Yes / The major components of human diet both past and present may be estimated by measuring the carbon and nitrogen isotope ratios (δ13C and δ15N) of the collagenous proteins in bone and tooth dentine. However, the results from these two tissues differ substantially: bone collagen records a multi-year average whilst primary dentine records and retains timebound isotope ratios deriving from the period of tooth development. Recent studies harnessing a sub-annual temporal sampling resolution have shed new light on the individual dietary histories of our ancestors by identifying unexpected radical short-term dietary changes, the duration of breastfeeding and migration where dietary change occurs, and by raising questions regarding factors other than diet that may impact on δ13C and δ15N values. Here we show that the dentine δ13C and δ15N profiles of workhouse inmates dating from the Great Irish Famine of the 19th century not only record the expected dietary change from C3 potatoes to C4 maize, but when used together they also document prolonged nutritional and other physiological stress resulting from insufficient sustenance. In the adults, the influence of the maize-based diet is seen in the δ13C difference between dentine (formed in childhood) and rib (representing an average from the last few years of life). The demonstrated effects of stress on the δ13C and δ15N values will have an impact on the interpretations of diet in past populations even in slow-turnover tissues such as compact bone. This technique also has applicability in the investigation of modern children subject to nutritional distress where hair and nails are unavailable or do not record an adequate period of time. / This study was supported by an Arts and Humanities Research Council grant funding to JB under AHRC Studentship AH/I503307/1.

Yes / This paper presents a case study of a young infant, from a larger isotopic and osteological investigation of Bronze/Iron Age (14th-4th century BC) skeletal assemblages from Croatia and Slovenia. The osteological analysis of this infant identified pathological lesions including abnormal porosity and new bone formation consistent with malnutrition and phases of recovery. The distribution and appearance of these pathological lesions (i.e. diffuse micro-porosities and plaques of subperiosteal new bone formation on the skull and long bones) led to the conclusion that this infant probably suffered from scurvy (vitamin C deficiency). The diet and nitrogen balance of this individual were investigated by incremental dentine sampling and stable carbon and nitrogen isotope analysis. This sampling method provided a high resolution record of dietary and metabolic changes from pre-birth to around the time of death. The resulting isotope data exhibited unusually high δ13C values for this region and time period (between -11.3‰ and -12.6‰), while δ15N values were observed to be c. 3‰ above that of rib collagen sampled from contemporary adults recovered from the same site. The isotope profiles generated from the incremental dentine analysis show that δ13C and especially δ15N continue to increase until death. The evidence from the skeletal remains and high resolution isotopic data support the hypothesis that this infant suffered from severe malnutrition and an increasingly negative nitrogen balance. The paper discusses some scenarios which could have resulted in these unusual isotope ratios, whilst considering the diagnosis of possible metabolic disease. The paper also addresses the need for context when interpreting isotopic results. The isotope data should not be viewed in isolation, but rather as part of a multidisciplinary approach, considering the multiple causes of isotopic variability.

Yes / The carbon (δ13C) and nitrogen (δ15N) isotope ratios of human bone collagen have been used extensively over the last 40 years to investigate the diet of past populations. It has become apparent that bone collagen can give an unreliable temporal dietary signature especially in juveniles. With higher temporal resolution sampling of collagen from tooth dentine, it is possible to identify short‐term changes in diet previously invisible in bone. This paper discusses the inherent problems of using bone collagen for dietary studies and suggests better sample choices which can make our interpretations more robust, using breastfeeding and weaning as an example. / The modern data was collected and analysed using funding from the Rank Prize Funds New Investigator Award and sponsorship from DB Orthodontics, Bradford. The Tooth Fairy team acknowledges the support of the National Institute for Health Research Clinical Research Network (NIHR CRN). / Research Development Fund Publication Prize Award winner, February 2020.

