Orientador: Carlos Alberto de Souza Costa / Co-orientador: Fernanda Gonçalves Basso / Banca: Gelson Luis Adabo / Banca: Carla Raquel Fontana Mendonça / Resumo:A idadedo indivíduo e a presença de inflamação podem influenciar o metabolismo celular, retardando o processo de reparo tecidual. Diferentes tratamentos, como a laserterapia de baixa potência (LBP) e a aplicação de fatores de crescimento são capazes promover bioestimulação celular através do aumento de sua proliferação e metabolismo. O objetivo deste estudo foi avaliar as atividades de fibroblastos obtidos da gengiva de indivíduos jovens e idosos (Estudo 1), a capacidade de resposta destas células expostas ao fator de necrose tumoral alfa (TNF-α) (Estudo 2), bem como o efeito da LBP ou do fator de crescimento epidérmico (EGF) sobre a viabilidade e metabolismo destas células em cultura (Estudo 3). Culturas primárias de fibroblastos de gengiva de 6 pacientes (3 jovens - J; e 3 idosos - I) foram obtidas e semeadas em meio de cultura completo (DMEM), de acordo com cada protocolo proposto. No Estudo 1, fibroblastos em DMEM completo foram cultivados em placas de 24 compartimentos pelos períodos de 24, 48 ou 72 horas e submetidos a análise da viabilidade celular (MTT, n=36), produção de proteína total (PT, n=36) e síntese de colágeno Sirius Red (SR, n=36). No Estudo 2, após 24 horas do cultivo celular em placas de 96 compartimentos, o meio de cultura completo foi substituído por DMEM sem soro fetal bovino (SFB) acrescido ou não de TNF-α (100 ng/mL) e mantido em contato com as células por 24 horas adicionais. Então, as células foram analisadas quanto a produção de espécies reativas de oxigênio (EROs, n=36), produção de óxido nítrico (NO, n=36) e síntese de quimiocina CCL5 (ELISA, n=36). No Estudo 3, fibroblastos cultivados em placas de 24 compartimentos foram tratados com EGF (100 μM) ou irradiados com laser de baixa potência (LASERTable, InGaAsP - 780 ± 3nm; 25mW). A viabilidade celular (Alamar Blue®, n=12), migração celular (Wound Healing, n=12)...(Resumo completo clicar acesso eletrônico) / Abstract: The age as well as the presence of inflammation can influence the cellular metabolism, slowing the tissue healing process. Different therapies, such as the application of growth factors and low level laser therapy (LLLT) are capable of biostimulating cells by increasing their proliferation and metabolism. The aim of this study was to evaluate: the functions of gingival fibroblasts obtained from young and elderly individuals (Study 1); the response of these cells exposed to tumor necrosis factor alpha - TNF-α (Study 2); and the effect of LLLT or epidermal growth factor - EGF on the viability and metabolism of these connective cell type (Study 3). Primary culture of gingival fibroblasts of 6 patients (3 young - Y and 3 elderly - E) were collected and cultured in complete culture medium (DMEM). In the Study 1 fibroblasts were seeded in 96-well plates in complete DMEM containing 10% of fetal bovine serum (FBS) for 24, 48 or 72 hours and subjected to evaluation of cell viability (MTT, n=36), total protein production (TP, n=36) and collagen synthesis (SR, n=36). In the Study 2, after 24-hour cell seeding, the complete DMEM was replaced by DMEM without FBS supplemented or not with TNF-α (100 ng/mL) which was kept in contact with cells for additional 24 hours. Then, the production of reactive oxygen TNF-α species (ROS, n=36), nitric oxide (NO, n=36) and synthesis of CCL5 chemokine (ELISA, n=36) were assessed. In the Study 3 fibroblasts were seeded in 24-well plates for 24 hours, and then exposed to EGF (100 μM) or submitted to low power laser irradiation (LASERTable device, InGaAsP - 780 ± 3nm; 25mW). Cell viability (Alamar Blue®, n=12), cell migration (Wound Healing, n=12), collagen synthesis (SR, n=12), and synthesis of vascular endothelial growth factor - VEGF (ELISA, n=12) were assessed 72 hours after the treatments. The data were statistical...(Complete abstract electronic access below) / Mestre

Made available in DSpace on 2015-09-17T15:25:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-03-18. Added 1 bitstream(s) on 2015-09-17T15:46:37Z : No. of bitstreams: 1 000846277.pdf: 2026141 bytes, checksum: 0bb21e6aa2c7922e2a387ecf5fd901d7 (MD5) / The age as well as the presence of inflammation can influence the cellular metabolism, slowing the tissue healing process. Different therapies, such as the application of growth factors and low level laser therapy (LLLT) are capable of biostimulating cells by increasing their proliferation and metabolism. The aim of this study was to evaluate: the functions of gingival fibroblasts obtained from young and elderly individuals (Study 1); the response of these cells exposed to tumor necrosis factor alpha - TNF-α (Study 2); and the effect of LLLT or epidermal growth factor - EGF on the viability and metabolism of these connective cell type (Study 3). Primary culture of gingival fibroblasts of 6 patients (3 young - Y and 3 elderly - E) were collected and cultured in complete culture medium (DMEM). In the Study 1 fibroblasts were seeded in 96-well plates in complete DMEM containing 10% of fetal bovine serum (FBS) for 24, 48 or 72 hours and subjected to evaluation of cell viability (MTT, n=36), total protein production (TP, n=36) and collagen synthesis (SR, n=36). In the Study 2, after 24-hour cell seeding, the complete DMEM was replaced by DMEM without FBS supplemented or not with TNF-α (100 ng/mL) which was kept in contact with cells for additional 24 hours. Then, the production of reactive oxygen TNF-α species (ROS, n=36), nitric oxide (NO, n=36) and synthesis of CCL5 chemokine (ELISA, n=36) were assessed. In the Study 3 fibroblasts were seeded in 24-well plates for 24 hours, and then exposed to EGF (100 μM) or submitted to low power laser irradiation (LASERTable device, InGaAsP - 780 ± 3nm; 25mW). Cell viability (Alamar Blue®, n=12), cell migration (Wound Healing, n=12), collagen synthesis (SR, n=12), and synthesis of vascular endothelial growth factor - VEGF (ELISA, n=12) were assessed 72 hours after the treatments. The data were statistical...(Complete abstract electronic access below)