published_or_final_version / Dentistry / Master / Master of Dental Surgery

The purpose of this study was to determine duration time, breakage and apical displacement, whilst using six different endodontic filing systems to prepare molar teeth. A total of 96 molar teeth were used in the study, divided equally, ie 16 teeth per system selected randomly, totalling 48 canals per system. A standardised access cavity was prepared for all the teeth before selection. The canals were filed according to the manufacturers&rsquo / guidelines. The result showed that PROTAPER&reg / , K3&trade / and the combination of: HERO Shaper&reg / , HERO Apical&reg / and Endoflare&reg / (Referred from hereon as HERO System for convenience) were statistically faster than PROFILE&reg / and FlexMaster&reg / , which were in turn faster than AETTM. Although breakage did occur in K3&trade / and HERO System this was not deemed statistically significant. Apical displacement occurred in the form of Type 1 in the AETTM, PROFILE&reg / and HERO System, but once again this was not statistically significant. It was concluded that more aggressive cutting features such as a positive rake<br /> angle, pyramidal shaped tip, progressive taper and absence of radial lands, if present, could have enabled K3&trade / , HERO System and PROTAPER&reg / to have faster times, and in addition these features did not compromise these systems with regard to apical foramina transportation and breakage.

As proteínas são importantes componentes da matriz extracelular da polpa dentária e possuem diferentes funções nos tecidos. A literatura odontológica não relata como os proteoglicanos se distribuem e agem na matriz extracelular de dentes decíduos durante o processo fisiológico de rizólise. Foi objetivo do trabalho analisar as expressões dos proteoglicanos biglican e decorin e relacioná-las com as diversas fases do processo de rizólise. Para isso foram utilizados dentes decíduos humanos extraídos e livres de lesões de cárie, apresentando vários estágios de reabsorção radicular, divididos em três grupos: com dois terços ou mais do comprimento radicular médio, um terço ou mais do comprimento radicular médio e menos de um terço do comprimento radicular médio. Foi utilizada a técnica da imunoistoquímica, com o método da estreptavidina-biotina-peroxidase, e anticorpos contra as proteínas anteriormente citadas. Os resultados mostraram que os proteoglicanos estudados apresentaram imunorreatividade na matriz extracelular da polpa e da dentina dos dentes nas três fases de reabsorção. Foi possível concluir que houve diferença na distribuição e no padrão de expressão dos proteoglicanos biglican e decorin apenas na área de reabsorção, nos dentes decíduos hígidos nas três fases de rizólise o que sugere um papel regulador destes proteoglicanos no processo de reabsorção fisiológica nos dentes decíduos hígidos. / Proteins are important components in pulp extracellular matrix and perform several different roles in the body tissues. The distribution and functions of some proteins have already been described but there aren?t studies showing how the proteoglycans are distributed and act in the extracellular matrix of deciduous teeth at the physiological resorption process. The aim of the present work was to study the expression and distribution of the following non-collagenous components of the extracellular matrix: biglycan and decorin in deciduous teeth dental tissues at the physiological root resorption. Hygid human deciduous teeth that were extracted for orthodontic reasons were grouped together according to root length: group I - two thirds or more of average length root, group II - one third or more of average length root and group III ? less than one third of average length root. The streptavidin-biotinperoxidase method of immunohistochemistry was used with antibodies against the antigens previously quoted. The results showed that the proteoglycans had been found in pulp and dentin extracellular matrix in all groups. In conclusion, biglycan and decorin were differentially found only in the resorption area, between tissues in resorption process and its adjacent zone, which make us believe that these proteoglycans act regulating the physiological resorption process in hygid deciduous teeth.

