Tesi sul tema "Periodontal cells"
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Worapamorn, Wilairat. "Cell-surface proteoglycan expression by periodontal cells /". St. Lucia, Qld, 2001. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe16097.pdf.
Gay, Isabel C. "Isolation and characterization of human periodontal ligament stem cells". Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2007. http://www.mhsl.uab.edu/dt/2007m/gay.pdf.
Stoianovici, Charles. "Directing Mesenchymal Stem Cells for Periodontal Regeneration". VCU Scholars Compass, 2018. https://scholarscompass.vcu.edu/etd/5335.
Winning, Lewis. "The osteogenic potential of periodontal ligament stem cells". Thesis, Queen's University Belfast, 2018. https://pure.qub.ac.uk/portal/en/theses/the-osteogenic-potential-of-periodontal-ligament-stem-cells(e5fdef0e-d55b-42b6-acb5-75a617b43edd).html.
Campbell, Lauren Dee. "The role of CD4+ T cells in periodontal disease". Thesis, University of Glasgow, 2017. http://theses.gla.ac.uk/8241/.
Åsman, Björn. "Juvenile periodontitis generation of free oxygen radicals and elastase by peripheral PMN cells /". Stockholm : Kongl. Carolinska Medico Chirurgiska Institutet, 1988. http://catalog.hathitrust.org/api/volumes/oclc/18171198.html.
Engman, Sara. "Expression and Regulation of the Cell Surface Proteins CD47 and SIRPα in Resident Periodontal Cells". Thesis, Umeå universitet, Institutionen för odontologi, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-129258.
Bou, Chebel Najib. "Periodontal bacterial-DNA initiated immuno-inflammatory responses in human osteoblastic cells". VCU Scholars Compass, 2010. http://scholarscompass.vcu.edu/etd/97.
Moore, Edward Andrew. "Cell attachment and spreading on physical barriers used in periodontal guided tissue regeneration /". Oklahoma City : [s.n.], 2002. http://library.ouhsc.edu/epub/theses/Moore-William-A.pdf.
Wescott, David Clark, e n/a. "Osteogenic gene expression by human periodontal ligament cells under cyclic mechanical tension". University of Otago. School of Dentistry, 2008. http://adt.otago.ac.nz./public/adt-NZDU20081202.131453.
Nuersailike, Abuduwali [Verfasser]. "Characterization of Parathyroid Hormone 1 Receptor in Periodontal Ligament Cells / Abuduwali Nuersailike". Bonn : Universitäts- und Landesbibliothek Bonn, 2012. http://d-nb.info/1044080973/34.
Almeida, Luciana Salles Branco de. "Influência da fluoxetina sobre a resposta imuno-inflamatória relacionada à doença periodontal". [s.n.], 2011. http://repositorio.unicamp.br/jspui/handle/REPOSIP/288518.
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: A fluoxetina é um droga inibidora seletiva da recaptação de serotonina que apresenta propriedades imunomoduladoras e antiinflamatórias. O objetivo desse estudo foi avaliar os efeitos da fluoxetina sobre a resposta imuno-inflamatória relacionada à doença periodontal (DP). In vitro, avaliou-se a influência da fluoxetina sobre a capacidade das células dendríticas (DCs) em apresentar antígeno aos linfócitos T. As DCs foram obtidas da medula óssea de camundongos C57BL/6 e diferenciadas utilizando-se GM-CSF (20 ng/mL). As DCs foram tratadas com a fluoxetina (concentrações: 0,01, 0,1 ou 1 ?M) para análise da produção de citocinas e quimiocinas, bem como da expressão de MHC-II e moléculas co-estimuladoras (CD80, CD86, PD-L1, ICOS-L) aos linfócitos T, utilizando-se ensaios de ELISA e citometria de fluxo respectivamente. Culturas de DCs e linfócitos T reativos ao Aggregatibacter actinomycetemcomitans (×Aa-T) foram utilizadas para avaliação de proliferação/ativação de linfócitos T. A desipramina, um inibidor seletivo da recaptação de norepinefrina, também foi incluída nos ensaios in vitro para comparação. In vivo, avaliou-se os efeitos da fluoxetina sobre a resposta inflamatória e a destruição tecidual utilizando-se modelo de DP induzida por ligadura. Ratos Wistar machos (SPF) foram submetidos à colocação de ligadura em torno dos primeiros molares inferiores e divididos em 3 grupos experimentais (n=10 animais/grupo): 1) ratos sem ligadura e sem tratamento (grupo controle); 2) ratos com ligadura e tratados com solução salina (grupo ligadura); 3) ratos com ligadura e tratados com a fluoxetina (20 mg/kg/dia, grupo ligadura + fluoxetina). Análises de reabsorção óssea na região de furca (lâminas coradas com H&E) e de colágeno no tecido conjuntivo da mesial dos primeiros molares (coloração de picrosirius) foram realizadas nos ratos submetidos a 15 dias de indução da DP. Tecidos gengivais de ratos submetidos a 3 dias de indução da DP foram submetidos às seguintes análises: expressão de IL-1?, COX-2, MMP-9 e iNOS utilizando-se RT-PCR e atividade da MMP-9 utilizando-se zimografia. Como resultados, a fluoxetina diminuiu a produção de IL-12, IL-1?, TNF-?, RANTES e MIP-1??pelas DCs estimuladas com LPS (P < 0,05, ANOVA, teste t de Student), bem como diminuiu significativamente a expressão de ICOSL. Além disso, reduziu a proliferação de ×Aa-T estimulados com Aa pelas DCs. A serotonina (5- HT) aumentou a proliferação de ×Aa-T, indicando que os efeitos da fluoxetina são independentes da 5-HT. A desipramina apresentou perfil semelhante à fluoxetina nos ensaios in vitro. No estudo in vivo, a fluoxetina reduziu a perda óssea em região de furca quando comparada ao grupo ligadura (P < 0,05 ANOVA, teste t de Student) e manteve a porcentagem de fibras colágenas com níveis similares ao grupo controle (P > 0,05). Ainda, a fluoxetina reduziu a expressão de IL-1??e COX-2 e a atividade da MMP-9 quando comparada ao grupo ligadura (P < 0,05). Em conjunto, os dados demonstram que a fluoxetina diminuiu a capacidade de apresentação de antígeno das DCs, bem como a resposta inflamatória, a reabsorção óssea e a perda de colágeno na DP, indicando que ela pode constituir uma abordagem terapêutica promissora como moduladora da resposta do hospedeiro na DP
Abstract: Fluoxetine is a selective serotonin reuptake inhibitor presenting immunomodulatory and anti-inflammatory properties. The aim of this study was to evaluate the fluoxetine effects on immunoinflammatory response associated with periodontal disease (PD). The in vitro study evaluated the effects of fluoxetine on antigen-presentation capacity of dendritic cells (DCs). Bone marrow DCs obtained from C57BL/6 wild type mice were differentiated using GM-CSF (20 ng/mL). DCs were treated with fluoxetine (concentrations of 0.01, 0.1 or 1 ?M) for subsequent cytokine/chemokine assays (ELISA) and analysis of expression of MHC-class II and co-stimulatory molecules (CD80, CD86, PD-L1, ICOS-L) to T cell activation using flow cytometry. Fluoxetine was also applied to cultures with both DCs and Aggregatibacter actinomycetemcomitans (Aa)-reactive T cells (×Aa-T), which were used for analysis of T cells proliferation/activation using thymidine and ELISA assays. Desipramine, a selective norepinephrine reuptake inhibitor, was also tested in vitro for comparison to fluoxetine. In vivo, male Wistar rats received ligature placement around mandibular first molars and were randomly assigned into three experimental groups (n=10/group): 1) Control rats (without ligature); 2) rats with ligature + placebo (saline; oral gavage); 3) rats with ligature + fluoxetine (20 mg/kg/day in saline; oral gavage). Histometric and histological analyses were performed for measurement of loss of bone in furcation region (H&E stain) and collagen fibers (picrosirius red stain) in the connective tissue of rats submitted to 15 days of PD induction. Gingival tissues were collected from animals submitted to 3 days of PD induction for analyses of mRNA expression of IL-1?, COX-2, MMP-9 and iNOS using RT-PCR, measurement of total protein concentration and MMP-9 activity using zymogram. Fluoxetine suppressed IL 12, IL-1?, TNF-?, RANTES and MIP-1??production by LPS-stimulated DCs (P < 0.05, ANOVA, Student's t test), as well as significantly reduced the expression of ICOS-L. Fluoxetine suppressed the proliferation of ×Aa-T stimulated with Aa-antigen presentation by DCs from co-cultures. When applied to ×Aa-T/DCs co-cultures, serotonin (5-HT) increased T cell proliferation, indicating that fluoxetine effects are independent of 5-HT. Desipramine effects were similar to those of fluoxetine. In the in vivo study, fluoxetine reduced alveolar bone loss as compared to ligature group (P < 0.05, ANOVA, Student's t test) and maintained collagen fibers levels similarly to control group (P > 0.05). Fluoxetine reduced IL-1??and COX-2 expression, as well as MMP-9 activity, from gingival tissues when compared to ligature group (P < 0.05). Altogether, data showed that fluoxetine can modulate the antigenpresentation capacity of DCs and reduce inflammatory response and loss of bone and collagen associated with PD. In conclusion, fluoxetine can modulate both immune and inflammatory responses on PD, suggesting that it may constitute a new therapeutic approach for modulation of host response in periodontal therapy
Doutorado
Farmacologia, Anestesiologia e Terapeutica
Doutor em Odontologia
Guimarães, Gustav [UNESP]. "Influência da extensão da doença periodontal no perfil sanguíneo de ratos Wistar". Universidade Estadual Paulista (UNESP), 2016. http://hdl.handle.net/11449/144733.
