Orientador: Regina Maria Puppin-Rontani / Tese (doutorado) - Universidade Estadual de Campina, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-12T09:08:02Z (GMT). No. of bitstreams: 1 Pascon_FernandaMiori_D.pdf: 20418378 bytes, checksum: 123cef34dc5e16d51cd9c6553ae586c5 (MD5) Previous issue date: 2008 / Resumo: A utilização de substâncias químicas auxiliares, durante o tratamento endodôntico, pode alterar a composição química e as propriedades físicas da estrutura dentinária corono-radicular. Além disso, pode interferir no processo de união do material restaurador às estruturas dentinárias da câmara pulpar. Dessa maneira, pesquisas devem ser realizadas possibilitando a indicação de uma substância, que apresente o mínimo efeito nas propriedades mecânicas dentinárias, enquanto exerce o efeito de debridamento desejado. No intuito de facilitar a apresentação desta Tese, a mesma foi dividida em três capítulos, como descrito nas proposições seguintes. Capítulo 1: revisar sistematicamente a literatura a respeito dos efeitos da solução de hipoclorito de sódio nas propriedades mecânicas da dentina do canal radicular quando a mesma é utilizada durante o tratamento endodôntico. Capítulo 2: verificar os efeitos de diferentes substâncias químicas auxiliares (soro fisiológico, hipoclorito de sódio, clorexidina gel, peróxido de uréia - Endo-PTC® e EDTA) na microdureza (realizada por meio do teste de microdureza Vickers) e morfologia (realizada por meio de análise em Microscopia Eletrônica de Varredura) da dentina da câmara pulpar de dentes anteriores decíduos e permanentes. Capítulo 3: avaliar o efeito de substâncias químicas auxiliares (soro fisiológico, hipoclorito de sódio, clorexidina gel, peróxido de uréia - Endo-PTC® e EDTA) na rugosidade superficial da dentina da câmara pulpar de dentes decíduos e permanentes. Baseando-se na revisão de literatura pôde-se concluir que há forte evidência de que o hipoclorito de sódio altera as propriedades mecânicas da dentina radicular, quando o mesmo é utilizado como solução irrigadora durante o tratamento endodôntico. O uso de substâncias químicas auxiliares diminuiu os valores de microdureza dentinária Vickers tanto para dentes decíduos quanto para os permanentes. Observaram-se resultados não conclusivos com o uso do EDTA 17% na dentina da câmara pulpar dos dentes decíduos e permanentes uma vez que a microdureza superficial não pôde ser mensurada. A solução de hipoclorito de sódio 1% e 5,25% produziu os menores valores de microdureza Vickers para dentes decíduos e permanentes, comparados à clorexidina gel 2% e grupo controle (soro fisiológico). Alterações morfológicas foram observadas quando a dentina da câmara pulpar foi irrigada com solução de hipoclorito de sódio 1% ou 5,25% associados ao EDTA 17%, EDTA 17% utilizado isoladamente e clorexidina gel 2% associada ou não ao EDTA 17%. Em relação à rugosidade superficial pôde-se concluir que a irrigação da dentina da câmara pulpar aumentou a rugosidade superficial dos dentes permanentes e decíduos tratados com hipoclorito de sódio 1% e 5,25% associados ao EDTA 17%. / Abstract: Irrigation solutions, endodontic auxiliary chemical substance, and chelating agents used on root canal treatment might yield changes in the chemical composition and physical properties of dentin surface, and affect its interaction with materials used for coronal sealing. Thus, studies are required to select a suitable chemical agent, which has minimal effects on the mechanical properties of the tooth whilst achieving the desired debridement effect. In order to facilitate the accomplishment of this Thesis, it was divided into three chapters, as described on the following descriptions. Chapter 1: to present a systematic review on the effect of sodium hypochlorite on the mechanical properties of root canal dentin, considering its use as an irrigation solution. Chapter 2: to verify the effects of endodontic irrigants, auxiliaries chemicals substances and chelating agent on the microhardness (Vickers Hardness test) and morphology (Scanning Electron Microscopy) of the pulp chamber dentin in primary and permanent teeth. Chapter 3: to evaluate the effect of endodontic irrigants, auxiliaries chemicals substances and chelating agent on the roughness of the pulp chamber dentin of primary and permanent teeth. Based on literature review it could be concluded that there is strong evidence showing that sodium hypochlorite alters the mechanical properties of root canal dentin, when used as an endodontic irrigant. The use of irrigation solutions decreased the Vickers Hardness Number values for both permanent and primary teeth. Inconclusive results were observed when 17% EDTA was used, since it could not be measured. Sodium hypochlorite provides lower Vickers Hardness Number for primary and permanent teeth, compared with 2% chlorhexidine and control group (saline solution). Morphological alterations were observed when pulp chamber irrigation was performed by 1% or 5.25% sodium hypochlorite associated with 17% EDTA, 17% EDTA, and 2% chlorhexidine gel associated or no with 17% EDTA. Regarding surface roughness, it was concluded that pulp chamber irrigation with 1% and 5.25% sodium hypochlorite associated with 17% EDTA increases pulp chamber roughness of primary and permanent teeth. / Doutorado / Odontopediatria / Doutor em Odontologia

Prenatal assessment of fetal health is routinely offered throughout pregnancy to ensure that the most effective management can be provided to maintain fetal and maternal well-being. Currently, invasive testing is used for definitive diagnosis of fetal aneuploidy, which is associated with a 1% risk of iatrogenic fetal loss. Developing non-invasive prenatal testing (NIPT) is a key area of research and methods to increase the level of cell-free fetal DNA (cffDNA) within the maternal circulation have been discussed to improve accuracy of such tests. In this study, three strategies; co-amplification at lower denaturation temperature polymerase chain reaction (COLD-PCR), inverse-PCR and Pippin Prep™ gel electrophoresis, were analysed to identify a novel approach to selectively enrich shorter cffDNA fragments from larger maternal cell-free DNA (cfDNA). The sensitivity of droplet digital PCR (ddPCR) against real-time PCR (qPCR) was compared for fetal sex and RHD genotyping. In addition RHD zygosity testing was carried out for non-maternal samples. Consequently, Pippin Prep™ gel electrophoresis was combined with ddPCR analysis for the NIPD of Down Syndrome (DS) in pseudo-maternal samples. The results revealed that the Pippin Prep™ gel electrophoresis enrichment approach successfully demonstrated 2-fold to 5-fold increases in the cffDNA fraction. However, further optimisation assays of COLD-PCR and inverse-PCR using actual maternal samples were required. The spike experiments for DS detection revealed that with the present assay IV overrepresentation of the chromosome 21 target could be significantly detected for samples with ≥15% ‘cffDNA fraction’. In conjunction with the Pippin Prep™ enrichment method, this would have enabled assessment of all 10 maternal samples. Alternatively, fetal sex and RHD genotyping results determined that ddPCR provides a more sensitive platform compared to qPCR approaches, particularly for samples that express low cffDNA fractions (<2%). The ddPCR platform also proved to be a rapid and accurate system for the determination of RHD zygosity. This study highlights that ddPCR could be used as opposed to qPCR for accurate determination of fetal sex and RHD status. While sequencing approaches currently provide the most sensitive platforms for NIPT of fetal aneuploidy, high costs (>£400) prevent universal application. The combination of cffDNA enrichment with ddPCR analysis could provide a cheaper and more widely available platform for NIPD. However, further large scale validation studies using actual maternal samples are required.