In vivo evaluation of porous titanium implants with biomimetic coating

The osseointegration of porous titanium implants was evaluated in the present work. Implants were fabricated from ASTM grade 2 titanium by a powder metallurgy method. Part of these implants were submitted to chemical and thermal treatment in order to deposit a biomimetic coating, aiming to evaluate its influence on the osseointegration of the implants. The implants were characterized by Scanning Electron Microscopy (SEM), Electron Dispersive X-Ray Spectroscopy (EDS) and Raman Spectroscopy. Three coated and three control (uncoated) implants were surgically inserted into thirty albino rabbits' left and right tibiae, respectively. Tibiae samples were submitted to histological and histomorphometric analyses, utilizing SEM, optical microscopy and mechanical tests. EDS results indicated calcium (Ca) and phosphorous (P) at the surface and Raman spectra exhibited an intense peak, characteristic of hydroxyapatite (HA). Bone neoformation was detected at the bone-implant interface and inside the pores, including the central ones. The mean bone neoformation percentage in the coated implants was statistically higher at 15 days, compared to 30 and 45 days. The mechanical tests showed that coated implants presented higher resistance to displacement, especially after 30 and 45 days.

In vivo biological behavior of calcium phosphate cements with different Ca/P ratio

Bioceramics with different Ca/P ratio were prepared from a mechanical mixture of NaPO3, CaCO3, Ca(OH)2 and phosphate buffer solution and implanted in rats subcutaneous tissues. The cements were characterized by Thermo gravimetric analysis (TG-TDA), X-ray diffraction and 31P-NMR. The implant sites were excised after 1, 4 and 16 weeks, fixed, dehydrated, included in paraffin wax for serial cutting and examined under the light transmitted microscope. They were biocompatible and biodegradable when implanted in rat subcutaneous. None of the materials induced ectopic osteogenesis. According to the results, the studied materials seem to be able for manufacturing reabsorbable bone implants.

Glucocorticoids in vivo induce both insulin hypersecretion and enhanced glucose sensitivity of stimulus-secretion coupling in isolated rat islets

Although glucocorticoids are widely used as antiinflammatory agents in clinical therapies, they may cause serious side effects that include insulin resistance and hyperinsulinemia. To study the potential functional adaptations of the islet of Langerhans to in vivo glucocorticoid treatment, adult Wistar rats received dexamethasone (DEX) for 5 consecutive days, whereas controls (CTL) received only saline. The analysis of insulin release in freshly isolated islets showed an enhanced secretion in response to glucose in DEX-treated rats. The study of Ca2 2+ signals by fluorescence microscopy also demonstrated a higher response to glucose in islets from DEX-treated animals. However, no differences in Ca2 2+signals were found between both groups with tolbutamide or KCl, indicating that the alterations were probably related to metabolism. Thus, mitochondrial function was explored by monitoring oxidation of nicotinamide dinucleotide phosphate autofluorescence and mitochondrial membrane potential. Both parameters revealed a higher response to glucose in islets from DEX-treated rats. The mRNA and protein content of glucose transporter-2, glucokinase, and pyruvate kinase was similar in both groups, indicating that changes in these proteins were probably not involved in the increased mitochondrial function. Additionally,weexplored the status of Ca2 2+-dependent signaling kinases. Unlike calmodulin kinase II, we found an augmented phosphorylation level of protein kinase Cα as well as an increased response of the phospholipase C/inositol 1,4,5-triphosphate pathway in DEX-treated rats. Finally, an increased number of docked secretory granules were observed in the β-cells of DEX animals using transmission electron microscopy. Thus, these results demonstrate that islets from glucocorticoid-treated rats develop several adaptations that lead to an enhanced stimulus-secretion coupling and secretory capacity. Copyright © 2010 by The Endocrine Society.

