GTPases RhoA and Rac1 are important for amelogenin and DSPP expression during differentiation of ameloblasts and odontoblasts

Morphogenesis and cytodifferentiation are distinct processes in tooth development. Cell proliferation predominates in morphogenesis; differentiation involves changes in form and gene expression. The cytoskeleton is essential for both processes, being regulated by Rho GTPases. The aim of this study was to verify the expression, distribution, and role of Rho GTPases in ameloblasts and odontoblasts during tooth development in correlation with actin and tubulin arrangements and amelogenin and dentin sialophosphoprotein (DSPP) expression. RhoA, Rac1, and Cdc42 were strongly expressed during morphogenesis; during cytodifferentiation, RhoA was present in ameloblasts and odontoblasts, Rac1 and its effector Pak3 were observed in ameloblasts; and Cdc42 was present in all cells of the tooth germ and mesenchyme. The expression of RhoA mRNA and its effectors RockI and RockII, Rac1 and Pak3, as analyzed by real-time polymerase chain reaction, increased after ameloblast and odontoblast differentiation, according to the mRNA expression of amelogenin and DSPP. The inhibition of all Rho GTPases by Clostridium difficile toxin A completely abolished amelogenin and DSPP expression in tooth germs cultured in anterior eye chamber, whereas the specific inhibition of the Rocks showed only a partial effect. Thus, both GTPases are important during tooth morphogenesis. During cytodifferentiation, Rho proteins are essential for the complete differentiation of ameloblasts and odontoblasts by regulating the expression of amelogenin and DSPP. RhoA and its effector RockI contribute to this role. A specific function for Rac1 in ameloblasts remains to be elucidated; its punctate distribution indicates its possible role in exocytosis/endocytosis.

Prevalence and microbiological diversity of Archaea in peri-implantitis subjects by 16S ribosomal RNA clonal analysis

Background and Objective: This study evaluated the prevalence and the molecular diversity of Archaea in the subgingival biofilm samples of subjects with peri-implantitis. Material and Methods: Fifty subjects were assigned into two groups: Control (n = 25), consisting of subjects with healthy implants; and Test (n = 25), consisting of subjects with peri-implantitis sites, as well as a healthy implant. In the Test group, subgingival biofilm samples were taken from the deepest sites of the diseased implant. In both groups, subgingival biofilm was collected from one site with a healthy implant and from one site with a periodontally healthy tooth. DNA was extracted and the 16S ribosomal RNA gene was amplified with universal primer pairs for Archaea. Amplified genes were cloned and sequenced, and the phylotypes were identified by comparison with known 16S ribosomal RNA sequences. Results: In the Control group, Archaea were detected in two and three sites of the implant and the tooth, respectively. In the Test group, Archaea were detected in 12, 4 and 2 sites of diseased implants, healthy implants and teeth, respectively. Diseased implants presented a significantly higher prevalence of Archaea in comparison with healthy implants and natural teeth, irrespective of group. Over 90% of the clone libraries were formed by Methanobrevibacter oralis, which was detected in both groups. Methanobacterium congelense/curvum was detected in four subjects from the Test group and in two subjects from the Control group. Conclusion: Although M. oralis was the main species of Archaea associated with both healthy and diseased implant sites, the data indicated an increased prevalence of Archaea in peri-implantitis sites, and their role in pathogenesis should be further investigated.

Induction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strain

The oral pathogen Streptococcus mutans expresses a surface protein, P1, which interacts with the salivary pellicle on the tooth surface or with fluid-phase saliva, resulting in bacterial adhesion or aggregation, respectively. P1 is a target of protective immunity. Its N-terminal region has been associated with adhesion and aggregation functions and contains epitopes recognized by efficacious antibodies. In this study, we used Bacillus subtilis, a gram-positive expression host, to produce a recombinant N-terminal polypeptide of P1 (P1(39-512)) derived from the S. mutans strain UA159. Purified P1(39-512) reacted with an anti-full-length P1 antiserum as well as one raised against intact S. mutans cells, indicating preserved antigenicity. Immunization of mice with soluble and heat-denatured P1(39-512) induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P1(39-512) antiserum was as effective at blocking saliva-mediated aggregation of S. mutans cells and better at blocking bacterial adhesion to saliva-coated plastic surfaces compared with the anti-full-length P1 antiserum. In addition, adsorption of the anti-P1 antiserum with P1(39-512) eliminated its ability to block the adhesion of S. mutans cells to abiotic surfaces. The present results indicate that P1(39-512), expressed and purified from a recombinant B. subtilis strain, maintains important immunological features of the native protein and represents an additional tool for the development of anticaries vaccines.

