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1.
J. appl. oral sci ; 24(5): 509-517, Sept.-Oct. 2016. tab, graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: lil-797983

RESUMO

ABSTRACT Tooth bleaching is a technique of choice to obtain a harmonious smile, but bleaching agents may damage the dental pulp. Objective: This study evaluated the inflammatory responses of human dental pulp after the use of two bleaching techniques. Material and Methods: Pulp samples were collected from human third molars extracted for orthodontic reasons and divided into three groups: control - no tooth bleaching (CG) (n=7); at-home bleaching with 15% carbamide peroxide (AH) (n = 10), and in-office bleaching with 38% hydrogen peroxide (IO) (n=12). Pulps were removed and stained with hematoxylin-eosin for microscopic analysis of inflammation intensity, collagen degradation, and pulp tissue organization. Immunohistochemistry was used to detect mast cells (tryptase+), blood vessels (CD31+), and macrophages (CD68+). Chi-square, Kruskal-Wallis, and Mann Whitney tests were used for statistical analysis. The level of significance was set at p<.05. Results: The inflammation intensity and the number of macrophages were significantly greater in IO than in AH and CG (p<0.05). The results of CD31+ (blood vessels per mm2) were similar in CG (61.39±20.03), AH (52.29±27.62), and IO (57.43±8.69) groups (p>0.05). No mast cells were found in the pulp samples analyzed. Conclusion: In-office bleaching with 38% hydrogen peroxide resulted in more intense inflammation, higher macrophages migration, and greater pulp damage then at-home bleaching with 15% carbamide peroxide, however, these bleaching techniques did not induce migration of mast cells and increased the number of blood vessels.


Assuntos
Humanos , Pulpite/induzido quimicamente , Clareamento Dental/efeitos adversos , Polpa Dentária/efeitos dos fármacos , Clareadores Dentários/toxicidade , Peróxidos/toxicidade , Pulpite/patologia , Fatores de Tempo , Clareamento Dental/métodos , Ureia/análogos & derivados , Ureia/toxicidade , Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/patologia , Imuno-Histoquímica , Antígenos de Diferenciação Mielomonocítica , Distribuição Aleatória , Antígenos CD , Contagem de Células , Colágeno/efeitos dos fármacos , Estatísticas não Paramétricas , Molécula-1 de Adesão Celular Endotelial a Plaquetas , Polpa Dentária/patologia , Peróxido de Hidrogênio/toxicidade
2.
J. appl. oral sci ; 22(3): 218-227, May-Jun/2014. tab, graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: lil-711715

RESUMO

Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. Objective: This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. Material and Methods: Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR. Results: MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (p<0.001, p<0.001, p=0.031; respectively). Conclusions: The isolation, cultivation, and differentiation of BM-MSCs from BR1 makes this animal eligible as a useful large-animal model for stem cell-based studies in Brazil. .


Assuntos
Animais , Masculino , Células da Medula Óssea/citologia , Células-Tronco Mesenquimais/citologia , Modelos Animais , Porco Miniatura , Engenharia Tecidual/métodos , Antígenos CD/análise , Brasil , Técnicas de Cultura de Células , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Citometria de Fluxo , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos
3.
J. appl. oral sci ; 22(2): 131-137, Mar-Apr/2014. tab, graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: lil-704194

RESUMO

Central giant cell lesion (CGCL) and peripheral giant cell lesion (PGCL) are non-neoplastic proliferative processes of the jaws. PGCL is a reactive process induced by irritant local factors and CGCL is an intra-osseous lesion of unknown etiology. Both lesions exhibit similar histologic features showing abundant mononuclear cells, admixed with a large number of multinucleated giant cells and a rich vascularized stroma with extravasated erythrocytes, hemosiderin deposition, and blood-filled pools. Recent studies have linked fatty acid synthase (FASN) with angiogenesis. Objective: To evaluate angiogenesis and lymphangiogenesis and their relationship with FASN expression in CGCL and PGCL. Material and Methods: Thirteen CGCL and 14 PGCL of the jaws were selected for immunoexpression of FASN; CD34 and CD105 (to assess blood microvessel density [MVD] and microvessel area [MVA]); and D2-40 (to assess lymphatic MVD and MVA). Results: Within PGCL and CGCL, MVD-CD34 was signifcantly higher than MVD-CD10S, followed by MVD-D2-40. Moreover, a signifcantly higher number of FASN-positive multinucleated giant cells than mononuclear cells were observed. Between PGCL and CGCL, only MVD-CD34 and all MVA were signifcantly higher in PGCL. Positive correlation between MVA-CD10S with FASNpositive mononuclear cells in both lesions was observed. Conclusions: Our results show both lesions exhibiting similar levels of FASN expression and neoangiogenesis, suggesting constitutive processes that regulate tissue maintenance. .