pH-changes during intracoronal bleaching: An in vivo study

Objectives: This study aimed to measure pH changes during 14 days intracoronal bleaching with hydrogen peroxide/sodium perborate and carbamide peroxide/sodium perborate. Materials and methods: Twenty patients presenting endodontically treated central maxillary incisors with color alterations were divided in two groups (n = 10): Group CP + SP: 37% carbamide peroxide + sodium perborate paste; Group HP + SP: 30% hydrogen peroxide + sodium perborate paste. The pH values were measured using a digital microprocessor at different times: Baseline, 2, 7 and 14 days. Data were analyzed with two-way ANOVA followed by Tukey's test (α = 0.05). Results: ANOVA showed p < 0.00 which indicated significant difference between the groups. The mean values (± sd) and the results of the Tukey's test were: HP + SP/14 days-7.98 (±0.58)a; HP + SP/7 days-8.59 (±0.18)b; HP + SP/2 days-8.83 (±0.32)bc; HP + SP/Baseline-8.83 (±0.01)bc; CP + SP/Baseline-8.89 (±0.01)bc; CP + SP/14 days-9.11 (±0.58)cd; CP + SP/7 days-9.54 (±0.16)de; CP + SP/2 days-9.66 (±0.08) de. The group HP + SP resulted in significantly lower pH values compared with group CP + SP. Conclusion: It can be concluded that both associations showed alkaline pH values; however, there was significant reduction in the pH values of the 30% hydrogen peroxide associated with sodium perborate after 14 days. Clinical Significance: The association of hydrogen peroxide and carbamide peroxide with sodium perborate paste presented alkaline characteristics during the 14-day evaluated period. Thus, regarding pH changes, both associations can be considered safe as intracoronal bleaching agents.

Evaluation of Agaricus blazei in vivo for antigenotoxic, anticarcinogenic, phagocytic and immunomodulatory activities

The development of various types of cancer results from the interaction among endogenous, environmental and hormonal factors, where the most notable of these factors is diet. The aim of the present study was to determine the antigenotoxic, anticarcinogenic, phagocytic and immunomodulatory activities of Agaricus blazei. The test antigenotoxicity (Comet Assay) and anticarcinogenic (Test of Aberrant Crypt Foci) assess changes in DNA and/or intestinal mucosa that correlate to cancer development. Tests of phagocytosis in the spleen and differential count in blood cells allow the inference of modulation of the immune system as well as to propose a way of eliminating cells with DNA damage. Supplementation with the mushroom was carried out under pre-treatment, simultaneous treatment, post-treatment and pre-treatment. +. continuous conditions. Statistical analysis demonstrated that the mushroom did not have genotoxic activity but showed antigenotoxic activity. Supplementation caused an increase in the number of monocytes and in phagocytic activity, suggesting that supplementation increases a proliferation of monocytes, consequently increasing phagocytic capacity especially in the groups pre-treatment, simultaneous and pre-treatment. +. continuous. The data suggest that A. blazei could act as a functional food capable of promoting immunomodulation which can account for the destruction of cells with DNA alterations that correlate with the development of cancer, since this mushroom was demonstrated to have a preventive effect against pre-neoplastic colorectal lesions evaluated by the aberrant crypt foci assay. According to these results and the literature, it is believed that supplementation with A. blazei can be an efficient method for the prevention of cancer as well as possibly being an important coadjuvant treatment in chemotherapy. © 2011 Elsevier Inc.

Use of bayesian inference to correlate in vitro embryo production and in vivo fertility in Zebu Bulls

The objective of this experiment was to test in vitro embryo production (IVP) as a tool to estimate fertility performance in zebu bulls using Bayesian inference statistics. Oocytes were matured and fertilized in vitro using sperm cells from three different Zebu bulls (V, T, and G). The three bulls presented similar results with regard to pronuclear formation and blastocyst formation rates. However, the cleavage rates were different between bulls. The estimated conception rates based on combined data of cleavage and blastocyst formation were very similar to the true conception rates observed for the same bulls after a fixed-time artificial insemination program. Moreover, even when we used cleavage rate data only or blastocyst formation data only, the estimated conception rates were still close to the true conception rates. We conclude that Bayesian inference is an effective statistical procedure to estimate in vivo bull fertility using data from IVP. © 2011 Mateus José Sudano et al.