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O objetivo deste trabalho foi investigar a influência da extensão da doença periodontal no perfil sanguíneo de ratos Wistar. Foram utilizados 30 ratos divididos em 3 grupos de 10 animais: C- ratos controle; DP1- ratos com doença periodontal em 3 molares superiores; DP2 – ratos com doença periodontal em 3 molares superiores e 3 inferiores. A doença periodontal foi induzida por meio da confecção de ligadura em torno do colo dentário dos molares selecionados de acordo com cada grupo. Após 30 dias, os animais foram anestesiados e, por meio de uma punção cardíaca, foi coletado 5ml de sangue para as análises dos parâmetros do hemograma, creatinina, triglicérides e colesterol. Em seguida, os animais foram sacrificados e as maxilas removidas e processadas para análise histopatológica em coloração de H.E. para caracterizar o perfil da doença periodontal em cada animal. Os resultados obtidos a partir do tecido hematológico foram analisados estaticamente por meio dos testes de ANOVA e Tukey, com nível de significância de 5%. Pode-se observar nos grupos DP1 e DP2 presença de infiltrado inflamatório crônico, perda de inserção conjuntiva, perda de estrutura óssea cortical e alveolar, presença de reabsorções dentárias, presença de biofilme e sequestro ósseo. Quanto ao perfil sanguíneo, pode-se observar diferença estatisticamente significante entre o grupo controle e os grupos DP1 e DP2 em relação à quantidade de leucócitos e linfócitos (p<0,05). Ainda, o grupo DP2 apresentou maior quantidade de leucócitos e neutrófilos em relação ao grupo DP1 e controle (p<0,05). Pode-se concluir que a presença da doença periodontal eleva a quantidade de leucócitos, neutrófilo e linfócitos no sangue de ratos Wistar, e que, a extensão da doença periodontal influencia na quantidade de leucócitos e neutrófilos.
The aim of this study was to evaluate the influence of the periodontal disease extension on the blood profile of Wistar rats. Thirty rats were divided into 3 groups of 10 animals each: C control rats; DP1- rats with periodontal disease in 3 upper molars; DP2 - rats with periodontal disease in 3 upper and 3 lower molars. Periodontal disease was induced by ligature around the tooth according to each group. After 30 days, the animals were killed and 5ml of blood was collected for analyzes of blood count parameters, creatinine, triglycerides and cholesterol. Then the animals were sacrificed and the jaws removed and processed for histopathological analysis to characterize, in H.E. staining, the profile of periodontal disease in each animal. The results from the hematologic tissue were analyzed statistically by ANOVA and Tukey test, with 5% significance level. It can be observed in groups DP1 and DP2 presence of chronic inflammatory infiltrates, connective tissue attachment loss and alveolar bone cortical structure loss, presence of resorptions, presence of biofilm and bone sequestration. As for the blood profile, it is can observe a statistically significant difference between the control group and the groups DP1 and DP2 to the amount of leukocytes and lymphocytes (p <0.05). Further, DP2 group had a higher amount of leukocytes and neutrophils compared to DP1 and control group (p <0.05). It can be concluded that the presence of periodontal disease increases the amount of leukocytes, neutrophils, and lymphocytes in the blood of Wistar rats, and the extent of periodontal disease affects the quantification of leukocytes and neutrophils.
Salmon, Richard J. "Prevalence of mast cells within the periodontal ligament of the developing rat molar /". Title page, contents and summary only, 2000. http://web4.library.adelaide.edu.au/theses/09DC/09dcs172.pdf.
Second copy has title: Prevalence and determination of mast cell type within the periodontal ligament of the developing rat tooth. Bibliography: leaves 109-119.
Adams, A. M. "In vitro biocompatibility studies of dental restorative materials using human periodontal ligament cells". Thesis, University of Newcastle Upon Tyne, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.287331.
Martinez, Catalina. "The Effects of Dynamic Culturing Environments on Cell Populations Relevant to Heart Valve Tissue Engineering". FIU Digital Commons, 2011. http://digitalcommons.fiu.edu/etd/505.
Oliveira, Guilherme Henrique Costa 1988. "Avaliação do papel do marcador de superfície CD166 na diferenciação osteoblástica/cementoblástica de células mesenquimais indiferenciadas do ligamento periodontal de humanos". [s.n.], 2015. http://repositorio.unicamp.br/jspui/handle/REPOSIP/290297.
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: Células mesenquimais isoladas do ligamento periodontal (PDLSCs) de dentes humanos mostraram ser multipotentes e possuem a capacidade de se diferenciar em adipócitos osteoblastos in vitro. Além disso, possuem a capacidade de formar tecido semelhante ao cemento e ao ligamento periodontal quando transplantadas para animais imunossuprimidos. Por outro lado foi mostrado na literatura uma grande heterogeneidade destas linhagens de células e muito tem sido feito na tentativa de isolar grupos com fenótipos mais favoráveis a diferenciação em tecido mineralizado. A literatura aponta que existe uma modulação na expressão do marcador de superfície CD166 durante a diferenciação osteoblástica e a partir destes achados levanta-se a hipótese de sua participação neste processo. Em acréscimo, células positivas para este marcador mostraram alta capacidade de diferenciar-se em fenótipo osteoblástico. Deste modo, este estudo tem como objetivo avaliar se um grupo de células enriquecido para o marcador CD166 através de separação magnética (PDL/CD166+) apresenta maior potencial de diferenciação osteoblástica/cementoblástica quando comparado ao pool de células não separadas (PDL) e ao grupo de células que não foram retidas na coluna magnética (PDL/CD166-). Este estudo ainda se propõe a avaliar se o marcador de superfície CD166 é regulado no processo de diferenciação osteoblástica. Após a separação magnética de três populações celulares, os grupos PDL/CD166+ foram submetidos a citometria de fluxo para que fosse comprovado a alta expressão de CD166. Em seguida os três grupos, PDL, PDL/CD166+ e PDL/CD166- das três populações, foram submetidos aos ensaio de Alizarina para avaliar a capacidade formação de nódulos minerais in vitro, foram ainda caracterizados por real-time PCR para avaliação da expressão de genes relacionados ao fenótipo osteoblástico (fator de transcrição relacionado a Runt-2 -Runx-2; a fosfatase alcalina ¿ ALP ¿ e a osteocalcina ¿ OCN). Nos grupos PDL e PDL/CD166+ também foi avaliado a expressão de CD166 por qPCR e citometria de fluxo. Os grupos enriquecidos apresentaram uma expressão média de 91,5% (±4,3%) de CD166. Apenas um grupo PDL/CD166- de uma população não apresentou depósitos minerais in vitro e foi identificado diferença estatisticamente significante entre os grupos PDL/CD166+ e PDL/CD166- (p<0,05). Quanto a expressão dos marcadores biológicos do tecido ósseo, foi identificado uma tendência a aumento da expressão destes marcadores quando as células foram cultivadas sob indução e ao longo do tempo de cultura. Os grupos cultivados em meio osteogênico apresentaram maior expressão de CD166 por PCRq e também foi identificado que o grupo PDL/CD166+ mostrou uma expressão significativamente maior que o PDL, ambos sob indução (p<0,05). Por citometria de fluxo, foi identificado um acréscimo na expressão de CD166 no terceiro dia de diferenciação nas células PDL/CD166+ e foi estatisticamente significativo comparado ao meio padrão (p<0,05). Ao longo do tempo de cultivo sob indução, a expressão deste marcador foi diminuída. Através dos resultados deste estudo é possível concluir que o grupo enriquecido na expressão de CD166 apresenta um alto potencial de formação de nódulos minerais in vitro. Além disso, foi notado que os três grupos apresentam uma tendência a maior expressão de Runx-2, ALP e OCN quando cultivados em meio osteogênico e que a grande heterogeneidade destas três populações dificulta a identificação de diferenças significativas tanto na análise intergrupo como intragrupo e compromete a identificação de um subgrupo com fenótipo mais favorável a diferenciação osteoblástica. A expressão gênica de CD166 é aumentada sob indução e ao longo do tempo de cultura e por citometria de fluxo observa-se que as células cultivadas em OM tendem a reduzir a expressão deste marcador
Abstract: Mesenchymal stem cells from periodontal ligament (PDLSCs) isolated from human teeth were shown to be multipotent and have the capacity to differentiate into adipocytes and osteoblasts in vitro. Moreover, they have the ability to form cement and periodontal ligament-like tissues when transplanted into immunosuppressed animals. On the other hand, it has been shown in literature a great heterogeneity of these cell lines and much has been done in the attempt to isolate groups with a phenotype more prone to differentiate into mineralized tissues. The literature suggests that there is a modulation of the CD166 surface marker expression during osteoblast differentiation and from these findings it is hypothesized of its participation in this process. In addition, cells positive for this marker have shown strong ability to differentiate into osteoblastic phenotype. Thus, this study aims to assess whether an enriched cell group for the CD166 marker (PDL/CD166+) has greater potential for osteoblastic/cementoblastic differentiation when compared to the not separated cell pool (PDL) and cell group which are not enriched (PDL/CD166-). Then, the three groups PDL, PDL/CD166+ and PDL/CD166- from three populations were submitted to Alizarin red-based assay to evaluate the ability of mineral nodule formation in vitro, groups were further characterized by qPCR for the evaluation of the expression of genes related to the osteoblastic phenotype (transcription factor related to Runt-2 -Runx-2; alkaline phosphatase - ALP - and osteocalcin - OCN). In the PDL and PDL/CD166+ groups the CD166 expression was also assessed by qPCR and flow cytometry. Enriched groups showed an average expression of 91.5% (± 4.3%) of CD166. Only a PDL/CD166- group from one population showed no mineral deposits in vitro and it was identified statistically significant difference between the groups PDL/CD166+ and PDL/CD166- (p <0.05). Regarding the expression of bone tissue biomarkers, it was noted a tendency to increased expression of these markers when cultured under osteogenic induction and along time in culture. Groups cultured in osteogenic medium showed higher expression of CD166 by qPCR and it was also identified that the PDL/CD166+ group showed a significantly higher expression than the PDL group, both under osteogenic induction (p <0.05). By flow cytometry, it was noticed an increase in CD166 expression on the third day of differentiation in PDL/CD166+ cells and it was statistically significant compared to the standard medium (p <0.05). Along time in culture, the expression of this marker was decreased. Through the results of this study it is possible to conclude that the group enriched in CD166 expression has a high potential to form mineral nodules in vitro. In addition, it was noted that the three groups have a tendency to increased Runx-2, ALP and OCN expression when cultured in osteogenic medium and the great heterogeneity of these three populations difficult to identify significant differences both intra and inter-group analysis what may jeopardize the identification of a subgroup with a phenotype more prone to osteoblastic differentiation. Gene expression for CD166 is increased under induction e through culture time and by flow cytometry it is observed that cells cultured in OM tend to reduce the expression of this marker
Mestrado
Periodontia
Mestre em Clínica Odontológica
Pelaez, Daniel. "Role of Mechanical Strain on the Cardiomyogenic Differentiation of Periodontal Ligament Derived Stem Cells". Scholarly Repository, 2011. http://scholarlyrepository.miami.edu/oa_dissertations/573.
