Susceptibility of peritoneal macrophage from different species of neotropical primates to Ex vivo Leishmania (L.) infantum chagasi-infection

Este estudo examinou a susceptibilidade do macrófago peritoneal (PM) dos primatas neotropicais: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus e Callimico goeldii para a infecção ex vivo por Leishmania (L.) infantum chagasi, o agente etiológico da leishmaniose visceral americana (LVA), como método de triagem para avaliar o potencial desses primatas como modelo de estudo da LVA. A susceptibilidade do PM para a infecção foi investigada através do índice de infecção do PM (PMI) a intervalos de 24, 72 horas e, ainda, pela média dessas taxas (FPMI), assim como, pelas respostas do TNF-α, IL-2 (ELISA de captura) e óxido nítrico (NO) (método de Griess). Às 24hs da infecção experimental, o PMI do primata A. azarae infulatus (128) foi maior que aqueles de C. penicillata (83), C. goeldii (78), S. sciureus (77) e C. jacchus (55). Às 72hs, houve uma redução significativa do PMI de quatro primatas: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) e C. jacchus (55/12), com exceção de C. goeldii (78/54). O FPMI dos primatas A. azarae infulatus (82.5) e C. goeldii (66) foi maior que do primata C. jacchus (33.5), porém, não foi maior que dos primatas C. penicillata (60.5) e S. sciureus (57.5). A resposta do TNF-α foi mais regular nos quatro primatas que reduziram o PMI no intervalo de 24-72hs: C. jacchus (145/122 pg/µL), C. penicillata (154/130 pg/µL), S. sciureus (164/104 pg/µL) e A. azarae infulatus (154/104 pg/µL), com exceção de C. goeldii (38/83 pg/µL). A resposta de IL-12 foi, principalmente, marcante nos primatas A. azarae infulatus e C. goeldii, os quais apresentaram as maiores taxas do FPMI, e a resposta do NO foi maior no primata C. goeldii, em especial no intervalo de 72hs. Estes achados sugerem, fortemente, que estes primatas neotropicais parecem ter desenvolvido mecanismos resistentes de resposta imune inata capaz de controlar o crescimento intracelular da infecção por L. (L.) i. chagasi no macrófago, o que não encoraja o uso destes primatas como modelo de estudo da LVA.

Efeito de agentes fluoretados e suplementados com trimetafosfato de sódio sobre a erosão e abrasão do esmalte dentário bovino: estudos in situ e ex vivo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Application of two segmentation protocols during the processing of virtual images in rapid prototyping: ex vivo study with human dry mandibles

The aim of this study was to evaluate the accuracy of virtual three-dimensional (3D) reconstructions of human dry mandibles, produced from two segmentation protocols (outline only and all-boundary lines).Twenty virtual three-dimensional (3D) images were built from computed tomography exam (CT) of 10 dry mandibles, in which linear measurements between anatomical landmarks were obtained and compared to an error probability of 5 %.The results showed no statistically significant difference among the dry mandibles and the virtual 3D reconstructions produced from segmentation protocols tested (p = 0,24).During the designing of a virtual 3D reconstruction, both outline only and all-boundary lines segmentation protocols can be used.Virtual processing of CT images is the most complex stage during the manufacture of the biomodel. Establishing a better protocol during this phase allows the construction of a biomodel with characteristics that are closer to the original anatomical structures. This is essential to ensure a correct preoperative planning and a suitable treatment.

Effect of fluoride varnish supplemented with sodium trimetaphosphate on enamel erosion and abrasion: An in situ/ex vivo study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expansão ex vivo de células uroteliais em scaffolds 3D