Absence of genotoxic effects of the coumarin derivative 4-methylesculetin in vivo and its potential chemoprevention against doxorubicin-induced DNA damage

4-Methylesculetin (4-ME) is a synthetic derivative of coumarin that displays a potent reactive oxygen species (ROS) scavenger and metal chelating agent and therefore has been produced to help reduce the risk of human disease. The main objective of this study was to investigate the in vivo genotoxicity of 4-ME and initially to verify its potential antigenotoxicity on doxorubicin (DXR)-induced DNA damage. Different doses of 4-ME (500, 1000 and 2000mgkg -1 body weight) were administered by gavage only or with a simultaneous intraperitoneal (i.p.) injection of DXR (80mgkg -1). The following endpoints were analyzed: DNA damage in peripheral blood, liver, bone marrow, brain and testicle cells according to an alkaline (pH>13) comet assay and micronucleus induction in bone marrow cells. Cytotoxicity was assessed by scoring polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). No differences were observed between the negative control and the groups treated with a 4-ME dose for any of the endpoints analyzed, indicating that it lacks genotoxic and cytotoxic effects. Moreover, 4-ME demonstrated protective effects against DXR-induced DNA damage at all tested doses and in all analyzed cell types, which ranged from 34.1% to 93.3% in the comet assay and 54.4% to 65.9% in the micronucleus test.

Genotoxicity of the medicinal plant Maytenus robusta in mammalian cells in vivo.

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

Comparative in vivo study of commercially pure Ti implants with surfaces modified by laser with and without silicate deposition: Biomechanical and scanning electron microscopy analysis

The purpose of this study was to evaluate commercially pure titanium implant surfaces modified by laser beam (LS) and LS associated with sodium silicate (SS) deposition, and compare them with machined surface (MS) and dual acid-etching surfaces (AS) modified. Topographic characterization was performed by scanning electron microscopy-X-ray energy dispersive spectroscopy (SEM-EDX), and by mean roughness measurement before surgery. Thirty rabbits received 60 implants in their right and left tibias. One implant of each surface in each tibia. The implants were removed by reverse torque for vivo biomechanical analysis at 30, 60, and 90 days postoperative. In addition, the surface of the implants removed at 30 days postoperative was analyzed by SEM-EDX. The topographic characterization showed differences between the analyzed surfaces, and the mean roughness values of LS and SS were statistically higher than AS and MS. At 30 days, values removal torque LS and SS groups showed a statistically significant difference (p < 0.05) when compared with MS and AS. At 60 days, groups LS and SS showed statistically significant difference (p < 0.05) when compared with MS. At 90 days, only group SS presented statistically higher (p < 0.05) in comparison with MS. The authors can conclude that physical chemistry properties and topographical of LS and SS implants increases bone-implant interaction and provides higher degree of osseointegration when compared with MS and AS. © 2012 Wiley Periodicals, Inc.

EIF5A dimerizes not only in vitro but also in vivo and its molecular envelope is similar to the EF-P monomer

The protein eukaryotic initiation factor 5A (eIF5A) is highly conserved among archaea and eukaryotes, but not in bacteria. Bacteria have the elongation factor P (EF-P), which is structurally and functionally related to eIF5A. eIF5A is essential for cell viability and the only protein known to contain the amino acid residue hypusine, formed by post-translational modification of a specific lysine residue. Although eIF5A was initially identified as a translation initiation factor, recent studies strongly support a function for eIF5A in the elongation step of translation. However, the mode of action of eIF5A is still unknown. Here, we analyzed the oligomeric state of yeast eIF5A. First, by using size-exclusion chromatography, we showed that this protein exists as a dimer in vitro, independent of the hypusine residue or electrostatic interactions. Protein-protein interaction assays demonstrated that eIF5A can form oligomers in vitro and in vivo, in an RNA-dependent manner, but independent of the hypusine residue or the ribosome. Finally, small-angle X-ray scattering (SAXS) experiments confirmed that eIF5A behaves as a stable dimer in solution. Moreover, the molecular envelope determined from the SAXS data shows that the eIF5A dimer is L-shaped and superimposable on the tRNAPhe tertiary structure, analogously to the EF-P monomer. © 2012 Springer-Verlag.

In vitro and in vivo investigation of the biological and mechanical behaviour of resin-modified glass-ionomer cement containing chlorhexidine