AFM analysis of bleaching effects on dental enamel microtopography

The purpose of this in vitro study was to test a new methodology to evaluate the effects of 35% hydrogen peroxide agent on the microtopography of sound enamel using an atomic force microscope (AFM). The buccal sound surfaces of three extracted human lower incisors were used, without polishing the surfaces to maintain them with natural morphology. These unpolished surfaces were subjected to bleaching procedure with 35% hydrogen peroxide that consisted of 4 applications of the bleaching agent on enamel surfaces for 10 min each application. Surface images were obtained in a 15 mu m x 15 mu m area using an AFM. The roughness (Ra and RMS) and the power spectral density (PSD) were obtained before and after the bleaching treatment. As results we could inquire that the PSD analyses were very suitable to identifying the morphological changes on the surfaces, while the Ra and RMS parameters were insufficient to represent the morphological alterations promoted by bleaching procedure on enamel. The morphological wavelength in the range of visible light spectrum(380-750 nm) was analyzed, showing a considerable increase of the PSD with the bleaching treatment. (C) 2009 Elsevier B. V. All rights reserved.

Study of environmental burden of lead in children using teeth as bioindicator

The south region of Sao Paulo city hosts the Guarapiranga dam, responsible for water supply to 25% of the city population. Their surroundings have been subject to intense and irregular occupation by people from very low socioeconomics classes. Measurements undertaken on sediment and particulate materials in the dam revealed concentrations of lead. copper, zinc and cadmium above internationally accepted limits. Epidemiological and toxicological studies undertaken by the World Health Organization in individuals exhibiting lead concentrations in blood, near or below the maximum recommended (10 mu g dl(-1)), surprisingly revealed that toxic effects are more intense in individuals belonging to low socioeconomics classes. Motivated by these facts, we aimed at the investigation of chronic incorporation of lead. as well as the use of our BIOKINETICS code, which is based on an accepted ICRP biokinetics model for lead, in order to extrapolate the results from teeth to other organs. The focus of our data taking was children from poor families, living in a small, restrict and allegedly contaminated area in Sao Paulo city. Thus, a total of 74 human teeth were collected. The average concentration of lead in teeth of children 5 to 10 years old was determined by means of a high-resolution inductively coupled plasma mass spectrometer (ICP-MS). For standardization of the measurements, an animal bone certified material (H-Animal Bone), from the International Atomic Energy Agency, was analyzed. The amount of lead in children living in the surroundings of the dam, was approximately 40% higher than those from the control region, and the average lead concentration was equal to 1.3 mu g g(-1) approximately. Grouping the results in terms of gender, tooth type and condition, it was concluded that a carious molar of boys is a much more efficient contamination pathway for lead, resulting in concentrations 70% higher than in the control region. We also inferred the average concentrations of lead in other organs of these children, by making use of our BIOKINETIC code. (C) 2008 Elsevier Ltd. All rights reserved.