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Adulto Jovem , Ácido Graxo Sintase Tipo I/análise , Células Gigantes/patologia , Doenças Maxilomandibulares/patologia , Linfangiogênese/fisiologia , Neovascularização Patológica/patologia , Antígenos CD/análise , /análise , Biópsia , Imuno-Histoquímica , Microvasos/patologia , Receptores de Superfície Celular/análise , Estudos Retrospectivos , Estatísticas não Paramétricas
4.
Bauru; s.n; 2014. 106 p. ilus, tab, graf.
Tese em Português | BBO - Odontologia | ID: biblio-867115

RESUMO

Os carcinomas de cabeça e pescoço representam um problema na saúde pública, sendo a oitava causa no mundo de morte por câncer. A taxa de crescimento do tumor, o seu local de expansão, bem como a metástase das células cancerígenas depende muito da vascularização do tumor, sendo que esta é a responsável pelo fornecimento constante de nutrientes e de oxigênio para o crescimento tumoral. Sendo assim a angiogênese é considerada um processo essencial dentro do processo neoplásico. A avaliação dos vasos sanguíneos tumorais neoformados em carcinomas espinocelulares de boca, usando o anticorpo CD105, mostra um crescimento significativo da densidade microvascular. Baseado nestes, o objetivo do trabalho é avaliar através da imunoistoquímica a possível correlação de aumento no número de vasos sanguíneos correlacionando com diferentes grupos de processo inflamatório divididos em: pouco, moderado e intenso infiltrado inflamatório no front tumoral. Na literatura os autores de um modo geral correlacionam infiltrados inflamatórios e angiogênese, neste trabalho tentamos correlacionar se um maior ou menor infiltrado inflamatório tem influência nessa angiogênese. As lâminas foram avaliadas microscopicamente por dois profissionais de forma que eles não sabiam da classificação dada pelo outro e só quando ambos estavam em comum acordo essas lâminas foram classificadas. Para a análise estatística foi realizado a comparação múltipla entre os 03 grupos através da Análise de variância (comparação das três médias) e também o teste de Tukey, onde se observou diferença entre os grupos I e III e nos grupos II e III, porém entre os grupos I e II não houve diferença significativa. Com isso os resultados nos mostram uma correlação positiva entre a presença de maior quantidade de vasos sanguíneos, onde se encontra maior quantidade de infiltrado inflamatório, quando comparado com áreas de menor infiltrado inflamatório.


Carcinomas of the head and neck represent a public health problem, being the eighth leading cause of worldwide cancer deaths. The growth rate of the tumor, its location expansion and metastasis of cancer cells depends greatly on tumor vascularity, and this is responsible for the constant supply of oxygen and nutrients for tumor growth. Thus angiogenesis is considered an essential process within the neoplastic process. The evaluation of newly formed tumor blood vessels in oral squamous cell carcinoma using the CD105 antibody, shows a significant increase in microvascular density. Based on these, the goal is to evaluate by immunohistochemistry the possible correlation of an increased number of blood vessels correlate with different groups of inflammatory process divided into: minor, moderate and intense inflammatory infiltrate in the tumor front. In the literature, authors generally correlated angiogenesis and inflammatory infiltrates, in this work we try to correlate to a greater or lesser inflammatory infiltrate that affects angiogenesis. The slides were evaluated microscopically by two professionals so that they did not know the rating given by others and only when both were in agreement these slides were classified. For statistical analysis, the multiple comparisons between the three groups was performed by analysis of variance (comparison of three average) and also the Tukey test, where a difference was observed between groups I and III and groups II and III, but between groups I and II significativa.Com that there was no difference in the results show a positive correlation between the presence of a larger amount of blood vessels, where it is most inflammatory infiltrate when compared to areas of lesser amounts of inflammation.


Assuntos
Humanos , Carcinoma de Células Escamosas/patologia , Neoplasias Bucais/patologia , Neovascularização Patológica/patologia , Análise de Variância , Antígenos CD/análise , Biópsia , Imuno-Histoquímica , Biomarcadores Tumorais , Processos Neoplásicos , Receptores de Superfície Celular/análise
5.
Braz. j. oral sci ; 9(4): 427-433, Oct.-Dec. 2010. ilus, graf
Artigo em Inglês | LILACS, BBO - Odontologia | ID: lil-582272

RESUMO

Aim: This study was developed to compare the morphological, proliferative and immunophenotypic profiles of pulp cells from permanent and primary teeth, obtained by two isolation methods. Methods: Normal human impacted third molars and exfoliated primary teeth were collected and cut around the cementoenamel junction. Pulp cells cultures were established by two approaches: enzyme digestion (3 mg/mL type I colagenase and 4 mg/mL dispase), or culture of the tissue explants in cell culture dishes. Morphological and proliferative analyses, as well as immunophenotype characterization with monoclonal antibodies against CD117, CD34 and CD45 surface receptors were performed. Results: For the permanent teeth, on the 4th day of culture, the cell number was significantly higher for the outgrowth method. By the end of the studied period (14th day), the enzymatic method was more efficient in promoting culture growth. On the other hand, for primary teeth, enzymatic digestion always promoted a higher cell proliferation. The immunophenotypic profiles were CD117+/ CD34-/ CD45- and CD117+/ CD34+/ CD45- for cells from permanent and primary teeth, respectively. Conclusions: The findings of this study indicate that both isolation methods can be efficiently used. The cell population displayed an immunophenotype compatible to the one of stem cells, with remarkable positive expression of CD117.


