Regeneração óssea guiada em implantodontia: relato de caso

Objective: The use of methods for tissue regeneration has been widely applied in Implantology, in clinical situations with disabilities or anatomical limitations that prevent the placement of osseointegrated dental implants. The evolution of the development of biomaterials revolutionized this therapeutic modality, facilitating the resolution of clinical cases with tissue deficiencies. Thus, this study aimed to describe a clinical case approaching the methods, techniques, and materials used in guided bone regeneration applied to Implantology. Case report: A clinical case of a patient who received a Morse taper dental implant (region 15) is described. The use of biomaterial and membrane on the buccal wall of the socket was required. After the osseointegration period, a reopening surgery was performed, and an immediate provisional implant was produced. After 2 months of follow-up, the final prosthesis was made involving other adjacent elements. Final considerations: The guided bone regeneration technique employed showed satisfactory performance. The patient was positive regarding esthetics and function. However, more controlled studies with longer follow-up period are needed for analyses of predictability

Movimentação dentária e aplicabilidade na clínica ortodôntica

A Ortodontia apresenta como um de seus pilares de sustentação as reações biológicas provenientes da indução mecânica. Desde seu início, vários estudos foram realizados para esclarecer os eventos biológicos decorrentes da movimentação dentária sob indução mecânica. Entretanto, mesmo sabendo-se que entre os procedimentos mecânicos e os fenômenos biológicos que acontecem no cotidiano ortodôntico não deve ocorrer disparidade, frequentemente observa-se uma preponderância para a busca do tipo de aparelhagem e o emprego de forças controladas, principalmente pelos fios. Esses aspectos deixam transparecer, principalmente para o profissional menos experiente, que somente o controle mecânico rege a movimentação dentária. Um exemplo que enfatiza exclusivamente o procedimento mecânico nos cursos de Ortodontia é o typodont. Convém salientar que essa metodologia de ensino é válida para simular a movimentação dentária no aspecto técnico-mecânico, porém, o conhecimento biológico da movimentação deve ter o mesmo nível de importância e não pode haver dicotomia; caso contrário, o resultado da movimentação ortodôntica pode causar prejuízos nas estruturas radiculares, óssea e periodontais, bem como comprometer as diversas estruturas em longo prazo.

A colocação de implantes em medicina dentária

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Efeito da geometria do implante dentário e do protocolo de fresagem óssea na estabilidade primária e na osseointegração inicial: estudo experimental em cães

