Protective effects of the galectin-1 protein on in vivo and in vitro models of ocular inflammation

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

pH-cycling models for in vitro evaluation of the efficacy of fluoridated dentifrices for caries control: strengths and limitations

Despite a plethora of in situ studies and clinical trials evaluating the efficacy of fluoridated dentifrices on caries control, in vitro pH cycling models are still broadly used because they mimic the dynamics of mineral loss and gain involved in caries formation. This paper critically reviews the current literature on existing pH-cycling models for the in vitro evaluation of the efficacy of fluoridated dentifrices for caries control, focusing on their strengths and limitations. A search was undertaken in the MEDLINE electronic journal database using the keywords "pH-cycling", "demineralization", "remineralization", "in vitro", "fluoride", "dentifrice". The primary outcome was the decrease of demineralization or the increase of remineralization as measured by different methods (e. g.: transverse microradiography) or tooth fluoride uptake. Inclusion of studies, data extraction and quality assessment were undertaken independently and in duplicate by two members of the review team. Disagreements were solved by discussion and consensus or by a third party. One hundred and sixteen studies were included, of which 42 addressed specifically the comparison of dentifrices using different pH-cycling models. The other studies included meta-analysis or reviews, data about the effect of different fluoride sources on de-remineralization, different methods for analysis de-remineralization and chemical variables and characteristics of dental hard tissues that might have influence on de-remineralization processes. Generally, the studies presented ability to detect known results established by clinical trials, to demonstrate dose-related responses in the fluoride content of the dentifrices, and to provide repeatability and reproducibility between tests. In order to accomplish these features satisfactorily, it is mandatory to take into account the type of substrate and baseline artificial lesion, as well as the adequate response variables and statistical approaches to be used. This critical review of literature showed that the currently available pH-cycling models are appropriate to detect dose-response and pH-response of fluoride dentifrices, and to evaluate the impact of new active principles on the effect of fluoridated dentifrices, as well as their association with other anti-caries treatments.

Effect of green Coffea arabica L. seed oil on extracellular matrix components and water-channel expression in in vitro and ex vivo human skin models

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

Antifungal Efficacy during Candida krusei Infection in Non-Conventional Models Correlates with the Yeast In Vitro Susceptibility Profile

The incidence of opportunistic fungal infections has increased in recent decades due to the growing proportion of immunocompromised patients in our society. Candida krusei has been described as a causative agent of disseminated fungal infections in susceptible patients. Although its prevalence remains low among yeast infections (2-5%), its intrinsic resistance to fluconazole makes this yeast important from epidemiologic aspects. Non mammalian organisms are feasible models to study fungal virulence and drug efficacy. In this work we have used the lepidopteran Galleria mellonella and the nematode Caenorhabditis elegans as models to assess antifungal efficacy during infection by C. krusei. This yeast killed G. mellonella at 25, 30 and 37°C and reduced haemocytic density. Infected larvae melanized in a dose-dependent manner. Fluconazole did not protect against C. krusei infection, in contrast to amphotericin B, voriconazole or caspofungin. However, the doses of these antifungals required to obtain larvae protection were always higher during C. krusei infection than during C. albicans infection. Similar results were found in the model host C. elegans. Our work demonstrates that non mammalian models are useful tools to investigate in vivo antifungal efficacy and virulence of C. krusei. © 2013 Scorzoni et al.

Anti-inflammatory mechanisms of the annexin A1 protein and its mimetic peptide Ac2-26 in models of ocular inflammation in vivo and in vitro

Annexin A1 (AnxA1) is a protein that displays potent anti-inflammatory properties, but its expression in eye tissue and its role in ocular inflammatory diseases have not been well studied. We investigated the mechanism of action and potential uses of AnxA1 and its mimetic peptide (Ac2-26) in the endotoxin-induced uveitis (EIU) rodent model and in human ARPE-19 cells activated by LPS. In rats, analysis of untreated EIU after 24 and 48 h or EIU treated with topical applications or with a single s.c. injection of Ac2-26 revealed the anti-inflammatory actions of Ac2-26 on leukocyte infiltration and on the release of inflammatory mediators; the systemic administration of Boc2, a formylated peptide receptor (fpr) antagonist, abrogated the peptide's protective effects. Moreover, AnxA1-/- mice exhibited exacerbated EIU compared with wild-type animals. Immunohistochemical studies of ocular tissue showed a specific AnxA1 posttranslational modification in EIU and indicated that the fpr2 receptor mediated the anti-inflammatory actions of AnxA1. In vitro studies confirmed the roles of AnxA1 and fpr2 and the protective effects of Ac2-26 on the release of chemical mediators in ARPE-19 cells. Molecular analysis of NF-κB translocation and IL-6, IL-8, and cyclooxygenase-2 gene expression indicated that the protective effects of AnxA1 occur independently of the NF-κB signaling pathway and possibly in a posttranscriptional manner. Together, our data highlight the role of AnxA1 in ocular inflammation, especially uveitis, and suggest the use of AnxA1 or its mimetic peptide Ac2-26 as a therapeutic approach. Copyright © 2013 by The American Association of Immunologists, Inc.