Pöllänen, Marja. "Epithelial cell degeneration in the presence of bacteria and host derived factors associated with periodontal disease". Turku : Turun Yliopisto, 2000. http://books.google.com/books?id=xOlpAAAAMAAJ.
Lin, Deborah G. "Storage conditions of avulsed teeth affect the phenotype of cultured human periodontal ligament cells". Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0003/MQ45972.pdf.
Lagerholm, Sara. "Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament of Healthy Teeth". Thesis, Malmö universitet, Odontologiska fakulteten (OD), 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:mau:diva-19683.
ABSTRACT:Isolation and Characterization of Mesenchymal Stem Cells from the Periodontal Ligament ofHealthy TeethAIM: To isolate and culture viable cells from the periodontal ligament and confirming theiridentity as mesenchymal stem cells.METHODS AND MATERIALS: Healthy premolars were collected at the time oforthodontic extractions. The middle 1/3 of the periodontal ligament was scraped andsubsequent cell isolation was performed using an enzymatic method; yielding single cellisolates. Cells were cultured and maintained under standard culture conditions. Cellcharacterization was performed by flow cytometry using two sets of cell surface markers; oneknown to be present and one known to be absent in mesenchymal stem cells. Ability of thecells for in vitro differentiation into adipogenic and osteogenic lineages was tested usingspecifically formulated media supplements.RESULTS: Cells were successfully isolated from 11 of 13 teeth and were maintained asadherent cultures for up to 8 generations. Cellular expression of positive markers; CD73, CD90and CD44 were confirmed by flow cytometry. For the negative marker panel, expression ofCD45, CD34, CD11b, CD19 and HLA class II were not detectable. The expression of CD105was inconclusive. As determined by phenotypic changes, cells appeared to have undergoneadipogenic and osteocytic differentiation at 21 days.CONCLUSION: This study has resulted in successful isolation and partial characterization ofmesenchymal stem cells from the periodontal ligament of healthy teeth. Non-invasive accessto these cells, provides an excellent tool for future studies, potentially leading to beneficialknowledge transferable to the dental clinical situation.
Brito, Victor Gustavo Balera [UNESP]. "Papel dos mastócitos sobre o metabolismo ósseo local e sistêmico de ratos normotensos e hipertensos com doença periodontal". Universidade Estadual Paulista (UNESP), 2018. http://hdl.handle.net/11449/154428.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Introdução: A doença periodontal (DP) é uma desordem inflamatória dos tecidos de suporte dos dente, iniciada pelo acumulo de biofilme bacteriano, apresentando consequências locais e sistêmicas. A coexistência de doenças sistêmicas, como a hipertensão, pode levar a uma inflamação exacerbada, maior reabsorção óssea e dano sistêmico pronunciado. Além disso, células imunes residentes têm importante papel na progressão da DP, porém, a participação dos mastócitos (MC) ainda não é bem compreendida. Objetivos: Avaliar o papel dos MC sobre o metabolismo ósseo local (mandíbula) e sistêmico (fêmur), em modelo animal normotenso (ratos Wistar) e hipertenso (ratos SHR) com DP. Métodos: Ratos machos Wistar e SHR (10 semanas) foram utilizados. A depleção de MC foi conduzida pelo pré-tratamento com o composto 48/80 e a DP foi induzida por ligadura bilateral nos primeiros molares inferiores, mantida por 15 dias. Foi realizada a identificação de MC no tecido gengival, por coloração com Azul de Toluidina. A perda óssea alveolar e parâmetros de arquitetura óssea na mandíbula e fêmur foram avaliados por microtomografia computadorizada, a expressão gênica de marcadores de formação, remodelamento e reabsorção na mandíbula e fêmur foram avaliada por RT-PCR em tempo real, e a produção de citocinas nos tecidos de interesse foi avaliada por ELISA. Principais resultados: Observamos significativa perda óssea induzida por DP, principalmente no SHR, em comparação ao Wistar, e a depleção de MC foi capaz de prevenir esta perda. A DP levou a expressão aumentada de Opn, Opg, Rankl e Rank, Trap, Ctsk, Vtn, Itga5 e Itgb5, além de aumento na produção de TNF-α, IL-6, IL-10 e CXCL3 na mandíbula, o que foi reduzido em animais depletados de MC. No fêmur, a DP não levou a alterações da arquitetura óssea, mas foi capaz de aumentar a expressão de Runx2, Opn, Opg, Rankl, Rank, Trap, Mmp2, Mmp9, Vtn, Itga5 e Itgb5, o que também foi significativamente reduzido pela depleção de MC. Conclusões: Nossos dados sugerem um importante papel dos MC nas consequências ósseas locais (mandíbula) da DP, especialmente nos animais SHR, além dos efeitos sistêmicos (fêmur) onde os MC possivelmente têm participação na modulação da expressão de marcadores ósseo, alterados pela DP, o que foi mediado por vias diferentes das observadas na resposta local.
Introduction: Periodontal disease (PD) is an inflammatory disorder of the tissues supporting the teeth, which start from the bacterial biofilm accumulation, and results in local and systemic damage. The coexistence of systemic diseases, such as hypertension, can lead to exacerbated inflammation, increased alveolar bone resorption and increased systemic damage. In addition, resident immune cells have an important role in PD progression, however the mast cells (MC) participation is still not well understood. Aims: To evaluate the role of MC in the local (mandible) and systemic (femur) bone metabolism in a normotensive (Wistar) and hypertensive (SHR) rats with PD. Methods: Males Wistar and SHR rats (10-week old) were used. MC depletion was conducted by compound 48/80 treatment and PD was induced by bilateral ligature placed in the lower first molars, which was maintained for 15 days. The identification of MC in the gingival tissue was done by Toluidine Blue staining, alveolar bone loss and bone architecture parameters of mandible and femurs were evaluated by micro-computed tomography, bone markers gene expression on the mandible and femur were evaluated by real time RT-PCR, and the cytokines production was evaluated by ELISA. Key results: We observed a more significant PD-induced bone loss in SHR, compared to Wistar, and MC depletion was able to prevent this loss. PD increased the expression of Opn, Opg/ Rankl/Rank, Trap, Ctsk, Vtn, Itga5 and Itgb5, as well as increased production of TNF-α, IL-6, IL-10 and CXCL3 in the mandible, what was prevented by MC depleted animals. In the femur, PD did not change bone architecture parameters, but was able to increase expression of Runx2, Opn, Opg/Rankl/Rank, Trap, Mmp2, Mmp9, Vtn, Itga5 and Itgb5, which also was significantly reduced by MC depletion. Conclusions: Our data suggest an important role of MC in the local bone consequences (mandible) of PD, especially in SHR, as well the systemic effects (femur), where MC may have a role in bone dynamics by modulating the expression of bone markers, altered by PD, through different mechanism from those observed in the local response.
15/03965-2
Aidoukovitch, Alexandra. "Effects of strontium on osteogenic capacity and proliferation of human periodontal ligament cells and osteoblasts". Thesis, Malmö högskola, Odontologiska fakulteten (OD), 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:mau:diva-19681.
Strontium (Sr2+) is the active substance of pharmaceuticals used for reducing fracture risk in osteoporotic patients. Lately, Sr2+ is combined with biomaterials to enhance osteogenesis, which has been vaguely studied considering periodontal tissue regeneration. Despite extensive use, the mechanisms of action of Sr2+ are not fully understood. The present study assesses the impact of Sr2+ on primary human periodontal ligament cells (PDL cells) and human osteoblasts in regard to proliferation and pro-osteogenic activity. Cultured human PDL cells and osteoblast cell lines MG63 and hFOB 1.19 were treated with SrCl2 (0.1-10 mM) or vehicle for 72 h. Cells were counted manually using a Bürker chamber. Total protein content was determined by colorimetric analysis using Bio-Rad protein assay. Alkaline phosphatase activity was determined enzymatically and normalized to total protein content. SrCl2 had no significant effect on PDL cells (p>0.05), but a tendency towards induced osteogenic characteristics was observed. In contrast, 5 mM SrCl2 enhanced total MG63 cell protein content by 37% (p<0.01), compared to vehicle, whereas a lower concentration (0.1 mM) did not. 5 mM SrCl2 increased MG63 cell number by 38% (p<0.001), while a higher concentration (10 mM) did not have a significant additional effect over the 5 mM (+54%, compared to vehicle, p<0.05). The results demonstrate that 72 h administration of ≥ 5 mM SrCl2 exerts a pro-proliferative effect on human osteoblast-like MG63 cells and display a tendency to induce osteogenic characteristics in primary human PDL cells.
Carrió, Bertrán Neus. "Comparación del perfil proteico del ligamento periodontal versus el hueso alveolar con el complejo periodontal diferenciado a partir de las DPPSCs". Doctoral thesis, Universitat Internacional de Catalunya, 2017. http://hdl.handle.net/10803/461770.
Farag, Amro Ahmed Mahmoud. "Decellularized Tissue Engineered Constructs Using Cell Sheet Technology". Thesis, Griffith University, 2016. http://hdl.handle.net/10072/367720.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Dentistry and Oral Health
Griffith Health
Full Text
Saito, Miki Taketomi 1986. "Caracterização do fenótipo de células mesenquimais indiferenciadas do ligamento periodontal de humanos com potencial para diferenciação osteoblásticas/cementoblástica". [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/289493.