Les méthodes osseuses de diagnose sexuelle donnent d’excellents résultats, avec des taux de bonnes prédictions du sexe supérieurs à 90%. Néanmoins, les éléments osseux ne sont pas toujours disponibles et, lorsqu’ils le sont, selon les causes de la mort et/ou les conditions de conservation du corps, ils peuvent avoir été lourdement dégradés. Pour parvenir à déterminer le sexe d’un individu dans ces situations là, des techniques dentaires de diagnose sexuelle sont nécessaires.Contrairement à la détermination du sexe, la problématique de la détermination de l’âge a fait l’objet de nombreuses études et d’autant de propositions de techniques différentes, signe d’une insuffisance de l’ensemble de ces méthodes, que la littérature n’a pas manqué de mettre en exergue.Le premier objectif de ce travail est de proposer une technique dentaire de détermination du sexe simple et précise, de façon à pouvoir répondre à cette problématique anthropologique de façon fiable en l’absence d’autres éléments du squelette.Le second est de développer une méthode d’estimation de l’âge de mise en œuvre facile, rapide et ne nécessitant ni un plateau technique lourd, ni la détérioration du matériel, pour une application aussi bien anthropologique que médico-légale. L’échantillon d’apprentissage était composé de 210 scanners d’individus présentant chacun 4 canines saines. L’échantillon d’application était composé de 55 scanners d’individus, présentant chacun au moins 1 canine saine. Les volumes pulpaire et total de chaque dent ont été modélisés et calculés à l’aide du logiciel Mimics. Une régression logistique binaire a été utilisée pour déterminer sept modèles de prédiction du sexe, en fonction du nombre et du type de canines exploitables. La comparaison des aires sous la courbe ROC a mis en évidence une plus grande performance du modèle utilisant les volumes des 4 canines. La méthode des moindres carrés pondérés a été utilisée pour déterminer les équations d’estimation de l’âge pour les sept mêmes modèles. Une plus grande performance du modèle utilisant les volumes des 2 canines maxillaires a été mise en évidence. Toutes ces régressions ont été testées sur l’échantillon d’application pour procéder à leur validation externe.Il s’avère que dans un contexte scientifique historique où les canines mandibulaires constituaient l’outil dentaire de référence dans la diagnose sexuelle, la modélisation tridimensionnelle souligne l’intérêt potentiel de leurs homologues maxillaires. Par ailleurs, il semblerait que l’évaluation de la réduction pulpaire physiologique, imputable à l’apposition naturelle de dentine secondaire sur les canines uniquement, soit un critère performant dans la détermination de l’âge.A défaut d’être parfaits, les modèles proposés à l’issue cette étude permettent donc toutefois de disposer d’éléments fiables, qui ont toute leur place dans l’indispensable faisceau de preuves, nécessaire à la détermination de l’âge et du sexe dans le cadre d’une identification estimative. / The bone methods of sexual diagnosis give excellent results, with rates of good predictions over 90%. Nonetheless, bones are not always available and, when they are, according to the causes of death and/or storage conditions of the body, they may have been severely degraded. To be able to determine the sex of an individual in these situations, dental techniques of sexual diagnosis are needed.In contrast to sex determination, the problem of age estimation has been the topic of many studies and so many different techniques, evidence of failure of all these methods, which the literature did not fail to highlight.The first aim of this work was to propose a simple and accurate dental technique for determining sex, in order to resolve this anthropological issue, reliably in the absence of other elements of the skeleton.The second was to develop a method for estimating the age of easy and fast implémentation, neither requiring a heavy technical support nor damage to material, for an application as well in forensics as in anthropology.The training sample consisted of 210 subjects’ CT-scans, each with four canines healthy. The validation sample was composed of 55 CT-scans, each with at least one healthy canine. Pulp and total volumes of each tooth were modeled and calculated using the software Mimics . A binary logistic regression was used to determine seven prediction models of sex, according to the number and type of canine available in practice. The comparison of the areas under the ROC curve showed a greater performance of the model using the volumes of the four canines. The weighted least squares method was used to determine the equations for estimating the age for the same seven models. Greater performance of the model using the volumes of maxillary canines has been demonstrated. All these regressions were tested on the validation sample to perform their external validation.It turns out that in a scientific context where historical mandibular canines were the reference tool in the sex diagnosis, three-dimensional modeling emphasizes the potential value of maxillary counterparts. Moreover, it appears that the evaluation of the physiological pulp reduction, due to the natural apposition of secondary dentine on canines only, is a performance criterion in determining age.The models proposed in this study can therefore have reliable evidence, which all have all their place in the necessary body of evidence for determining age and sex, in estimating identification.