CHAPTER 1 - This study histologically evaluated two implant designs: a classic thread design versus another specifically designed for healing chamber formation placed with two drilling protocols. Forty dental implants (4.1 mm diameter) with two different macrogeometries were inserted in the tibia of 10 Beagle dogs, and maximum insertion torque was recorded. Drilling techniques were: until 3.75 mm (regular-group); and until 4.0 mm diameter (overdrillinggroup) for both implant designs. At 2 and 4 weeks, samples were retrieved and processed for histomorphometric analysis. For torque and BIC (bone-to-implant contact) and BAFO (bone area fraction occupied), a general-linear model was employed including instrumentation technique and time in vivo as independent. The insertion torque recorded for each implant design and drilling group significantly decreased as a function of increasing drilling diameter for both implant designs (p<0.001). No significant differences were detected between implant designs for each drilling technique (p>0.18). A significant increase in BIC was observed from 2 to 4 weeks for both implants placed with the overdrilling technique (p<0.03) only, but not for those placed in the 3.75 mm drilling sites (p>0.32). Despite the differences between implant designs and drilling technique an intramembranous-like healing mode with newly formed woven bone prevailed. CHAPTER 2 - The objective of this preliminary histologic study was to determine whether the alteration of drilling protocols (oversized, intermediate, undersized drilling) present different biologic responses at early healing periods of 2 weeks in vivo in a beagle dog model. Ten beagle dogs were acquired and subjected to surgeries in the tibia 2 weeks before euthanasia. During surgery, 3 implants, 4 mm in diameter by 10 mm in length, were placed in bone sites drilled to 3.5 mm, 3.75 mm, and 4.0 mm in final diameter. The insertion and removal torque was recorded for all samples. Statistical significance was set to 95% level of confidence and the number of dogs was considered as the statistical unit for all comparisons. For the torque and BIC and BAFO, a general linear model was employed including instrumentation technique and time in vivo as independent. Overall, the insertion torque increased as a function of drilling diameter from 4.0 mm, to 3.75 mm, to 3.5 mm, with a significant difference in torque levels between all groups (p<0.001). Statistical assessment of BIC and BAFO showed significantly higher values for the 3.75 mm (recommended) drilling group was observed relative to the other two groups (p<0.001). Different drilling dimensions resulted in variations in insertion torque values (primary stability) and different pattern of healing and interfacial remodeling was observed for the different groups. CHAPTER 3 - The present study evaluated the effect of different drilling dimensions (undersized, regular, and oversized) in the insertion and removal torques of dental implants in a beagle dog model. Six beagle dogs were acquired and subjected to bilateral surgeries in the radii 1 and 3 weeks before euthanasia. During surgery, 3 implants, 4 mm in diameter by 10 mm in length, were placed in bone sites drilled to 3.2 mm, 3.5 mm, and 3.8 mm in final diameter. The insertion and removal torque was recorded for all samples. Statistical analysis was performed by paired t tests for repeated measures and by t tests assuming unequal variances (all at the 95% level of significance). Overall, the insertion torque and removal torque levels obtained were inversely proportional to the drilling dimension, with a significant difference detected between the 3.2 mm and 3.5 mm relative to the 3.8 mm groups (P < 0.03). Although insertion torque–removal torque paired observations was statis- tically maintained for the 3.5 mm and 3.8 mm groups, a significant decrease in removal torque values relative to insertion torque levels was observed for the 3.2 mm group. A different pattern of healing and interfacial remodeling was observed for the different groups. Different drilling dimensions resulted in variations in insertion torque values (primary stability) and stability maintenance over the first weeks of bone healing.

Aplicação de células estaminais dentárias na Medicina e na Medicina Dentária

A temática relativa às células estaminais inicia-se na década de 60 com a descoberta da primeira fonte viável destas células: a medula óssea. Diversos estudos permitiram definir a sua função de renovação tecidular e regeneração pós-dano, assim como a sua caraterização num grupo heterogéneo de células indiferenciadas, clonogénicas, definidas pela capacidade de auto-renovação e diferenciação em células maduras. Nos últimos anos, estas células ganharam popularidade face à alternativa terapêutica que representam para muitas doenças, tais como: diabetes, anomalias congénitas, danos do tecido nervoso, Parkinson, Alzheimer e outras alterações degenerativas, exposições pulpares, defeitos periodontais e perda do órgão dentário. Apesar do seu potencial terapêutico, apresentam vários efeitos adversos, especialmente em relação ao seu envolvimento direto (via transformação maligna das MSCs) e indireto (via efeito modulatório das MSCs) no desenvolvimento do cancro. Preconiza-se o seu uso no âmbito da Engenharia Tecidular, introduzindo o processo de regeneração tecidular através da utilização combinada de biomateriais e mediadores biológicos, a fim de proporcionar novas ferramentas para a medicina regenerativa. Mais tarde, tornou-se possível identificar cinco populações de células estaminais de origem dentária (DPSCs, SHEDs, DFPCs, SCAPs e PDLCs) que, para além da sua multipotência e capacidade de diferenciação, constituem fontes acessíveis para recolha. O isolamento destas células constitui ainda uma prática relativamente recente, na qual se torna preponderante isolar células com fenótipo pré-determinado e cultivá-las em meios de cultura adequados. Estudos comprovam que o método de isolamento e as condições de cultura utilizados podem dar origem a diferentes linhas celulares. A conservação é uma prática baseada na convicção de que a medicina regenerativa é o caminho mais promissor para o desenvolvimento da medicina personalizada. Informação adicional relativa à terapia com células estaminais é ainda necessária. Esta utiliza princípios de biomimética altamente desejáveis, pelo que os resultados obtidos têm vindo a despoletar grandes expetativas e a sua implementação na Engenharia Tecidular apresenta-se promissora.

