Miriam F. Zaccaro Scelza

A multiparametric assay to compare the cytotoxicity of endodontic sealers with primary human osteoblasts.

AIM   To compare the cytotoxicity of four endodontic sealers (Sealapex, Pulp Canal Sealer EWT, Real Seal and MTA Fillapex) either 1 or 7 days after mixing, when assessed through a multiparametric analysis employing human primary cells closely related to periapical tissues. METHODOLOGY   Extracts of each sealer were prepared following 24-h exposure to culture media, at either 24 h or 7 days after mixing. Primary human osteoblasts were exposed to extracts for 24 h, at 37 °C with 5% CO(2) , and cell viability was evaluated by a multiparametric assay assessing sequentially, on the same cells, mitochondrial activity (XTT), membrane integrity (neutral red test) and total cell density (crystal violet dye exclusion test). Results from each test and experimental time were compared by 2-way analysis of variance (anova). RESULTS   All endodontic sealers had strong cytotoxicity 24 h after mixing, according to all parameters evaluated. At a longer setting period (7 days), viability for Sealapex was significantly increased (P < 0.05) and Pulp Canal Sealer achieved levels of cytocompatibility similar to the control group. The anova indicated a general correlation between the cytotoxicity of the materials and the time after mixing, with some level of dependence on the cell viability assay employed. CONCLUSIONS   All materials had high cytotoxic levels for human primary cells, mostly on a time-dependent basis, as shown by three different cell viability tests.

Comparative sem evaluation of three solvents used in endodontic retreatment: an ex vivo study

This study compared, by scanning electron microscopy (SEM), the efficacy of three solvents on the removal of filling materials from dentinal tubules during endodontic retreatment. Forty human maxillary canines with straight canals were prepared according to a crown-down technique and enlarged to a#30 apical file size, before obturation with gutta-percha and a zinc-oxide-eugenol based sealer. The samples were stored for 3 months before being randomly assigned to four groups: chloroform (n=10), orange oil (n=10), eucalyptol (n=10) and control (n=10). Solvents were applied to a reservoir created on the coronal root third using Gates Glidden drills. The total time for retreatment using the solvents was 5 minutes per tooth. Following retreatment the roots were split longitudinally for SEM evaluation. SEM images were digitized, analyzed using Image ProPlus 4.5 software, and the number of dentinal tubules free of filling material from the middle and apical thirds was recorded. No significant difference was found among the solvent groups regarding the number of dentinal tubules free of root filling remnants in the middle and apical root thirds (p>0.05). However, the control group had fewer dentinal tubules free of filling material (p<0.05). Under the tested conditions, it may be concluded that there was no significant difference among the solvents used to obtain dentinal tubules free of filling material remnants.

Antimicrobial activity of calcium hydroxide pastes on Enterococcus faecalis cultivated in Root Canal Systems

The effectiveness of calcium hydroxide pastes: Calen and PMCC-Calen associated to chemo-mechanical preparation was assessed on Enterococcus faecalis grown within root canals. Seventy incisors were inserted into TSB medium, sterilized and contaminated with E. faecalis. Culture medium was replaced each 24 h and incubated at 37 degrees C for 72 h. After chemo-mechanical preparation, root canals were filled with Calen or PMCC-Calen (7 or 14 days). Pastes were removed and teeth were inserted into test tubes containing Enterococcosel broth. Calen paste (maintained for 7 and 14 days) induced 70% elimination of enterococci and PMCC-Calen 100% elimination only after maintenance for 14 days. These medications were significantly more effective (p<0.001) than chemo-mechanical protocol alone and PMCC-Calen maintained for 7 days, both incapable to eliminate the viability of enterococci. Calcium hydroxide pastes demonstrated important adjuvant effects in the elimination of enterococci during chemo-mechanical preparation of root canal systems. When associated with PMCC, calcium hydroxide pastes should be maintained for at least 14 days.

Comparative evaluation of antimicrobial action of MTA, calcium hydroxide and Portland cement

The present study aimed to evaluate and compare the antimicrobial effect of MTA Dentsply, MTA Angelus, Calcium Hydroxide and Portland cement. Four reference bacterial strains were used: Pseudomonas aeruginosa, Escherichia coli, Bacteroides fragilis, and Enterococcus faecalis. Plates containing Mueller-Hinton agar supplemented with 5% sheep blood, hemin, and menadione were inoculated with the bacterial suspensions. Subsequently, wells were prepared and immediately filled with materials and incubated at 37°C for 48 hours under anaerobic conditions, except P. aeruginosa. The diameters of inhibition zones were measured, and data analyzed using ANOVA and the Tukey test with 1% level of significance. MTA Dentsply, MTA Angelus and Portland cement inhibited the growth of P. aeruginosa. Calcium Hydroxide was effective against P. aeruginosa and B. fragillis. Under anaerobic conditions, which may hamper the formation of reactive oxygen species, the materials failed to inhibit E. faecalis, and E. coli.

