture and fluorescein diacetate and ethidium bromide fluorescent staining method. Results from the different pHs and experimental times, including those from different techniques measuring fungal viability, were compared using the chi-square and Fisher's exact tests (α=0.05). The yeasts became completely inviable after 48 h of contact with the calcium hydroxide solution. On the other hand, when exposed to the alkaline culture broth, the yeasts were found to be viable at pHs 9.5 and 10.5 for up to 7 days. In conclusion, C. albicans can only be completely inhibited by direct contact with saturated calcium hydroxide aqueous solution after 48 h of exposure.

Agonistas do GnRH, antagonistas do GnRH e a reprodução assistida: Análogos do GnRH x fertilização in vitro

The agonists of gonadotropin-releasing hormone (GnRH) were introduced in ovarian stimulation for in vitro fertilization to avoid a premature surge of luteinizing hormone. Although they are accompanied by some disadvantages, GnRH agonists have become well accepted in clinical practice, and their use is associated with increased rates of pregnancy. The development of GnRH antagonists capable of blocking the pituitary immediately offered a therapeutic option. Comparative studies between the two analogs have suggested that the use of antagonists is associated with a shorter duration of ovulatory stimulus and a decreased incidence of ovarian hyperstimulation syndrome, while the type of GnRH analogues used show no significant effects on the rates of pregnancy and live birth. However, GnRH agonists have other applications in assisted reproductive technology cycles than the pituitary downregulation.