Edvaldo Antonio Ribeiro Rosa

Relationships between subgingival microbiota and GCF biomarkers in generalized aggressive periodontitis.

AIM To examine relationships between subgingival biofilm composition and levels of gingival crevicular fluid (GCF) cytokines in periodontal health and generalized aggressive periodontitis (GAP). MATERIALS AND METHODS Periodontal parameters were measured in 25 periodontally healthy and 31 GAP subjects. Subgingival plaque and GCF samples were obtained from 14 sites from each subject. Forty subgingival taxa were quantified using checkerboard DNA-DNA hybridization and the concentrations of eight GCF cytokines were measured using Luminex. Cluster analysis was used to define sites with similar subgingival microbiotas in each clinical group. Significance of differences in clinical, microbiological and immunological parameters among clusters was determined using the Kruskal-Wallis test. RESULTS GAP subjects had statistically significantly higher GCF levels of interleukin-1beta (IL-1beta) (p<0.001), granulocyte-macrophage colony-stimulating factor (GM-CSF) (p<0.01) and IL-1beta/IL-10 ratio (p<0.001) and higher proportions of Red and Orange complex species than periodontally healthy subjects. There were no statistically significant differences in the mean proportion of cytokines among clusters in the periodontally healthy subjects, while the ratio IL-1beta/IL-10 (p<0.05) differed significantly among clusters in the aggressive periodontitis group. CONCLUSIONS Different subgingival biofilm profiles are associated with distinct patterns of GCF cytokine expression. Aggressive periodontitis subjects were characterized by a higher IL-1beta/IL-10 ratio than periodontally healthy subjects, suggesting an imbalance between pro- and anti-inflammatory cytokines in aggressive periodontitis.

Improvement of XTT assay performance for studies involving Candida albicans biofilms

2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay has been used to study Candida biofilm formation. However, considering that the XTT reduction assay is dependent on cell activity, its use for evaluating mature biofilms may lead to inaccuracies since biofilm bottom cell layers tend to be relatively quiescent at later stages of biofilm formation. The aim of this study was to improve XTT reduction assay by adding glucose supplements to the standard XTT formulation. Candida albicans ATCC 90028 was used to form 24-, 48- and 72-h biofilms. The oxidative activity at 90, 180 and 270 min of incubation was evaluated. The control consisted of standard XTT formulation without glucose supplements, and was modified by the addition of 50, 100 and 200 mM of glucose. The XTT assay with 200 mM glucose showed more accurate and consistent readings correlating with biofilm development at 24, 48 and 72 h. Biofilm growth yield after 180 min incubation, when evaluated with the 200 mM glucose supplemented XTT, produced the most consistent readings on repetitive testing. It may be concluded that glucose supplementation of XTT could minimize variation and produce more accurate data for the XTT assay.

Hospital gowns as a vehicle for bacterial dissemination in an intensive care unit

The microbiota from the uniforms of 31 professionals from the general intensive care unit was analyzed. The samples were collected in duplicate at the beginning and at the end of the work period. Total viable counts of microorganisms were determined; there was a significant increase in the counts at the end of the period, when compared with those obtained at the beginning. No significant difference was observed between the first and second counts obtained from the cuffs. However, differences were observed for the samples from the abdominal region. Among the isolated pathogens 11/18 were Staphylococcus aureus, 2/18 were Acinetobacter baumannii, 2/18 were Klebsiela pneumoniae and 1/18 were Serratia rubidae. Some of these isolates were multi-resistant to antibiotics. Emphasis should be placed on reducing the spread of these pathogens in the hospital, making sure that biosafety protocols are followed by the staff.

Sliding resistance with esthetic ligatures: an in-vitro study.

