Thesis (M.S.) -- Worcester Polytechnic Institute. / Keywords: tension; failure strain; fibroblasts; regenerative medicine; serum-free; proteoglycans; glycosaminoglycans; collagen; wound-healing model; cell-remodeled matrix; cell-derived matrix; biomechanical characterization; fibrin gel; growth factors; tissue growth; total protein content; tissue formation. Includes bibliographical references (p.44-53).

Thesis (M.S.)--University of Delaware, 2005. / Principal faculty advisor: Mary C. Farach-Carson, Dept. of Biological Sciences. Includes bibliographical references.

Thesis (M. Med. Sc.)--University of Hong Kong, 2006.

Made available in DSpace on 2014-06-11T19:35:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-07-18Bitstream added on 2014-06-13T20:26:09Z : No. of bitstreams: 1 silva_rb_dr_botfmvz.pdf: 496820 bytes, checksum: 055e35414ee23830cc537670a4ae215b (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Durante o desenvolvimento ovariano fetal, a formação folicular inicial é decisiva para a fertilidade da fêmea, pois define sua reserva gametogênica. Tem sido proposto que a progesterona e o estradiol desempenham papel regulatório na foliculogênese pré-antral, de forma que sua produção reduzida em ovários fetais bovinos antecede o surgimento de folículos primordiais e primários. Recentemente, os FGFs 10 e 18 foram reportados em folículos ovarianos bovinos como redutores dos níveis de esteróides, o que parece envolver a inibição da expressão de enzimas necessárias à esteroidogênese. Em adição, a expressão do FGF10 foi observada durante o desenvolvimento ovariano fetal bovino, e esteve positivamente associada ao aumento no número de folículos primários. O presente estudo investigou primeiramente o padrão de expressão do RNAm das enzimas esteroidogênicas (StAR, CYP11A1, 3β-HSD, CYP17A1, CYP19A1 e 17β-HSD) em ovários de fetos bovinos em idades gestacionais específicas (60, 75, 90, 120, 150 e 210 dias). Todos os genes investigados se mostraram expressos e regulados ao longo da gestação. Os níveis de RNAm da CYP19A1 diminuíram dos 60 para os 90 dias, sugerindo envolvimento desta enzima com a produção decrescente de estradiol observada previamente durante este período gestacional. A expressão das demais enzimas foi elevada ao longo da gestação, coincidente com o aumento da competência esteroidogênica descrito preliminarmente durante o desenvolvimento folicular inicial. Em adição, foi investigada a participação dos FGFs 10 e 18 na esteroidogênese ovariana fetal bovina. A expressão do FGF18 e de seus receptores (FGFR2C, FGFR3C e FGFR4) foi detectada em ovários fetais bovinos ao longo da gestação (60, 75, 90, 120, 150 e 210 dias). A abundância de RNAm do FGF18 aumentou... / During fetal ovarian development, early follicular formation is essential to female fertility, when the gametogenic reserve is defined. It has been proposed that progesterone and estradiol play regulatory role on preantral folliculogenesis, once its reduced production in bovine fetal ovaries precedes primordial and primary follicle assembly. Recentlly, FGFs 10 and 18 were reported in bovine ovarian follicles as reducers of steroids levels, and this seems to involve the inhibition of enzymes necessary to steroidogenesis. In addition, FGF10 expression was observed during bovine fetal ovary development, and it was positively associated with the elevation on primary follicles number. The present study first investigated the mRNA expression patterns for steroidogenic enzymes (StAR, CYP11A1, HSD3B1, CYP17A1, CYP19A1 and HSD17B1) in bovine fetal ovaries at specific gestational ages (60, 75, 90, 120, 150 e 210 days). Expression of all investigated genes was detected and regulated through gestation. Messenger RNA levels of CYP19A1 decreased from days 60 to 90 of gestation, suggesting involvement of this enzyme on decrescent estradiol production previously observed during this gestational period. The expression of other enzymes was elevated during gestational period, which was coincident with the enhance of steroidogenic competence previously described during early follicular development. In addition, the participation of FGFs 10 and 18 on steroidogenesis in bovine fetal ovaries was investigated. The expression of FGF18 and its receptors (FGFR2C, FGFR3C and FGFR4) was detected in bovine fetal ovaries through gestation (60, 75, 90, 120, 150 e 210 days). The mRNA abundance of FGF18 enhanced between 90 and 120 days and decreased at 210 days. The expression of FGFR2C and FGFR4 did not vary during... (Complete abstract click electronic access below)