Modelação do risco de perda dentária por doença periodontal

O prognóstico da perda dentária é um dos principais problemas na prática clínica de medicina dentária. Um dos principais fatores prognósticos é a quantidade de suporte ósseo do dente, definido pela área da superfície radicular dentária intraóssea. A estimação desta grandeza tem sido realizada por diferentes metodologias de investigação com resultados heterogéneos. Neste trabalho utilizamos o método da planimetria com microtomografia para calcular a área da superfície radicular (ASR) de uma amostra de cinco dentes segundos pré-molares inferiores obtida da população portuguesa, com o objetivo final de criar um modelo estatístico para estimar a área de superfície radicular intraóssea a partir de indicadores clínicos da perda óssea. Por fim propomos um método para aplicar os resultados na prática. Os dados referentes à área da superfície radicular, comprimento total do dente (CT) e dimensão mésio-distal máxima da coroa (MDeq) serviram para estabelecer as relações estatísticas entre variáveis e definir uma distribuição normal multivariada. Por fim foi criada uma amostra de 37 observações simuladas a partir da distribuição normal multivariada definida e estatisticamente idênticas aos dados da amostra de cinco dentes. Foram ajustados cinco modelos lineares generalizados aos dados simulados. O modelo estatístico foi selecionado segundo os critérios de ajustamento, preditibilidade, potência estatística, acurácia dos parâmetros e da perda de informação, e validado pela análise gráfica de resíduos. Apoiados nos resultados propomos um método em três fases para estimação área de superfície radicular perdida/remanescente. Na primeira fase usamos o modelo estatístico para estimar a área de superfície radicular, na segunda estimamos a proporção (decis) de raiz intraóssea usando uma régua de Schei adaptada e na terceira multiplicamos o valor obtido na primeira fase por um coeficiente que representa a proporção de raiz perdida (ASRp) ou da raiz remanescente (ASRr) para o decil estimado na segunda fase. O ponto forte deste estudo foi a aplicação de metodologia estatística validada para operacionalizar dados clínicos na estimação de suporte ósseo perdido. Como pontos fracos consideramos a aplicação destes resultados apenas aos segundos pré-molares mandibulares e a falta de validação clínica.

Lesões endo-periodontais: diagnóstico, tratamento e prognóstico

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Piezocirurgia no consultório de Medicina Dentária

A presente Tese de Mestrado em Medicina Dentária tem como tema a Piezocirurgia e, sobre ela, pretende-se saber a sua aplicabilidade no consultório do Médico Dentista, nomeadamente na área da Cirurgia oral. A pesquisa bibliográfica foi realizada através de bases de dados, como a Medline, Science Direct e a Pub-Med, entre outras. Limitou-se a pesquisa, temporalmente, entre 2000 e 2016, e nas línguas Portuguesa e Inglesa. Excluiu-se todas as referências a estudos animais e, todas as que, em nada, contribuiríam para o conteúdo especificado. A cirurgia Piezoeléctrica é uma técnica de osteotomia atraumática, quando comparada a técnicas convencionais, e que revoluciona a área da Cirurgia Oral, através do seu corte preciso e altamente selectivo, mas também através do fenómeno de cavitação que a caracteriza, e que garante um campo operatório livre, uma óptima visibilidade, e, consequentemente, uma maior segurança do operador, culminando tudo isto no sucesso do tratamento. A sua maior desvantagem é o tempo de trabalho que requere, contudo este tende a diminuir, e a ser ainda menor que o tempo de trabalho exigido na cirurgia convencional. Inúmeras são as aplicações da Piezocirurgia, mas na área da Medicina Dentária, nomeadamente na Cirurgia Oral, destaca-se o levantamento do seio maxilar, o aumento da crista óssea, a extracção dos terceiros molares impactados e a cirurgia ortognática.