Changes on degree of conversion of dual-cure luting light-cured with blue LED

The indirect adhesive procedures constitute recently a substantial portion of contemporary esthetic restorative treatments. The resin cements have been used to bond tooth substrate and restorative materials. Due to recently introduction of the self-bonding resin luting cement based on a new monomer, filler and initiation technology has become important to study the degree of conversion of these new materials. In the present work the polymerization reaction and the filler content of dual-cured dental resin cements were studied by means of infra-red spectroscopy (FT-IR) and thermogravimetry (TG). Twenty specimens were made in a metallic mold (8 mm diameter x 1 mm thick) from each of 2 cements, PanaviaA (R) F2.0 (Kuraray) and RelyX (TM) Unicem Applicap (3M/ESPE). Each specimen was cured with blue LED with power density of 500 mW/cm(2) for 30 s. Immediately after curing, 24 and 48 h, and 7 days DC was determined. For each time interval 5 specimens were pulverized, pressed with KBr and analyzed with FT-IR. The TG measurements were performed in Netzsch TG 209 under oxygen atmosphere and heating rate of 10A degrees C/min from 25 to 700A degrees C. A two-way ANOVA showed DC (%) mean values statistically significance differences between two cements (p < 0.05). The Tukey`s test showed no significant difference only for the 24 and 48 h after light irradiation for both resin cements (p > 0.05). The Relx-Y (TM) Unicem mean values were significantly higher than PanaviaA (R) F 2.0. The degree of conversion means values increasing with the storage time and the filler content showed similar for both resin cements.

Dentin hypersensitivity clinical study comparing LILT and LEDT keeping the same irradiation parameters

Dentin hypersensitivity is a common condition associated with high dental pain. A new LED-based (light emitting diode) light source has been used as an experimental tool in some studies. Purpose: The main objective was to compare these two light sources emitting in the same spectral band (red from 625 to 660 nm) to promote pain relief. Material and methods: A total of 6 sessions were accomplished, being three irradiation sessions and three follow-up sessions. This single-blind study compared a control group (Placebo) and two other groups with different equipments: low laser intensity treatment (LILT) and a light emitting diode system treatment (LEDT). Results: The results showed that there is no statistical difference between LILT and LEDT groups, however, both were better than control group (p=0.01) in terms of treatment efficiency; there is no difference between the second and the third sessions for both treatment, it means that the third session was not necessary; finally, the improvement at the end of the entire research (follow up care of 30 days) was very expressive in comparison to pre-treatment situation for all teeth (p=0.01). Conclusion: LILT and LEDT were equally effective to treat dentine hypersensitivity, a 3rd treatment session was not necessary/two sessions are enough.

Chondrichthyans from the base of the Irati Formation (Early Permian, Parana Basin), Sao Paulo, Brazil

Three species of chondrichthyans are reported from conglomeratic sandstone at the base of the Lower Permian (Artinskian) Irati Formation (Parana Basin) near Rio Claro, Sao Paulo State, Brazil. The fossils include: 1) dispersed teeth of the petalodont ltapyrodus punctatus Silva Santos 1990, first described from the Permian (Artinskian) Pedra do Fogo Formation, Parnaiba Basin, Northeast Brazil; 2) a new species of tooth of the Order Orodontiformes; and 3) a new species of finspine of the Order Ctenacanthiformes. These fossils occur in an allochthonous assemblage of vertebrate remains including other Chondrichthyes, Xenacanthiformes and cladodonts, paleonisciform bony fish, and tetrapods. This discovery is a significant contribution to the sparse record of South American Chondrichthyes from the Early Permian and raises questions regarding the paleoenvironmental adaptations among these fish within Paleozoic basins of Brazil at this time. (C) 2010 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Placas maxilo-dentárias em Rincossauros Hyperodapedon Huxley, 1859. Histologia e morfogênese maxilo-dental: nova abordagem.