Made available in DSpace on 2015-06-17T19:34:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-03-14. Added 1 bitstream(s) on 2015-06-18T12:47:03Z : No. of bitstreams: 1 000830081_20160101.pdf: 121876 bytes, checksum: 342f21a15b1d2e738af168b1e2467da5 (MD5) Bitstreams deleted on 2016-01-04T10:26:29Z: 000830081_20160101.pdf,. Added 1 bitstream(s) on 2016-01-04T10:28:25Z : No. of bitstreams: 1 000830081.pdf: 809941 bytes, checksum: ea53cd5ef0cec7fb993a2745e82dad53 (MD5) / The antimicrobial peptides LL-37, hBD-1, hBD-2 and hBD-3 are considered an endogenous antibiotic, with important role in the prevention of periodontal diseases due to their ability to regulate the immune response. However those peptides could be degraded by periodontal pathogens. Therefore, therapies able to up regulate the secretion of those peptides by human cells, and the association of antimicrobial peptides with natural compounds, which may act in synergism to modulate the immune response, may be a novel approach for effectively controlling periodontal diseases. The aim of this in vitro study were: i) investigate the ability of green tea extract and EGCG to induce hBD-1 and hBD-2 secretion and gene expression by gingival epithelial cells (B11) and to protect hBDs from degradation by P. gingivalis, ii) A 3D co-culture model of gingival epithelial cells and fibroblasts stimulated with A. actinomycetemcomitans LPS (1 μg/ml) were used to investigated the anti-inflammatory properties of the hBD-3, LL-37, ACPACs and EGCG and to determine whether LL-37 acts in synergy with AC-PACs, EGCG and hBD-3. Gingival epithelial cells were stimulated with green tea extract or EGCG in the presence and absence of specific inhibitors. The secretion and gene expression of hBD-1 and hBD-2 was respectively measured by ELISA and qPCR. The ability of green tea extract and EGCG to prevent hBDs degradation by P. gingivalis present in a bacterial culture supernatant was evaluated by ELISA. A 3D co-culture model composed of gingival fibroblasts embedded in a collagen matrix overlaid with gingival epithelial cells had a synergistic effect with respect to the secretion of IL-6 and IL-8 in response to A. actinomycetemcomitans LPS stimulation compared to fibroblasts and epithelial cells individually. The 3D co-culture model was stimulated with noncytotoxic concentrations of: i) hBD-3 (10 and 20 μM) ...(Complete abstract electronic access below)

Made available in DSpace on 2014-06-11T19:27:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-29Bitstream added on 2014-06-13T19:56:59Z : No. of bitstreams: 1 grandi_f_me_botfm.pdf: 914284 bytes, checksum: 7e8beca0af748a53dac236ca291fea62 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O melanoma é uma das neoplasias cutâneas mais comumente diagnosticadas no homem e nos cães. O microambiente tumoral é composto pelas células neoplásicas e células estromais interagindo constantemente para garantir a progressão tumoral. Os fibroblastos associados ao câncer (FAC) representam uma população heterogênea de células caracterizadas pela expressão de diversos marcadores incluindo a proteína α-SMA e S100A4. Acredita-se que estas células originem-se de diversas fontes incluindo a transição endotelial fibroblástica. Neste trabalho, quantificamos a imunoexpressão da prpteína S100A4 nos fibroblastos associados ao câncer em melanomas cutâneos caninos, verificamos a potencial contribuição das células endoteliais na gênese desta população e correlacionamos os achados com parâmetros patológicos, incluindo a microdensidade vascular. Quarenta e oito casos de melanoma dermais caninos (21 epitelióides, 14 fusiformes e 13 mistos) previamente categorizados nos níveis de Clark 4, 5 e até 4 foram submetidos a imunofluorescência dupla utilizando os anticorpos primários α-SMA, fator de Von Willebrand (vWF) e S100A4 objetivando-se caracterizar os fibroblastos associados ao câncer e a contribuição da transição endotelial mesenquimal. Os melanomas não pigmentados foram caracterizados pela imunoistoquímica utilizando-se os anticorpos vimentina, pancitoqueratina, S100 e Melan A. A densidade microvascular foi analisada utilizando-se a técnica de imunofluorescência e o anticorpo primário anti-fator de Von Willebrand. O cálculo do número de vasos foi realizado selecionando-se cinco campos microscópicos de 200x contendo o maior número de vasos. O percentul de expressão da proteína S100A4 foi calculado através da técnica de segmentação de conglomerados de cor. Apenas um caso demonstrou... / Skin melanoma is one of the most common skin neoplasm seen in humans and dogs. Tumor microenvironment is composed by cancer cells and stromal cells that interacts to guarantee tumor progression. Cancer associated fibroblasts (CAF) represents a heterogeneous cell population characterized by expression of several markers including α-SMA and S100A4 proteins. These cells are thought to derive from different sources including endothelial-to-fibroblast transition. Here we characterize CAF in canine skin melanomas, verify the potential contribution of the endothelial cells to this population and correlate findings to pathological parameters, including microvascular density (MVD). Forth-eight cases of canine dermal melanomas (21 epithelioid, 14 spindle and 13 mixed) classified under Clark´s level 4 and 5 were submitted to a double immunofluorescence assay using primary antibodies α-SMA, Von Willebrand Factor (vWF) and S100A4 in order to characterize cancer associated fibroblasts and verify the contribution of endothelial to fibroblast transition. Non-pigmented samples were characterized by immunohistochemistry using primary antibodies pan-cytokeratin, vimentin, S100 and Melan A. Microvascular density was evaluated by immunofluorescent assay using vWF and by calculating total number of vessels in five 200x fields (“hotspots”).S100A4 imunoexpression was calculated using K-means clustering segmentation method. Only one case showed α-SMA and vWF co-expression restricted to myofibroblasts in tumor stroma. The cells were predominantly peritumoral and periadnexal. S100A4 expression was significantly different among three histotypes with mixed melanomas displaying lesser percentage of positive cells. Some neoplastic cells mainly in spindle cell melanomas were also positive for S100A4. There were no significant differences between MVD/histotypes... (Complete abstract click electronic access below)