Avaliou-se a proliferação das células tronco da polpa de dentes decíduos esfoliados humanos (SHED) após aplicação única do laser de baixa potência. Foi realizada a análise da viabilidade das SHED cultivadas sob déficit nutricional e em condições ideais após irradiação com o laser de baixa potência vermelho de Indio Gálio Alumínio e Fósforo - InGaAlP (660nm, 40mW e 10J/cm2) e infravermelho (780nm, 40mW e 10J/cm2) durante 4 e 8 segundos, nos períodos de 24, 48 e 72 horas através dos ensaios de redução do MTT e do ensaio colorimétrico de Busatti e Gomes. Para análise estatística utilizou-se o teste ANOVA complementado pelo teste de Tukey com nível de significância de 5% (p< 0,05). Observou-se tanto com o MTT quanto com o ensaio colorimétrico de Busatti e Gomes uma tendência de aumento da proliferação celular diretamente relacionada à dose do LBP, estatisticamente significante nos períodos de 24, 48 e 72 horas. Ao analisar os resultados e considerando os parâmetros utilizados e o protocolo de LBP, pode-se concluir que o LBP promoveu a proliferação das SHED tanto a 660 nm quanto a 780nm, pode influenciar a viabilidade e a proliferação das SHED nas doses e comprimentos de onda utilizados e os ensaios do MTT e colorimétrico de Busati e Gomes demonstraram dentro de suas limitações ser eficientes para determinar a viabilidade e proliferação das SHED. / It was evaluated the proliferation of stem cells from human exfoliated deciduous teeth (SHED) after a single application of low power laser. The viability of SHED grown under ideal conditions and under nutritional deficit after irradiation with red laser (660/780nm, 10J/cm2 and 40mW) during periods of 4 and 8 seconds was analyzed through the MTT reduction assays and rapid colorimetric assay of Busatti and Gomes. Statistical analysis was performed using the ANOVA and Tukey´s multiple comparisons test with a significance level of 5% (p < 0.05). It was observed with the MTT assay and Busatti and Gomes assay a trend of cell proliferation increase directly releated to the irradiation dose, statistically significant. After 24, 48 and 72 hours, all the groups showed higher cell proliferation when compared to control. Analyzing the results and considering the used parameters and LBP protocol, it can be concluded that LBP promoted the proliferation of SHED both 660nm and 780nm according to the dosage and wavelengths used, and MTT assay and colorimetric Busatti and Gomes demonstrated within their limitations to be effective in determining the viability and proliferation of SHED.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Um dos principais objetivos do tratamento endodÃntico dos canais radiculares de dentes com polpa necrÃtica consiste em eliminar os microorganismos localizados no sistema de canais radiculares. Dessa forma, esta dissertaÃÃo, constituÃda por um artigo, propÃe-se a avaliar o efeito antibacteriano do preparo quÃmico-mecÃnico e de uma pasta à base de hidrÃxido de cÃlcio sobre bactÃrias presentes em canais radiculares de dentes decÃduos necrosados apÃs trauma bem como verificar a presenÃa dos microorganismos Fusobacterium nucleatum e bacilo pigmentado negro nestes dentes. Seguindo os critÃrios de inclusÃo, a amostra consistiu de 18 dentes, totalizando 14 pacientes. As coletas microbiolÃgicas foram realizadas apÃs a abertura coronÃria (C1) e 72h apÃs a remoÃÃo da medicaÃÃo intracanal (C3), sendo que, para 10 dentes, realizou-se outra coleta apÃs a instrumentaÃÃo (C2). As coletas foram realizadas introduzindo-se sequencialmente 3 cones de papel absorvente estÃril, de diÃmetro visualmente compatÃvel com o do canal radicular, no interior deste por aproximadamente 1 minuto. ApÃs este intervalo, os cones foram removidos, transferidos para um tubo contendo um fluido reduzido para transporte e levados ao laboratÃrio para processamento