Brenda Paula Figueiredo de Almeida Gomes

In vitro antimicrobial activity of propolis and Arnica montana against oral pathogens

Arnica and propolis have been used for thousands of years in folk medicine for several purposes. They possess several biological activities such as anti-inflammatory, antifungal, antiviral and tissue regenerative, among others. Although the antibacterial activity of propolis has already been demonstrated, very few studies have been done on bacteria of clinical relevance in dentistry. Also, the antimicrobial activity of Arnica has not been extensively investigated. Therefore the aim here was to evaluate in vitro the antimicrobial activity, inhibition of adherence of mutans streptococci and inhibition of formation of water-insoluble glucan by Arnica and propolis extracts. Arnica montana (10%, w/v) and propolis (10%, w/v) extracts from Minas Gerais State were compared with controls. Fifteen microorganisms were used as follows: Candida albicans--NTCC 3736, F72; Staphylococcus aureus--ATCC 25923; Enterococcus faecalis--ATCC 29212; Streptococcus sobrinus 6715; Strep. sanguis--ATCC 10556; Strep. cricetus--HS-6; Strep. mutans--Ingbritt 1600; Strep. mutans--OMZ 175; Actinomyces naeslundii--ATCC 12104, W 1053; Act. viscosus OMZ 105; Porphyromonas gingivalis; Porph. endodontalis and Prevotella denticola (the last three were clinical isolates). Antimicrobial activity was determined by the agar diffusion method and the zones of growth inhibition were measured. To assess cell adherence to a glass surface, the organisms were grown for 18 h at 37 degrees C in test-tubes at a 30 degree angle. To assay water-insoluble glucan formation, a mixture of crude glucosyltransferase and 0.125 M sucrose was incubated for 18 h at 37 degrees C in test-tubes at a 30 degree angle. Arnica and propolis extracts (20 microl) were added to these tubes to evaluate the % of inhibition of cell adherence and water-insoluble glucan formation. The propolis extract significantly inhibited all the microorganisms tested (p < 0.05), showing the largest inhibitory zone for Actinomyces spp. The Arnica extract did not demonstrate significant antimicrobial activity. Cell adherence and water-insoluble glucan formation were almost completely inhibited by the propolis extract at a final concentration of 400 microg/ml and 500 microg/ml, respectively. The Arnica extract showed slight inhibition of the adherence of the growing cells (19% for Strep. mutans and 15% for Strep. sobrinus) and of water-insoluble glucan formation (29%) at these same concentrations. Thus, the propolis extract showed in vitro antibacterial activity, inhibition of cell adherence and inhibition of water-insoluble glucan formation, while the Arnica extract was only slightly active in those three conditions.

In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite

The aim of this study was to investigate in vitro the antimicrobial activity of 0.2%, 1%, and 2% chlorhexidine gluconate (CHX gel and CHX liquid), against endodontic pathogens and compare the results with the ones achieved by 0.5%, 1%, 2.5%, 4%, and 5.25% sodium hypochlorite (NaOCl). A broth dilution test was performed, and the timing for irrigants to kill microbial cells was recorded and statistically analyzed. Both 2.0% gel and liquid formulations eliminated Staphylococcus aureus and Candida albicans in 15 seconds, whereas the gel formulation killed Enterococcus faecalis in 1 minute. All tested irrigants eliminated Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in 15 seconds. The timing required for 1.0% and 2.0% CHX liquid to eliminate all microorganisms was the same required for 5.25% NaOCl. The antimicrobial action is related to type, concentration, and presentation form of the irrigants as well as the microbial susceptibility.

In vitro assessment of the antimicrobial action and the mechanical ability of chlorhexidine gel as an endodontic irrigant.

The objective of this study was to assess the chlorhexidine gluconate gel as an endodontic irrigant. First the ability of chlorhexidine gel to disinfect root canals contaminated in vitro with Enterococcus faecalis was investigated. A scanning electron microscope was also used to evaluate its cleansing ability compared with endodontic irrigants commonly used, such as sodium hypochlorite and chlorhexidine gluconate liquid. The results indicated that the chlorhexidine gel produced a cleaner root canal surface and had an antimicrobial ability comparable with that obtained with the other solutions tested. It was concluded that chlorhexidine gluconate in gel form has potential for use as an endodontic irrigant.