O estudo das estruturas da placas maxilo-dentárias dos rincossauros Hyperodapedon Huxley, 1859, do Triássico da Formação Santa Maria, em nova abordagem histológica e ontogênica resultou na identificação da natureza do esmalte aprismático verdadeiro, de variados elementos histológicos dentinários e osteológicos, e de um centro de ossificação periosteal primário. Também foram encontradas evidências histológicas dos mecanismos de fusionamento maxilo-dentinário e amelo-maxilar. Com estes elementos, inferimos os modelos de organogênese dental, da ontogênese maxilar e dos mecanismos de fusionamento maxilodental. Encontrou-se uma singular e raríssima coroa dental, imatura e ainda não erupcionada, na região posterior da placa dental e assim evidenciou-se a correta posição da margem odontogenicamente ativa. Adicionalmente inferiu-se a localização da posição da lâmina dentária embrionária. Constatou-se a não formação de alvéolos dentários, de cemento radicular e do espaço necessário à formação do ligamento periodontal e, assim, se deduziu a não formação do folículo dental embriônico. As presenças de especiais elementos anatômicos e histológicos nos tecidos ósseos periapicais evidenciam o crescimento radicular contínuo, enquanto a forma e o fusionamento radicular imediato depõe a favor de uma função dentária fisiológica diferenciada para as baterias dentárias maxilares dos Rincossauros do gênero Hyperodapedon. Os mecanismos que possibilitaram o controle embriônico para a deposição das lamelas de tecido ósseo coronal e seu preciso fusionamento sobre o esmalte dentário, declinam por modificações nas funções tardias do órgão reduzido do esmalte e pela presença de uma membrana oral com funções osteogênicas e também protetivas, situada nas porções posteriores da placa maxilo-dentária em desenvolvimento. Mudanças heterocrônicas no tempo de diferenciação das células da crista neural embriônica e em seus derivados, como a lâmina dentária e órgãos dentários embrionários ou correlacionadas com a organogênese das placas maxilo-dentárias e seus anexos periodontais, todos como condições plesiomórficas para Diápsidas Triássicos, poderiam ser as causas responsáveis pela origem e evolução deste estranho aparelho estomatognático nos clados de Hyperodapedon sp..

Análise da dentina de dentes decíduos submetidos ao capeamento pulpar indireto

O objetivo do estudo foi avaliar as características da dentina cariada remanescente de molares decíduos quanto à coloração, consistência e microdureza, após um período médio de 3 anos e 8 meses da realização do capeamento pulpar indireto. O estudo clínico constou de 27 dentes que apresentavam lesões cariosas ativas com expressão radiográfica em metade interna de dentina. Aleatoriamente, foi realizado o capeamento pulpar indireto e utilizado como material capeador o hidróxido de cálcio (HC) e o cimento de ionômero de vidro resinoso modificado (CIVRM). As crianças foram acompanhadas através de exames clínicos e radiográficos, e ao longo de um período médio de 3 anos e 8 meses, observou-se um índice de sucesso de 89% para o grupo do HC e de 93% para o CIVRM, não havendo diferença estatisticamente significante entre os dois grupos (p=0,62). Os dentes considerados como sucesso no estudo clínico foram agrupados em um único grupo (grupo teste), que constou de 13 dentes que sofreram esfoliação natural ou exodontia por motivos ortodônticos. Além disso, foram selecionados 15 molares decíduos hígidos (grupo controle positivo) e 15, portadores de lesões cariosas ativas em metade interna de dentina (grupo controle negativo). As amostras do grupo teste tiveram suas respectivas restaurações removidas, a profundidade medida e a dentina remanescente avaliada por um operador calibrado, seguindo critérios descritivos, quanto à consistência e a coloração. Nos dentes do grupo controle positivo, foram realizados preparos cavitários oclusais em até 4 mm de profundidade, enquanto que, no grupo controle negativo, o mesmo operador do estudo clínico realizou a remoção parcial de tecido cariado in vitro. A partir daí, todos os dentes foram preparados para análise de microdureza, que foi realizada por um examinador calibrado, que empregou o princípio de cegamento. Na análise da consistência, todos os dentes do grupo teste (n=13) apresentaram-se endurecidos, enquanto que 9, apresentaram coloração amarela-clara (8 do CIVRM e 1 do HC) e 4, castanho-escura (1 do CIVRM e 3 do HC). Para o teste de microdureza no grupo teste, obteve-se uma média KHN de 40,81 (±16,28) MPa, enquanto que nos grupos controles positivo e negativo, foram alcançados valores médios de 62,73 (±11,24) MPa e 19,15 (±6,99) MPa, respectivamente. A análise estatística mediante o teste ANOVA indicou que houve diferença significativa entre os 3 grupos. Foi constatada a remineralização da dentina de dentes decíduos em que foi realizada a técnica do capeamento pulpar indireto após um período médio de 3 anos e 8 meses, através de critérios clínico (consistência) e laboratorial (análise da microdureza).