hSSB1 interacts directly with the MRN complex stimulating its recruitment to DNA double-strand breaks and its endo-nuclease activity

hSSB1 is a recently discovered single-stranded DNA binding protein that is essential for efficient repair of DNA double-strand breaks (DSBs) by the homologous recombination pathway. hSSB1 is required for the efficient recruitment of the MRN complex to sites of DSBs and for the efficient initiation of ATM dependent signalling. Here we explore the interplay between hSSB1 and MRN. We demonstrate that hSSB1 binds directly to NBS1, a component of the MRN complex, in a DNA damage independent manner. Consistent with the direct interaction, we observe that hSSB1 greatly stimulates the endo-nuclease activity of the MRN complex, a process that requires the C-terminal tail of hSSB1. Interestingly, analysis of two point mutations in NBS1, associated with Nijmegen breakage syndrome, revealed weaker binding to hSSB1, suggesting a possible disease mechanism.

rac-3-exo-Ammonio-7-anti-carboxytricyclo-[2.2.10.2,6]heptane-3-endo-carboxylate

The racemic title compound, C9H11NO4 . H2O, a tricyclic rearranged aminonorbornane dicarboxylic acid is a conformationally rigid analogue of glutamic acid and exists as an ammonium-carboxylate zwitterion, with the bridghead carboxylic acid group anti-related. In the crystal, intermolecular N-H...O and O-H...O hydrogen-bonding interactions involving the ammonium, carboxylic acid and water donor groups with both water and carboxyl O-atom acceptors give a three-dimensional framework structure.

1H and 13C NMR spectral studies of C-2 substituted isomeric exo- and endo-5-methyl-bicyclo[3.2.1]octane-6,8-diones

A detailed analysis of the 1H and 13C NMR spectra of C-2 aryl and alkyl/desalkyl substituted isomeric exo- and endo-5-methylbicyclo[3.2.1]octane-6,8-diones is presented. The chemical shift of the C-5 angular methyl, the C-2 alkyl/olefinic (C-10)/C-2 methine protons, the aromatic proton shieldings and the characteristic AMX and ABX spectral pattern of the ketomethylene and bridgehead protons were found to be sensitive to the phenyl ring orientation (anisotropy). These distinctive features could be used for configurational distinction for this class of compounds. With increasing ortho-methoxy substitution on the phenyl ring, considerable deshilelding of the bridgehead proton was observed (ca. 0.6 ppm). Absence of the C-2 alkyl group in the desalkyl isomers resulted in substantial changes in the chemical shifts of different protons. A study of the NMR spectra of the corresponding bicyclic compounds with C-2 methoxy/hydroxy substitution instead of the aryl group revealed that the anisotropy of the phenyl ring and the electronegative oxygen substituents have opposite effects. The 13C NMR spectral assignment of each carbon resonance of C-2 aryl and alkyl/desalkyl substituted isomeric exo- and endo-5-methylbicyclo[3.2.1]octane-6,8-diones and the corresponding C-2 methoxy/hydroxy/chloro and methyl bicyclic compounds are reported. Additional ortho-methoxy substitution on the phenyl ring was found to produce considerable high field shifts of the C-10 and C-1 carbon resonances. A high-field shift was observed for the C-6 and C-8 carbonyl carbons, presumably due to 1,3-dicarbonyl interactions. The chemical shifts of C-1 aromatic, C-10 alkyl and C-2 carbons, which are sensitive to exo/endo isomerism, could be utilized in differentiating a pair of isomers.