Made available in DSpace on 2014-10-09T12:50:15Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:01:54Z (GMT). No. of bitstreams: 1 10826.pdf: 3720317 bytes, checksum: 26d9f64b9d44684f557ae44f37e86002 (MD5) / Dissertação (Mestrado Profissionalizante em Lasers em Odontologia) / IPEN/D-MPLO / Intituto de Pesquisas Energeticas e Nucleares, IPEN/CNEN-SP; Faculdade de Odontologia, Universidade de Sao Paulo

Made available in DSpace on 2014-10-09T12:50:14Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:45Z (GMT). No. of bitstreams: 1 10825.pdf: 4609998 bytes, checksum: abaefc3f2b2d697034a947707dec7944 (MD5) / Dissertacao (Mestrado Profissionalizante em Lasers em Odontologia) / IPEN/D-MPLO / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP; Faculdade de Odontologia - USP

Orientador: Denise Madalena Palomari Spolidório / Banca:Juliana Rico Pires / Banca: Shelon Cristina Souza Pinto / Banca: Luciene Cristina Souza Pinto / Banca: Joni Augusto Cirelli / Resumo:Os peptídeos antimicrobianos como por exemplo a catelicina (LL-37) e as defensinas humanas (hBD-1, hBD-2 e a hBD-3) são considerados antibióticos endógenos com importante papel na prevenção das doenças periodontais, devido a sua capacidade de regulação da resposta imune, sendo que os mesmos podem ser degradados pelos periodontopatógenos. Terapias que aumentem a produção destes peptídeos pelas próprias células do organismo, assim como a associação destes peptídeos com compostos naturais os quais possam agir em sinergismo na regulação da resposta imune, podem ser considerados novas estratégias para o melhor controle das doenças periodontais. Portanto os objetivos deste estudo in vitro foram: i) Avaliar a capacidade do extrato do chá verde (Camellia sinensis) e do seu polifenol, o EGCG, sobre a expressão gênica de hBD-1 e hBD-2 pelas células epiteliais gengivais (B11), sobre a degradação das mesmas frente ao P. gingivalis, ii) Através da utilização do modelo 3D de co-cultura celular, avaliar a capacidade antiinflamatória dos peptídeos hBD-3 e LL-37 quando em associação sobre a produção de citocinas, quimiocinas e fatores de crescimento, iii) Avaliar a capacidade anti-inflamatória da associação do EGCG e do polifenol proveniente do cranberry, o AC-PACs, com o peptídeo LL-37 sobre a produção de citocinas, quimiocinas e fatores de crescimento em modelo de co-cultura celular. As células epiteliais gengivais (B11) foram estimuladas com o extrato do chá verde e com o EGCG na presença e ausência de inibidores específicos. A produção e expressão gênica de hBD-1 e hBD-2 foram quantificados respectivamente pelas técnica de ELISA e qPCR. A capacidade do extrato do chá verde e do EGCG em proteger a degradação de hBDs pelo P. gingivalis foi mensurado através da técnica de ELISA. Foi desenvolvido um modelo em 3D de co-cultura de fibroblastos gengivais embebidos em....(Resumo completo, clicar acesso eletrôni / Abstract: The antimicrobial peptides LL-37, hBD-1, hBD-2 and hBD-3 are considered an endogenous antibiotic, with important role in the prevention of periodontal diseases due to their ability to regulate the immune response. However those peptides could be degraded by periodontal pathogens. Therefore, therapies able to up regulate the secretion of those peptides by human cells, and the association of antimicrobial peptides with natural compounds, which may act in synergism to modulate the immune response, may be a novel approach for effectively controlling periodontal diseases. The aim of this in vitro study were: i) investigate the ability of green tea extract and EGCG to induce hBD-1 and hBD-2 secretion and gene expression by gingival epithelial cells (B11) and to protect hBDs from degradation by P. gingivalis, ii) A 3D co-culture model of gingival epithelial cells and fibroblasts stimulated with A. actinomycetemcomitans LPS (1 μg/ml) were used to investigated the anti-inflammatory properties of the hBD-3, LL-37, ACPACs and EGCG and to determine whether LL-37 acts in synergy with AC-PACs, EGCG and hBD-3. Gingival epithelial cells were stimulated with green tea extract or EGCG in the presence and absence of specific inhibitors. The secretion and gene expression of hBD-1 and hBD-2 was respectively measured by ELISA and qPCR. The ability of green tea extract and EGCG to prevent hBDs degradation by P. gingivalis present in a bacterial culture supernatant was evaluated by ELISA. A 3D co-culture model composed of gingival fibroblasts embedded in a collagen matrix overlaid with gingival epithelial cells had a synergistic effect with respect to the secretion of IL-6 and IL-8 in response to A. actinomycetemcomitans LPS stimulation compared to fibroblasts and epithelial cells individually. The 3D co-culture model was stimulated with noncytotoxic concentrations of: i) hBD-3 (10 and 20 μM) ...(Complete abstract electronic access below) / Doutor