L’activation du système complément, qui se produit à la suite d’une infection ou d’un trauma, génère de puissants signaux moléculaires capables d’initier la réaction inflammatoire. Parmi ces signaux, le fragment actif C5a permet de recruter sur le site lésé les cellules qui expriment son récepteur (le C5aR/CD88). Bien que le C5aR/CD88 soit initialement connu pour être exprimé par les cellules inflammatoires, il est établi que de nombreuses cellules non immunitaires expriment ce récepteur indiquant son implication dans d’autres processus. Nos résultats ont permis de démontrer que l’activation du système du complément au niveau de la pulpe dentaire est réalisée non seulement à partir des protéines plasmatiques mais aussi des protéines synthétisées par les fibroblastes pulpaires. Ainsi, l’activation locale du système du complément, produit à la suite d’une infection, d’un trauma ou de l’application de biomatériaux, génère du C5a qui induit la migration des cellules progénitrices. Ce travail démontre pour la toute première fois l’implication du fragment actif C5a dans le recrutement de progénitrices pulpaires, étape clef au processus de régénération pulpo-Dentinaire. Ces travaux pourraient donc constituer une piste sérieuse dans l’établissement de nouvelles thérapies permettant de cibler les cellules progénitrices au cours du processus de régénération. / After tissue injury or infection, Complement activation provides powerful signals initiating the inflammatory reaction. These events are mediated by biologically active fragments such as C5a which attracts cells expressing its receptor (C5aR/CD88) to the injury site. Besides inflammatory cells as the main C5aR-Expressing cells, various tissue cells have been reported to express this receptor suggesting its involvement in other processes. In order to investigate the possible relationship between complement activation and pulp regeneration, we investigated Complement activation in the dental pulp and progenitor cell migration from their perivascular niches to the pulp injury site to initiate the regeneration process.Our results indicate that complement activation in the dental pulp is the result of both plasma and fibroblast secreted complement proteins. Thus upon local complement activation, which can occur after pathological injury or biomaterials application, C5a induces pulp progenitors’ migration which is critical in initiating the regenerative processes. To our knowledge, this is the first work to demonstrate the involvement of C5a biologically active fragment in the recruitment human pulp progenitor cells. This may provide a useful future therapeutic tool in targeting the progenitor cells in a dentin/pulp regeneration process.

Este estudo teve como objetivos: 01.Avaliar ex vivo o efeito de diferentes sistemas clareadores na microdureza dentinária em dentes bovinos submetidos ao clareamento dental interno. 02.Avaliar ex vivo o efeito de diferentes sistemas clareadores na morfologia superficial da dentina bovina. Fragmentos de 4 x 4mm, contendo esmalte e dentina, foram obtidos de coroas de incisivos bovinos extraídos. Os espécimes foram submetidos ao clareamento dental interno com peróxido de hidrogênio a 35% e peróxido de carbamida a 37% utilizando sistemas convencionais (Opalescence Endo® and Whiteness Super Endo®) e fotoativados (Opalescence Xtra® and Whiteness HP Maxx®). Os controles foram tratados com perborato de sódio misturado com peróxido de hidrogênio a 10% ou nenhum tratamento foi realizado. A microdureza dentinária foi mensurada antes e após os tratamentos clareadores, e os valores de dureza Knoop (KHN) foram submetidos à análise estatística (two-way ANOVA, Tukey\'s port-test). Os espécimes foram observados e fotografados sob microscópio eletrônico de varredura e avaliados com relação às alterações morfológicas da superfície da dentina. Houve uma redução significante na microdureza dentinária para todos os grupos testados quando comparados aos grupos controles. Opalescence Xtra® (-11.36 ± 8.14 KHN), Opalescence Endo® (-13.71 ± 8.02 KHN), Whiteness HP Maxx® (-15.18 ± 9.58 KHN) e Whiteness Super Endo® (-16.97 ± 6.55 KHN) foram semelhantes estatisticamente. O grupo do perborato de sódio misturado com peróxido de hidrogênio 10% (2.10 ± 8.58 KHN) e