Avaliação do padrão de clonalidade e virulencia de S. mutans isolados de individuos livres de carie e carie-ativos

Orientador: Reginaldo Bruno GonçalvesDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaMestrad

Osteoclastogenesis induction RANK-independent by a lipid fraction from Porphyromonas gingivalis

Orientador: Reginaldo Bruno GonçalvesTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Porphyromonas gingivalis (Pg) sintetiza diversas classes de fosfolipídeos. Entretanto, pouco se sabe sobre os efeitos biológicos que estes lipídeos exercem sobre a reabsorção óssea. Células precursoras de osteoclastos de camundongos, incluindo RAW264.7 e células da medula óssea, foram estimuladas com um lipídeo recém caracterizado isolado de Pg (F24), LPS de Pg, RANKL e um isômero estrutural isolado de mamíferos (DMPE), seguida de coloração das células por fosfatase ácida (TRAP) para detecção de osteoclastos. A formação de lacunas de reabsorção em discos de dentina foi realizados para identificar a presença de osteoclastos maduros. Proteínas de células precursoras RAW264.7 com afinidade a F24 foram identificadas como sendo nucleolin através de espectometria de massa, e confirmada sua presença através de ensaio em microscopia confocal. Utilizando Western-blot e RNAi, a sinalização intracelular desencadeada pela F24 durante a osteoclastogênese foi analisada. A F24 mas não o LPS ou DMPE induziu números significantemente maiores de células multinucleadas TRAP+ quando comparado ao RANKL, assim como a formação de lacunas de desmineralização em discos de dentina, o qual não foi inibida pelo repressor do RANKL, a OPG. LPS de Pg, mas não a F24, induziu a produção de TNF-a, IL-1ß e IL-6 pelas células RAW264.7. Microscopia confocal mostrou que a F24 está co-localizada com este receptor na superfície de células RAW264.7. A F24 induziu a fosforilação de maior intensidade do p38 e JNK comparado ao RANKL. Este novo fosfolipídeo isolado de P. gingivalis, distinto do LPS, mostrou-se capaz de induzir osteoclastogênese utilizando nucleolin como a molécula receptora através de uma via independente do RANKAbstract: Porphyromonas gingivalis (Pg) synthesizes several classes of complex phospholipids in addition to LPS. However, little is known about the biological effects of these phospholipids on bone resorption. Mouse osteoclast precursors cells including, RAW264.7 and bone marrow cells, were stimulated with phospholipids isolated from Pg (F24), Pg LPS, RANKL and a mammalian structural isomer (DMPE). Tartrate-resistant acid phosphatase (TRAP) staining and a dentine-pit formation assay were used for the identification of activated mature osteoclasts. F24¿s counter-ligand expressed on RAW264.7 cells were identified by affinity purification and mass-spectrometry based proteomics, and counter-examined by a confocal microscopy. Using Western-blot, signaling pathways triggered by F24 during osteoclastogenesis were examined with RNAi technology. F24, but not LPS or DMPE, induced significantly higher number of TRAP+ multinuclear cells from osteoclast precursors than RANKL, along with dentine-pit formation, which was not inhibited by RANKL decoy receptor OPG. Pg LPS, but not F24, induced TNF-a, IL-1ß and IL-6 by RAW264.7 cells. Proteomics analyses identified nucleolin as a ligand for F24. Confocal microscopy revealed the co-localization of F24 and its ligand on the surface of RAW264.7 cells. F24 induced stronger phosphorylation of p38 MAP kinase than RANKL. A novel P. gingivalis phospholipid that is distinct from LPS represents a new class of RANK-independent osteoclast differentiation factorDoutoradoMicrobiologia e ImunologiaDoutor em Biologia Buco-Denta

