In vitro effects of Crotalus durissus terrificus and Bothrops jararaca venoms on Giardia duodenalis trophozoites

Considering the snake venoms' pharmacological properties and chemotherapeutic potential as well as the need for new alternatives for Giardia infection treatment, the present study was carried out aiming to evaluate the in vitro effects of crude Crotalus durissus terrificus and Bothrops jararaca venoms on the growth and adherence of Giardia duodenalis trophozoites. Trophozoites (10(6)) were exposed to serial twofold dilutions of C. durissus terrificus and B. jararaca venoms that ranged from 3.125 to 200 mu g/ml and from 5 to 320 mu g/ml, respectively. The two venoms inhibited the growth of trophozoites, and the level of inhibition varied according to the assayed concentrations and incubation times. The highest reduction of parasite growth was observed with increasing concentrations of the venoms. No effect on parasite adherence was observed. Light microscope observations revealed changes of the pear-shape aspect of the cell and reduction of flagellar beating frequency in the great part of the trophozoites. This is the first attempt to evaluate the in vitro effects of snake venoms on Giardia trophozoites. The findings stress the need for more investigations and prompt us to perform further studies to identify active principles with antigiardial effect.

In vitro study of CD133 human stem cells labeled with superparamagnetic iron oxide nanoparticles

Superparamagnetic iron oxide nanoparticles (SPIONs) are applied in stem cell labeling because of their high magnetic susceptibility as compared with ordinary paramagnetic species, their low toxicity, and their ease of magnetic manipulation. The present work is the study of CD133(+) stem cell labeling by SPIONs coupled to a specific antibody (AC133), resulting in the antigenic labeling of the CD133+ stem cell, and a method was developed for the quantification of the SPION content per cell, necessary for molecular imaging optimization. Flow cytometry analysis established the efficiency of the selection process and helped determine that the CD133 cells selected by chromatographic affinity express the transmembrane glycoprotein CD133. The presence of antibodies coupled to the SPION, expressed in the cell membrane, was observed by transmission electron microscopy. Quantification of the SPION concentration in the marked cells using the ferromagnetic resonance technique resulted in a value of 1.70 x 10 (13) mol iron (9.5 pg) or 7.0 x 10 (6) nanoparticles per cell ( the measurement was carried out in a volume of 2 mu L containing about 6.16 x 10 5 pg iron, equivalent to 4.5 x 10 (11) SPIONs). (c) 2008 Elsevier B.V. All rights reserved.

Influence of acute pancreatitis on the in vitro responsiveness of rat mesenteric and pulmonary arteries

Background: Acute pancreatitis is an inflammatory disease characterized by local tissue injury and systemic inflammatory response leading to massive nitric oxide (NO) production and haemodynamic disturbances. Therefore, the aim of this work was to evaluate the vascular reactivity of pulmonary and mesenteric artery rings from rats submitted to experimental pancreatitis.Male Wistar rats were divided into three groups: saline (SAL); tauracholate (TAU) and phospholipase A(2) (PLA(2)). Pancreatitis was induced by administration of TAU or PLA(2) from Naja mocambique mocambique into the common bile duct of rats, and after 4 h of duct injection the animals were sacrificed. Concentration-response curves to acetylcholine (ACh), sodium nitroprusside (SNP) and phenylephrine (PHE) in isolated mesenteric and pulmonary arteries were obtained. Potency (pEC(50)) and maximal responses (E(MAX)) were determined. Blood samples were collected for biochemical analysis.Results: In mesenteric rings, the potency for ACh was significantly decreased from animals treated with TAU (about 4.2-fold) or PLA(2) (about 6.9-fold) compared to saline group without changes in the maximal responses. Neither pEC(50) nor E(MAX) values for Ach were altered in pulmonary rings in any group. Similarly, the pEC(50) and the E(MAX) values for SNP were not changed in both preparations in any group. The potency for PHE was significantly decreased in rat mesenteric and pulmonary rings from TAU group compared to SAL group (about 2.2- and 2.69-fold, for mesenteric and pulmonary rings, respectively). No changes were seen in the E(MAX) for PHE. The nitrite/nitrate (NO(x)(-)) levels were markedly increased in animals submitted to acute pancreatitis as compared to SAL group, approximately 76 and 68% in TAU and PLA(2) protocol, respectively.Conclusion: Acute pancreatitis provoked deleterious effects in endothelium-dependent relaxing response for ACh in mesenteric rings that were strongly associated with high plasma NO(x)(-) levels as consequence of intense inflammatory responses. Furthermore, the subsensitivity of contractile response to PHE in both mesenteric and pulmonary rings might be due to the complications of this pathological condition in the early stage of pancreatitis.