Malformations and possible damages to the urogenital system can be originated in the embryonic period. Moreover, fire guns, knives and accidents, where there is the disruption of the urethra, also cause these lesions. The objective was to analyze the contribution of tissue engineering in the construction of neo-urethra, developed by bioengineering. We performed an urothelial ex vivo expansion of cells in 3D scaffolds (platelet gel matrix and acellular porcine aorta) to assess the contribution of this technique in the construction of a neo-urethra. Mechanical dissociation was made of the inner wall of 10 North Folk rabbit’s bladder, weighing 2.5 to 3.0 kg. After dissociation the cell content was centrifuged and obtained a pellet of urothelial cells. The pellet was ressuspended in culture medium DMEM F12 and cells were maintained in culture for 15 days. Immunohistochemical analysis characterized the urothelial culture. The cells were then implanted in the scaffold - platelet gel. In a second experiment using aortic porcine acellular matrix were implanted urothelial cells alone and urothelial cells on platelet gel, on the inner wall of the scaffold - aorta, with space for setting bordered by a urethral probe. The complex probe - cells - aorta and probe - cells in platelet gel - aorta, were sealed with suture material and culture were maintained in a humidified 37ºC incubator with 5% CO2 in air for 12 days to subsequent histological analysis of urothelium cell adhesion to the scaffolds. By observation under an optical microscope, we could see the growth of cells in the scaffold platelet gel, from a monolayer in to a three-dimensional structure. In the acellular porcine aortic matrix containing the platelet gel, we could observe a few quantity of urothelial cells adhered. However with the acellular porcine aortic matrix in which was implanted only the urothelial cells, we have obtained adhesion to the wall

Preliminary in vitro and ex vivo evaluation of afzelin, kaempferitrin and pterogynoside action over free radicals and reactive oxygen species

Biological activities of flavonoids have been extensively reviewed in literature. The biochemical profile of afzelin, kaempferitrin, and pterogynoside acting on reactive oxygen species was investigated in this paper. The flavonoids were able to act as scavengers of the superoxide anion, hypochlorous acid and taurine chloramine. Although flavonoids are naturally occurring substances in plants which antioxidant activities have been widely advertised as beneficial, afzelin, kaempferitrin, and pterogynoside were able to promote cytotoxic effect. In red blood cells this toxicity was enhanced, depending on flavonoids concentration, in the presence of hypochlorous acid, but reduced in the presence of 2,20 -azo-bis(2-amidinopropane) free radical. These flavonoids had also promoted the death of neutrophils, which was exacerbated when the oxidative burst was initiated by phorbol miristate acetate. Therefore, despite their well-known scavenging action toward free radicals and oxidants, these compounds could be very harmful to living organisms through their action over erythrocytes and neutrophils.

Digital radiography for determination of primary tooth length: in vivo and ex vivo studies

Background. Methods for determining the root canal length of the primary tooth should yield accurate and reproducible results. In vitro studies show some limitations, which do not allow their findings to be directly transferred to a clinical situation. Aim. To compare the accuracy of radiographic tooth length obtained from in vivo digital radiograph with that obtained from ex vivo digital radiograph. Method. Direct digital radiographs of 20 upper primary incisors were performed in teeth (2/3 radicular resorption) that were radiographed by an intraoral sensor, according to the long-cone technique. Teeth were extracted, measured, and mounted in a resin block, and then radiographic template was used to standardise the sensor-target distance (30 cm). The apparent tooth length (APTL) was obtained from the computer screen by means of an electronic ruler accompanying the digital radiography software (CDR 2.0), whereas the actual tooth length (ACTL) was obtained by means of a digital calliper following extraction. Data were compared to the ACTL by variance analysis and Pearson’s correlation test. Results. The values for APTL obtained from in vivo radiography were slightly underestimated, whereas those values obtained from ex vivo were slightly overestimated. No significance was observed between APTL and ACTL. Conclusion. The length of primary teeth estimated by in vivo and ex vivo comparisons using digital radiography was found to be similar to the actual tooth length.

Caracterização radiográfica ex vivo de biomateriais utilizados para regeneração óssea em mandíbulas de porcos