o grupo sem tratamento clareador (-2.71 ± 2.40 KHN) também foram estatisticamente semelhantes entre si. Ocorreu uma grande variação no padrão de alterações da morfologia superficial da dentina com os sistemas clareadores utilizados. Todos os produtos testados apresentaram redução significativa da microdureza dentinária, exceto o grupo de perborato de sódio misturado com peróxido de hidrogênio a 10%. Ambos, pH e oxidação do peróxido de hidrogênio apresentam o papel de alterar a estrutura da dentina durante o clareamento interno. A utilização de produtos alcalinos com um tempo reduzido de aplicação (técnicas fotoativadas) pode diminuir as alterações morfológicas na dentina. / The aim of this study was: 01. To evaluate in vitro the effect of different in-office bleaching systems on the surface morphology of bovine dentin. 02. To evaluate ex vivo the effect of different bleaching systems on the microhardness of bovine dentine. Tooth fragments measuring 4 x 4mm, containing enamel and dentin, were obtained from the crowns of extracted bovine incisors. Samples were submitted to simulated intracoronal bleaching techniques with 35% hydrogen peroxide and 37% carbamide peroxide using conventional (Opalescence Endo® and Whiteness Super Endo®) and light activated systems (Opalescence Xtra® and Whiteness HP Maxx®). Controls were treated either with sodium perborate mixed with 10% hydrogen peroxide or no bleaching agent. Dentine microhardness values were measured before and after bleaching procedures, recorded as KHN (Knoop Hardness Number), and the differences between them analyzed (two-way ANOVA, Tukey\'s port-test). The samples were observed under SEM and the recorded images were evaluated for topographic alterations. Significant reductions of dentine microhardness were observed for all treatments when compared to the control groups. Opalescence Xtra® (-11.36 ± 8.14 KHN), Opalescence Endo® (-13.71 ± 8.02 KHN), Whiteness HP Maxx® (-15.18 ± 9.58 KHN) and Whiteness Super Endo® (- 16.97 ± 6.55 KHN) presented similar differences. The walking bleach technique (2.10 ± 8.58 KHN) and the untreated groups (-2.71 ± 2.40 KHN) were statistically alike.The ultrastructural alterations of dentin observed in this study varied greatly between groups, according to the products used. Apparently, higher pH products associated to in-office techniques yielded to better maintenance of dentin ultrastructure. The in-office products tested in the present study caused a significant reduction in dentine microhardness. The walking bleach technique did not affect dentine microhardness. Both low pH and hydrogen peroxide oxidation play a role in altering the ultrastructure of dentin during internal dental bleaching. The use of alkaline products with reduced time of application (in-office techniques) may decrease such morphological alterations.

O objetivo do presente estudo foi avaliar in situ o efeito da associação do verniz fluoretado ao laser de Er:YAG e ao laser Nd:YAG na permeabilidade da dentina radicular erodida. Quarenta e oito fragmentos de dentina radicular bovina, com dimensões 2x2x2 mm, foram submetidos a um desafio erosivo inicial com ácido cítrico (0,3%, pH 3,2), por duas horas, sob agitação e armazenados em saliva artificial por vinte e quatro horas. Posteriormente, os espécimes foram divididos em relação aos tratamentos: verniz fluoretado e não fluoretado e subdivididos conforme a irradiação: laser de Er:YAG (100mJ, 3Hz), laser de Nd:YAG (70mJ, 15 Hz) e não irradiado. Após um período de lead in (2 dias), os voluntários (n=8) utilizaram dispositivos palatinos contendo três espécimes que foram submetidos a desafios erosivos ex vivo, quatro vezes ao dia, com ácido cítrico (0,3%, pH 3,2), durante 90s, por cinco dias. Na primeira fase do experimento, metade dos voluntários utilizou dispositivos contendo fragmentos tratados com verniz fluoretado, verniz fluoretado + laser de Er:YAG e verniz fluoretado + laser de Nd:YAG. A outra metade utilizou verniz não fluoretado, verniz não fluoretado + laser de Er:YAG e verniz não fluoretado + laser de Nd:YAG. Depois de um período de wash-out (15 dias), os voluntários foram cruzados quanto aos tratamentos, caracterizando um experimento cross-over 2x2. Ao final de cada fase experimental, a permeabilidade