INTRODUCTION This study was developed to evaluate in vitro the properties related to sliding resistance of esthetic ligatures. METHODS Frictional force of 6 ligatures--2 conventional, 2 specially coated elastomeric, Teflon-coated (Dupont, Wilmington, Del) stainless steel, and stainless steel (control) ligatures--were studied by sliding 0.019 x 0.025-in stainless steel wire through the 0.22-in slot of stainless steel bracket. Elastomeric ligatures were tested for frictional and tensile forces under 3 experimental conditions: recent stretching, after 21 days of simulated stretching in artificial saliva, and a demineralizing/remineralizing regimen. Statistical analysis was conducted with ANOVA and Games-Howell tests. RESULTS There was high correlation between frictional and tensile forces of elastomeric ligatures, with reduction of both after 21 days. The demineralizing/remineralizing regimen reduced the frictional forces of ligatures to the same level as the ligatures in artificial saliva. Teflon-coated and stainless steel ligatures showed the lowest initial frictional forces, but there was no difference in friction of stainless steel and post-stretched elastomeric ligatures. CONCLUSIONS Frictional forces generated by esthetic elastomeric ligatures under simulated oral environments are not stable and are more related to tensile force than to surface characteristics of the ligatures.

Influence of cigarette smoke condensate on cariogenic and candidal biofilm formation on orthodontic materials

INTRODUCTION An experimental analysis was made to quantify the adherence rates and the biofilm formation capacity of Streptococcus mutans ATCC25175 and Candida albicans SC5314 on orthodontic material surfaces in the presence of cigarette smoke condensate (CSC). METHODS Metal brackets, bands, acrylic resin, and polyurethane elastic rings were coated with stimulated saliva and submitted to adhesion and biofilm formation tests with and without CSC in a dynamic system. RESULTS The CSC increased the adhesion of S mutans ATCC25175 to the acquired pellicle (P <0.05) for bands (4.08 times), acrylic resin (2.89 times), and brackets (3.37 times) and reduced it in polyurethane elastic (2.66 times; P <0.05). S mutans ATCC25175 biofilm biomass was increased by CSC only on brackets (1.60 times; P <0.05). In the presence of CSC, the adhesion of C albicans SC5314 increased (P <0.05) on bands (1.81 times), brackets (9.61 times), elastics (29,133 times), and acrylic resin (177 times). Greater formation of C albicans SC5314 biofilm caused by CSC (P <0.05) was observed on acrylic resin (2.13 times) and brackets (2.32 times). CONCLUSIONS The results indicated that cigarette tobacco smoke can interfere with the adhesion and biofilm formation of these microorganisms to various orthodontic materials.

Reaction kinetics of sodium ascorbate and dental bleaching gel.

OBJECTIVE The aim of this study was to establish the reaction kinetics of 35% hydrogen peroxide and sodium ascorbate and to determine the mass of antioxidant required to neutralize the bleaching gel. METHODS The method used to quantify sodium ascorbate was based on the United States Pharmacopeia (1995)(26). Oxidation-reduction titration was used to confirm the concentration of hydrogen peroxide and sodium ascorbate and to determine the reaction kinetics between them. RESULTS The results indicated a direct correlation between the mass of hydrogen peroxide and that of the antioxidant agent. In addition, 5 min of contact was sufficient to neutralize the hydrogen peroxide used. CONCLUSION This in vitro study showed that the amount of sodium ascorbate required for reduction of hydrogen peroxide is directly related to the concentration of the latter. In addition, the reaction kinetics between oxidant and antioxidant showed that a longer application time for sodium ascorbate does not influence the effectiveness of the reaction and that 5 min is sufficiently long for this antioxidant to exert an antioxidant effect.

Proposal of a low-cost protocol for colorimetric semi-quantification of secretory phospholipase by Candida albicans grown in planktonic and biofilm phases

Biofilms are aggregates of microorganisms living in multilayered structures inside polymeric matrices onto surfaces. These biofilms may subvert the physiological properties of adjacent tissues causing morphofunctional failure. Many studies have shown that the expression of virulence attributes is maximized when microbes form such communities. This study evaluated the differential phospholipasic activity of Candida albicans SC5314 grown in planktonic phase and in biofilm. We propose two distinct protocols for the colorimetric evaluation of phosphatidylcholine hydrolysis in neutral and acidic conditions. The results showed that both protocols are suitable for the proposed intention and that 72 h-old planktonic cultures of C. albicans SC5314 secrete higher quantities of neutral (6.42-fold) and acidic (3.85-fold) phospholipases than biofilms.