Assuntos
Criança , Adolescente , Células-Tronco/citologia , Técnicas In Vitro , Imunofenotipagem/métodos , Polpa Dentária/citologia , Técnicas de Cultura de Células/métodos , Antígenos CD , Dente Serotino , Proliferação de Células , Reprodutibilidade dos Testes , Fatores de Tempo , Dente Decíduo , Dente Impactado
6.
Bauru; s.n; 2009. 208 p. ilus, tab, graf.
Tese em Português | LILACS, BBO - Odontologia | ID: lil-578024

RESUMO

O sistema imune serve como uma barreira contra os patógenos e ao crescimento anormal de células. Para impedir as respostas imunes excessivas ou indiscriminadas que podem comprometer a sobrevivência do organismo, diversos mecanismos regulatórios são ativados visando manter o delicado balanço entre início e término de uma resposta imune. As celular T reguladoras (Treg) parecem desempenhar papel central na regulação da resposta imune em infecções crônicas e durante o desenvolvimento de tumores. Outro mecanismo importante no controle da resposta imune é desempenhado por moléculas co-estimulatórias, dentre as quais estão CTLA-4 e PD-1, todas associadas à função das células T reguladoras. Um aspecto importante é q a sobrevida de tecido tumoral e de transplantes tem sido associada à função das células T reguladoras. Assim, buscamos definir o envolvimento de células T reguladoras e PD-1 na modulação da resposta imune L. braziliensis, ao fungo P. brasiliensis, à doença periodontal e ao tumor de cabeça e pescoço. Baseado nos resultados já publicados e em dados preliminares, as hipóteses são que: (a) a interação do parasita (ou célula tumoral) com o hospedeiro leva à migração de linfócitos T e efetores e células T reguladoras para o local da lesão; (b) a dinâmica do acúmulo dessas células em tais sítios determina a eficiência da eliminação do patógeno ou tumor. No caso das parasitoses, há o desenvolvimento de imunidade concomitante; (c) as células T regulam a resposta imune local de forma contato dependente e modulando a função de APC através da liberação de IL-10 e/ou TGF-β; (d) infecção e progressão tumoral levam à modulação da expressão de PD-1 nos leucócitos e seus ligantes nos órgãos; (e) a interação PD-PDL-1 regula a resposta imune local de forma a favorecer a persistência do patógeno e os mecanismos de escape tumoral.


The immune system serves as a barrier against pathogens and abnormal cellular growth. To avoid tissue and organ damage during immune response several regulatory mechanisms are activated to limit, terminate and attenuate T-cells response. Regulatory T cells (Treg) play a central role in the regulation of the immune response in chronic infections and tumor-specific immunity. Programmed death-1 (PD-1) is a transmembrane protein that acts as a negative regulator in effector T cells, modulating the delicate balance between effective antimicrobial immune defenses and immune-mediated tissue damage. However, recent data suggest that the PD-1:PD-L1 pathway can also block antitumor immune responses even when tumor antigens can be recognized. An important aspect it that the survival of tumor and transplant tissues has been associated with the function or regulatory T cells. Thus, we discuss the role of Treg cells and PD-1 molecules in the modulation of the immune response to L. braziliensis, P. brasiliensis, periodontal disease and head and neck tumors. Based on published results and preliminary data, the hypotheses are that: (a) the interaction of the parasite (or tumoral cells) with the host leads to the migration of effector T lymphocytes and Treg cells to the local; (b) the dynamics of cells accumulation in such sites determinate the elimination efficiency of tumors. In infectious disease, there is the development of concomitant immunity; (c) Treg cells regulate the local immune response, modulating the APC function through the release of IL-10 and/or TGF-β; (d) infection and tumor progression leads to the modulation of PD-1 expression in the leukocytes and their ligands in the tissue; (e) PD-PDL-1 interactions regulate the immune response and may mediate the persistence of pathogen and contribute to immune evasion by cancers.T.