Biomechanical in vitro evaluation of three stable internal fixation techniques used in sagittal osteotomy of the mandibular ramus: a study in sheep mandibles

Among the osteotomies performed in orthognathic surgery, the sagittal osteotomy of the mandibular ramus (SOMR) is the most common, allowing a great range of movements and stable internal fixation (SIF), therefore eliminating the need of maxillomandibular block in the postoperative period. Objectives: The purpose of this study was to evaluate the biomechanical resistance of three national systems used for SIF in SOMR in sheep mandibles. Material and methods: The study was performed in 30 sheep hemi-mandibles randomly divided into 3 experimental groups, each containing 10 hemi-mandibles. The samples were measured to avoid discrepancies and then subjected to SOMR with 5-mm advancement. In group I, 2.0x12 mm screws were used for fixation, inserted in an inverted "L" pattern (inverted "L" group). In group II, fixation was performed with two 2.0x12 mm screws, positioned in a linear pattern and a 4-hole straight miniplate and four 2.0x6.0 mm monocortical screws (hybrid group). In group III, fixation was performed with two-hole straight miniplates and eight 2.0x6.0 mm monocortical screws (mini plate group). All materials used for SIF were supplied by Osteosin - SIN. The hemimandibles were subjected to vertical linear load test by Kratos K2000MP mechanical testing unit for loading registration and displacement. Results: All groups showed similar resistance during mechanical test for loading and displacement, with no statistically significant differences between groups according to analysis of variance. Conclusion: These results indicate that the three techniques of fixation are equally effective for clinical fixation of SOMR.

In vitro effects of calcium hydroxide and polymyxin B on endotoxins in root canals

Objectives. To evaluate the effects of intracanal medicaments on endotoxins in root canals.Methods. Seventy-five freshly extracted maxillary incisors were used in this study. The crowns of teeth were sectioned near the CEJ in order to standardize the root length to 14 mm. The root canals were instrumented to an apical size #50 file and irrigated with 1% sodium hypochlorite solution and sterilized with 60 Co gamma irradiation. Standardized suspension containing Escherichia coli endotoxin was inoculated into the 60 root canals. The specimens were randomly assigned to 5 groups (n=15), according to the intracanal medicament used: (G1) calcium hydroxide; (G2) polymyxin B; (0) combination neomycin-potymyxin B-hydrocortisone; (G4) positive control (no intracanal medicament); (G5) negative control (no endotoxin and no intracanal medicament). After 7 days, the detoxification of endotoxin was evaluated by Limulus lysate assay and antibody production in B-tymphocytes culture.Results. Groups 1, 2 and 5 presented the best results by Limulus lysate and were significantly different to groups 3 and 4 (p<0.05). Stimulation of antibodies production in cell culture by groups 1 and 6 was smaller and statistically different than groups 2, 3, 4 and 5 (p<0.05). Groups 2 and 5 induced a small increase in the antibodies production in relation to the groups 1 and 6. Groups 3 and 4 induced a significant increase of antibodies production (p<0.05).Conclusions. The calcium hydroxide and polymyxin B intracanal medicaments detoxified endotoxin in root canals and altered the properties of LPS to stimulate the antibody production by B-Lymphocytes. The combination neomycin-polymyxin B-hydrocortisone did not detoxified endotoxin. (C) 2004 Elsevier Ltd. All rights reserved.