microbiolÃgico. Na C1, os microorganismos foram isolados em 17/18 (94,4%) dos canais radiculares, sendo a mÃdia de CFUs de 5.4 x 105, na C2, em apenas 1/10 (10%) canal radicular, com uma mÃdia de 4.3 x 102, e na C3 em 15/18 (83.3%), com mÃdia de 1.5 x 105. Houve uma diferenÃa estatisticamente significante entre C1 e C2, o mesmo nÃo ocorrendo entre C1 e C3 e entre C2 e C3. O microorganismo Fusobacterium nucleatum e o bacilo pigmentado negro foram observados em 55.5% (10/18) e 11.1% (2/18), respectivamente, na C1, nÃo sendo detectados na C2, e na C3 estavam presentes em 16.6% (3/18) e 5.5% (1/18), respectivamente. Na C1, observou-se uma predominÃncia de cocos gram-negativos (15/18) e bacilos gram-negativos (14/18), representando 83.3% e 77.8%, respectivamente. Na C2, os Ãnicos morfotipos detectados foram cocos gram-positivos (1/10), presente em 10% das amostras positivas, e na C3, os cocos-gram positivos predominaram (66.7%). Pode-se concluir que o preparo quÃmico-mecÃnico desempenhou sua funÃÃo antibacteriana ao reduzir significativamente o nÃmero de microorganismos do canal principal, porÃm o hidrÃxido de cÃlcio possui um efeito antibacteriano limitado, nÃo sendo capaz de prevenir o recrescimento de bactÃrias apÃs seu uso como medicaÃÃo intracanal. / One of the main objectives of endodontic treatment of roots canals with necrotic pulps consists in eliminating the microorganisms spread throughout the ramifications of root canal system. This study, comprised by one manuscript, had the objective to evaluate the antiseptic efficacy of biomechanical preparation and a calcium hydroxide-paste in root canals of human primary teeth with necrotic pulp after trauma and to detect the microorganisms Fusobacterium nucleatum and black-pigmented bacilli in this teeth. According to stringent inclusion criteria, 18 primary teeth with necrotic pulp were selected. Bacterial samples were taken after crown access (S1) and 72h after the removal of dressing with a calcium hydroxide paste (S3), but to 10 teeth were taken a other bacterial sample after chemomechanical preparation with 0.5% NaOCl as an irrigant (S2). Bacteriological samples were collected by introducing 3 sequential sterile absorbent paper points, of a size visually compatible with the root canal diameter. After approximately 1 min, the paper points were removed and placed in a test tube containing reduced transport fluid and were sent for microbiological evaluation. In the S1, the microorganisms were found in 7/18 (94,4%) of the samples, with a colony forming units (CFUâs) media of 5.4 x 105. In the S2, bacteria were cultured in only 1/10 (10%) root canal, with the CFUâs media of 4.3 x 102, and in the S3 bacteria were cultured in 15/18 (83.3%), with the CFUâs media of 1.5 x 105. A statistically significant reduction in bacterial counts was observed between S1 and S2, however no statistically significant difference was observed for comparisons involving S1 and S3, and S2 and S3. The microorganisms Fusobacterium nucleatum and black-pigmented bacilli were detected in 55.5% (10/18) and 11.1% (2/18), respectively, in the S1, no were found in the S2, and in the S3 were found in 16.6% (3/18) and 5.5% (1/18), respectively. In the S1, the gram-negative cocci (15/18) and gram-negative rods (14/18) were the most prevalent groups (83.3% and 77.8%, respectively). In the S2, the gram-positive cocci was the only group of the bacteria observed (1/10), and in the S3, the gram-positive cocci was the group most commonly recovered (66.7%). It was conclude that biomechanical preparation were important in the antisepsis of the root canal because reduced significantly the number of bacteria in the main canal however the calcium hydroxide paste had a antibacterial efficacy limited, no prevent the regrowing bacterial after used as dressing intracanal.