Introduction: The radiographic characteristics of a biomaterial, such as its density, may influence the evaluation of the results obtained following its clinical use. Objective: The aim of this study was to evaluate the radiographic density of biomaterials used as bone substitutes, inserted into dental sockets and bone defects in created in the jaws of pigs. The influence of a soft tissue simulator on the results was also evaluated. Material and method: Two and three-millimeter-deep bone defects were created in the pigs mandible and the right first molar extraction socket were used. Commercial samples of five biomaterials were tested: Hydroxyapatite, Lyophilized Bovine Bone, 45S5 bioglass (generic), PerioGlass and β-Tri-Calcium Phosphate, and compared to a positive (mandibular bone) and negative (empty alveolar bone defects) controls. Radiographic images were acquired with and without a 10 mm thick soft-tissue simulator. Result: The results for the extraction sockets showed no differences between the biomaterials and the negative control. For the bone defects, the depth of the defect density influenced the density, both in the negative control (p < 0.01) and biomaterials (p < 0.05) groups. The soft- tissue simulator did not alter the results. Conclusion: The type of the evaluated defect can interfere in the radiographic features presented by each biomaterial, while the simulation of soft tissues was not statistically significant.

Modelo de conductancia hidráulica de la dentina humana ex vivo

The main objective of this work was to mount and test an experimental model to measure the hydraulic conductance of ex vivo dentin. Seventeen healthy third molars, with indication of extraction of healthy donors aged between 15 and 30 years were obtained by informed consent. After cleaning them, disinfecting them, including them in resin epoxy and cutting them, there were 17 samples of dentin, corresponding to a disk of resin with a coronal section of tooth showing the dentin exposed on both sides of it. Three machines to measure the hydraulic conductance of the dentin were assembled according to the description of the model of Pashley. Samples were installed in a Chamber of diffusion, connected by using silicone tubes to a graduated transfer pipette and a 20cm water column. Through the displacement of a bubble of water in the inside of the pipette, the hydraulic conductance of each sample was measured 3 times on the 14th, 21st, 28th and 35th day post extraction. The data were tabulated and analyzed statistically. There is no SS difference in the rate of flow of a measured sample in the three machines (p=0.5937). There is no SS difference in measurements of the hydraulic conductance of 13 samples of human dentin measured in days 14, 21, 28 and 35 postextraction (p=0.0704). It is possible to mount an experimental model to study the hydraulic conductance of dentin ex vivo, based on the model of Pashley. The model seems to be reliable, but more research is needed in order to validate its reliability.

Effect of toothpastes with different abrasives on eroded human enamel: an in situ/ex vivo study

The aim of the present study was to investigate the abrasive effect of CaCO3 and SiO2-based fluoride-free experimental toothpastes on eroded human permanent dental enamel and evaluate the effectiveness of waiting periods between acid exposure and tooth brushing. Twelve volunteers wore palatal appliances containing human enamel blocks for two periods of five days each. The appliances were immersed in a soft drink for five minutes four times a day (9:00 am, 11:00 am, 2:00 pm and 4:00 pm). On two occasions, two blocks were not submitted to additional treatment; two blocks were brushed (30 s) either with a CaCO3 or SiO2 toothpaste immediately after erosion and two blocks were brushed 1 h after erosion. Thus, the sample was divided into six groups: erosion alone (CaCO3 and SiO2 control); brushing with fluoride- free toothpaste (CaCO3 immediate and 1 h after erosion; SiO2 immediate and 1 h after erosion). Significant differences in wear depth were found between the enamel blocks in the CaCO3 immediate and 1 h after erosion groups and the blocks in the CaCO3 control group (p=0.001; p=0.022). No significant differences were found regarding the change in roughness and wear depth between blocks submitted to immediate abrasion and 1 h after erosion (CaCO3 and SiO2). The data revealed that surface roughness and wear depth is increased when erosion is combined with dental abrasion, regardless of the abrasive used. Waiting for 1 h to brush the eroded blocks offered no protective effect.

Perfil fenotípico de potenciais células iniciadoras tumorais no tumor venéreo transmissível canino ex vivo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ex vivo torsional properties of the Targon Vet Nail System in canine femurs: comparison with the 2.4 mm LC-DCP Plate

Pós-graduação em Cirurgia Veterinária - FCAV

Desenvolvimento e validação de método bioanalítico para quantificação de tiazolilhidrazona-9 (TZH-9) e determinação da estabilidade em plasma ex vivo