In vitro evaluation of the action of irrigating solutions associated with intracanal medications on Escherichia coli and its endotoxin in root canals

Objective: The purpose of this study was to evaluate the efficacy of auxiliary chemical substances and intracanal medications on Escherichia coli and its endotoxin in root canals. Material and Methods: Teeth were contaminated with a suspension of E. coli for 14 days and divided into 3 groups according to the auxiliary chemical substance used: G1) 2.5% sodium hypochlorite (NaOCl); G2) 2% chlorhexidine gel (CLX); G3) pyrogen-free solution. After, these groups were subdivided according to the intracanal medication (ICM): A) Calcium hydroxide paste (Calen (R)), B) polymyxin B, and C) Calcium hydroxide paste+2% CLX gel. For the control group (G4), pyrogen-free saline solution was used without application of intracanal medication. Samples of the root canal content were collected immediately after biomechanical preparation (BMP), at 7 days after BMP, after 14 days of intracanal medication activity, and 7 days after removal of intracanal medication. The following aspects were evaluated for all collections: a) antimicrobial activity; b) quantification of endotoxin by the limulus Amebocyte lysate test (LAL). Results were analyzed by the kruskal-wallis and Dunn's tests at 5% significance level. Results: The 2.5% NaOCl and CLX were able to eliminate E. coli from root canal lumen and reduced the amount of endotoxin compared to saline. Conclusions: It was concluded that 2.5% NaOCl and CLX were effective in eliminating E. coli. Only the studied intracanal medications were to reduce the amount of endotoxin present in the root canals, regardless of the irrigant used.

In vitro adherence of Candida albicans isolated from patients with chronic periodontitis

Adherence is considered an extremely important virulence factor in yeast. Objective: The aim of this study was to analyze the adherence to epithelial cells of C. albicans isolated from patients with chronic periodontitis in comparison to healthy patients. Material and methods: Candida albicans cells isolated from individuals with chronic periodontitis (n=25) and healthy controls (n=25) were included in this study. Suspensions of C. albicans (10(6) cells/rnL) and epithelial cells (10(5) cells/mL) were mixed and incubated at 37 degrees C for 1 h. The number of yeasts adhered to 25 epithelial cells was counted. Results: The number of C. albicans cells adhered to epithelial cells was statistically higher in the chronic periodontitis group than in the control group (Student's t-test, p=0.000). Conclusion: The results of the present study suggest a higher Candida adherence of samples isolated from patients with chronic periodontitis.

IN VITRO INCIDENCE of ROOT CANAL BIFURCATION IN MANDIBULAR INCISORS BY RADIOVISIOGRAPHY

Objective: The aim of this study was to verify, in vivo and in vitro, the prevalence of root canal bifurcation in mandibular incisors by digital radiography. Material and Methods: Four hundred teeth were analyzed for the in vivo study. Digital radiographs were taken in an orthoradial direction from the mandibular incisor and canine regions. The digital radiographs of the canine region allowed visualizing the incisors in a distoradial direction using 20 degrees deviation. All individuals agreed to participate by signing an informed consent form. The in vitro study was conducted on 200 mandibular incisors positioned on a model, simulating the mandibular dental arch. Digital radiographs were taken from the mandibular incisors in both buccolingual and mesiodistal directions. Results: The digital radiography showed presence of bifurcation in 20% of teeth evaluated in vitro in the mesiodistal direction. In the buccolingual direction, 17.5% of teeth evaluated in vivo and 15% evaluated in vitro presented bifurcation or characteristics indicating bifurcation. Conclusions: Digital radiography associated with X-ray beam distally allowed detection of a larger number of cases of bifurcated root canals or characteristics of bifurcation.

In vitro evaluation of the effectiveness of irrigants and intracanal medicaments on microorganisms within root canals

Aim To evaluate in vitro the effectiveness of sodium hypochlorite (NaOCl). chlorhexidine (CHX) and live intracanal medicaments on microorganisms within root canals.Methodology Ninety-six human single-rooted extracted teeth were used. After removing the crowns, canal preparation was completed and the external root Surfaces were coated with epoxy resin. Following sterilization. The teeth were contaminated with Candida albicans and enterococcus faecalis. and were incubated at 37 +/- 1 degreesC for 7 days. The teeth were divided according to the irrigant solution or intracanal medicament: group 1. sterile physiologic solution (SPS) and calcium hydroxide (Ca(OH)(2)) paste: group 2. SPS and camphorated paramonochlorophenol (CPMC): group 3.SPS and tricresol formalin: group 4, SPS and CaOH2 + CPMC paste: group 5, SPS and PMC furacin; group 6.2.5%, NaOCl without intracanal medication: group 7, 2.0% CHX without intracanal medication and group 8, SPS Without intracanal medication (control group). Microbiological samples were collected with sterile paper points, and bacterial growth was determined. The data were submitted to the analysis of variance (ANOVA. P = 0.05).Results For C. albicans, groups 3 and S were statistically less effective than groups 1, 2. 4 and 5 (Kruskal-Wallis (K-W) = 65.241; gl = 7; P = 0.001). For E. faecalis, groups 6 and 8 were statistically less effective than groups 1-4 and 7 (K-W = 61.048; gl = 7; P = 0.001).Conclusions Ca(OH)(2) + CPMC paste was the most effective intracanal medicament for the elimination of the two microorganisms; 2.0% CHX solution was more effective than 2.5% NaOCl against E. faecalis.