Orientador: Noeme Sousa Rocha / Banca: Hélio Amante Miot / Banca: Bruno Cogliati / Resumo: O melanoma é uma das neoplasias cutâneas mais comumente diagnosticadas no homem e nos cães. O microambiente tumoral é composto pelas células neoplásicas e células estromais interagindo constantemente para garantir a progressão tumoral. Os fibroblastos associados ao câncer (FAC) representam uma população heterogênea de células caracterizadas pela expressão de diversos marcadores incluindo a proteína α-SMA e S100A4. Acredita-se que estas células originem-se de diversas fontes incluindo a transição endotelial fibroblástica. Neste trabalho, quantificamos a imunoexpressão da prpteína S100A4 nos fibroblastos associados ao câncer em melanomas cutâneos caninos, verificamos a potencial contribuição das células endoteliais na gênese desta população e correlacionamos os achados com parâmetros patológicos, incluindo a microdensidade vascular. Quarenta e oito casos de melanoma dermais caninos (21 epitelióides, 14 fusiformes e 13 mistos) previamente categorizados nos níveis de Clark 4, 5 e até 4 foram submetidos a imunofluorescência dupla utilizando os anticorpos primários α-SMA, fator de Von Willebrand (vWF) e S100A4 objetivando-se caracterizar os fibroblastos associados ao câncer e a contribuição da transição endotelial mesenquimal. Os melanomas não pigmentados foram caracterizados pela imunoistoquímica utilizando-se os anticorpos vimentina, pancitoqueratina, S100 e Melan A. A densidade microvascular foi analisada utilizando-se a técnica de imunofluorescência e o anticorpo primário anti-fator de Von Willebrand. O cálculo do número de vasos foi realizado selecionando-se cinco campos microscópicos de 200x contendo o maior número de vasos. O percentul de expressão da proteína S100A4 foi calculado através da técnica de segmentação de conglomerados de cor. Apenas um caso demonstrou... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Skin melanoma is one of the most common skin neoplasm seen in humans and dogs. Tumor microenvironment is composed by cancer cells and stromal cells that interacts to guarantee tumor progression. Cancer associated fibroblasts (CAF) represents a heterogeneous cell population characterized by expression of several markers including α-SMA and S100A4 proteins. These cells are thought to derive from different sources including endothelial-to-fibroblast transition. Here we characterize CAF in canine skin melanomas, verify the potential contribution of the endothelial cells to this population and correlate findings to pathological parameters, including microvascular density (MVD). Forth-eight cases of canine dermal melanomas (21 epithelioid, 14 spindle and 13 mixed) classified under Clark's level 4 and 5 were submitted to a double immunofluorescence assay using primary antibodies α-SMA, Von Willebrand Factor (vWF) and S100A4 in order to characterize cancer associated fibroblasts and verify the contribution of endothelial to fibroblast transition. Non-pigmented samples were characterized by immunohistochemistry using primary antibodies pan-cytokeratin, vimentin, S100 and Melan A. Microvascular density was evaluated by immunofluorescent assay using vWF and by calculating total number of vessels in five 200x fields ("hotspots").S100A4 imunoexpression was calculated using K-means clustering segmentation method. Only one case showed α-SMA and vWF co-expression restricted to myofibroblasts in tumor stroma. The cells were predominantly peritumoral and periadnexal. S100A4 expression was significantly different among three histotypes with mixed melanomas displaying lesser percentage of positive cells. Some neoplastic cells mainly in spindle cell melanomas were also positive for S100A4. There were no significant differences between MVD/histotypes... (Complete abstract click electronic access below) / Mestre