Microbial Analysis of Canals of Root-filled Teeth with Periapical Lesions Using Polymerase Chain Reaction

The objective of the present study was to investigate the presence of nine bacterial species in root-filled teeth associated with periapical lesions using a polymerase chain reaction analysis and to correlate these species with clinical features of the cases. DNA was extracted from 45 canal samples of root-filled teeth with periapical lesions. A PCR assay using species-specific primers of 16S rDNA and the downstream intergenic spacer region was used for microbial detection. Enterococcus faecalis was the most prevalent species, detected in 77.8% of the study teeth, followed by Peptostreptococcus micros, detected in 51.1%. Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, and Prevotella nigrescens were detected in 35.6%, 22.2%, 11.1%, and 11.1% of the sampled teeth, respectively. Moreover, PCR detected Filifactor alocis in 26.7%, Treponema denticola in 24.4%, and Tannerella forsythia in 4.4% of the samples. T. denticola and P. micros were statistically associated with tenderness to percussion (p < 0.05). P. nigrescens was associated with the presence of spontaneous pain and abscess (p < 0.05). P. endodontalis and P. nigrescens were associated with purulent exudates (p < 0.05). Synergistic relationship was also observed between some species. The results of this study indicated that E. faecalis was the most frequently identified test species by PCR in teeth with failing endodontic treatment.

Evaluation of Cytotoxicity and Physicochemical Properties of Calcium Silicate-based Endodontic Sealer MTA Fillapex

INTRODUCTION The aim of the study was to evaluate the cytotoxicity, radiopacity, pH, and flow of a calcium silicate-based and an epoxy resin-based endodontic sealer, MTA Fillapex (Angelus, Londrina, PR, Brazil) and AH Plus (Dentsply, Konstanz, Germany), respectively. METHODS Cytotoxicity, radiopacity, and flow evaluation were performed following ISO requirements. The pH level was measured at periods of 3, 24, 72, and 168 hours. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to check the Balb/c 3T3 cells viability at 1- to 4-week periods. Data were statistically analyzed by analysis of variance and the Tukey test with a significance level of 5%. RESULTS In all tested periods, MTA Fillapex was more cytotoxic than AH Plus (P < .05). Although AH Plus presented higher radiopacity than MTA Fillapex (P < .05), both sealers showed minimum required values. MTA Fillapex presented alkaline pH in all experimental times, whereas AH Plus cement showed a slightly neutral pH and a flow significantly lower than that of MTA Fillapex (P < .05). CONCLUSIONS Although MTA Fillapex was more cytotoxic than AH Plus, it showed suitable physicochemical properties for an endodontic sealer.

Microbial Susceptibility to Calcium Hydroxide Pastes and Their Vehicles

The aim of this study was to investigate the susceptibility of some microorganisms commonly isolated from root canals to calcium hydroxide in combination with several vehicles by the agar diffusion method. Stainless-steel cylinders were placed on each inoculated agar medium. The test medications and their controls were placed inside the cylinders. The zones of growth inhibition were measured and recorded after the incubation period for each plate, and the results were analyzed statistically. Enterococcus faecalis was most resistant, whereas the anaerobic Porphyromonas endodontalis was more susceptible to all medications, followed by P. gingivalis and Prevotella intermedial intermedia. Ca(OH)2 + CMCP + glycerin showed significantly larger mean zones of inhibition when compared with the other medications. We conclude that anaerobic Gram-negative bacteria are more susceptible to calcium hydroxide pastes than facultative Gram-positive microorganisms.

Quantification of Endotoxins and Cultivable Bacteria in Root Canal Infection before and after Chemomechanical Preparation with 2.5% Sodium Hypochlorite

This clinical study was conducted to quantify endotoxins and cultivable bacteria in teeth with pulp necrosis and apical periodontitis before and after chemomechanical preparation with 2.5% sodium hypochlorite (NaOCl) and to investigate the possible correlation of endotoxin and cultivable bacteria with the presence of clinical symptomatology. Twenty-four root canals were selected. Samples were collected before (s1) and after chemomechanical preparation (s2). Culture techniques were used to determine the colony-forming unit. A limulus amebocyte lysate (LAL) assay was used to quantify endotoxins (lipopolysaccharide, LPS). LPS and bacteria were detected in 100% of the initial samples (s1), with a median concentration of 139 endotoxin units/mL and 2.64 x 10(5) colony-forming units/mL, respectively. Higher levels of LPS were found in teeth with clinical symptomatology (p < .05). At s2, mean endotoxin reduction of 59.99% and mean bacterial load reduction of 99.78% were found. Our findings indicated that chemomechanical preparation with 2.5% NaOCl was moderately effective against bacteria but less effective against endotoxins in root canal infection. Furthermore, a statistically significant association was found between higher levels and clinical symptomatology.