dentinária foi avaliada. A ANOVA dois critérios e o teste complementar de Duncan revelaram uma diferença significativa entre os espécimes tratados com o verniz fluoretado (p=0,005), verniz fluoretado + laser de Er:YAG (p=0,014) e verniz fluoretado + laser de Nd:YAG (p=0,025), em comparação aos tratados com verniz não fluoretado. Observou-se que, independentemente da associação aos lasers (Er:YAG ou Nd:YAG), o verniz fluoretado foi capaz de promover a redução da permeabilidade da dentina radicular erodida. Não se observou efeito adicional com a associação dos tratamentos. / The present study aimed to evaluate in situ the effect of associating the fluoride varnish to Er:YAG or Nd:YAG laser at permeability of eroded root dentin. Forty-eight specimens of bovine root dentin (2x2x2mm) were subjected to initial erosive challenge with citric acid (0.3%, pH 3.2), for 2 hours under agitation and stored at artificial saliva, at 37&deg C, for 24 hours, followed by a remineralization period in artificial saliva. After that, specimens were divided according to the treatment: fluoride varnish and non-flouride varnish, and subdivided according to the irradiation: Er:YAG laser (100mJ, 3Hz), Nd:YAG laser (70mJ, 15Hz) and non-irradiated. After a lead-in period (2 days), 8 volunteers (n=8) wore a palatal device containing 3 specimens that was subjected to erosive challenges ex vivo, four times a day with citric acid (0.3%, pH3.2), for 90s, during 5 days. At the first experimental phase, half of volunteers wore devices containing specimens treated with fluoride varnish, fluoride varnish + Er:YAG laser and fluoride varnish + Nd:YAG laser, and the other half wore the specimens treated with non-fluoride varnish, non-fluoride varnish + Er:YAG laser and non-flouride varnish + Nd:YAG laser. After a wash-out period (15 days), volunteers were crossed to different treatment, characterizing a 2x2 cross-over experiment. At the end of each experimental phase, the dentinal permeability was evaluated. Two-way ANOVA and Duncans test revealed a significant difference between specimens treated with fluoride varnish (p=0,005), fluoride varnish + Er:YAG laser (p=0,014) and fluoride varnish + Nd:YAG (p=0,025) compared to specimens treated with no-fluoride varnish. It was observed that regardless of association to laser (Er:YAG or Nd:YAG), fluoride varnish was able to promote, in situ, the reduction of permeability of eroded root dentin, but no additional effect was observed when the treatments were associated.

Este estudo teve como objetivos: 01.Avaliar ex vivo o efeito de diferentes sistemas clareadores na microdureza dentinária em dentes bovinos submetidos ao clareamento dental interno. 02.Avaliar ex vivo o efeito de diferentes sistemas clareadores na morfologia superficial da dentina bovina. Fragmentos de 4 x 4mm, contendo esmalte e dentina, foram obtidos de coroas de incisivos bovinos extraídos. Os espécimes foram submetidos ao clareamento dental interno com peróxido de hidrogênio a 35% e peróxido de carbamida a 37% utilizando sistemas convencionais (Opalescence Endo® and Whiteness Super Endo®) e fotoativados (Opalescence Xtra® and Whiteness HP Maxx®). Os controles foram tratados com perborato de sódio misturado com peróxido de hidrogênio a 10% ou nenhum tratamento foi realizado. A microdureza dentinária foi mensurada antes e após os tratamentos clareadores, e os valores de dureza Knoop (KHN) foram submetidos à análise estatística (two-way ANOVA, Tukey\'s port-test). Os espécimes foram observados e fotografados sob microscópio eletrônico de varredura e avaliados com relação às alterações morfológicas da superfície da dentina. Houve uma redução significante na microdureza dentinária para todos os grupos testados quando comparados aos grupos controles. Opalescence Xtra® (-11.36 ± 8.14 KHN), Opalescence Endo® (-13.71 ± 8.02 KHN), Whiteness HP Maxx® (-15.18 ± 9.58 KHN) e Whiteness Super Endo® (-16.97 ± 6.55 KHN) foram semelhantes estatisticamente. O grupo do perborato de sódio misturado com peróxido de hidrogênio 10% (2.10 ± 8.58 KHN) e o grupo sem tratamento clareador (-2.71 ± 2.40 KHN) também foram estatisticamente semelhantes entre si. Ocorreu uma grande variação no padrão de alterações da morfologia superficial da dentina com os sistemas clareadores utilizados. Todos os produtos testados apresentaram redução significativa da microdureza dentinária, exceto o grupo de perborato de sódio misturado com peróxido de hidrogênio a 10%. Ambos, pH e oxidação do peróxido de hidrogênio apresentam o papel de alterar a estrutura da dentina durante o clareamento interno. A utilização de produtos alcalinos com um tempo reduzido de aplicação (técnicas fotoativadas) pode diminuir as alterações morfológicas na dentina. / The aim of this study was: 01. To evaluate in vitro the effect of different in-office bleaching systems on the surface morphology of bovine dentin. 