The antimicrobial activity of gray-colored mineral trioxide aggregate (GMTA) and white-colored MTA (WMTA) under aerobic and anaerobic conditions.

OBJECTIVE The objective of this study was to test the hypothesis that the antimicrobial activity of gray-colored MTA (GMTA) and white-colored MTA (WMTA) is related to the induction of reactive oxygen species (ROS). STUDY DESIGN In vitro antimicrobial activity of MTA was carried out using E. coli mutant (AB2463-RecA13), triple mutant (BW535), and wild type (AB1157). The diffusion method on Müller-Hinton agar (MHA) was used. A base layer was made using MHA agar and 4 wells were made by removing agar at equidistant points. MTA was placed into the wells immediately after manipulation. The plates were incubated at 37 degrees C for 48 hours under aerobic and anaerobic conditions. Afterward the zones of inhibition were measured. Also the damage caused to the plasmid DNA in the presence of the cement was evaluated through 0.8% agarose gel electrophoresis. RESULTS It was possible to observe a zone of inhibition under aerobic conditions promoted by both cements on both of the mutant E. coli but not in the wild-type E. coli. On the other hand, both cements were not able to induce any bacterial inhibition under anaerobic conditions, suggesting that the inhibitory action is a result of ROS production. Both cements promoted damage in the DNA plasmid when compared with distilled water (control). CONCLUSION We concluded that MTA under aerobic conditions provoked antimicrobial activity by induction of ROS. Furthermore, the antimicrobial activity occurred by damage caused directly to the bacterial DNA.

The influence of medication on salivary flow of the elderly: preliminary study.

OBJECTIVE This study was to evaluate the influence of medications on unstimulated and stimulated salivary flow in elderly men and women. BACKGROUND Several diseases and conditions are associated with decreasing salivary flow. Medications can be risk factors for hyposalivation due to the effect particular drug categories. METHODS Seventy-five elderly of both gender (sixty years old or over) from the Geriatric Dental Clinic at the Federal Fluminense University Dental School, were interviewed about their health status and chronic use of medication. After the interview, unstimulated and stimulated saliva were collected from each subject. The collection time was five minutes, and the flow rate was calculated as ml/min. RESULTS The mean unstimulated salivary flow was 0.25 ml/min for women and 0.30 ml/min for men, while the mean for stimulated salivary flow was 1.23 ml/min for women and 1.31 ml/min for men, without both differences being non-significant (p > 0.05). The difference between the mean production of unstimulated and stimulated salivary flow was statistically significant, regardless of gender (p < 0.01). A significant reduction of stimulated salivary flow was observed with the usage of cardiovascular agents (p < 0.05). CONCLUSION Drugs used in cardiovascular disease influence the flow of stimulated saliva.

Cytotoxicity evaluation of two root canal sealers and a commercial calcium hydroxide paste on THP1 cell line by Trypan Blue assay

Objective: The aim of this investigation was to evaluate the cytotoxicity of two brands of root canal sealers, epoxy-resin based and zinc oxide-eugenol based, and one commercial calcium hydroxide paste on a monocyte cell line THP-1. Material and methods: Undiluted (crude extract) and diluted extracts to 10%, 1%, 0.1%, 0.01%, 0.001% and 0.0001% of the sealers were tested for cytotoxicity to THP-1 cells using the trypan blue assay. Extracts were obtained according to ISO standard. Data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests at 5% significance level. Results: Crude extract of AH Plus and Fill Canal killed approximately 90% of THP-1 cells versus 36% of THP-1 cells killed by L&C crude extract (p<0.05). Ten-fold dilutions of L&C, Fill Canal and AH Plus killed 24, 35 and 61% of THP-1 cells (p<0.05), respectively. Dilutions lesser than 1% caused minimal cell death as compared to the control groups (p>0.05), except for L&C 1% extract. Conclusions: The results revealed that the L&C paste crude extract was less cytotoxic to THP-1 cells than AH Plus or Fill Canal crude extracts.

