As células da polpa dental têm o potencial de responder a estímulos externos e reparar o tecido dentário lesionado. Nos dentes, as lesões cariosas profundas e as lesões traumáticas são frequentes e resultam em alterações pulpares. Com o desenvolvimento da ciência e a busca de formas inovadoras de tratamento utilizando biomateriais de tamanho cada vez menor, verificar a biocompatibilidade desses materiais antes da aplicação clínica se mostra necessário e promissor. Assim, a finalidade deste estudo foi avaliar a biocompatibilidade de uma nova nanopartícula de hidroxiapatita utilizando células pulpares humanas em cultura. As Nanopartículas de Hidroxiapatita (HAp) utilizadas em nosso estudo são um material novo e ainda não disponível para a prática clínica. Como parâmetro comparativo, foi utilizado o material já consagrado na Odontologia para tratamentos endodônticos, o Agregado Trióxido Mineral (MTA). Para tanto, foram realizados testes de biocompatibilidade (MTS (3-(4,5-dimetiltiazol-2-y)5-(3-carboximetoxifenil)-2-(4-sulfofenil-2H-tetrazolio)). Foi preparada uma solução de material na concentração de 0,1g/ml diluído em meio de cultura, proporção preconizada pela norma ISO 10.993-12 (ISO, 1996). Esta solução foi diluída em 3 diferentes concentrações (1/10, 1/100 e 1/1000). Também foram realizados testes de migração celular e testes de capacidade de diferenciação, na concentração de 1/100. Como resultados pudemos verificar que as células da polpa foram viáveis quando expostas a diferentes concentrações de HAp, proliferaram mais quando tratadas com HAp na concentração de 1/100, migraram em direção aos materiais estudados e diferenciaram-se em tecido mineralizado quando em contato direto com ambos materiais estudados. Concluímos que o novo material testado é biocompatível e capaz de induzir mineralização em células de polpa dental sugerindo que as HAp podem ter aplicações clínicas futuras em tecidos mineralizados como osso e dente. / The dental pulp cells have the potential to respond to external stimuli and repair the dental tissue injury. On teeth, deep carious lesions and traumatic injuries are common and result in pulp changes. With the development of science and the search for innovative treatment forms using reduced size biomaterials, to verify the biocompatibility of these materials before clinical application has become necessary and promising. Thus, the purpose of this study was to evaluate the biocompatibility of a new hydroxyapatite nanoparticle using human pulp cells in culture. The nanoparticles of hydroxyapatite (HAp) used in our study are new material and not yet available for clinical practice. As a comparator, we used the material already used in dentistry to root canal treatment, the Mineral Trioxide Aggregate (MTA). To do so, biocompatibility (MTS (3-(4,5-dimetiltiazol-2-y)5-(3-carboximetoxifenil)-2-(4-sulfofenil-2H-tetrazolio)). A solution of the material in a concentration of 0.1 g / ml diluted in culture medium recommended by ISO 10993-12 (ISO 1996) standard was prepared. This solution was diluted in three different concentrations (1/10, 1/100 and 1/1000). Were also performed cell migration and differentiation capacity assays at a concentration of 1/100. As results, we found that the pulp cells were viable when exposed to different concentrations of HAp, they proliferated more when treated with 1/100 HAp concentration, migrated toward the studied materials and differed into mineralized tissue when there was direct contact with both materials studied. We conclude that the new material tested is biocompatible and able to induce mineralization of dental pulp cells suggesting that the HAp may have future clinical applications in mineralized tissues such as bone and tooth.

No description available.

Thesis (M.S.)--University of Michigan, 1968.

Thesis (M.S.)--University of Michigan, 1968.