Computational analyses of the catalytic and heparin-binding sites and their interactions with glycosaminoglycans in glycoside hydrolase family 79 endo-d-glucuronidase (heparanase)

Mammalian heparanase is an endo-β-glucuronidase associated with cell invasion in cancer metastasis, angiogenesis and inflammation. Heparanase cleaves heparan sulfate proteoglycans in the extracellular matrix and basement membrane, releasing heparin/heparan sulfate oligosaccharides of appreciable size. This in turn causes the release of growth factors, which accelerate tumor growth and metastasis. Heparanase has two glycosaminoglycan-binding domains; however, no three-dimensional structure information is available for human heparanase that can provide insights into how the two domains interact to degrade heparin fragments. We have constructed a new homology model of heparanase that takes into account the most recent structural and bioinformatics data available. Heparin analogs and glycosaminoglycan mimetics were computationally docked into the active site with energetically stable ring conformations and their interaction energies were compared. The resulting docked structures were used to propose a model for substrates and conformer selectivity based on the dimensions of the active site. The docking of substrates and inhibitors indicates the existence of a large binding site extending at least two saccharide units beyond the cleavage site (toward the nonreducing end) and at least three saccharides toward the reducing end (toward heparin-binding site 2). The docking of substrates suggests that heparanase recognizes the N-sulfated and O-sulfated glucosamines at subsite +1 and glucuronic acid at the cleavage site, whereas in the absence of 6-O-sulfation in glucosamine, glucuronic acid is docked at subsite +2. These findings will help us to focus on the rational design of heparanase-inhibiting molecules for anticancer drug development by targeting the two heparin/heparan sulfate recognition domains.

Endo-polygalacturonate lyase of Cytophaga johnsonii

An extracellular endopolygalacturonate lyase of Cytophaga johnsonii was purified from the culture filtrate. It appeared to be homogeneous as judged by polyacrylamide gel electrophoresis at pH 8.6 as well as pH 4.3. The purified enzyme had a pH optimum around 9.0 and required Ca++ ions for its maximum activity. The apparent Kmfor polygalacturonic acid was found to be 0.22%. Both paper and column chromatography indicated formation and accumulation of an unsaturated monomer along with unsaturated di-, tri-, tetra- and pentamers from polygalacturonic acid by the enzyme action, indicating that the enzyme cleaved the substrate randomly in a non-hydrolytic manner. The glycosidic linkage next to the non-reducing end of polygalacturonic acid was not resistant to attack by this enzyme unlike in other known polygalacturonate lyases.

Purification and characterization of endo-1,4-β-xylanase from Paecilomyces varioti Bainier

An endo-xylanase (1,4-β-d-xylanxylanohydrolase EC 3.2.1.8) was isolated from the culture filtrate of Paecilomyces varioti Bainier. The enzyme was purified 3.2 fold with a 60% yield by gel filtration and ion exchange chromatography. The purified enzyme had a molecular weight of 25,000 with a sedimentation coefficient of 2.2 S. The isoelectric point of the enzyme was 3.9. The enzyme was obtained in crystalline form. The optimum pH range was 5.5–7.0 and the temperature, 65°C. The Michaelis constant was 2.5 mg larchwood xylan/ml. The enzyme was found to degrade xylan by an endo mechanism producing arabinose, xylobiose, xylo- and arabinosylxylo-oligosaccharides, during the initial stages of hydrolysis. On prolonged incubation, xylotriose, arabinosylxylotriose and xylobiose were the major products with traces of xylotetraose, xylose and arabinose.

A comparison of the DNMR behaviour of methyl 5,6-bis(2-methoxyphenyl)-1,4-dimethyl-7-oxobicyclo[2.2.1]hept-5-en-2-endo-carboxylate and its 7-oxa analogue

Methyl 5,6-Bis(2-methoxyphenyt)-1,4-dimethyl-7-oxobicyclo[2.2.1]hept-5-en-2-endo-carboxylate, a moderately crowded norbornenone ester, exhibits complex VT-DNMR behaviour. A similar behaviour is not seen in its 7-oxa analogue, showing that conformational transmission from position 7 has a crucial influence on the distance parameters that govern the dynamic processes involving the substituents on the bicycloheptene framework.