As proteínas são importantes componentes da matriz extracelular da polpa dentária e possuem diferentes funções nos tecidos. A literatura odontológica não relata como os proteoglicanos se distribuem e agem na matriz extracelular de dentes decíduos durante o processo fisiológico de rizólise. Foi objetivo do trabalho analisar as expressões dos proteoglicanos biglican e decorin e relacioná-las com as diversas fases do processo de rizólise. Para isso foram utilizados dentes decíduos humanos extraídos e livres de lesões de cárie, apresentando vários estágios de reabsorção radicular, divididos em três grupos: com dois terços ou mais do comprimento radicular médio, um terço ou mais do comprimento radicular médio e menos de um terço do comprimento radicular médio. Foi utilizada a técnica da imunoistoquímica, com o método da estreptavidina-biotina-peroxidase, e anticorpos contra as proteínas anteriormente citadas. Os resultados mostraram que os proteoglicanos estudados apresentaram imunorreatividade na matriz extracelular da polpa e da dentina dos dentes nas três fases de reabsorção. Foi possível concluir que houve diferença na distribuição e no padrão de expressão dos proteoglicanos biglican e decorin apenas na área de reabsorção, nos dentes decíduos hígidos nas três fases de rizólise o que sugere um papel regulador destes proteoglicanos no processo de reabsorção fisiológica nos dentes decíduos hígidos. / Proteins are important components in pulp extracellular matrix and perform several different roles in the body tissues. The distribution and functions of some proteins have already been described but there aren?t studies showing how the proteoglycans are distributed and act in the extracellular matrix of deciduous teeth at the physiological resorption process. The aim of the present work was to study the expression and distribution of the following non-collagenous components of the extracellular matrix: biglycan and decorin in deciduous teeth dental tissues at the physiological root resorption. Hygid human deciduous teeth that were extracted for orthodontic reasons were grouped together according to root length: group I - two thirds or more of average length root, group II - one third or more of average length root and group III ? less than one third of average length root. The streptavidin-biotinperoxidase method of immunohistochemistry was used with antibodies against the antigens previously quoted. The results showed that the proteoglycans had been found in pulp and dentin extracellular matrix in all groups. In conclusion, biglycan and decorin were differentially found only in the resorption area, between tissues in resorption process and its adjacent zone, which make us believe that these proteoglycans act regulating the physiological resorption process in hygid deciduous teeth.

O objetivos deste estudo foi avaliar a influência da radiação ionizante na vitalidade do tecido pulpar mensurada através dos níveis de saturação de oxigênio (%SpO2) em pacientes com tumores malígnos intraoral ou de orofaringe, passado de 4-6 anos da radioterapia (RT). Noventa pacientes com tumores malígnos intraoral ou de orofaringe, submetidos de 4-6 anos anteriores a RT foram selecionados para este estudo. Os níveis de oxigenação e sensibilidade pulpar, avaliados através do oxímetro de pulso e do spray refrigerante TFE (tetrafluoretano), foram analisados nos dentes anteriores (superior e inferior) de cada paciente selecionado (n=693),, indiferente do quadrante e da área irradiada. Como grupo controle foram selecionados noventa pacientes saudáveis (nunca submetidos a RT) e os mesmos testes foram empregados (n=693). Todos os dentes foram considerados vitais. 100% mostraram resposta positva ao teste térmico e as médias de %SpO2 foram de 92.7% no grupo dos pacientes irradiados (SD ± 1.83) e 92.6% no grupo dos não-irradiados (SD ± 1.80), sem diferença estatística observada. Houve uma tendência de valores de %SpO2 menores em dentes caninos comparados aos incisivos, entretanto sem diferença estatística significante. Passados de 4-6 anos da RT, as %SpO2 da polpa dental estão dentro dos padrões considerados normais para uma polpa vital e podese assumir que a RT não têm influência danosa a longo prazo sobre a vitalidade do tecido pulpar, assim sugerindo que este tecido pode ser apto a retornar o fluxo sanguíneo normal após a RT. As mudanças observadas na microcirculação pulpar devido a RT parecem ser temporárias, então o tratamento endodôntico preventivo ou a extração dental em pacientes que receberam radiação ionizante são desnecessários. / The aim of this study was to evaluate the influence of radioation on pulp vitality through the measurement of pulpal oxigenation levels (%SpO2) in patients with malignant intraoral and oropharyngeal tumors at 4-6 years after radiotherapy (RT). Ninety patients with malignant tumors in the oral cavity or oropharynx, submitted to RT 4-6 years prior to the study were selected. Pulp oxygenation levels and pulp sensitivity, measured by pulse oximetry and by cold refrigerant spray TFE (tetrafluoroethane), were analyzed in the anterior teeth (upper and lower) of each patient selected (n=693), regardless of the quadrant and the irradiated area. As a control group were selected ninety healthy patients (never submitted to RT) and the same tests were performed (n=693). All teeth were considered vital. 100% showed a positive response to the thermal test and the %SpO2 rates were 92.7% in irradiated group (SD ± 1.83) and 92.6% in non-irradiated group (SD ± 1.80), without statistical difference. There was a trend for lower %SpO2 values in canine teeth compared to incisors, however it was not statistically significant. After 4-6 years of RT the dental pulp %SpO2 are within the normal range considered for a vital pulp and it can be assumed that RT did not have a long term influence on the pulp vitality, therefore suggesting that the pulp tissue may be able to recover normal blood flow after RT. The changes observed in the pulp microcirculation due to RT seems to be temporary, so preventive endodontic treatment or tooth extraction in patients who will receive RT may not be necessary.