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: Vários estudos têm sido conduzidos com o intuito de isolar e caracterizar células com fenótipo mesenquimal indiferenciado a partir do ligamento periodontal de humanos e avaliar o seu potencial em promover a neoformação dos tecidos periodontais. A partir destes estudos, sabe-se que há uma grande heterogeneidade celular no ligamento periodontal. Contudo, ainda não está claro na literatura se apenas um tipo de célula progenitora é capaz de se diferenciar em todos os tecidos presentes no periodonto ou se há um fenótipo celular mais favorável à regeneração periodontal. Neste contexto, o objetivo deste estudo foi isolar e caracterizar subclones celulares que apresentam maior comprometimento para aquisição do fenótipo osteoblástico/cementoblástico a partir de uma população de células do ligamento periodontal caracterizadas como mesenquimais indiferenciadas (CD105+ CD34- CD45-). Utilizando-se a técnica do cilindro de clonagem, subclones celulares foram isolados e avaliados quanto ao seu potencial de diferenciação osteoblástica/cementoblástica (ensaio de Von Kossa), à capacidade proliferativa (ensaio de MTS), e expressão da proteína STRO-1 pela técnica da imunofluorescência. Adicionalmente, os subclones celulares que apresentaram potencial de diferenciação osteoblástica/cementoblástica foram caracterizados pelo PCRq quanto à expressão de genes relacionados ao fenótipo osteoblástico (fator de transcrição relacionado à Runt- 2 - RUNX2 - e fosfatase alcalina - ALP), e modulação dos marcadores específicos para células mesenquimais indiferenciadas (CD105, CD166 e OCT-4) durante o processo de indução osteogênica. Os resultados mostraram que dos seis subclones isolados, três apresentavam potencial de diferenciação osteoblástica/cementoblástica, (grupo C-O), e os outros três não possuíam capacidade de formar matriz mineralizada (grupo C-F). O grupo C-O apresentou capacidade proliferativa significativamente menor comparada ao grupo C-F (p?0,05) e ambos os grupos apresentaram marcação positiva para proteína STRO-1. Durante o processo de indução osteogênica do grupo C-O, foi observado um aumento significativo (p?0,05) da expressão de RUNX2 e CD166, mas não dos outros marcadores avaliados (ALP, CD105 e OCT-4). Os achados deste estudo mostraram que as células mesenquimais indiferenciadas do ligamento periodontal de humanos CD105+ CD34- CD45- constituem uma população celular heterogênea, compreendendo um grupo de células mais proliferativas, mas sem potencial para depositar matriz mineralizada (grupo C-F), e outro grupo de células com menor potencial proliferativo, mas que possuem capacidade de diferenciação osteoblástica/cementoblástica (grupo C-O). Adicionalmente, foi observado que a expressão do marcador de superfície CD166 é modulada durante o processo de indução à diferenciação osteoblástica/cementoblástica no grupo C-O
Abstract: Several studies have been conducted in order to isolate and characterize cells from human periodontal ligament with mesenchymal stem cell phenotype, and evaluate its potential to promote periodontal tissues neoformation. From these studies, it was observed that there is a great heterogeneity in periodontal ligament cells. However, it is not clear in the literature whether only one type of progenitor cell is able to differentiate into all tissues of the periodontium or whether there is a cell phenotype more favorable for periodontal regeneration. In this context, the aim of this study was to isolate and characterize subclones that show greater commitment to acquisition of an osteoblastic/cementoblastic phenotype from a population of periodontal ligament cells characterized as mesenchymal stem cells (CD105+ CD34- CD45-). Cell subclones were isolated by ring-cloning technique and evaluated for their osteoblastic/cementoblastic differentiation potential (Von Kossa assay), proliferative capacity (MTS assay), and expression of STRO-1 protein by immunofluorescence technique. Additionally, the subclones that showed potential to osteoblastic/cementoblastic differentiation were characterized by qPCR for expression of genes related to osteoblastic phenotype (runt-related transcriptor factor-2 - RUNX2 and alkalin phosphatase - ALP) and modulation of specific markers of undifferentiated mesenchymal cells (CD105, CD166 and OCT-4) during osteogenic induction. Six subclones were isolated, and three of them presented osteoblastic/cementoblastic differentiation potential (C-O group), and the other three did not present this potential (C-F group). The C-O group showed significantly lower proliferative capacity compared to the C-F group (p ?0.05), and both groups were positively stained for protein STRO-1. During the osteogenic induction of C-O group, there was a significant increase in the expression of RUNX2 and CD166 (p ?0.05), but not in the other assessed markers (ALP, CD105 and OCT-4). The findings of the present study showed that CD105+ CD34- CD45- mesenchymal stem cells from human periodontal ligament are a heterogeneous cell population, comprising a group of more proliferative cells without potential to deposit mineralized matrix (C-F group), and another group of cells with lower proliferative potential with capacity of osteoblastic/cementoblastic differentiation (C-O group). Additionally, it was observed that the expression of CD166 is modulated during osteoblastic/cementoblastic induction process in C-O group
Mestrado
Periodontia
Mestra em Clínica Odontológica
Rincon, Julio Cesar. "Expression of non-collagenous proteins by the epithelial rest cells of Malassez /". [Brisbane, Qld.], 2004. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe18072.pdf.
Saoji, Nachiket A. "Effect of bisphosphonate on osteogenic differentiation of pulp and PDL cells". Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2008. https://www.mhsl.uab.edu/dt/2009r/saoji.pdf.
李晓 e Xiao Li. "Macrophage migration inhibitory factor and circulating progenitor cells: relevance and implications inperiodontal medicine". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2011. http://hub.hku.hk/bib/B45894267.
Mota, Patrícia Joana Soares. "Aplicação de células estaminais em Medicina Dentária". Master's thesis, [s.n.], 2012. http://hdl.handle.net/10284/3459.
Nos últimos anos têm-se realizado pesquisas sobre a utilidade de células estaminais em diversas áreas da saúde. Estes avanços científicos na área da bioengenharia tecidual têm levado ao desenvolvimento de novas terapias e curas de doenças que, até aos dias de hoje, não teriam cura. Recentes pesquisas têm revelado que os dentes são ótimas fontes de células estaminais. Tanto os dentes decíduos como permanentes têm células estaminais com grande potencial proliferativo e apresentam as caraterísticas de células estaminais: capacidade de autorrenovação e diferenciação em várias linhagens celulares. Mediante este contexto, o objetivo geral desta monografia foi realizar uma revisão de literatura em foco para atualizar o conhecimento acerca das células estaminais e a sua aplicação em medicina dentária. Tendo como objetivos específicos: analisar as células estaminais dentárias, ampliando o conhecimento sobre as possibilidades do seu potencial regenerativo, como fonte de células estaminais e sistematizar os conhecimentos, avanços científicos, limitações e perspetivas relativas à aplicação de células estaminais dentárias. Com esta revisão bibliográfica, é possível esclarecer determinadas dúvidas sobre células estaminais. O futuro passará em usar estas células como tratamento terapêutico, de maneira a regenerar muitos tecidos, fazendo com que os órgãos lesados possam voltar à sua vitalidade completamente saudável. As células estaminais são, assim, de uma enorme utilidade. Stem cells have been the target of abundant research focusing their potential in regenerative medicine. Scientific advances in bioengineered tissue has allowed the development of new therapies for diseases that, until recently, would not be possible. Recent researches have revealed teeth as great sources of stem cells. Both deciduous or permanent teeth have stem cells with high proliferative potential, exhibiting the characteristics of other studied stem cells: self-renewal capacity and differentiation into various cell lines. Under this context, the general objective of this thesis was to conduct a literature review focused on the stem cells knowledge and its application in dentistry. Having specific objectives: the study the dental stem cells, considering their regenerative potential as a source of stem cells and systematize the knowledge, scientific advances, limitations and prospects for the application of stem cells decay. This literature review, enables the clarification of certain doubts about stem cells. The future use of these cells as a therapeutic treatment in order to regenerate different tissues, causing the damaged organs can return to its vitality completely healthy. The stem cells are therefore extremely useful.
Belibasakis, Georgios N. "Cellular and molecular responses of periodontal connective tissue cells to Actinobacillus actinomycetemcomitans cytolethal distending toxin". Doctoral thesis, Umeå : Univ, 2004. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-345.
El-Hossary, Wafaa Hassanein Hassan. "The effect of cyclic tensile strain on gene expression of cultured human periodontal ligament cells". Thesis, University of Leeds, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.509041.
Al-Taweel, Firas Bashir Hashim. "The interaction of Porphyromonas gingivalis with host epithelial cells and its relevance to periodontal disease". Thesis, University of Sheffield, 2017. http://etheses.whiterose.ac.uk/17729/.
Vernon, Lauren Louise. "A Comparison of the Osteogenic Tissue Engineering Potential of Dental-Derived Stem Cell Lines: Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs) vs. Periodontal Ligament Stem Cells (PERIOS)". Scholarly Repository, 2010. http://scholarlyrepository.miami.edu/oa_theses/19.
Freire, Isabelle Rodrigues [UNESP]. "Estudo dos mecanismos envolvidos na resposta tecidual de camundongos diabéticos com doença periodontal: papel dos mastócitos". Universidade Estadual Paulista (UNESP), 2009. http://hdl.handle.net/11449/95484.
O objetivo do presente estudo foi investigar o papel dos Mastócitos (MAST) na resposta tecidual de camundongos com Diabetes Mellitus (DM) submetidos a Doença Periodontal (DP). Os camundongos foram pré-tratados com uma dose única de estreptozotocina (STZ) para indução do DM. Para avaliar o papel dos MAST no controle da DP, os camundongos foram depletados de MAST pelo tratamento com composto 48/80. Subsequentemente foi realizada a indução da DP nos camundongos com DM e controles pela ligadura dos primeiros molares homólogos. Após um período de 7 e 14 dias os animais foram sacrificados para coleta das amostras para ensaios subseqüentes. Os níveis de reabsorção óssea dos camundongos diabéticos e normais foram avaliados radiograficamente para confirmação da presença da DP. O recrutamento de Neutrófilos (NE) foi avaliado pela produção da enzima Mieloperoxidase (MPO) no tecido gengival. Os níveis de IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 e Linfotactina/ XCL1 nos tecidos gengivais e IFN-g e IL-4 no plasma foram avaliados pelo método imunoenzimático (ELISA). Os resultados mostram que animais diabéticos com DP apresentaram após 14 dias da indução da DP uma perda óssea significativa quando comparado ao grupo controle, diferentemente do grupo de 7 dias. Esta perda foi potenciada nos animais diabéticos com DP depletados de MAST. Verificou-se elevados níveis de MPO nos animais normais e com DM após 14 dias da indução da DP. Nos animais com DM e com DP tratados com 48/80 foi observada uma redução parcial dos níveis de MPO. A produção de IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 e Lin-fotactina/XCL1 foi observada nos animais diabéticos independente da indução da DP após o período 7 e 14 dias. Conclui-se então que o DM favoreceu o aumento da perda óssea e o recrutamento de neutrófilos na DP. Como também induziu a produção de altos níveis dos mediadores...