In vitro study of the Nd : YAG laser effect on human dental enamel: Optical and scanning electron microscope analysis

Objective: the Nd:YAG laser irradiation of dental enamel was evaluated in enamel demineralization experiments in a Streptococcus mutans culture media. Summary Background Data: Previous studies had shown that a continuous wave Nd:YAG laser at an energy of approximately 67 mJ may induce an increased acid resistance in human dental enamel when exposed to severe demineralization conditions. Methods: Enamel windows of 3 x 4 cm in the buccal surface were irradiated with a continuous wave Nd:YAG laser at a wavelength of 1,064 Ecm using energy densities of from 83.75 to 187.50 J/cm(2), Enamel windows of 3 x 4 cm on the lingual surface served as control (without the laser irradiation). The enamel windows were then exposed to a Streptococcus mutans culture media at a temperature of 37 degrees C for 15 and 21 days. The laser effects and demineralization were examined both by optical microscopy and scanning electron microscopy (SEM), Results: A comparison between the lased and the unlased windows of enamel showed fusion and recrystalization of the enamel and increased acid-resistance in all groups irradiated with the Nd:YAG laser, on the other hand, the 3 x 4 delimited enamel surfaces from the control group (not irradiated with the Nd:YAG laser) showed 100% deminerization, Conclusions: These findings are consistent with the finding that laser irradiation of dental results in significant reduction of the effective solubility of enamel mineral.

In vitro evaluation of apical sealing in root apex treated with demineralization agents and retrofiled with mineral trioxide aggregate through marginal dye leakage

O objetivo deste estudo foi avaliar o selamento de ápices radiculares tratados com diferentes agentes desmineralizantes e retrobturados com agregado de trióxido mineral (MTA), mediante infiltração marginal por corante. Cinqüenta e seis dentes unirradiculares humanos extraídos foram instrumentados, obturados e seccionados apicalmente. Os preparos cavitários apicais foram confeccionados com pontas ultra-sônicas e os agentes desmineralizantes foram aplicados previamente à retrobturação com Pro Root MTA. Os espécimes foram divididos aleatoriamente em 4 grupos (n=14): grupo 1 (sem agente desmineralizante); grupo 2 (ácido fosfórico 35% durante 15 s); grupo 3 (solução de EDTA 17%, pH 7, durante 3 min); grupo 4 (gel de EDTA 24%, pH 7, durante 4 min). A extensão da infiltração de corante (rodamina B 2% a 37°C, por 24 h) foi avaliada em milímetros utilizando-se um estereomicroscópio. Os resultados foram analisados estatisticamente por meio de análise de variância a um critério e do teste Tukey com nível de significância de 5%. Dentre os grupos experimentais, a menor extensão de infiltração do corante foi verificada no grupo 1 (1,89 mm), seguido pelos grupos 2 (2,18 mm), 4 (2,54 mm) e 3 (2,64 mm). Não houve diferenças estatisticamente significante (p>0.05) na infiltração marginal pelo corante entre os grupos 1, 2 e 4 e os grupos 2, 3 e 4. Com base nos resultados obtidos, pode-se concluir que a aplicação de agentes desmineralizantes não pode ser recomendada quando da utilização do MTA em cirurgias parendodônticas.