Thesis (M.S.)--University of Alabama at Birmingham, 2008. / Title from PDF title page (viewed on Feb. 3, 2010). Includes bibliographical references (p. 38-42).

Thesis (M.D.S.)--University of Hong Kong, 2001. / Includes bibliographical references.

A major challenge facing the tissue engineering discipline is cartilage tissue repair and engineering, because of the highly specialized structure and limited repair capacity that cartilage possesses. Dental pulp stem cells (DPSCs) were identified about a decade ago as a potential candidate for cell based therapy and tissue engineering applications. The present study aimed to utilize gene therapy with isolated DPSCs to induce chondrogenic transgene expression and chondrogenic lineage differentiation, with the ultimate goal of engineering cartilage tissue-like constructs. We isolated DPSCs from human teeth extracted for orthodontic treatment. We further enriched the isolated population using immunomagnetic bead selection, which increased stem cell markers: Stro-1 and CD146, compared to unselected population. The DPSCs showed the ability to differentiate into the chondrogenic lineage when induced with recombinant hTGFβ-3 and when transduced with hTGFβ-3 transgene. We successfully constructed the recombinant adeno-associated viral vector encoding the human TGFβ-3, and determined the best multiplicity of infection for DPSCs. The transduced DPSCs highly expressed hTGFβ-3 for up to 60 days. Expression of chondrogenic markers; Collagen IIa1, Sox9, and aggrecan was verified by immunohistochemistry and mRNA. We successfully fabricated an electrospun nano-fiber scaffold upon which morphology, proliferation and viability of the DPSCs were examined. DPSCs attached and proliferated on nano-fiber scaffolds demonstrating better viability compared to micro-fiber scaffolds. Transduced cells expressed hTGFβ-3 protein up to 48 days. Cells seeded on nanofiber scaffolds showed higher expression levels compared to micro-fiber scaffolds or culture plate. Scaffolds seeded with DPSCs were implanted in nude mice. Immunohistochemistry for TGFβ-3 DPSCs constructs (n=5/group) showed cartilage-like matrix formation with glucoseaminoglycans as shown by Alcian blue. Immunostaining showed positivity for Collagen IIa1, Sox9 and aggrecan. Semi-thin sections of the transduced DPSCs constructs examined by transmission electron microscopy (TEM) showed chondrocytic cellular and intra-cellular features, as well as extracellular matrix formation (n=2/group). In vivo constructs with the TGFβ-3 DPSCs showed higher collagen type II and Sox9 mRNA expression relative to non-transduced DPSCs constructs (n=5/group). Western blot analysis confirmed this expression pattern on the protein level (n=3/group). Engineered constructs mechanical properties were examined and compared to patellar bovine cartilage to assess functionality (n=5/group). TGFβ-3 transduced DPSCs constructs showed a higher equilibrium elastic modulus compared to nontransduced constructs. Micro-fiber scaffolds constructs showed a higher elastic modulus (0.11 MPa, 18% of bovine cartilage), compared to nano-fiber constructs modulus (0.032 MPa, 6% of bovine cartilage). Nano-fiber based constructs showed a similar Poisson‘s ration to bovine cartilage, while that of micro-fiber scaffolds was lower. As an alternative gene delivery method, electroporation parameters for DPSCs transfection were optimized, and compared to commonly used chemical transfection methods. TGFβ-3 transfected DPSCs showed a significantly higher relative TGFβ-3 mRNA and protein expression compared to non transfected control and to eGFP transfected DPSCs. Transfected DPSCs showed increased relative expression of chondrogenic markers; Collagen II, Sox9 and aggrecan, compared to non transfected DPSCs. Successful chondrogenic differentiation of DPSCs gene therapy with TGFβ-3 transgene, and seeding them on PLLA/PGA scaffolds makes it a potential candidate for cartilage tissue engineering and cell based therapy. / published_or_final_version / Dentistry / Doctoral / Doctor of Philosophy