Avaliação da saúde bucal e anomalias dentárias em pacientes tratados com leucemia na infância e adolescência

Objetivo: o objetivo deste estudo foi avaliar as condições de saúde bucal e a ocorrência de anomalias dentárias em crianças tratadas para leucemia linfoblástica aguda (LLA) no Serviço de Oncologia Pediátrica (SOP) do Hospital de Clínicas de Porto Alegre (HCPA), comparadas com um grupo crianças saudáveis. Amostra: foram selecionadas 56 crianças com diagnóstico de LLA e analisadas as presenças de anomalias dentárias e os índices CPO-D (cariado, perdido, obturado-dente), IPV (índice de placa visível), ISG (índice de sangramento gengival) e fluxo salivar. As crianças tratadas apresentaram uma média de idade, na época do diagnóstico da LLA, de 5,3 ± 2,6, e 11,8 ± 4,2 na avaliação, sendo 32 masculinos e 24 femininos. Os pacientes tratados para LLA foram divididos em três grupos: crianças tratadas somente com quimioterapia, com quimio e radioterapia, e com quimio, radio e transplante de medula óssea. Resultados: os resultados revelaram 80,4% de anomalias dentárias nas crianças tratadas, ou seja, 45 destas apresentaram pelo menos uma alteração, e o grupo de pacientes tratados com quimio, radio e submetidos ao transplante de medula óssea foi o que revelou a maior média de anormalidades dentárias por indivíduo (15,37 ± 15,03), não ocorrendo diferença estatística entre os gêneros. As crianças tratadas para LLA obtiveram CPO-D de 1,9 ± 4,0, ISG de 26,5%, IPV de 72,0%, e índice de fluxo salivar médio de 0,19 mL/min criança. Já o grupo de crianças sadias apresentou CPO-D de 1,52 ± 3,5, ISG de 11,1%, IPV de 53,8% e índice de fluxo salivar médio de 0,27 mL/min. Conclusão: o tratamento para a cura da LLA provoca um aumento significativo no número de anomalias dentárias, sendo mais freqüente nos pacientes menores de cinco anos de idade, principalmente nos pacientes submetidos a quimio e radioterapia associadas ao transplante de medula óssea, o que requer, por parte do cirurgião-dentista, intervenções clínicas diferenciadas e cuidadosas nestes pacientes, tendo-se em vista também os índices aumentados de ISG e IPV. As alterações sofridas pelas glândulas salivares durante o tratamento não são permanentes sob o aspecto do fluxo salivar, que retorna à normalidade. A orientação e o acompanhamento adequados destes pacientes por parte da equipe de saúde bucal podem mantê-los com o índice de CPO-D dentro dos padrões preconizados pela Organização Mundial de Saúde.