A doença de Chagas é uma doença parasitária causada pelo protozoário Trypanosoma cruzi (T. cruzi), que acomete o sistema digestivo e, principalmente, o cardíaco. É uma doença endêmica principalmente nos países da América Latina. A busca por novos fármacos para o tratamento dessa doença é de suma importância já que os existentes podem causar severos efeitos adversos e são efetivos somente na fase aguda. A molécula tiazolilhidrazona-9 (TZH-9) tem se destacado como uma excelente candidata a fármaco para tratamento da doença de Chagas e a continuidade de seu desenvolvimento deve envolver estudos de farmacocinética e metabolismo. Para a realização destes estudos é necessário o desenvolvimento e validação de um método bioanalítico para a determinação do composto em plasma, assim como avaliar a estabilidade do fármaco nesta matriz biológica com o intuito de verificar a ação de enzimas plasmáticas. Neste trabalho, foi desenvolvido e validado o método bioanalítico para quantificar o TZH-9 em plasma de ratos e humano que apresentou limites de confiança adequados. Também, avaliou-se a estabilidade do TZH-9 nessas matrizes biológicas, em plasma de ratos, o composto apresentou instabilidade após 2 horas na concentração inferior, sugerindo ação de enzimas plasmáticas no seu metabolismo; e em plasma de humanos, apresentou instabilidade após 15 minutos em ambas concentrações, sugerindo a susceptibilidade a ação das enzimas plasmáticas

Desenvolvimento e validação de método bioanalítico para quantificação de tiazolilhidrazona-9 (TZH-9) e determinação da estabilidade em plasma ex vivo

A doença de Chagas é uma doença parasitária causada pelo protozoário Trypanosoma cruzi (T. cruzi), que acomete o sistema digestivo e, principalmente, o cardíaco. É uma doença endêmica principalmente nos países da América Latina. A busca por novos fármacos para o tratamento dessa doença é de suma importância já que os existentes podem causar severos efeitos adversos e são efetivos somente na fase aguda. A molécula tiazolilhidrazona-9 (TZH-9) tem se destacado como uma excelente candidata a fármaco para tratamento da doença de Chagas e a continuidade de seu desenvolvimento deve envolver estudos de farmacocinética e metabolismo. Para a realização destes estudos é necessário o desenvolvimento e validação de um método bioanalítico para a determinação do composto em plasma, assim como avaliar a estabilidade do fármaco nesta matriz biológica com o intuito de verificar a ação de enzimas plasmáticas. Neste trabalho, foi desenvolvido e validado o método bioanalítico para quantificar o TZH-9 em plasma de ratos e humano que apresentou limites de confiança adequados. Também, avaliou-se a estabilidade do TZH-9 nessas matrizes biológicas, em plasma de ratos, o composto apresentou instabilidade após 2 horas na concentração inferior, sugerindo ação de enzimas plasmáticas no seu metabolismo; e em plasma de humanos, apresentou instabilidade após 15 minutos em ambas concentrações, sugerindo a susceptibilidade a ação das enzimas plasmáticas

Bactericidal effects of two parameters of Er:YAG laser intracanal irradiation: ex-vivo study

The success of endodontic treatment depends on the complete elimination of microorganisms from the root canal system, thus the search for new procedures to eliminate them is justified. The aim of this study was to assess bacterial reduction after intracanal irradiation with the Er:YAG laser. The canals of 70 extracted human maxillary canines were prepared up to file #40 using 1% NaOCl, irrigated with 17% EDTA, and then washed with physiological solution activated by ultrasound. The roots were sterilized by autoclaving, inoculated with 10 mu l of a suspension containing 1.5 x 10(8) CFU/ml of Enterococcus faecalis ATCC 29212 and incubated at 37A degrees C for 72 h. The canals were irradiated with the Er:YAG laser using two energy settings: 60 mJ and 15 Hz, and 100 mJ and 10 Hz. The remaining bacteria were counted immediately and 48 h after laser irradiation. The results showed a high bacterial reduction at both time points. With 60 mJ and 15 Hz there was an immediate reduction of 99.73% and the reduction was 77.02% after 48 h, and with 100 mJ and 10 Hz there was an immediate reduction of 99.95% and the reduction was 84.52% after 48 h. Although the best results were observed with 100 mJ of energy, the difference between the two settings was not statistically significant. The count performed 48 h after irradiation showed that E. faecalis were able to survive, and can grow even from small numbers.