Biological response to wollastonite doped α-tricalcium phosphate implants in hard and soft tissues in rats

The biological response following subcutaneous and bone implantation of β-wollastonite(β-W)-doped α-tricalcium phosphate bioceramics in rats was evaluated. Tested materials were: tricalcium phosphate (TCP), consisting of a mixture of α- and β-polymorphs; TCP doped with 5 wt. % of β-W (TCP5W), composed of α-TCP as only crystalline phase; and TCP doped with 15 wt. % of β-W (TCP 15), containing crystalline α-TCP and β-W. Cylinders of 2×1 mm were implanted in tibiae and backs of adult male Rattus norvegicus, Holtzman rats. After 7, 30 and 120 days, animals were sacrificed and the tissue blocks containing the implants were excised, fixed and processed for histological examination. TCP, TCP5W and TCP15W implants were biocompatible but neither bioactive nor biodegradable in rat subcutaneous tissue. They were not osteoinductive in connective tissue either. However, in rat bone tissue β-W-doped α-TCP implants (TCP5W and TCP 15W) were bioactive, biodegradable and osteoconductive. The rates of biodegradation and new bone formation observed for TCP5W and TCP15W implants in rat bone tissue were greater than for non-doped TCP.

Bothropstoxin-I reduces evoked acetylcholine release from rat motor nerve terminals: Radiochemical and real-time video-microscopy studies

Understanding the biological activity profile of the snake venom components is fundamental for improving the treatment of snakebite envenomings and may also contribute for the development of new potential therapeutic agents. In this work, we tested the effects of BthTX-I, a Lys49 PLA2 homologue from the Bothrops jararacussu snake venom. While this toxin induces conspicuous myonecrosis by a catalytically independent mechanism, a series of in vitro studies support the hypothesis that BthTX-I might also exert a neuromuscular blocking activity due to its ability to alter the integrity of muscle cell membranes. To gain insight into the mechanisms of this inhibitory neuromuscular effect, for the first time, the influence of BthTX-I on nerve-evoked ACh release was directly quantified by radiochemical and real-time video-microscopy methods. Our results show that the neuromuscular blockade produced by in vitro exposure to BthTX-I (1 μM) results from the summation of both pre- and postsynaptic effects. Modifications affecting the presynaptic apparatus were revealed by the significant reduction of nerve-evoked [3H]-ACh release; real-time measurements of transmitter exocytosis using the FM4-64 fluorescent dye fully supported radiochemical data. The postsynaptic effect of BthTX-I was characterized by typical histological alterations in the architecture of skeletal muscle fibers, increase in the outflow of the intracellular lactate dehydrogenase enzyme and progressive depolarization of the muscle resting membrane potential. In conclusion, these findings suggest that the neuromuscular blockade produced by BthTX-I results from transient depolarization of skeletal muscle fibers, consequent to its general membrane-destabilizing effect, and subsequent decrease of evoked ACh release from motor nerve terminals. © 2012 Elsevier Ltd.

Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n=64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies. © 2013 Wiley Periodicals, Inc.

In vitro enamel remineralization by low-fluoride toothpaste with calcium citrate and sodium trimetaphosphate

The objective of this study was to evaluate in vitro the effect of a low fluoride toothpaste (450 μgF/g, NaF) combined with calcium citrate (Cacit) and sodium trimetaphosphate (TMP) on enamel remineralization. Bovine enamel blocks had the enamel surface polished sequentially to determine the surface hardness. After production of artificial carious lesions, the blocks selected by their surface hardness were submitted to remineralization pH cycling and daily treatment with dentifrice suspensions (diluted in deionized water or artificial saliva): placebo, 275, 450, 550 and 1,100 μgF/g and commercial dentifrice (positive control, 1,100 μgF/g). Finally, the surface and cross-section hardness was determined for calculating the change of surface hardness (%SH) and mineral content (%ΔZ). Fluoride in enamel was also determined. The data from %SH, %ΔZ and fluoride were subjected to two-way analysis of variance followed by Student-Newman-Keuls's test (p<0.05). The mineral gain (%SH and %ΔZ) was higher for toothpastes diluted in saliva (p<0.05), except for the 450 mgF/g dentifrice with Cacit/TMP (p>0.05). The 450 Cacit/ TMP toothpaste and the positive control showed similar results (p>0.05) when diluted in water. A dose-response was observed between fluoride concentration in toothpastes and fluoride present in enamel, regardless of dilution. It was concluded that it is possible to enhance the remineralization capacity of low F concentration toothpaste by of organic (Cacit) and inorganic (TMP) compounds with affinity to hydroxyapatite.