The aim of this study was to investigate the role of Mast cells (MAST) on the tissue response in mice with Diabetes Mellitus (DM) submitted to Periodontal Disease (PD). The mice were pretreated with a single dose of streptozotocin (STZ) for the induction of DM. To evaluate the role of MAST in the PD, the mice were depleted of MAST by a pretreatment with compound 48/80. Subsequently, PD was induced in diabetic and normoglycemics mice by using a ligature around the first molars homologous. Seven and fourteen days after the surgery, the animals were sacrificed and the samples were collected for the subsequent experiments. The levels of bone resorption in diabetic and normoglycemic mice with PD were radiographically evaluated to confirm the presence of the PD. Neutrophil migration (NE) was quantified by the presence of the MPO enzyme in the gingival tissue. The levels of IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 and Lymphotactin/XCL1 from gingival tissues and plasma were evaluated by Enzyme- Linked Immunosorbent Assay (ELISA). The results showed that diabetic mice had a significant bone resorption 14 days after the induction of PD when compared to normoglycemics mice. This bone resorption was higher in the diabetic MAST-cell depleted mice with PD. The level of MPO was higher in diabetic and normoglycemic mice 14 days after the induction of PD. Furthermore, it was observed a partial reduction of MPO levels in diabetic mice with PD treated with compound 48/80. The level of IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 and Lymphofotactin/XCL was observed in diabetic mice independently of the induction of PD after 7 or 14 days. In conclusion, DM increased bone resorption and neutrophil recruitment in the PD mice, as well as it induced the production of high levels of IFN-g, IL -4, RANTES/CCL5, KC/CXCL1 and Lymphofotactin/XCL1. The MAST depletion increased bone resorption and reduced NE recruitment... (Complete abstract click electronic access below)
Soares, Diego Moura. "Influ?ncia da laserterapia na prolifera??o de c?lulas-tronco do ligamento periodontal humano". Universidade Federal do Rio Grande do Norte, 2013. http://repositorio.ufrn.br:8080/jspui/handle/123456789/17838.
Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior
Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm? - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm? promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm?) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm? exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm?. No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration
O laser de baixa intensidade (LBI) tem sido utilizado na Odontologia com a finalidade de promover cicatriza??o e regenera??o dos tecidos. A literatura mostra um efeito positivo do LBI na prolifera??o celular, por?m pouco se sabe sobre a sua efic?cia na prolifera??o de c?lulas-tronco. O objetivo deste estudo foi avaliar o efeito da irradia??o do LBI na taxa proliferativa de c?lulas -tronco do ligamento periodontal humano. Extratos de ligamento periodontal foram isolados de dois terceiros molares h?gidos removidos por indica? ?o cir?rgica e/ou ortod?ntica. Ap?s a digest?o enzim?tica, as c?lulas foram cultivadas em meio de cultura α-MEM suplementado com antibi?ticos e 15% de soro fetal bovino. No terceiro subcultivo, as c?lulas foram irradiadas com um laser diodo InGaAlP, utilizando-se duas diferentes densidades de energia (0,5J/cm 2 - 16 segundos e 1,0J/cm? - 33 segundos), comprimento de onda de 660nm e pot?ncia de 30mW. Uma nova irradia??o, utilizando os mesmos par?metros, foi realizada 48 h ap?s a primeira. Um grupo controle (n?o irradiado) foi mantido nas mesmas condi??es experimentais de cultivo. O m?todo de exclus?o por azul de Tripan e a atividade mitocondrial das c?lulas medida atrav?s do ensaio de MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetraz?lio], nos intervalos de 0, 24, 48 e 72 h p?s-irradia??o, foram utilizados a fim de avaliar a prolifera??o celular. Os dados das contagens celulares foram submetidos a testes estat?sticos n?o param?tricos de Kruskal-Wallis e Mann-Whitney, considerando um intervalo de confian?a de 95%. Com o objetivo de verificar poss?veis altera??es morfol?gicas nucleares induzidas pelo laser, as c?lulas foram submetidas ? marca??o com DAPI (4 -6-Diamidino-2-phenylindole) no intervalo de 72 h. Os resultados do presente estudo mostraram que a densidade de energia de 1,0 J/cm? promoveu maior prolifera??o das c?lulas em compara??o com os outros grupos (controle e laser 0,5 J/cm?) nos intervalos de 48 e 72 h. A atividade mitocondrial, medida pelo ensaio de MTT, apresentou resultados semelhantes ?s contagem celulares com azul de Tripan, com o grupo irradiado com 1,0 J/cm? exibindo uma atividade significativamente maior do MTT nos intervalos de 48 e 72 h, quando comparado com o grupo irradiado com 0,5 J/cm?. Nenhuma altera??o morfol?gica nuclear foi observada, tanto das c?lulas do grupo controle quanto nas c?lulas irradiadas. Conclui-se que o LBI apresenta efeitos estimulantes sobre a prolifera??o de c?lulas-tronco do ligamento periodontal humano. Portanto, a aplica??o da laserterapia neste tipo celular pode ser importante para futuros avan?os na regenera??o periodontal
Junior, Samuel de Barros Ferreira. "Modulação da severidade da doença periodontal experimental por células CCR5+". Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/25/25142/tde-02072009-112551/.
The periodontal diseases (PD) affect the supportive tissues of the teeth and are triggered by periodontopathogens present in the dental biofilm. The clinical outcome is highly influenced by the host inflammatory and immune response with participation of many cellular types, that act in the local microenvironment modulating the host response to control the infection. Inflammatory cytokines, chemokines and its receptors are thought to be involved in the cellular migration to the periodontal tissues, but there is little knowledge about the mechanisms of determination of resistance or susceptibility to the PD and in the triggering of tissue damage by immune response components. This study evaluated the role of CCR5+ cells in the experimental PD induced by oral inoculation of Aggregatibacter actinomycetemcomitans in C57BL/6 wild type mice and CCR5-knockout mice. The phenotypic analysis of inflammatory infiltrate demonstrated that the most of CCR5+ cells coexpress CD3 and CXCR3, suggesting a phenotype compatible with Th1-type cells, and also co-express RANKL. In the absence of CCR5+ cells there was a significant overall reduction of inflammatory cells and RANKL+ cells influx to the periodontal tissues, reduction in the alveolar bone resorption, reduction in the levels of pro-inflammatory cytokines TNFα-, IL-1β and IFN-γ expression, as a reduction in the expression of MMP-1, MMP-2 and MMP-13. The absence of CCR5+ cells did not impair the control of periodontal infection, despite the reduction of iNOS levels. In conclusion, these data demonstrate that the most of CCR5+ cells are Th1 cells, which act as important modulators of TNFα-, IL-1β and IFN-γ, MMP-1, MMP- 2 and MMP-13 levels, and which also express and modulate the expression of RANKL, playing an important role in the immunopathogenesis of experimental PD, without impairing the control of periodontal infection. These facts point to CCR5+ cells as potentials targets to therapeutic interventions aimed to control periodontal diseases.
Freire, Isabelle Rodrigues. "Estudo dos mecanismos envolvidos na resposta tecidual de camundongos diabéticos com doença periodontal : papel dos mastócitos /". Araçatuba : [s.n.], 2009. http://hdl.handle.net/11449/95484.
Banca: João Eduardo Gomes Filho
Banca: Ana Paula Campanelli
Resumo: O objetivo do presente estudo foi investigar o papel dos Mastócitos (MAST) na resposta tecidual de camundongos com Diabetes Mellitus (DM) submetidos a Doença Periodontal (DP). Os camundongos foram pré-tratados com uma dose única de estreptozotocina (STZ) para indução do DM. Para avaliar o papel dos MAST no controle da DP, os camundongos foram depletados de MAST pelo tratamento com composto 48/80. Subsequentemente foi realizada a indução da DP nos camundongos com DM e controles pela ligadura dos primeiros molares homólogos. Após um período de 7 e 14 dias os animais foram sacrificados para coleta das amostras para ensaios subseqüentes. Os níveis de reabsorção óssea dos camundongos diabéticos e normais foram avaliados radiograficamente para confirmação da presença da DP. O recrutamento de Neutrófilos (NE) foi avaliado pela produção da enzima Mieloperoxidase (MPO) no tecido gengival. Os níveis de IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 e Linfotactina/ XCL1 nos tecidos gengivais e IFN-g e IL-4 no plasma foram avaliados pelo método imunoenzimático (ELISA). Os resultados mostram que animais diabéticos com DP apresentaram após 14 dias da indução da DP uma perda óssea significativa quando comparado ao grupo controle, diferentemente do grupo de 7 dias. Esta perda foi potenciada nos animais diabéticos com DP depletados de MAST. Verificou-se elevados níveis de MPO nos animais normais e com DM após 14 dias da indução da DP. Nos animais com DM e com DP tratados com 48/80 foi observada uma redução parcial dos níveis de MPO. A produção de IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 e Lin-fotactina/XCL1 foi observada nos animais diabéticos independente da indução da DP após o período 7 e 14 dias. Conclui-se então que o DM favoreceu o aumento da perda óssea e o recrutamento de neutrófilos na DP. Como também induziu a produção de altos níveis dos mediadores... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The aim of this study was to investigate the role of Mast cells (MAST) on the tissue response in mice with Diabetes Mellitus (DM) submitted to Periodontal Disease (PD). The mice were pretreated with a single dose of streptozotocin (STZ) for the induction of DM. To evaluate the role of MAST in the PD, the mice were depleted of MAST by a pretreatment with compound 48/80. Subsequently, PD was induced in diabetic and normoglycemics mice by using a ligature around the first molars homologous. Seven and fourteen days after the surgery, the animals were sacrificed and the samples were collected for the subsequent experiments. The levels of bone resorption in diabetic and normoglycemic mice with PD were radiographically evaluated to confirm the presence of the PD. Neutrophil migration (NE) was quantified by the presence of the MPO enzyme in the gingival tissue. The levels of IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 and Lymphotactin/XCL1 from gingival tissues and plasma were evaluated by Enzyme- Linked Immunosorbent Assay (ELISA). The results showed that diabetic mice had a significant bone resorption 14 days after the induction of PD when compared to normoglycemics mice. This bone resorption was higher in the diabetic MAST-cell depleted mice with PD. The level of MPO was higher in diabetic and normoglycemic mice 14 days after the induction of PD. Furthermore, it was observed a partial reduction of MPO levels in diabetic mice with PD treated with compound 48/80. The level of IFN-g, IL-4, RANTES/CCL5, KC/CXCL1 and Lymphofotactin/XCL was observed in diabetic mice independently of the induction of PD after 7 or 14 days. In conclusion, DM increased bone resorption and neutrophil recruitment in the PD mice, as well as it induced the production of high levels of IFN-g, IL -4, RANTES/CCL5, KC/CXCL1 and Lymphofotactin/XCL1. The MAST depletion increased bone resorption and reduced NE recruitment... (Complete abstract click electronic access below)
Mestre
Yilmaz, Ozlem. "Epithelial cell sensing and responses to P. gingivalis /". Thesis, Connect to this title online; UW restricted, 2002. http://hdl.handle.net/1773/6389.