02. To evaluate ex vivo the effect of different bleaching systems on the microhardness of bovine dentine. Tooth fragments measuring 4 x 4mm, containing enamel and dentin, were obtained from the crowns of extracted bovine incisors. Samples were submitted to simulated intracoronal bleaching techniques with 35% hydrogen peroxide and 37% carbamide peroxide using conventional (Opalescence Endo® and Whiteness Super Endo®) and light activated systems (Opalescence Xtra® and Whiteness HP Maxx®). Controls were treated either with sodium perborate mixed with 10% hydrogen peroxide or no bleaching agent. Dentine microhardness values were measured before and after bleaching procedures, recorded as KHN (Knoop Hardness Number), and the differences between them analyzed (two-way ANOVA, Tukey\'s port-test). The samples were observed under SEM and the recorded images were evaluated for topographic alterations. Significant reductions of dentine microhardness were observed for all treatments when compared to the control groups. Opalescence Xtra® (-11.36 ± 8.14 KHN), Opalescence Endo® (-13.71 ± 8.02 KHN), Whiteness HP Maxx® (-15.18 ± 9.58 KHN) and Whiteness Super Endo® (- 16.97 ± 6.55 KHN) presented similar differences. The walking bleach technique (2.10 ± 8.58 KHN) and the untreated groups (-2.71 ± 2.40 KHN) were statistically alike.The ultrastructural alterations of dentin observed in this study varied greatly between groups, according to the products used. Apparently, higher pH products associated to in-office techniques yielded to better maintenance of dentin ultrastructure. The in-office products tested in the present study caused a significant reduction in dentine microhardness. The walking bleach technique did not affect dentine microhardness. Both low pH and hydrogen peroxide oxidation play a role in altering the ultrastructure of dentin during internal dental bleaching. The use of alkaline products with reduced time of application (in-office techniques) may decrease such morphological alterations.

O objetivo do presente estudo foi avaliar in situ o efeito da associação do verniz fluoretado ao laser de Er:YAG e ao laser Nd:YAG na permeabilidade da dentina radicular erodida. Quarenta e oito fragmentos de dentina radicular bovina, com dimensões 2x2x2 mm, foram submetidos a um desafio erosivo inicial com ácido cítrico (0,3%, pH 3,2), por duas horas, sob agitação e armazenados em saliva artificial por vinte e quatro horas. Posteriormente, os espécimes foram divididos em relação aos tratamentos: verniz fluoretado e não fluoretado e subdivididos conforme a irradiação: laser de Er:YAG (100mJ, 3Hz), laser de Nd:YAG (70mJ, 15 Hz) e não irradiado. Após um período de lead in (2 dias), os voluntários (n=8) utilizaram dispositivos palatinos contendo três espécimes que foram submetidos a desafios erosivos ex vivo, quatro vezes ao dia, com ácido cítrico (0,3%, pH 3,2), durante 90s, por cinco dias. Na primeira fase do experimento, metade dos voluntários utilizou dispositivos contendo fragmentos tratados com verniz fluoretado, verniz fluoretado + laser de Er:YAG e verniz fluoretado + laser de Nd:YAG. A outra metade utilizou verniz não fluoretado, verniz não fluoretado + laser de Er:YAG e verniz não fluoretado + laser de Nd:YAG. Depois de um período de wash-out (15 dias), os voluntários foram cruzados quanto aos tratamentos, caracterizando um experimento cross-over 2x2. Ao final de cada fase experimental, a permeabilidade dentinária foi avaliada. A ANOVA dois critérios e o teste complementar de Duncan revelaram uma diferença significativa entre os espécimes tratados com o verniz fluoretado (p=0,005), verniz fluoretado + laser