In vitro evaluation of dental bleaching effectiveness using hybrid lights activation

OBJETIVO: Avaliar se fontes de luz aumentam a eficácia do peróxido de hidrogênio na técnica de clareamento profissional. METODOLOGIA: Foram empregados 60 dentes incisivos bovinos, com dimensões coronárias e radiculares padronizadas a partir do limite amelo-cementário, sendo descartada a porção lingual. Os corpos-de-prova (cp) foram limpos em ultra-som por 20 min e a dentina condicionada com H3PO4 a 38% por 15 s, sendo os (cp) imersos em solução de café solúvel a 25% por duas semanas. A dentina foi impermeabilizada com esmalte e os (cp) divididos em 5 grupos, sendo a cor inicial mensurada através do espectofotômetro-EasyShade (VITA). Todos os (cp) receberam três aplicações por 10 min do gel clareador Opalescence Xtra-Boost (Ultradent) conforme segue: Grupo 1 - controle, não recebeu fotoativação, Grupo 2 - ativado com luz halôgena, Grupo 3 - ativado com LED azul/LASER, Grupo 4 - ativado com LED verde/LASER e Grupo 5 - ativado com LED vermelho. Após o clareamento foi mensurada a variação de cor E, a*, b*e L* e as referentes à escala de cor Vita Clássico. Os dados foram submetidos à análise de variância, teste de Tukey e de Dunn (α=5%). RESULTADOS: A diferença geral da cor foi reduzida quando se empregou LED Azul e Luz Halógena, sendo que o desempenho do peróxido de hidrogênio a 38% foi intensificado dependendo da fonte de luz utilizada. A avaliação quantitativa de cor, obtida por espectrofotômetro e pela escala de cor Vita Clássico, foram coincidentes. CONCLUSÃO: O tipo de fonte de luz empregada interfere na eficácia do agente clareador.

In Vitro Assessment of Chemical Activation Efficiency During In-office Dental Bleaching

Purpose: This study compared five types of chemical catalyzing agents added to 35% hydrogen peroxide gel, with regard to their capacity of intensifying in-office dental bleaching results.Methods: One-hundred and twenty bovine incisors were used, of which the crowns and roots were cut in the incisor-apical direction, to acquire the dimensions of a human central incisor. The specimens were sectioned in the mesiodistal direction by means of two longitudinal cuts, the lingual halves being discarded. The vestibular halves received prophylaxis with a bicarbonate jet, ultrasound cleaning and acid etching on the dentinal portion. Next, the specimens were stored in receptacles containing a 25% instant coffee solution for two weeks. After the darkening period, initial measurement of the shade obtained was taken with the Easy Shade appliance, which allowed it to be quantified by the CIELab* method. The samples were divided into six groups, corresponding to the chemical activator used: a) none (CON); b) ferric chloride (CF); c) ferrous sulphate (SF); d) manganese gluconate (GM); e) manganese chloride (CM); f) mulberry root extract (RA). Each group received three 10-minute applications of the gels containing the respective activating agents. Next, a new shade measurement was made.Results: The Analysis of Variance and Tukey tests (alpha=5%) showed statistically significant differences for the shade perception values (p=0.002). Groups GM, CM and RA showed significantly higher means than the control group.Conclusion: The presence of some chemical activators is capable of resulting in a significant increase in tooth shade variation.

Relationship between structural features and in vitro release of doxorubicin from biocompatible anionic microemulsion

In this work structural features of anionic microemulsions, containing the pharmaceutical biocompatible components soya phosphatidylcholine (SPC), eumulgin HRE 40 (EU) and sodium oleate (SO) as surfactant, cholesterol (CHO) as oil phase and aqueous buffer were studied. Microemulsions were formulated with and without the antitumor drug doxorubicin (DOX). The various microstructures characterized in the pseudo-temary phase diagram were analyzed by polarized light microscopy, small-angle X-ray scattering (SAXS) and X-ray diffraction (XRD) as well as by their ability to incorporate and release DOX. The experimental results demonstrated a correlation between the composition, the structural features and drug delivery. It was found that at higher cholesterol contents, the crystallization of CHO polymorph phases changed the mobility of DOX molecules. Droplets were formed with short-range spatial correlation from a microemulsion (ME) with a low surfactant:oil ratio. More ordered structures with lamellar arrangements formed by the increasing of the CHO proportions in the formulation may be due to CHO crystallization. The in vitro release of DOX showed that the presence of a high content of crystalline CHO prolongs the release of DOX from ME. The retention of DOX in the internal oil phase of the ME may modulate the drug release for a prolonged time. These results clearly demonstrate the potential of ME as a drug-delivery system. (c) 2007 Elsevier B.V. All rights reserved.