Coyac, Benjamin R. "Periodontal pathobiology and defective cell-autonomous mineralization in X-linked hypophosphatemia". Thesis, Sorbonne Paris Cité, 2017. http://www.theses.fr/2017USPCB006/document.
X-linked hypophosphatemia (XLH) is a rare X-linked dominant disorder caused by inactivating mutations in the PHEX gene. The impairment of PHEX protein leads to an increase in FGF23, a circulating factor that causes systemic loss of phosphate. The rachitic skeleton of patients with XLH displays short stature and osteomalacia. Dental defects include poorly mineralized dentin and spontaneous dental abscesses. Little is known about the periodontal condition of XLH and if patients are more prone to develop periodontitis, eventually leading to tooth loss. Although the exact function and substrate of PHEX are not known, it has been shown in vitro that PHEX could interact with SIBLING proteins such as MEPE or OPN, both involved in the regulation of bone and dentin mineralization, but it is not yet clear if the defects in the calcified extracellular matrices of XLH are caused by systemic hypophosphatemia only, or also by local consequences of the absence of PHEX. The aim of this doctoral dissertation was to explore the pathobiology of the XLH periodontium and to determine the impact of PHEX deficiency at the local level in a model of human biomineralization where phosphate supply could be adjusted and normalized. We first examined 34 adults with XLH in a case-control study and observed that periodontitis frequency and severity were increased in individuals with late or incomplete supplementation in phosphate and vitamin D analogs. The periodontium was then analyzed in XLH dental roots and further characterized in the Hyp mouse, the murine model of XLH. We performed a model of tooth movement adaptation leading to the formation of cellular cementum and a model of periodontal breakdown and repair to investigate the impact of XLH on the pathobiology of periodontal tissues. Our results showed strongly affected XLH/Hyp periodontal phenotype and impaired pathobiology and suggested that the key role played by OPN in bone could not be generalized to other periodontal mineralized tissues. In order to determine the role of PHEX in local human mineralization, dense collagen gels were seeded with primary human dental pulp cells harvested from XLH patients displaying PHEX mutations and age-matched healthy individuals. Cell-seeded gels were cultured up to 24 days under osteogenic conditions and controlled phosphate medium concentrations. Our results showed that despite normal phosphate concentrations, PHEX deficiency led to decreased quantity and quality of the mineral phase and a pathologic accumulation and processing of OPN. Overall the original contributions of this doctoral dissertation consist in the demonstration of a higher susceptibility of XLH patients to periodontitis and in the evidence of a local effect of PHEX deficiency in the pathologic intrinsic mineralization from XLH osteogenic cells
Albiero, Mayra Laino 1988. "Efeito do lipopolissacarídeo bacteriano sobre as propriedades biológicas das células mesenquimais indiferenciadas do ligamento periodontal de humanos". [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/289492.
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: O objetivo do presente estudo foi avaliar se a exposição das células mesenquimais indiferenciadas do ligamento periodontal (PDLMSCs) ao lipopolissacarídeo da Porphyromonas gingivalis (Pg) e da Escherichia coli (Ec), levaria a alterações biológicas que comprometessem as propriedades relacionadas ao fenótipo mesenquimal indiferenciado. Para testar essa hipótese, inicialmente foi verificado se as cinco populações de células mesenquimais indiferenciadas purificadas para o antígeno de superfície CD105 (células CD105+) expresssavam os receptores Toll-like 2 e 4 (TLR2 e 4). Em seguida, as células foram cultivadas na presença do LPS da Pg e da Ec, e avaliadas quanto a sua viabilidade e proliferação pelo ensaio do MTS, expressão dos genes para citocinas inflamatórias IL-1?, IL-6, IL-8 e TNF-? (técnica do PCRq), imunomarcação para STRO-1 e expressão do gene identificador de pluripotencialidade - Oct-4, e capacidade de diferenciação osteoblástica/cementoblástica através dos ensaios para identificação de nódulos minerais e expressão dos genes para RUNX2, ALP e OCN. Os resultados mostraram que todas as populações celulares apresentaram fenótipo mesenquimal indiferenciado com marcação positiva para STRO-1, além de se mostrarem positivas para os receptores TRL2 e 4. O ensaio de MTS revelou que a exposição às três concentrações de LPS da Pg e da Ec (100 ng, 1 ?g e 10 ?g/ml) não comprometeu a viabilidade celular, mantendo todas as populações de PDLMSCs proliferativas ao longo dos 10 dias de cultivo. Em paralelo, verificou que LPS da Pg não alterou os níveis de RNAm para citocinas estudadas enquanto que, o LPS da Ec promoveu um aumento significativo na expressão de IL-6 e IL-8, quando aplicado nas concentrações de 100 ng e 1 ?g/ml. Adicionalmente, os resultados revelaram que a exposição celular aos LPS bacterianos, ambos na concentração de 1?g/ml, não alterou o fenótipo mesenquimal indiferenciado destas populações, uma vez que a expressão do gene OCT-4 e a marcação positiva para STRO-1 mantiveram-se semelhantes às células do grupo controle. Além disso, as células mantiveram a capacidade de diferenciação em fenótipo osteoblástico/cementoblástico, confirmado pela produção de nódulos minerais (ensaio de vermelho de alizarina) e expressão dos genes para RUNX-2, ALP e OCN semelhante ao grupo controle (células cultivadas em meio osteogênico). Somente foi observado um aumento significativo (p<0,05) na produção de nódulos minerais quando as células foram cultivadas na presença de 1?g/ml do LPS de Ec. Contudo, esse aumento da matriz mineral não está relacionado ao aumento nos níveis de RNAm para os genes relacionados ao fenótipo osteogênico comparado ao grupo controle. Dentro das condições experimentais avaliadas, concluí-se que a exposição das PDLMSCs ao LPS da Porphyromonas gingivalis e da Escherichea coli não alterou as propriedades biológicas destas células, mantendo assim, as características que conferem a estas a condição de mesenquimal indiferenciada
Abstract: The aim of this study was to evaluate if the exposure of mesenchymal stem cells of the periodontal ligament (PDLMSCs) to lipopolysaccharide (LPS) of Porphyromonas gingivalis (Pg) and Escherichia coli (Ec), would lead to biological changes that compromised the properties related to undifferentiated mesenchymal phenotype. To test this hypothesis, initially it was checked whether the five populations of mesenchymal stem cells purified by surface antigen CD105 (CD105+ cells) expressed the Toll-like receptors 2 and 4 (TRL2 and 4). Then, cells were cultured in the presence of LPS from Pg and Ec, and evaluated for viability and proliferation by the MTS assay, expression of proinflammatory cytokines IL-1?, IL-6, IL-8 and TNF-? (PCRq technique), immunostaining for STRO-1 and OCT-4 gene expression, an identifier of pluripotency, and osteoblast/cementoblastic differentiation capacity through assays to identify minerals nodules, as well as the gene expression of RUNX2, ALP and OCN. All these assays were carried out also in the presence of LPS from Escherichia coli (Ec), which is not considered a periodontal pathogen. The results showed that all cell populations with undifferentiated mesenchymal phenotype had positive staining for STRO-1, and also showed to be positive for the receptors TLR2 and 4. The MTS assay revealed that exposure to the three concentrations of Pg and Ec LPS (100 ng, 1?g and 10?g/ml) did not affect cell viability, keeping all PDLMSCs populations proliferative over 10 days of culture. In parallel, it was found that the LPS Pg did not alter mRNA levels for the cytokines studied, while the Ec LPS caused a significant increase in IL-6 and IL-8, when applied concentrations of 100ng/ml and 1?g/ ml. Additionally, the results revealed that cellular exposure to both LPS in the concentration of 1?g/ml, did not alter the undifferentiated mesenchymal phenotype of these populations, since the expression of gene OCT-4 and positive STRO-1 staining remained similar to cells in the control group. In addition, these cells retained their ability to differentiate into osteoblastic/cementoblastic phenotype confirmed by production of mineral nodules (alizarin red assay) and expression of the genes for RUNX-2, ALP and OCN similar to cells control group (cultured in osteogenic medium only). It was only observed a significant increase (p<0.05) in the production of mineral nodules when cells were cultured in the presence of 1?g/ml Ec LPS. However, this increase in the mineral matrix it is not related to increased levels of mRNA for genes related to osteogenic phenotype compared to the control group. Within the experimental conditions, it can be concluded that exposure of PDLMSCs to LPS of Porphyromonas gingivalis and Escherichia coli did not alter the biological properties of these cells thereby, maintaining the characteristics that confer to these cells the condition of mesenchymal stem cells
Mestrado
Periodontia
Mestra em Clínica Odontológica
Pinkerton, Mark Neil, e n/a. "The molecular basis of orthodontic tooth movement : cytokine signaling by PDL cells in tension an in vitro study". University of Otago. School of Dentistry, 2007. http://adt.otago.ac.nz./public/adt-NZDU20071207.161056.
Shaheen, Ibrahim Ahmed. "Studies of the effects of recombinant human bone morphogenetic protein-2 on human periodontal ligament cells in vitro". Thesis, Queen Mary, University of London, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.267481.
VECCHIATINI, Renata. "Mesenchymal stem cells from Wharton's Jelly and periodontal ligament: reliable not controversial sources for osteogenic differentiation and regenerative medicine". Doctoral thesis, Università degli studi di Ferrara, 2011. http://hdl.handle.net/11392/2388818.
Costa, Camila Alves. "Células tronco mesenquimais de medula óssea na regeneração periodontal. Prova de princípios - in vitro e in vivo". Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/58/58132/tde-12072016-161505/.