As células da polpa dental têm o potencial de responder a estímulos externos e reparar o tecido dentário lesionado. Nos dentes, as lesões cariosas profundas e as lesões traumáticas são frequentes e resultam em alterações pulpares. Com o desenvolvimento da ciência e a busca de formas inovadoras de tratamento utilizando biomateriais de tamanho cada vez menor, verificar a biocompatibilidade desses materiais antes da aplicação clínica se mostra necessário e promissor. Assim, a finalidade deste estudo foi avaliar a biocompatibilidade de uma nova nanopartícula de hidroxiapatita utilizando células pulpares humanas em cultura. As Nanopartículas de Hidroxiapatita (HAp) utilizadas em nosso estudo são um material novo e ainda não disponível para a prática clínica. Como parâmetro comparativo, foi utilizado o material já consagrado na Odontologia para tratamentos endodônticos, o Agregado Trióxido Mineral (MTA). Para tanto, foram realizados testes de biocompatibilidade (MTS (3-(4,5-dimetiltiazol-2-y)5-(3-carboximetoxifenil)-2-(4-sulfofenil-2H-tetrazolio)). Foi preparada uma solução de material na concentração de 0,1g/ml diluído em meio de cultura, proporção preconizada pela norma ISO 10.993-12 (ISO, 1996). Esta solução foi diluída em 3 diferentes concentrações (1/10, 1/100 e 1/1000). Também foram realizados testes de migração celular e testes de capacidade de diferenciação, na concentração de 1/100. Como resultados pudemos verificar que as células da polpa foram viáveis quando expostas a diferentes concentrações de HAp, proliferaram mais quando tratadas com HAp na concentração de 1/100, migraram em direção aos materiais estudados e diferenciaram-se em tecido mineralizado quando em contato direto com ambos materiais estudados. Concluímos que o novo material testado é biocompatível e capaz de induzir mineralização em células de polpa dental sugerindo que as HAp podem ter aplicações clínicas futuras em tecidos mineralizados como osso e dente. / The dental pulp cells have the potential to respond to external stimuli and repair the dental tissue injury. On teeth, deep carious lesions and traumatic injuries are common and result in pulp changes. With the development of science and the search for innovative treatment forms using reduced size biomaterials, to verify the biocompatibility of these materials before clinical application has become necessary and promising. Thus, the purpose of this study was to evaluate the biocompatibility of a new hydroxyapatite nanoparticle using human pulp cells in culture. The nanoparticles of hydroxyapatite (HAp) used in our study are new material and not yet available for clinical practice. As a comparator, we used the material already used in dentistry to root canal treatment, the Mineral Trioxide Aggregate (MTA). To do so, biocompatibility (MTS (3-(4,5-dimetiltiazol-2-y)5-(3-carboximetoxifenil)-2-(4-sulfofenil-2H-tetrazolio)). A solution of the material in a concentration of 0.1 g / ml diluted in culture medium recommended by ISO 10993-12 (ISO 1996) standard was prepared. This solution was diluted in three different concentrations (1/10, 1/100 and 1/1000). Were also performed cell migration and differentiation capacity assays at a concentration of 1/100. As results, we found that the pulp cells were viable when exposed to different concentrations of HAp, they proliferated more when treated with 1/100 HAp concentration, migrated toward the studied materials and differed into mineralized tissue when there was direct contact with both materials studied. We conclude that the new material tested is biocompatible and able to induce mineralization of dental pulp cells suggesting that the HAp may have future clinical applications in mineralized tissues such as bone and tooth.

Le rachitisme vitamino-résistant hypophosphatémique (RVRH) est une maladie génétique rare causée par des mutations du gène PHEX. La perte de fonction de la protéine PHEX conduit à l’augmentation du FGF23, une hormone circulante qui agit sur le rein et entraîne une perte systémique de phosphate. Le squelette rachitique des patients atteints de RVRH présente des déformations osseuses et une ostéomalacie. La dentine hypominéralisée des patients est à l’origine d’abcès dentaires fréquents, mais le statut parodontal des patients RVRH est mal connu, de même que leur risque de développer une parodontite pouvant aboutir à la perte des dents. La fonction et le substrat de la protéine PHEX ne sont pas identifiés avec exactitude. Il a été montré in vitro que PHEX avait la capacité d’interagir et de dégrader des protéines membres de la famille des SIBLINGs comme MEPE ou OPN, toutes les deux impliquées dans la régulation de la minéralisation des tissus osseux et dentinaires, mais on ne sait pas si in vivo les défauts de minéralisation observés résultent principalement de l’hypophosphatémie systémique ou bien également des effets directs de l’absence de PHEX sur les protéines régulatrices de la minéralisation. L’objectif de cette thèse a consisté à s’intéresser à la physiopathologie du parodonte dans le RVRH ainsi qu’à déterminer quel était l’impact de la mutation de PHEX dans un modèle de biominéralisation humaine où les conditions de concentration en phosphate pouvaient être ajustées et normalisées. Nous avons d’abord analysé le statut parodontal de 34 patients RVRH dans une étude clinique cas-témoins et ainsi montré que les malades dont la supplémentation en phosphate et vitamine D était tardive ou incomplète présentaient une fréquence et une sévérité accrues de maladie parodontale. Le phénotype parodontal du RVRH a alors été étudié sur des échantillons humains et sur le modèle murin du RVRH, la souris HYP. Nous avons réalisé un modèle d’égression dentaire de façon à permettre une apposition du cément cellulaire, ainsi qu’un modèle de résorption et de réparation osseuses parodontales afin de caractériser l’impact du RVRH sur la physiopathologie parodontale. Nos résultats ont montré que le phénotype parodontal et sa physiopathologie étaient très perturbés dans le rachitisme vitamino-résistant hypophosphatémique et chez la souris HYP, nous avons aussi pu mettre en évidence que le rôle pathologique majeur joué par l’ostéopontine dans le tissu osseux au cours du RVRH ne pouvait pas être généralisé aux autres tissus minéralisés du parodonte. De façon à identifier le rôle de PHEX dans la minéralisation matricielle locale indépendamment de la phosphatémie systémique, nous avons ensemencé des matrices de collagène dense avec des cellules primaires humaines issues de patients RVRH comparés à des contrôles que nous avons cultivés pendant 24 jours en conditions ostéogéniques avec des concentrations en phosphate identiques. Nos résultats ont montré que malgré une concentration normale en phosphate, la perte de fonction de la protéine PHEX entraînait une diminution de la quantité et de la qualité de la phase minérale et une accumulation et une dégradation pathologiques de la protéine OPN. Les contributions originales de ce travail de thèse doctorale ont consisté à démontrer sur le plan clinique et biologique la susceptibilité accrue du rachitisme hypophosphatémique lié à l’X quant au risque de développer une maladie parodontale, ainsi qu’à apporter la preuve d’un rôle pathologique de l’absence de PHEX indépendant de la phosphatémie sur des cultures primaires humaines. / 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.