Propolis and its constituent caffeic acid suppress LPS-stimulated pro-inflammatory response by blocking NF-κB and MAPK activation in macrophages

Ethnopharmacological relevance Propolis is a bee product with numerous biological and pharmacological properties, such as immunomodulatory and anti-inflammatory activities. It has been used in folk medicine as a healthy drink and in food to improve health and prevent inflammatory diseases. However, little is known about its mechanism of action. Thus, the goal of this study was to verify the antioxidant activity and to explore the anti-inflammatory properties of propolis by addressing its intracellular mechanism of action. Caffeic acid was investigated as a possible compound responsible for propolis action. Materials and methods The antioxidant properties of propolis and caffeic acid were evaluated by using the 2,2-Diphenyl-1-picrylhydrazyl free radical (DPPH) scavenging method. To analyze the anti-inflammatory activity, Raw 264.7 macrophages were treated with different concentrations of propolis or caffeic acid, and nitric oxide (NO) production, a strong pro-inflammatory mediator, was evaluated by the Griess reaction. The concentrations of propolis and caffeic acid that inhibited NO production were evaluated on intracellular signaling pathways triggered during inflammation, namely p38 mitogen-activated protein kinase (MAPK), c-jun NH2-terminal kinase (JNK1/2), the transcription nuclear factor (NF)-κB and extracellular signal-regulated kinase (ERK1/2), through Western blot using specific antibodies. A possible effect of propolis on the cytotoxicity of hepatocytes was also evaluated, since this product can be used in human diets. Results Caffeic acid showed a higher antioxidant activity than propolis extract. Propolis and caffeic acid inhibited NO production in macrophages, at concentrations without cytotoxicity. Furthermore, both propolis and caffeic acid suppressed LPS-induced signaling pathways, namely p38 MAPK, JNK1/2 and NF-κB. ERK1/2 was not affected by propolis extract and caffeic acid. In addition, propolis and caffeic acid did not induce hepatotoxicity at concentrations with strong anti-inflammatory potential. Conclusions Propolis exerted an antioxidant and anti-inflammatory action and caffeic acid may be involved in its inhibitory effects on NO production and intracellular signaling cascades, suggesting its use as a natural source of safe anti-inflammatory drugs. © 2013 Elsevier B.V.

Efeitos da redução no conteúdo de sódio da dieta sobre o volume de água corporal e marcadores inflamatórios em pacientes com insuficiência renal crônica em tratamento hemodialítico

Pós-graduação em Fisiopatologia em Clínica Médica - FMB

Coleta, processamento, caracterização fenótipa das células-tronco mesenquimais e sua viabilidade de aplicação por via intratecal em equinos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cellular and Molecular Tissue Response to Triple Antibiotic Intracanal Dressing

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microparticulated and nanoparticulated zirconium oxide added to calcium silicate cement: Evaluation of physicochemical and biological properties

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biocompatibility of New Calcium Aluminate Cement: Tissue Reaction and Expression of Inflammatory Mediators and Cytokines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The physicochemical characterization and in vivo response of micro/nanoporous bioactive ceramic particulate bone graft materials