AVALIAÇÃO DA PRESENÇA DE LEPTINA NO FLUIDO PERI-IMPLANTAR DE PACIENTES DIABÉTICOS TIPO 2

A leptina é um hormônio produzido principalmente pelo tecido adiposo capaz de regular a energia necessária para manter a homeostasia dos tecidos, tendo efeito regulatório sobre o sistema reprodutivo, neuroendócrino, imune e sobre o metabolismo ósseo. Sistemicamente, a leptina atua como um importante modulador da resposta do hospedeiro frente a estímulos infecciosos e inflamatórios, desencadeando no sistema imune, o aumento da produção de citocinas pró-inflamatórias e a ativação de macrófagos. Essas citocinas pró-inflamatórias como, por exemplo, IL-6 e TNF, quando encontradas no plasma em elevadas concentração têm sido consideradas fatores de risco para problemas cardiovasculares, obesidade e diabetes. Apesar de alguns trabalhos investigarem o papel da leptina na progressão da doença periodontal, o papel dessa adipocina não foi investigado no fluido peri-implantar. Dessa forma, o objetivo desse estudo foi de avaliar e comparar os níveis de leptina no fluido peri-implantar de pacientes sadios (n=10), pacientes diabéticos tipo 2 controlados (n=10) e pacientes diabéticos tipo 2 não controlados (n=10). As amostras foram coletadas do interior do sulco peri-implantar clinicamente sadio dos participantes, com o auxílio de cones de papel estéreis, e em seguida analisadas pelo método do ELISA. Os resultados da concentração de leptina não foram diferentes estatisticamente entre os grupos de participantes segundo os testes ANOVA one-way, seguido do teste de Tukey (p0,05). Em conclusão, pode-se sugerir que a variação de taxa glicêmica encontrada entre os grupos (pacientes diabéticos e sadios) parece não interferir na condição de osseointegração e na taxa de leptina ao redor de implantes dentais sadios

15d-PGJ(2)-loaded solid lipid nanoparticles: physicochemical characterization and evaluation of pharmacological effects on inflammation

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist, has physiological properties including pronounced anti-inflammatory activity, though it binds strongly to serum albumin. The use of solid lipid nanoparticles (SLN) can improve therapeutic properties increasing drug efficiency and availability. 15d-PGJ(2)-SLN was therefore developed and investigated in terms of its immunomodulatory potential. 15d-PGJ(2)-SLN and unloaded SLN were physicochemically characterized and experiments in vivo were performed. Animals were pretreated with 15d-PGJ(2)-SLN at concentrations of 3, 10 or 30 mu g.kg(-1) before inflammatory stimulus with carrageenan (Cg), lipopolysaccharide (LPS) or mBSA (immune response). Interleukins (IL-1 beta, IL-10 and IL-17) levels were also evaluated in exudates. The 15d-PGJ(2)-SLN system showed good colloidal parameters and encapsulation efficiency of 96%. The results showed that the formulation was stable for up to 120 days with low hemolytic effects. The 15d-PGJ(2)-SLN formulation was able to reduce neutrophil migration in three inflammation models tested using low concentrations of 15d-PGJ(2). Additionally, 15d-PGJ(2)-SLN increased IL-10 levels and reduced IL-1 beta as well as IL-17 in peritoneal fluid. The new 15d-PGJ(2)-SLN formulation highlights perspectives of a potent anti-inflammatory system using low concentrations of 15d-PGJ(2).15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist, has physiological properties including pronounced anti-inflammatory activity, though it binds strongly to serum albumin. The use of solid lipid nanoparticles (SLN) can improve therapeutic properties increasing drug efficiency and availability. 15d-PGJ(2)-SLN was therefore developed and investigated in terms of its immunomodulatory potential. 15d-PGJ(2)-SLN and unloaded SLN were physicochemically characterized and experiments in vivo were performed. Animals were pretreated with 15d-PGJ(2)-SLN at concentrations of 3, 10 or 30 mu g.kg(-1) before inflammatory stimulus with carrageenan (Cg), lipopolysaccharide (LPS) or mBSA (immune response). Interleukins (IL-1 beta, IL-10 and IL-17) levels were also evaluated in exudates. The 15d-PGJ(2)-SLN system showed good colloidal parameters and encapsulation efficiency of 96%. The results showed that the formulation was stable for up to 120 days with low hemolytic effects. The 15d-PGJ(2)-SLN formulation was able to reduce neutrophil migration in three inflammation models tested using low concentrations of 15d-PGJ(2). Additionally, 15d-PGJ(2)-SLN increased IL-10 levels and reduced IL-1 beta as well as IL-17 in peritoneal fluid. The new 15d-PGJ(2)-SLN formulation highlights perspectives of a potent anti-inflammatory system using low concentrations of 15d-PGJ(2)118e0161796FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO2014/11016-8303555/2013-

Microbiological Sealing Analysis of a Tapered Connection and External Hexagon System

Considering the variety of implant connection systems available in the market and the contrasting literature regarding tapered connection systems in terms of bacterial leakage, the aim of this in vitro study was to compare the effectiveness of the bacterial seal at the implant/abutment interface between an external hexagon and a tapered connection system. Twelve sets of indexed tapered connection components and twelve sets of external hexagon connection components were used for microbiological analysis. In addition, for each model, an implant with its respective prosthetic abutment was used as a negative control and another as a positive control of microbial contamination. Failure of the abutment/implant interface seal was observed via turbidity or presence of deposits in the culture. Descriptive analysis of the data and relative frequency (percentage) as well as Fisher’s exact test were used at a significance level of 5%. Two of ten (20%) external hexagon specimens showed contamination against 0/10 (0%) tapered connection implants. In conclusion, both implant/abutment connections were able to prevent bacterial leakage in vitro