Clinical studies have shown satisfactory results in the application of the enamel matrix derivate (EMD) on root surfaces. However, the complete regeneration of all lost periodontal tissues remains a challenge. To achieve this aim the use of bone marrow mesenchymal stem cells (BM-MSCs) in an appropriate vehicle in tissue engineering has been extensively studied. An interesting application of the EMD would be like a specific biological agent for BM-MSCs in order to promote cell differentiation and proliferation and assist in periodontal regeneration processes. Therefore, the aim of this study is evaluate the influence of Emdogain (EMD) and its vehicle (Propylene Glycol Alginate - PGA) in proliferation, differentiation and mineralization of CD 45-90+ bone marrow mesenchymal stem cells (BM-MSCs). Rat bone marrow stromal cells (BMSCs) were colleted from iliac crest of rat Wistar-Kyoto, isolated by Ficoll method and separated into CD 45-90+ BM-MSCs by flow cytometer. For cellular characterization the presence of stem cells markers (Rex-1 gen and surface antigen CD 90), capacity to form colonies (CFU) and to express osteoblast and adipogenic phenotype was assessed. Four experimental groups was plated with CD 45-90+ BM-MSCs: Positive Control group: media containing 10% of fetal bovine serum (FBS); (2) Negative Control Group: media containing 2% FBS; (3) EMD Group: 25 μg/ml EMD in 2% FBS media; (4) PGA Group: 25 μg/ml PGA in 2% FBS media. Cellular proliferation was assed in 3, 7, 10 and 14 days with MTT assay. After osteogenic induction, alkaline phosphatase (ALP) activity and gene expression of osterix (OSX), osteopontin (OPN), ALP and RUNX2 by RT-PCR was assessed at 7, 10 and 14 days. Extracellular matrix mineralization was evaluated at 21 and 28 days qualitatively by Von-kossa and Alizarina Red staining, and quantitatively by calcium content. CD 45-90+ BM-MSCs presented higher Rex-1 gen expression, more surface antigen CD 90 and more CFU/well than BMSCs, and expressed osteoblast and adipogenic phenotype. EMD and PGA not influenced proliferation of CD45-90+ BM13 MSCs. ALP activity was not influenced by EMD and was reduced in PGA group. EMD and PGA not influenced expression of OSX and RUNX2 in 7 and 10 days and reduced in 14 days. EMD not influenced ALP and OPN expression, but the presence of the gel provided better results, with higher expressions of ALP and OPN genes. Both PGA and EMD groups presented less mineralized ECM in 28 days. It concludes that EMD not influenced the proliferation, differentiation and ALP activity of CD 45-90+ BM-MSCs, but the presence of the gel (PGA) seems optimize the differentiation of these cells. Both reduced ECM mineralization.
Bittencourt, Marília da Silva Pereira. "A influência do tratamento periodontal não cirúrgico sobre o perfil lipídico e células sangüíneas de pacientes portadores de periodontite crônica generalizada". Universidade do Estado do Rio de Janeiro, 2008. http://www.bdtd.uerj.br/tde_busca/arquivo.php?codArquivo=3483.
The aim of this study was to verify whether non-surgical periodontal treatment had any influence on lipid profiles, white and red blood cell elements, platelets and ESR of patients with generalized chronic periodontitis. Eighteen patients average aged 50,6 years ( 7,6) had 10ml of peripheral blood collected prior to the periodontal treatment and 30 days after it to be analyzed for lipidic profile, white and red blood cell elements and ESR. Seven of those patients average aged 47,4 years ( 5,9) were also reassessed 90 days after the end of the treatment. The clinical parameters used prior to the treatment and on the reassessment were Plaque Index (PI), Silness and Löe (1964), Gengival Index (GI), Löe (1967), Bleeding On Probing (BOP), Pocket Probing Depth (PPD), and Attachment Level (AL). Furcation involvement sites were also recorded and classified. The periodontal treatment consisted of non-surgical basic therapy. Thirty days after the end of the treatment all patients were reassessed and a significant improvement (P<0,05) in PI, GI, BOP, PPD values and in AL ≥6mm (P=0,05) was observed. Class I and II furcation involvement sites also showed a significant decrease. The seven patients reassessed at 30 days and at 90 days pos-treatment also showed significant (P<0,05) PI, GI, BOP and PPD value improvement between those phases. The 4-5 mm AL increased significantly (P=0,04) between pre-treatment and 90 days pos-treatment, while AL ≥6mm decreased significantly between the 30-day and 90-day follow-up reassessments (P=0,01 and P= 0,02 respectively). When the values at 30 days and those at 90 days were compared results similar to the ones described above were observed, including the 4-5 mm AL increase (P=0,02). An increased GI (P=0,07) is also verified. As for hematological values there was a significant decrease in band cell levels (P=0,05) and monocyte levels (P= 0,03) after the periodontal treatment (30 days), while both total cholesterol and LDL showed a trend toward an increase (P=0,09). The seven patients who underwent two follow-up reassessments showed a significant increase in total cholesterol when pre-treatment, 30 days (P=0,04) and 90 days (P=0,02) phases were compared as well as in LDL (P=0,04 and P=0,03, respectively ).When the 30-day and 90-day pos-treatment platelets values were compared, a trend toward a decrease (P =0,09) was observed. Castelli Index II (cholesterol/ HDL) shows a trend toward an increase (P=0,09) between the pre-treatment and the 30-day pos-treatment phases. These results allow us to conclude that the periodontal treatment influenced band cells and monocytes, by decreasing them, as well as on total cholesterol and LDL as shown by their value increase.
Fritz, Jason Ronald. "The Chondrogenesis of PDLs by Dynamic Unconfined Compression Is Dependent on p42/44 and Not p38 or JNK". Scholarly Repository, 2009. http://scholarlyrepository.miami.edu/oa_theses/225.
Philips, Julia Rachel. "B-1 And B-2 B Cell Responses To Lipopolysaccharide: Putative Roles In The Pathogenesis Of Periodontitis". Thesis, The University of Sydney, 2006. http://hdl.handle.net/2123/4395.
Periodontal disease is one of the most widespread diseases in humans and is characterised by chronic gingival inflammation and B cell accumulation and resorption of the crest of alveolar bone with subsequent loss of teeth. Porphyromonas gingivalis has been identified as a putative aetiological agent for periodontitis. The aim of the research presented in this thesis was to investigate, using in vitro systems, the responses of autoreactive B-1 and B-2 cells to enterobacterial and nonenterobacterial lipopolysaccharide (LPS) to shed light on the pathogenesis of chronic periodontitis and other diseases involving B cell accumulation and autoantibody production. The hypotheses tested were: (1) B cells respond differently to enterobacterial and non-enterobacterial LPS. (2) B-1 cells are activated by a lower concentration of LPS than B-2 cells. (3) LPS stimulation results in preferential accumulation of B-1 cells. Findings consistent with these hypotheses would provide new evidence for different roles for B-1 and B-2 cells in immune responses and that LPS stimulation could lead to B-1 cell accumulation in diseases thus characterised. Initial experiments investigated the responses of representative B-1 (CH12) and B-2 (WEHI-279) cell lines to preparations of P. gingivalis and Salmonella enteritidis LPS utilising flow cytometric and quantitative molecular methods. The cell lines responded differently to the two LPS preparations. There were significant but limited effects on viability and proliferation in the WEHI-279 cell line, but no significant changes in mRNA expression levels for genes including Toll-like receptors (TLR2, TLR4, RP105), immunoglobulin (IgM), cytokines (IL-6, IL-10), co-stimulatory molecules (CD80, CD86), and regulators of apoptosis (Bcl-2, Bax). In the CH12 cell line however, LPS stimulation had greater effect. Addition of S. enteritidis LPS from a threshold level of 100ng/mL was found to rescue the cells from death, reflected by the percentage viability and proliferation. Stimulation of CH12 cells with S. enteritidis LPS also led to a decrease in expression of RP105 mRNA, which may be part of a negative feedback loop. Interestingly, stimulation with low concentrations P. gingivalis LPS appeared to inhibit proliferation but high LPS concentrations stimulated proliferation of CH12 cells, although no further significant effects were noted in other analyses. Evidence was found that CH12 cells have a high basal level of activation. This suggests that this line is constitutively activated. Stimulation with P. gingivalis or S. enteritidis LPS did not affect the level of CD80 mRNA expression. It is possible that the CH12 line constitutively expresses a maximal level of CD80 (and possibly CD86) and further stimulation will not cause any increase. Since S. enteritidis LPS appeared to have more pronounced effects on both B cell populations, this LPS was used to further investigate B cell subset responses in a mixed splenocyte culture system. Experiments examining percentage viability and number of viable cells indicated that B-1 and B-2 B cells responded differently to LPS stimulation. A threshold level for B-2 cell response (significant increase in cell number) was found to be 100ng/mL LPS, in contrast to the B-1 B cell subset which were only significantly different to the unstimulated cells when stimulated with 50μg/mL LPS. By examining the expression of CD80, the majority of murine splenic B-1 cells were found to activated prior to any LPS stimulation in vitro. In contrast, the B-2 subset showed significant increase in CD80 expression only at high (≥10μg/mL) LPS concentrations. Studies of the division index of B-1 and B-2 cells showed a significant response in both subsets following stimulation with 1μg/mL and 10μg/mL LPS. However, overall, the results are inconsistent with LPS driving the preferential accumulation of B-1 cells in disease states. These experiments provided useful evidence that supported the idea that B-1 and B-2 cells respond differently to LPS. However, these studies were unable to directly address the role of P. gingivalis LPS in periodontitis. It may be that P. gingivalis LPS could have different effects to S. enteritidis LPS on primary B cells. It is still possible that B-1 cells may be more sensitive to P. gingivalis, as opposed to S. enteritidis LPS. Studies by other groups have suggested that the TH1/TH2 profile is skewed towards TH2 in chronic periodontitis and that P. gingivalis may drive this shift via its ability to signal through TLR2 (and modulate TLR4 signalling). Further, recent studies in our laboratories have found that P. gingivalis gingipains are able to polyclonally activate B cells and to break down both IFNγ and IL-12. Future studies should further examine the effects of B-1 and B-2 interactions in the mixed lymphocyte system together with subsequent studies utilising human periodontitis biopsies. The results presented in this thesis, together with work undertaken by other investigators, suggests that LPS could perturb the normal homeostatic mechanisms of the B-1 B cell-subset and increase polyclonal activation therefore contributing to the genesis of pathologies such as chronic periodontitis.
Philips, Julia Rachel. "B-1 and B-2 B cell responses to lipopolysaccharide putative roles in the pathogenesis of periodontitis /". University of Sydney, 2006. http://hdl.handle.net/2123/1852.