Different bacterial models for in vitro induction of non-cavitated enamel caries-like lesions: microhardness and polarized light miscroscopy analyses

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

Effect of mucoadhesive polymers on the in vitro performance of insulin-loaded silica nanoparticles: Interactions with mucin and biomembrane models

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

Simulação e análise econômica da produção in vivo e in vitro de embriões em bovinos

The objective of this work was to evaluate the effect of the variables number of recipients, synchronization protocol, reproductive efficiency indicators and pregnancy cost, in the economic effectiveness of in vivo and in vitro bovine embryo production. A simulation application was elaborated to allow the user to insert the input variable parameters. A basic scenario, from the efficiency traditional rates of in vivo (ET) and in vitro production (IVP) techniques of bovine embryos, was introduced in the software as a criterion to compare the results. This software was able to reproduce both ET and IVP scenarios. The embryo production was simulated through stochastic simulation. The optimal number of recipients using sensitivity analysis was determined. The net present value and cost per pregnancy were used as a decision parameter. The synchronization for fixed-time embryo transfer decreased the recipient idleness and, consequently, the final cost of pregnancy, in comparison to the traditional methodology. Foetal sexing must be associated to IVP of bovine embryos. In addition, the optimal recipient number per donor is variable and depends on data inserted in the system.

In vitro characterization of coevaporates containing chitosan for colonic drug delivery

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of phosphated cross-linked high amylose on in vitro release of different drugs

The influence of structural characteristics of high amylose cross-linked at different degrees on the release of drugs with important molecular differences, namely sodium diclophenac (SD) and nicotinamide (NI), was assessed in vitro from non-compacted systems. The release profiles were related with classical kinetic mathematical models for better understanding of the release mechanism. An increase in polymer cross-linking degree resulted in longer release time for both drugs, although SD generally was released slower than NI. SD release from samples cross-linked at 2% of basis was driven mainly by Fickian diffusion, while from samples cross-linked at 4% of basis follows anomalous mechanism. Inversely, anomalous mechanism was responsible for NI release from 2% samples and Fickian diffusion from 4% samples. Results suggest that the performance of cross-linked high amylose as excipient for controlled drug release not only depends on cross-linking degree but also is highly influenced by structural characteristics of the drug. (C) 2009 Elsevier Ltd. All rights reserved.

Prognostic value of canine frozen-thawed semen parameters on in vitro sperm-oocyte interactions

Combining the data from conventional semen analysis with oocyte penetration assays should improve the assessment of the fertilizing ability of a semen sample. Thus, the objective of the present study was to evaluate the prognostic value of various semen parameters on the in vitro interactions between frozen-thawed canine sperm and homologous oocytes. Ten ejaculates from five stud dogs (two ejaculates/dog) were collected by digital manipulation. Semen samples were evaluated, extended in Tris-egg yolk-glycerol, frozen and stored in liquid nitrogen, and thawed several weeks later. Samples were evaluated for motility and sperm populations by computer-aided semen analysis (CASA), plasma membrane integrity (carboxy-fluorescein diacetate and propidium iodide), and sperm morphology (Bengal Rose). Thawed spermatozoa were also incubated with homologous oocytes for 18 h in an atmosphere of 5% CO2 and 95% air at 38 degrees C and sperm-oocyte interactions were evaluated. Simple linear regression models were calculated, with sperm parameters as independent variables and sperm-oocyte interactions as the dependent variable. There were significant associations between: percentage of oocytes bound to spermatozoa and beat cross frequency (BCF; R-2 = 63%); percentage of oocytes that interacted with spermatozoa and BCF (R-2 = 73%); and number of penetrated spermatozoa and velocity average pathway (VAP; R-2 = 64%) and velocity straight line (VSL; R-2 = 64%). Although plasma membrane integrity and sperm morphology had little prognostic value for in vitro interactions between canine frozen-thawed sperm and homologous oocytes, some motility patterns (evaluated by CASA) were predictive of in vitro sperm-oocyte interactions. (c) 2005 Elsevier B.V. All rights reserved.

Effect of iron on enamel demineralization and remineralization in vitro

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

In Vitro Biomechanical Evaluation of the Use of Conventional and Locking Miniplate/Screw Systems for Sagittal Split Ramus Osteotomy

Purpose: The aim of this in vitro study was to assess the biomechanical stability of 9 different osteosynthesis methods after sagittal split ramus osteotomy by simulating the masticatory forces and using a 3-point biomechanical test method.Materials and Methods: Forty-five polyurethane hemimandibles with bone-like consistency were randomly assigned to 9 groups (n = 5) and subjected to sagittal split ramus osteotomy. After 4-mm advancement of the distal segment, the bone segments were fixed by different osteosynthesis methods using 2.0-mm miniplate/screw systems: group A, one 4-hole conventional straight miniplate; group B, one 4-hole locking straight miniplate; group C, one 4-hole conventional miniplate and one bicortical screw; group D, one 4-hole locking miniplate and 1 bicortical screw; group E, one 6-hole conventional straight miniplate; group F, one 6-hole locking straight miniplate; group (3: two 4-hole conventional straight miniplates; group H. two 4-hole locking straight miniplates; and group 1, 3 bicortical screws in an inverted-L. pattern. All models were mounted on a base especially constructed for this purpose. Using a 3-point biomechanical test model, the hemimandibles were loaded in compressive strength in an Instron machine (Norwood, MA) until a 3-mm displacement occurred between segments vertically or horizontally. Data were analyzed by analysis of variance and Tukey test (alpha = 1%).Results: The multiparametric comparison of the groups showed a statistically significant difference (P<.01) between groups that used 2 miniplates (groups G and H), 1 miniplate and 1 bicortical screw (groups C and D), and only bicortical screws (group D compared with groups that used only 1 miniplate with 2 screws per segment (groups A and B) and 3 screws per segment (groups E and F).Conclusion: The placement of 2.0-mm-diameter bicortical screws in the retromolar region, associated or not with conventional and locking miniplates with monocortical screws, promoted a better stabilization of bone segments. Locking miniplates presented a better performance in bone fixation in all groups. (C) 2010 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 68:724-730, 2010

Biomechanical evaluation of internal and external hexagon platform switched implant-abutment connections: An in vitro laboratory and three-dimensional finite element analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)