Caracterização clonal de cepas de Streptococcus mutans por eletroforese de enzimas codificadas por multilocus

Twenty-one Streptococcus mutans strains were clustered by Multilocus Enzyme Electrophoresis (MLEE). Six isoenzymes showed strong infra-specific discriminatory power (M1P, MPI, PLP, NSP, GOT, and LAP). MLEE is a robust technique that may be used to explore clonal diversity of S. mutans isolates in epidemiological surveys.Vinte e uma cepas de Streptococcus mutans foram agrupadas pela eletroforese de enzimas codificadas por multilocus (MLEE). Seis isoenzimas apresentaram forte poder discriminatório (M1P, MPI, PLP, NSP, GOT e LAP). A MLEE é uma técnica robusta que pode ser empregada no estudo da diversidade clonal de cepas de S. mutans, em estudos epidemiológicos.171

Peri-implantite e mucosite peri-implantar. Fatores de risco, diagnóstico e tratamento.

Introduction: The aim of this paper is to deepen the knowledge of peri-implant disease and existing treatments described in the literature. Material and methods: Literature review of peri-implant disease based on a search in databases like PubMed and Medline using keywords "periimplantitis, mucositis, tabaquism, oral microbiota, occlusal overload, surgical treatment, antimicrobial therapy, detoxification, regenerative therapy, bone defects". Development and discussion: Analysis of risk factors, diagnosis and treatment of peri-implant disease. Conclusions: Factors such as tabaquism or a history of periodontitis, along with poor oral hygiene are the main causes of peri-implant disease. A correct diagnosis of the cause and the appropriate choice of therapy may stop the progression of peri-implant disease. Following treatment done, it will be very important to do the periodic controls and maintenance of proper oral hygiene.Introdução: O objetivo deste trabalho é aprofundar o conhecimento da doença peri-implantar e os tratamentos existentes descritos na literatura. Material e método: Revisão bibliográfica das doenças peri-implantares, com base em uma pesquisa nos indexadores Pubmed, Medline e Cochrane Library, utilizando os uni termos " peri-implantite, mucosite, dependência de tabaco, microbiota oral, sobrecarga oclusal, tratamento cirúrgico, terapia antimicrobiana, desintoxicação,  terapia regenerativa, defeitos ósseos ". Desenvolvimento e discussão: Análise de fatores de risco, diagnóstico e tratamento de doenças peri-implantares. Conclusões: Fatores como tabaco ou histórico de periodontite, juntamente com falta de higiene bucal, são as principais causas da doença peri-implantar. Um diagnóstico correto da etiologia, bem como a escolha apropriada da terapêutica, pode interromper o avanço da doença peri-implantar. Após o tratamento realizado, será muito importante o monitoramento e manutenção regulares da higiene bucal adequada

Geopropolis from Melipona scutellaris decreases the mechanical inflammatory hypernociception by inhibiting the production of IL-1 beta and TNF-alpha

Ethnopharmacological relevance: The pharmacological activity of geopropolis collected by stingless bees (important and threatened pollinators), a product widely used in folk medicine by several communities in Brazil, especially in the Northeast Region, needs to be studied. Objective: The aim of this study was to evaluate the antinociceptive activity of Melipona scutellaris geopropolis (stingless bee) using different models of nociception. Material and methods: The antinociceptive activity of the ethanolic extract of geopropolis (EEGP) and fractions was evaluated using writhing induced by acetic acid, formalin test, carrageenan-induced hypernociception, and quantification of IL-1 beta and TNF-alpha. The chemical composition was assessed by quantification of total flavonoids and phenolic compounds. Results: EEGP and its hexane and aqueous fractions showed antinociceptive activity. Both EEGP and its aqueous fraction presented activity in the mechanical inflammatory hypernociception induced by the carrageenan model, an effect mediated by the inhibition of IL-1 beta and TNF-alpha. The chemical composition of EEGP and its hexane and aqueous fractions showed a significant presence of phenolic compounds and absence of flavonoids. Conclusion: Our data indicate that geopropolis is a natural source of bioactive substances with promising antinociceptive activity. (C) 2012 Elsevier Ireland Ltd. All rights reserved.FAPESP [2009/12352-3, 2010/20214-7