Periodontal disease is one of the most widespread diseases in humans and is characterised by chronic gingival inflammation and B cell accumulation and resorption of the crest of alveolar bone with subsequent loss of teeth. Porphyromonas gingivalis has been identified as a putative aetiological agent for periodontitis. The aim of the research presented in this thesis was to investigate, using in vitro systems, the responses of autoreactive B-1 and B-2 cells to enterobacterial and nonenterobacterial lipopolysaccharide (LPS) to shed light on the pathogenesis of chronic periodontitis and other diseases involving B cell accumulation and autoantibody production. The hypotheses tested were: (1) B cells respond differently to enterobacterial and non-enterobacterial LPS. (2) B-1 cells are activated by a lower concentration of LPS than B-2 cells. (3) LPS stimulation results in preferential accumulation of B-1 cells. Findings consistent with these hypotheses would provide new evidence for different roles for B-1 and B-2 cells in immune responses and that LPS stimulation could lead to B-1 cell accumulation in diseases thus characterised. Initial experiments investigated the responses of representative B-1 (CH12) and B-2 (WEHI-279) cell lines to preparations of P. gingivalis and Salmonella enteritidis LPS utilising flow cytometric and quantitative molecular methods. The cell lines responded differently to the two LPS preparations. There were significant but limited effects on viability and proliferation in the WEHI-279 cell line, but no significant changes in mRNA expression levels for genes including Toll-like receptors (TLR2, TLR4, RP105), immunoglobulin (IgM), cytokines (IL-6, IL-10), co-stimulatory molecules (CD80, CD86), and regulators of apoptosis (Bcl-2, Bax). In the CH12 cell line however, LPS stimulation had greater effect. Addition of S. enteritidis LPS from a threshold level of 100ng/mL was found to rescue the cells from death, reflected by the percentage viability and proliferation. Stimulation of CH12 cells with S. enteritidis LPS also led to a decrease in expression of RP105 mRNA, which may be part of a negative feedback loop. Interestingly, stimulation with low concentrations P. gingivalis LPS appeared to inhibit proliferation but high LPS concentrations stimulated proliferation of CH12 cells, although no further significant effects were noted in other analyses. Evidence was found that CH12 cells have a high basal level of activation. This suggests that this line is constitutively activated. Stimulation with P. gingivalis or S. enteritidis LPS did not affect the level of CD80 mRNA expression. It is possible that the CH12 line constitutively expresses a maximal level of CD80 (and possibly CD86) and further stimulation will not cause any increase. Since S. enteritidis LPS appeared to have more pronounced effects on both B cell populations, this LPS was used to further investigate B cell subset responses in a mixed splenocyte culture system. Experiments examining percentage viability and number of viable cells indicated that B-1 and B-2 B cells responded differently to LPS stimulation. A threshold level for B-2 cell response (significant increase in cell number) was found to be 100ng/mL LPS, in contrast to the B-1 B cell subset which were only significantly different to the unstimulated cells when stimulated with 50μg/mL LPS. By examining the expression of CD80, the majority of murine splenic B-1 cells were found to activated prior to any LPS stimulation in vitro. In contrast, the B-2 subset showed significant increase in CD80 expression only at high (≥10μg/mL) LPS concentrations. Studies of the division index of B-1 and B-2 cells showed a significant response in both subsets following stimulation with 1μg/mL and 10μg/mL LPS. However, overall, the results are inconsistent with LPS driving the preferential accumulation of B-1 cells in disease states. These experiments provided useful evidence that supported the idea that B-1 and B-2 cells respond differently to LPS. However, these studies were unable to directly address the role of P. gingivalis LPS in periodontitis. It may be that P. gingivalis LPS could have different effects to S. enteritidis LPS on primary B cells. It is still possible that B-1 cells may be more sensitive to P. gingivalis, as opposed to S. enteritidis LPS. Studies by other groups have suggested that the TH1/TH2 profile is skewed towards TH2 in chronic periodontitis and that P. gingivalis may drive this shift via its ability to signal through TLR2 (and modulate TLR4 signalling). Further, recent studies in our laboratories have found that P. gingivalis gingipains are able to polyclonally activate B cells and to break down both IFNγ and IL-12. Future studies should further examine the effects of B-1 and B-2 interactions in the mixed lymphocyte system together with subsequent studies utilising human periodontitis biopsies. The results presented in this thesis, together with work undertaken by other investigators, suggests that LPS could perturb the normal homeostatic mechanisms of the B-1 B cell-subset and increase polyclonal activation therefore contributing to the genesis of pathologies such as chronic periodontitis.
Alves, Luciana Bastos. "Expressão dos fenótipos fibroblástico e osteoblástico em culturas tridimensionais na presença de partículas de vidro bioativo". Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/58/58132/tde-22102012-090829/.
The aim of this study was to analyze the fibroblastic and osteoblastic phenotypes expression on three-dimensional cultures in the presence or not of bioactive glass particles. Fibroblasts derived from human periodontal ligament (hPDLF) and osteogenic cells from newborn rat calvaria were cultured on bi-dimensional surfaces - plastic coverslips ThermanoxTM (control), bi-dimensional collagen surfaces - ThermanoxTM coated with collagen I without bioactive glass particles (2D) and with bioactive glass particles (2D+BG), on three-dimensional collagen gel without bioactive glass (3D) and with particles (3D+BG). Were evaluated: Cell viability (MTT) in 3 days, 7 and 10 days; Phosphatase alkaline activity (ALP) normalized by total protein content at 7 and 14 days; Immunolocalization of non-matrix proteins collagen (ALP and OPN in hPDLF at 7 and 14 days, OPN and BSP in osteogenic cells at 7 days) by indirect immunofluorescence; Genes expression (real-time PCR) for Periostin (PRT), Calcium-Binding Protein (S100A4) and Fibromodulin (FBM): fibroblastic markers in hPDLF, and Alkaline phosphatase (ALP), Osteopontin (OPN), Bone sialoprotein (BSP), Osteocalcin (OC), Collagen I (COL I) and RUNX2: osteoblastic markers in both cell types; and Mineralization (staining with Alizarin red). The results obtained on hPDLF cultures showed that cell viability on 2D+BG was higher than on control at 3 days (p<0.05), and no significant difference between groups was observed at 7 and 10 days. The total protein content at 7 and 14 days was higher on 3D and 3D+BG cultures compared those on 2D+BG (p<0.05) and control (p<0.05) at 7 days. Significant difference was also observed between 3D and 2D at 14 days. The ALP activity at 7 days was higher on 2D and 2D+BG compared with 3D (p<0.05) and 3D+BG (p<0.05), it was also lower on 3D+BG than on control (p<0.05). However, at 14 days 3D and 3D+BG showed higher ALP activity than control (p<0.05). Immunolabeling for OPN and ALP were observed in cells on 3D at both periods and on 2D, 2D+ BG and control only at 14 days. At 7 days, the expression of mRNA for PRT was upregulated on 3D and 3D+BG compared with control (p<0.05), for the FBM it was higher on 3D and 2D than on 3D+BG (p<0,05), but it was lower on 3D+BG than on control (p<0.05). Cells on 2D exhibited higher levels of S100A4 mRNA expression than those grown on 2D+BG (p<0.05) and control (p<0.05). The mRNA levels expression for COL I and ALP on 2D+BG and 3D, and for RUNX2 and OPN on 3D and 2D, and for OC on 2D presented upregulated compared with control (p<0.05). Also, cells on 2D showed the level expression of OC higher than those on and 2D+BG (p<0.05). At 14 days, there was a decrease in all evaluated genes expression compared with 7 days analyses. Cells on 2D+BG expressed higher levels of PRT than on 3D+BG (p<0.05). 2D and 2D+BG showed the highest levels of mRNA expression for FBM. Gene expression of S100A4 on 2D was higher than on 3D (p<0.05) and 3D+BG (p<0.05), in which the levels of S100A4 were downregulated compared to control. COL I and ALP were downregulated on 3D (p<0.05) and on 3D+BG (p<0.05) compared with control. There was also a significant difference between 2D+BG and 3D+BG for mRNA ALP expression. RUNX2 expression was higher on 3D than on 3D+BG (p<0.05), and OC expression was higher on control. Calcified matrix formation was observed on BG cultures at 10 and 14 days. At 10 days, areas stained by Alizarin were no observed by microscopy, but the amount of mineralization on 2D+BG and 3D+BG was significantly higher than on control (p<0.05), 2D (p<0.05) and 3D (p<0.05). At 14 days, more extensive staining was observed on cultures with BG, but the mineralized nodules formation was independent of the particles. Calcium content on 2D+BG and 3D+BG was significantly higher than control (p<0.05) and 2D (p<0.05). Osteogenic cell cultures showed that cells grown on 3D and 3D+BG surfaces exhibited the lowest levels of cell viability. Significant differences were observed when 3D+BG was compared with 2D+BG (p<0.05) and control (p<0.05) and also between 2D and 3D (p<0.05). At 3 and 10 days, there were no significant differences for cell viabililty between the cultures. The total protein content was higher on 3D+BG than on the control (p<0.05) and 2D+BG (p<0.05) at 7 and 14 days. Significant differences were also observed between 3D and 2D (p<0.05) at 14 days. ALP activity at 7 days was higher on 2D+BG and 3D+BG. Significant differences were found between 2D and 2D+BG (p<0.05), 3D and 3D+BG (p<0.05), and between 3D and control (p<0.05). However, at 14 days 3D and 3D+BG had the lowest levels of ALP activity, significantly different from 2D (p<0.05) and 2D+BG (p<0.05). Immunolabeling for OPN and BSP were observed at 7 days on 2D, 2D+BG, 3D and control. At 7 days, the expression levels of mRNA for ALP, COL I and RUNX2 were higher on 3D and 3D+BG. OPN, BSP and OC exhibited higher expression levels on 2D+BG. COL I expression was higher on 3D+BG than on 2D+BG (p<0.05). Cells on 2D+BG showed higher expression levels of OPN and OC than those on 2D (p<0.05) and 3D+BG (p<0.05), in which both of these genes and BSP expression were downregulated compared with control. At 10 and 14 days areas stained with Alizarin red were observed all evaluated groups, especially on BG cultures. At 10 days the amount of calcium on 3D+BG was higher than on control (p<0.05), and it was also higher on 2D+BG compared with 2D (p<0.05) and control (p<0.05). At 14 days, 2D+BG and 3D+BG showed greater calcium amount than control (p<0.05) and 2D (p<0.05). In conclusion, this study demonstrated that in vitro 3D cultures of hPDLF and osteogenic cells from newborn rats calvaria were able of support cell viability, differentiation, and to contribute to expression of fibroblastic and osteoblastic phenotype in hPDLF. The BG particles also favored the viability, differentiation, mineralized matrix formation and phenotypic expression in both cell types.