In vitro antimicrobial activity of calcium hydroxide pastes and their vehicles against selected microorganisms

The aim of this study was to investigate in vitro the antimicrobial activity of calcium hydroxide in combination with several vehicles against some microorganisms commonly isolated from root canals. Antimicrobial activity was determined by the agar diffusion method. Stainless-steel cylinders were placed on each inoculated agar medium. The test medicaments and their controls were placed inside the cylinders. The zones of growth inhibition were measured and recorded after incubation for each plate and the results were analyzed statistically (ANOVA). The in vitro antimicrobial effects of the medications were ranked from strongest to weakest as follows: Ca(OH)2 + CMCP + glycerine, Ca(OH)2 + CMCP, Ca(OH)2 + glycerine, Ca(OH)2 + anesthetic, Ca(OH)2 + saline, Ca(OH)2 + H2O, Ca(OH)2 + polyethyleneglycol. The pastes with oily vehicles showed significantly larger mean zones of inhibition compared to those with aqueous or viscous vehicles. It was concluded that diffusion and antimicrobial activity of calcium hydroxide were affected by the type of vehicle used.

Chlorhexidine in Endodontics

Chemical auxiliary substances (CAS) are essential for a successful disinfection and cleanness of the root canals, being used during the instrumentation and if necessary, as antimicrobial intracanal medicaments. Different CAS have been proposed and used, among which sodium hypochlorite (NaOCl), chlorhexidine (CHX), 17% EDTA, citric acid, MTAD and 37% phosphoric acid solution. CHX has been used in Endodontics as an irrigating substance or intracanal medicament, as it possesses a wide range of antimicrobial activity, substantivity (residual antimicrobial activity), lower cytotoxicity than NaOCl whilst demonstrating efficient clinical performance, lubricating properties, rheological action (present in the gel presentation, keeping the debris in suspension); it inhibits metalloproteinase, is chemically stable, does not stain cloths, it is odorless, water soluble, among other properties. CHX has been recommended as an alternative to NaOCl, especially in cases of open apex, root resorption, foramen enlargement and root perforation, due to its biocompatibility, or in cases of allergy related to bleaching solutions. The aim of this paper is to review CHXs general use in the medical field and in dentistry; its chemical structure, presentation form and storage; mechanism of action; antimicrobial activity including substantivity, effects on biofilms and endotoxins, effects on coronal and apical microbial microleakage; tissue dissolution ability; interaction with endodontic irrigants; effects on dentin bonding, metalloproteinases and collagen fibrils; its use as intracanal medicament and diffusion into the dentinal tubules; its use as disinfectant agent of obturation cones; other uses in the endodontic therapy; and possible adverse effects, cytotoxicity and genotoxicity.

Antimicrobial effect and pH of chlorhexidine gel and calcium hydroxide alone and associated with other materials

The purposes of this study were to evaluate the effectiveness of 2% chlorhexidine (CHX) gluconate gel, calcium hydroxide [Ca(OH)2] and their combination with iodoform and zinc oxide powder as intracanal medications against select microorganisms, and to measure the pH changes caused by these medications. Antimicrobial activity was determined by the agar diffusion method. The zones of growth inhibition were measured and the results were analyzed statistically by Kruskal-Wallis test (p<0.05). The pH of the pastes was measured right after preparation, after 24 h and 1 week later. The largest mean zones of microbial inhibition were produced by 2% CHX gel, followed by Ca(OH)2 + 2% CHX gel + iodoform, Ca(OH)2 + 2% CHX gel, Ca(OH)2 + 2% CHX gel + zinc oxide, and Ca(OH)2 + water. The mean pH of all medications stayed above 12.0 during the whole experiment, except for CHX gel (pH=7.0). The results of this study showed that all medications had antimicrobial activity, but the most effective against the tested microorganisms were 2% CHX gel, followed by its combination with Ca(OH)2 and iodoform.