Evaluation of Inflammatory Response of EDTA, EDTA-T, and Citric Acid in Animal Model

INTRODUCTION The biocompatibility of chelating agents and organic acids have been explained by a variety of methods, and suggestions for use have been based more on clinical observations and physicochemical properties than on biological aspects. The present study aimed to evaluate the inflammatory response of 17% EDTA, 17% EDTA-T, and 10% citric acid in bony defect created in rat jaws. METHODS Mandibular through and through critical size defects were created bilaterally in 60 rats. Fibrinol (Baldacchi SA, São Paulo, Brazil), a cube-shaped compound of absorbable bovine fibrin foam and sodium chloride, was used as a carrier of the substances. One side had received Fibrinol (control), whereas the opposite side had received Fibrinol soaked with each substance on the 1st, on the 7th, on the 14th, and on the 28th day (n=5 for each day). Hemijaws were prepared for light microscopy, and samples were stained with hematoxylin and eosin. Digitized images were analyzed with a morphometric software (ImageJ; National Institute of Mental Health, Bethesda, MD). to obtain the number of inflammatory cells per area. Comparisons were performed by using the Kruskal-Wallis test (p=0.05). RESULTS For all days, 10% citric acid and 17% EDTA-T showed, respectively, the lowest and highest number of inflammatory cells per area. All tested substances and controls showed the highest inflammatory cell response on the 14th day. CONCLUSION Among the tested substances, 10% citric acid proved to be the less aggressive tested solution at 14 days. At 28 days, all solutions were similar, but EDTA-T kept showing the higher number of inflammatory cells.

Relation between oral health and nutritional condition in the elderly

Oral health is a prerequisite for a good chewing function, which may have an impact on food choices and nutritional well-being. Objective This study was designed to evaluate the relationship between oral health status and nutritional status in the elderly. Material and Methods In this cross-sectional study, 33 elderly people from the Group for the Elderly Interdisciplinary Geriatrics and Gerontology Program, at Fluminense Federal University, Niteroi, RJ, Brazil, completed a questionnaire to collect information on socioeconomic status, eating habits, physical activity and health habits, undertook a clinical oral examination, blood test, and anthropometric measurements, and were allocated into groups according to age. The oral health status was assessed using the index for decayed, missing and filled teeth (DMFT). The nutritional status was assessed using hemoglobin, hematocrit and albumin concentrations in blood, anthropometric values and the body mass index. Results Tooth loss was the biggest nuisance to the elderly subjects (57.6%), followed by the use of dentures (30.3%) and ill-fitting dentures (33.3%). 66.6% of patients had difficulty in chewing, and 54.5% reported this to be due to prostheses and 13.6% to the absence of teeth. A significant correlation was found between DMFT and the value of suprailiac skinfold thickness (rho=0.380, p=0.029). Conclusion The results support the temporal association between tooth loss and detrimental changes in anthropometry, which could contribute to increased risk of developing chronic diseases.

Cytotoxicity Evaluation of Two Bis-Acryl Composite Resins Using Human Gingival Fibroblasts

Bis-acryl resins are used for temporary dental restorations and have shown advantages over other materials. The aim of this work was to evaluate the in vitro cytotoxicity of two bis-acryl composite resins (Protemp 4 and Luxatemp Star), obtained at 1, 7 and 40 days after mixing the resin components, using a standardized assay employing human primary cells closely related to oral tissues. Human gingival fibroblast cell cultures were exposed for 24 h to either bis-acryl composite resins, polystyrene beads (negative control) and latex (positive control) extracts obtained after incubation by the different periods, at 37 °C under 5% CO2. Cell viability was evaluated using a multiparametric procedure involving sequential assessment (using the same cells) of mitochondrial activity (XTT assay), membrane integrity (neutral red test) and total cell density (crystal violet dye exclusion test). The cells exposed to the resin extracts showed cell viability indexes exceeding 75% after 24 h. Even when cells were exposed to extracts prepared with longer conditioning times, the bis-acryl composite resins showed no significant cytotoxic effects (p>0.05), compared to the control group or in relation to the first 24 h of contact with the products. There were no differences among the results obtained for the bis-acryl composite resins evaluated 24 h, 7 days and 40 days after mixing. It may be concluded that the bis-acryl resins Protemp 4 and Luxatemp Star were cytocompatible with human gingival fibroblasts, suggesting that both materials are suitable for use in contact with human tissues.