In this study, the physicochemical characteristics of calcium phosphate based bioactive ceramics of different compositions and blends presenting similar micro/nanoporosity and micrometer scale surface texture were characterized and evaluated in an in vivo model. Prior to the animal experiment, the porosity, surface area, particle size distribution, phase quantification, and dissolution of the materials tested were evaluated. The bone regenerative properties of the materials were evaluated using a rabbit calvaria model. After 2, 4, and 8 weeks, the animals were sacrificed and all samples were subjected to histologic observation and histomorphometric analysis. The material characterization showed that all materials tested presented variation in particle size, porosity and composition with different degrees of HA/TCP/lower stoichiometry phase ratios. Histologically, the calvarial defects presented temporal bone filling suggesting that all material groups were biocompatible and osteoconductive. Among the different materials tested, there were significant differences found in the amount of bone formation as a function of time. At 8 weeks, the micro/nanoporous material presenting similar to 55,TCP:45%,HA composition ratio presented higher amounts of new bone regeneration relative to other blends and a decrease in the amount of soft tissue infiltration. (C) 2014 Elsevier B.V. All rights reserved.

ChIP-seq reveals a role for CrzA in the Aspergillus fumigatus high-osmolarity glycerol response (HOG) signalling pathway

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Immediate human pulp response to ethanol-wet bonding technique

To evaluate the short-term response of human pulps to ethanol-wet bonding technique. Methods Deep class V cavities were prepared on 17 sound premolars and divided into three groups. After acid-etching, the cavities from groups 1 (G1) and 2 (G2) were filled with 100% ethanol or distilled water, respectively, for 60 s before the application of Single Bond 2. In group 3 (G3, control), the cavity floor was lined with calcium hydroxide before etching and bonding. All cavities were restored with resin composite. Two teeth were used as intact control. The teeth were extracted 48 h after the clinical procedures. From each tooth serial sections were obtained and stained with haematoxylin and eosin (H/E) and Masson's trichrome. Bacteria microleakage was assessed using Brown & Brenn. All sections were blindly evaluated for five histological features. Results Mean remaining dentine thickness was 463 ± 65 μm (G1); 425 ± 184 μm (G2); and 348 ± 194 μm (G3). Similar pulp reactions followed ethanol- or water-wet bonding techniques. Slight inflammatory responses and disruption of the odontoblast layer related to the cavity floor were seen in all groups. Stained bacteria were not detected in any cavities. Normal pulp tissue was observed in G3 except for one case. Conclusions After 48 h, ethanol-wet bonding does not increase pulpal damage compared to water-wet bonding technique. Clinical significance Ethanol-wet bonding may increase resin-dentine bond durability. This study reported the in vivo response of human pulp tissue when 100% ethanol was applied previously to an etch-and-rinse simplified adhesive system.

Repair of Bone Defects Filled with New Calcium Aluminate Cement (EndoBinder)

The aim of this study was to evaluate and compare the repair of bone defects filled with calcium aluminate cement (EndoBinder), mineral trioxide aggregate (MTA), and calcium hydroxide. Methods After mixing, the cements were inserted into bone defects (3.3 mm) mechanically created in the right and left tibias of 30 rats (Rattus norvegicus, Wistar). In the control group, the bone defects were filled with blood clot of the animal itself. After time intervals of 7, 30, and 90 days had elapsed, bone tissue biopsies (n = 5) were surgically obtained and submitted to laboratory processing. The response of bone tissue in contact with the materials was microscopically analyzed. The percentage of neoformed bone tissue in the defect was determined by means of planimetry counting points superimposed on the histologic image. Results Significant increase in the percentage of neoformed bone tissue was observed throughout the experimental periods in all groups (P < .05). For the cements EndoBinder and MTA (30 and 90 days), these percentage values were statistically higher than those of the control group (P < .05); however, they were similar to those of calcium hydroxide (P > .05). Conclusions EndoBinder and MTA allowed complete repair of bone defects created in rat tibias.

Response of human pulps to different in-office bleaching techniques: preliminary findings

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)