de Er:YAG (p=0,014) e verniz fluoretado + laser de Nd:YAG (p=0,025), em comparação aos tratados com verniz não fluoretado. Observou-se que, independentemente da associação aos lasers (Er:YAG ou Nd:YAG), o verniz fluoretado foi capaz de promover a redução da permeabilidade da dentina radicular erodida. Não se observou efeito adicional com a associação dos tratamentos. / The present study aimed to evaluate in situ the effect of associating the fluoride varnish to Er:YAG or Nd:YAG laser at permeability of eroded root dentin. Forty-eight specimens of bovine root dentin (2x2x2mm) were subjected to initial erosive challenge with citric acid (0.3%, pH 3.2), for 2 hours under agitation and stored at artificial saliva, at 37&deg C, for 24 hours, followed by a remineralization period in artificial saliva. After that, specimens were divided according to the treatment: fluoride varnish and non-flouride varnish, and subdivided according to the irradiation: Er:YAG laser (100mJ, 3Hz), Nd:YAG laser (70mJ, 15Hz) and non-irradiated. After a lead-in period (2 days), 8 volunteers (n=8) wore a palatal device containing 3 specimens that was subjected to erosive challenges ex vivo, four times a day with citric acid (0.3%, pH3.2), for 90s, during 5 days. At the first experimental phase, half of volunteers wore devices containing specimens treated with fluoride varnish, fluoride varnish + Er:YAG laser and fluoride varnish + Nd:YAG laser, and the other half wore the specimens treated with non-fluoride varnish, non-fluoride varnish + Er:YAG laser and non-flouride varnish + Nd:YAG laser. After a wash-out period (15 days), volunteers were crossed to different treatment, characterizing a 2x2 cross-over experiment. At the end of each experimental phase, the dentinal permeability was evaluated. Two-way ANOVA and Duncans test revealed a significant difference between specimens treated with fluoride varnish (p=0,005), fluoride varnish + Er:YAG laser (p=0,014) and fluoride varnish + Nd:YAG (p=0,025) compared to specimens treated with no-fluoride varnish. It was observed that regardless of association to laser (Er:YAG or Nd:YAG), fluoride varnish was able to promote, in situ, the reduction of permeability of eroded root dentin, but no additional effect was observed when the treatments were associated.

The fate of DNA from dead cells is an important issue when interpreting results from root canal infections analysed by the PCR technique. DNA from dead bacterial cells is known to be detectable long time after cell death and its stability is dependent on many different factors. This work investigated factors found in the root canal that could affect the recovery of microbial DNA. In an ex vivo experiment, DNA from non-viable gram-positive Enterococcus faecalis was inoculated in instrumented root canals and recovery of DNA was assessed by PCR over a two-year period. DNA was still recoverable two years after cell death in 21/25 teeth. The fate of DNA from the gram-negative bacteria Fusobacterium nucleatum and the gram-positive Peptostreptococcus anaerobius was assessed in vitro. DNA from dead F. nucleatum and P. anaerobius could be detected by PCR six months post cell death even though it was clear that the DNA was released from the cells due to lost of cell wall integrity during the experimental period. The decomposition rate of extracellular DNA was compared to cell-bound and it was evident that DNA still located inside the bacterium was much less prone to decay than extracellular DNA. Free (extracellular) DNA is very prone to decay in a naked form. Binding to minerals is known to protect DNA from degradation. The fate of extracellular DNA was assessed after binding to ceramic hydroxyapatite and dentine. The data showed that free DNA, bound to these materials, was protected from spontaneous decay and from enzymatic decomposition by nucleases. The main conclusions from this thesis were: i) DNA from dead bacteria can be detected by PCR years after cell death ex vivo and in vitro. ii) Cell-bound DNA is less prone to decomposition than extracellular DNA. iii) DNA is released from the bacterium some time after cell death. iv) Extracellular DNA bound to hydroxyapatite or dentine is protected from spontaneous decomposition and enzymatic degradation.