Assuntos
Humanos , Antígenos CD/química , Doenças Transmissíveis/imunologia , Linfócitos T Reguladores/imunologia , Neoplasias de Cabeça e Pescoço/imunologia , Carcinoma de Células Escamosas/imunologia , Leishmania braziliensis/imunologia , Paracoccidioides/imunologia , Periodontite Crônica/imunologia , Queilite/microbiologia
7.
Natal; s.n; 20080000. 105 p. ilus, tab. (BR).
Tese em Português | BBO - Odontologia | ID: biblio-864461

RESUMO

Nesta pesquisa buscou-se avaliar a expressão imunoistoquímica dos anticorpos CD105 e FvW na angiogênese do Carcinoma Epidermóide Oral (CEO), correlacionando-o com o estadiamento clínico pelo sistema TNM, visando uma melhor compreensão do seu comportamento biológico e utilização como indicador de prognóstico. A amostra foi composta por 30 casos de CE, sendo 10 de assoalho bucal, 10 da região retromolar e 10 de língua, além de 10 casos de granuloma piogênico, integrantes do grupo controle. Os resultados desta pesquisa mostraram que as médias da MVC foram correspondentemente mais elevadas no grupo do granuloma piogênico (CD105 = 57,26 vasos e FvW = 39,64) do que no grupo do CE (CD105 = 10,09 e FvW = 12,20) e as diferenças se revelaram estatisticamente significantes entre os grupos para cada um dos biomarcadores angiogênicos (p=0,002 para o CD105 e p<0,001 para o FvW ). O CD105 se mostrou com melhor positividade no granuloma piogênico (média = 57,26 vasos) e, para o CE, o FvW foi o que apresentou maior marcação (média = 12,20 vasos). Com relação ao CE, a faixa etária mais acometida foi entre 51 e 70 anos (n=14; 46,7%), apresentando uma MVC representativa para ambos os marcadores. Não se comprovou diferença estatisticamente significante entre os sexos para nenhum dos marcadores (p=0,967 para o CD105 e p=0,744 para o FvW). A média do CD105 foi bem mais elevada entre os pacientes com estadiamento T3 e T4 (17,13) e menos elevada entre os pacientes com estadiamento N+ (6,36). Quando se avaliou o FvW, a média foi mais elevada no grupo dos pacientes com T1 e T2 (12,23), sendo mais baixa nos pacientes com T3 e T4 (12,10), porém sem diferença estatisticamente significante. Em relação à localização anatômica, comprovou-se diferença estatisticamente significante entre as localizações assoalho bucal e retromolar (p=0,013) para o marcador FvW. Portanto, este estudo sugere que a marcação do CD105 na angiogênese do CEO, ao contrário de outros tipos de neoplasias malignas, pode não estar correlacionada com o prognóstico e agressividade do tumor, enquanto que o FvW se mostrou um anticorpo mais efetivo na marcação desta lesão (AU).


The purpose of this study was to assess the immunohistochemical expression of CD105 and FvW antibodies in the angiogenesis of oral epidermoid carcinoma (OEC), correlating it with the TNM clinical staging system, seeking a better understanding of its biological behavior and use as an indicator of prognosis.The sample consisted of 30 epidermoid carcinoma (EC) cases, 10 of the floor of the mouth, 10 of the retromolar region and 10 of the tongue, in addition to 10 cases of pyogenic granuloma, which made up the control group. The results showed that mean microvessel counts (MVC) were correspondingly higher in the pyogenic granuloma group (CD105 = 57.26 vessels and FvW = 39.64) than in the EC group (CD105 = 10.09 and FvW = 12.20) and that the differences were statistically significant between the groups for each of the angiogenic biomarkers (p = 0.002 for CD105 and p< 0.001 for FvW). CD105 had better positivity in the pyogenic granuloma group (mean = 57.26 vessels) and for EC, FvW had the highest expression (mean = 12.20 vessels). With respect to EC, the most affected age group was between 51 and 70 years (n = 14; 46.7%), with a representative MVC for both markers. No statistically significant difference was found between the sexes for any of the markers (p = 0.967 for CD105 and p = 0.744 for FvW). Mean CD105 levels were much higher in patients with stage T3 and T4 (17.13) and lower in those with stage N+ (6.36). Mean FvW levels were higher in the patients with stage T1 and T2 (12.23) and lower in patients with T3 and T4 (12.10), but without a statistically significant difference. In regard to anatomic location, a statistically significant difference was observed between FvW sites, with a statistically significant difference between floor of the mouth cases and those located in the retromolar region (p = 0.013). Therefore, this study suggests that CD105 expression in OEC angiogenesis, in contrast to other types of malignant neoplasias, may not be correlated with prognosis and tumor aggressiveness, whereas FvW was a more effective antibody for staining this lesion (AU).


Assuntos
Antígenos CD , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Granuloma Piogênico/patologia , Imuno-Histoquímica/métodos , Neovascularização Patológica/patologia , Distribuição de Qui-Quadrado , Estatísticas não Paramétricas , Neoplasias Bucais/patologia
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