Objectives: To evaluate: (1) the in vitro antibacterial, cytotoxic and mechanical properties of a resin-modified glass ionomer cement (RMGIC) containing different concentrations of chlorhexidine (CHX) and (2) the in vivo microbiologic action of the best concentration of CHX associated with the RMGIC applied on remaining dentine after indirect pulp treatment (IPT). Methods: For the in vitro studies, RMGIC was associated with 0.2, 0.5, 1.25 and 2.5% CHX. Microbiologic evaluation consisted of an agar diffusion test on cariogenic bacteria for 24 h. Odontoblast-like cell metabolism and morphology analyses measured the cytotoxic effects of the RMGIC groups after 24 h. The same groups were submitted to compressive and diametral tensile strength. The in vivo treatment consisted of IPT using an RMGIC associated with the best CHX concentration. Clinical and microbiologic evaluations were performed before and after 3 months. Results: The use of 1.25% CHX significantly improved the antibacterial effects of the evaluated RMGIC, without causing any detrimental effects to the odontoblast-like cells and on the mechanical properties. This RMGIC and CHX combination completely eliminated mutans streptococci after 3 months of IPT. Conclusion: The RMGIC and 1.25% CHX mixture showed great biological and mechanical behaviour and could be a good treatment against caries progression. Clinical significance: The association of CHX with a liner RMGIC opens a new perspective for arresting residual caries after IPT. © 2012 Elsevier Ltd. All rights reserved.

Use of grape polyphenols against carcinogenesis: Putative molecular mechanisms of action using in vitro and in vivo test systems

Polyphenols are present in foods and beverages and are related to sensorial qualities such as color, bitterness, and astringency, which are relevant in wine, tea, grape juice, and other products. These compounds occur naturally in forms varying from simple phenolic acids to complex polymerized tannins. Thus, it is reasonable to expect that grape-derived products elaborated in the presence of skins and seeds, such as wine and grape juice, are natural sources of flavonoids in the diet. Carcinogenesis is a multistep process that is characterized by genetic, epigenetic, and phenotypic changes. With increasing knowledge of these mechanisms, and the conclusion that most cases of cancer are preventable, efforts have focused on identifying the agents with potential anticancer properties. The use of grape polyphenols against the carcinogenesis process seems to be a suitable alternative for either prevention and/or therapeutic purposes. The aim of this article is to show the molecular data generated from the use of grape polyphenols against carcinogenesis using in vivo and in vitro test systems. © Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition.

Ex-vivo Effect of Intracanal Medications Based on Ozone and Calcium Hydroxide in Root Canals Contaminated with Enterococcus faecalis

This ex vivo study evaluated the antibacterial effect of intracanal medications in root canals contaminated with Enterococcus faecalis. Fifty single-rooted human teeth were contaminated with E. faecalis (ATCC 29212) and incubated at 37°C for 21 days. The specimens were randomly divided into 5 groups according to the intracanal medication used: OZ-PG: ozonized propylene glycol; CH/CPMC: calcium hydroxide/camphorated paramonochlorophenol; OZ-PG/CH ozonized PG/CH; PC: positive control group (no medication); and NC: negative control group (no contamination). The samples were collected after 7 days (post-medication) and 14 days (final). Bacterial growth was checked by counting the colony-forming units (CFU). OZ-PG and CH/CPMC reduced significantly the CFU counts compared with PC in the post-medication and final samples, with no statistically significant differences among them. On the other hand, OZ-PG/CH did not reduce significantly the number of bacteria compared with PC. In conclusion, among the evaluated medications OZ-PG and CH/CPMC were the most effective against E. faecalis.

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Morphology, sex ratio and gene expression of Day 14 in vivo and in vitro bovine embryos

The present study was designed to compare Day 14 bovine embryos that were produced entirely in vitro using the post-hatching development (PHD) system with in vivo-derived embryos without or with transient PHD culture from Day 7 to Day 14. Embryos on Day 14 were used for sex determination and gene expression analysis of PLAC8, KRT8, CD9, SLC2A1, SLC2A3, PGK1, HSF1, MNSOD, HSP70 and IFNT using real-time quantitative (q) polymerase chain reaction (PCR). First, Day 7 in vivo-and in vitro-produced embryos were subjected to the PHD system. A higher rate of survival was observed for in vitro embryos on Day 14. Comparing Day 14 embryos produced completely in vivo or completely in vitro revealed that the mean size of the former group was greater than that of the latter (10.29±1.83 vs 2.68±0.33mm, respectively). Expression of the HSP70 and SLC2A1 genes was down-and upregulated, respectively, in the in vitro embryos. The present study shows that in vitro embryos cultured in the PHD system are smaller than in vivo embryos and that of the 10 genes analysed, only two were differentially expressed between the two groups. These findings indicate that, owing to the poor survival rate, the PHD system is not reliable for evaluation of in vitro embryo quality. © 2013 CSIRO.