Periodontal diseases in a representative urban population in south Brazil

Background and Aims: There is little information about the epidemiology and risk factors of periodontal diseases in Latin America in general, and Brazil in particular. The principal aims of this study were to: 1) describe the prevalence and severity of periodontal attachment loss and gingival recession, and to assess the contribution of demographic, behavioral, and environmental exposures to the occurrence of periodontal disease outcomes in a sample representative of the urban population in the state of Rio Grande do Sul in south Brazil; and 2) report the epidemiology and risk indicators of aggressive periodontitis in this population. Methods: A representative sample consisting of 1,586 subjects 14-103 years of age (mean 38 y) and comprising 45.3% males and 54.7% females was selected using a multi-stage, probability, cluster sampling strategy. The subjects were interviewed using a structured questionnaire and underwent a full-mouth, six sites per tooth clinical examination in a mobile examination center. Results: Moderate and severe clinical attachment loss and gingival recession were widespread among adults in this population. The prevalence and extent of attachment loss ³5 and ³7 mm were 79% and 52% subjects, and 36% and 16% teeth; and for gingival recession ³3 mm and ³5 mm were 52% and 22% subjects, and 17% and 6% teeth, respectively. Aggressive periodontitis was diagnosed in 5.5% of subjects, which is significantly higher than the reported prevalence in most other populations. Among the main risk indicators for chronic as well as aggressive destructive periodontal diseases were: older age, low socioeconomic status, dental calculus, and smoking. Cigarette smoking accounted for an important part of periodontal disease burden, particularly in adults, and should be considered an important target in any prevention strategy aimed at reducing the burden of periodontal diseases. Partial recording methods consistently underestimated the prevalence of attachment loss in the population, and the extent of underestimation was dependent on the type of system used and the threshold of attachment loss. Conclusions: Destructive periodontal diseases are prevalent in this Brazilian population. Suitable disease prevention and health promotion programs should be established to improve the periodontal health in this population.

Technologies that motivates healthy toothbrushing: practices through social translucence

Fifty percent of the European Union’s population suffers from an oral disease. Studies have repeatedly shown that while acquiring healthy toothbrushing practices early on in one’s life is of significance, children and adults often fail to adhere to those. In this thesis we attempt to design and prototype interactive technologies that motivate healthy tooth brushing habits on individuals. Rather than focusing on the technologies’ persuasive power over individuals, we tap on the social mechanisms employed by families. In this sense, we think of these technologies as social translucent rather than persuasive, whose goal is to raise awareness within the family on each other’s habits and that aim at leveraging families’ existing social mechanisms for behavior change, rather than replacing them. More specifically, we aim to gain insights with respect to the following questions: a) What are the drivers and barriers towards adhering to healthy tooth brushing behaviors? b) Can we effectively measure toothbrushing behaviors? c) How can technologies leverage family communication practices in motivating proper toothbrushing behaviors? First, we present two studies about children and adults’ tooth brushing behaviors and how these are influenced by social interactions within the family. Secondly, we present the design and prototyping of two systems that sense toothbrushing practices and provide feedback, using the Social Translucence Framework as a design lens. We conclude with an overview of lessons learnt from the prototyping of these systems supported by an analysis of the strengths and pitfalls of the developed technologies.

Incidência de cárie dentária em adolescentes em município do Nordeste brasileiro, 2006

NORO, Luiz Roberto Augusto et al. Incidência de cárie dentária em adolescentes em município do Nordeste brasileiro, 2006, Cadernos de Saúde Pública, Rio de Janeiro, v. 25, n. 4, p. 783-790, abr. 2009.

Estudo de sistemas vítreos para infiltração em alumina

The system in-Ceram Alumina, produced by VITA, consists in a technique of prepare of a substructure of ceramics to dental crowns. First burning is made in the alumina decanted by slip casting process under a stone die that reproduces the tooth prepared to receive a crown. In a second burning, alumina is infiltrated by vitreous system, giving to this set a high mechanic resistance. In this work, it s made a study of the composition of a new infiltrating material more used nowadays, giving to alumina desirable mechanics proprieties to its using like substructure of support to ceramic s crown used in the market today. The addition of Lanthanum oxide (frit A) and calcium oxide (frit B) was made in attempt to increase the viscosity of LZSA and to reduce fusion temperature. The frits were put over samples of alumina and took to the tubular oven to 1400ºC under vacuum for two groups (groups 1 and 2). For another two groups (groups 3 and 4) it was made a second infiltration, following the same parameters of the first. A fifth group was utilized like group of control where the samples of pure alumina were not submitted to any infiltrating process. Glasses manifested efficient both in quality and results of analysis of mechanic resistance, being perfectly compatible with oral environment in this technical requisite. The groups that made a second infiltration had he best results of fracture toughness, qualify the use in the oral cavity in this technical question. The average of results achieved for mechanic resistance to groups 1, 2, 3, 4 and 5 were respectively 98 MPa, 90 MPa, 144 MPa, 236 MPa and 23 MPa