Enhancement of Secretory Aspartyl Protease production in biofilms of Candida albicans exposed to sub‐inhibitory concentrations of fluconazole

The production of Secretory Aspartyl Proteases (Sap) is an important virulence factor of Candida albicans. Many studies have shown that a challenge with sub‐inhibitory concentrations of antifungals lead species of Candida to the secretion of higher concentrations of Sap. Nevertheless, published studies only reported the secretion of such enzymes by cells growing in planktonic phase, with few mention of biofilms. The present study evaluated the alterations in the secretion of Sap by C. albicans grown in biofilms and exposed to sub‐inhibitory concentrations of fluconazole. The MICs for fluconazole of seven clinical strains were determined for planktonic cells. Biofilm and planktonic cells were grown in the presence of ½ MIC, ¼ MIC, and no medication (control). The relative metabolic activity, indirectly related to cell loads, were estimated by the absorbance of reduced XTT and the Sap activity was evaluated by bovine albumin test. It was observed that 72 h‐old biofilms under the influence of ½ MIC had fewer cells than ¼ MIC and control. The production of Sap was inversely proportional to the cell content, with higher secretion in ½ MIC, followed by ¼ MIC and control. Biofilms of C. albicans challenged by sub‐MICs of fluconazole tend to secrete higher quantities of Sap.

Phenotypic evaluation of the effect of anaerobiosis on some virulence attributes of Candida albicans

The current assumption that Candida albicans is a facultatively anaerobic organism has been widely accepted since its recovery from anoxic sites became common. However, the link between anaerobiosis and virulence remains uncertain. This study investigated the differential cell-surface hydrophobicity (CSH) using a hydrocarbon/water partition technique and analysed the differential secretion rates of secretory aspartyl proteases (Saps), esterase, chondroitinase and haemolysins of C. albicans strains recovered from periodontal pockets and non-periodontium-related intra-oral sites. For the enzymic tests, all strains from both sets were grown under aerobic and anaerobic conditions and the harvested cells were inoculated onto suitable normal or pre-reduced culture media in the presence or absence of molecular oxygen, respectively. The results showed that no variations were perceptible for CSH and chondroitinase (P>0.05). The secretion rates of esterase and haemolysins strongly decreased in an anoxic environment (P<0.0001). However, a consistent increment (P<0.0001) in Sap secretion was detected when cultures were grown under anaerobic conditions. Based on these results, it is suggested that the oxygen concentration in the atmosphere surrounding cells exerts a variable influence on the virulence attributes of C. albicans.

The role of candidal histolytic enzymes on denture-induced stomatitis in patients living in retirement homes

MATERIAL AND METHODS Fifty nine elders wearing complete dentures and living in retirement homes in Curitiba (southern Brazil), were divided into two groups: group #1, 26 patients with denture-induced stomatitis and group #2, 33 patients without denture-induced stomatitis. The two groups were evaluated in relation to the degree of denture-induced stomatitis, salivary fungal loads, and secretion of some histolytic enzymes. RESULTS Patients from group #1 showed higher degrees of colonisation by Candida albicans (p = 0.031). Candida krusei, Candida tropicalis, and Candida parapsilosis were also isolated, but there were no differences between the groups (p > 0.05). Secretory aspartyl protease (Sap) and chondroitinase did not show significant differences among the isolated Candida spp. in the two groups. Phospholipase secretion rates were higher among the strains of C. albicans from group #2 (p = 0.036). The same behaviour was not detected for non-albicans Candida species. CONCLUSIONS The results could infer that differences in the secretion rates of candidal histolytic enzymes should not be imputed as imperative for the progress of denture-induced stomatitis.