Neste trabalho, foi realizado um estudo crítico dos modelos metodológicos in vivo e in vitro sobre a avaliação microbiológica da clorexidina empregada como agente irrigante e medicação intracanal na terapia endodôntica. Para tanto, foi realizada uma busca sistemática nas bases de dados: Medline, Lilacs e BBO nos últimos dez anos sobre o assunto em questão. Foram selecionados os trabalhos que avaliaram a clorexidina de uma maneira não combinada com outras substâncias ou fármacos, abordando sua utilização tanto como irrigante endodôntico, e como medicação intracanal. Para uma melhor análise, os trabalhos foram agrupados de acordo com a forma de uso e metodologia com seus dados tabulados, respectivamente: Clorexidina como irrigante in vitro, Clorexidina como irrigante in vivo, Clorexidina como medicação intracanal in vitro e Clorexidina como medicação intracanal in vivo. Na análise, pode-se observar a efetividade antimicrobiana da clorexidina como irrigante nos ensaios laboratoriais e clínicos, sendo nas formas de gel e líquida com variação na concentração de 0.2 a 2%. Quanto o uso da clorexidina como medicação intracanal, pode ser constatada sua atividade antimicrobiana em ambos ambientes experimentais, embora com poucos trabalhos in vivo, seu desempenho foi observado apresentando largo espectro de ação quando utilizada a 2% em gel por um período de sete dias. / In this essay, it was realized a critical study of the methodological models, in vivo and in vitro, about the microbiological assessment of the chlorhexidine employed as root canal irrigants and dressing in the Endodontics. A systematic search was carried out on the databases: Medline, Lilacs and BBO from the last ten years concerning the respective subject. There were selected the papers which evaluated the chlorhexedine as an individual group, without joined to another substances or drugs, broaching its application as root canal irrigants as much as dressing. For improved analyzed, the investigations were divided according its way of use and methodology, with its data tabulated in the respective way: Chlorhexidine as irrigant in vitro, Chlorhexidine as irrigant in vivo, Chlorhexidine as intracanal dressing in vitro and Chlorhexidine as intracanal dressing in vivo. At the analysis, it can be observed the antimicrobial effectiveness of the chlorhexidine as irrigant in the clinical and laboratories tests, being the gel and liquid form with concentration changed from 0.2 to 2%. About the chlorhexidine as root canal dressing, it can be evidenced its antimicrobial activity in both experimental methods, although a few trials in vivo, its performance was observed showing a large spectral action when used in 2% gel for a period of seven days.

Orientador: Izabel Yoko Ito / Banca: Mario Roberto Leonardo / Banca: Lea Assed Bezerra da Silva / Banca: Reginaldo Bruno Gonçalves / Banca: Brenda Paula Figueiredo de Almeida Gomes / Resumo: No presente estudo foi monitorada a microbiota do canal radicular de 30 dentes de humanos portadores de lesão periapical crônica. Foram utilizados curativos de demora à base de hidróxido de cálcio entre sessões: Calen (Grupo I), Calen PMCC (Grupo II) e Calasept (Grupo III), que foram mantidos nos canais radiculares por aproximadamente 30 dias. O material colhido dos canais radiculares, antes e após a realização da terapêutica endodôntica, foi submetido a teste laboratorial baseado na biologia molecular: Reação em Cadeia da Polimerase (PCR). Como controle positivo da presença bacteriana, foi utilizado primer 16S rDNA. As seguintes espécies bacterianas foram monitoradas: Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens (bacilos pigmentados de negro), e Bacteroides forsythus, microrganismos marcantes na patologia de processos infecciosos dos tecidos pulpoperiapicais. Os resultados evidenciaram diminuição das espécies microbianas, aqui monitoradas de forma indireta à partir da presença de bandas, ou amplicons (produtos do PCR), quando do uso de curativos de demora à base de hidróxido de cálcio. Com relação à eficácia particular a cada material, os Grupos I, II e III não demonstraram entre si diferenças estatisticamente significantes. / Abstract: In the present study the root canal flora of 30 human teeth with chronic periapical lesion was monitored. Dressing pastes based on calcium hydroxide were used, among sessions namely: Calen (Group I), Calen/PMCC (Group II) and Calasept (Group III) that were used as a temporary dressings in root canals for approximately 30 days. The microbial material was colected from root canals, before and after the root canal treatment and submitted to laboratorial test based on molecular biology assay: Polymerase Chain Reaction (PCR). As a positive control of bacterial presence, a primer 16S rDNA was utilized. The following bacterial strains were monitored: Porphyromonas endodontalis, P. gingivalis, P. intermedia, P. nigrescens (black pigmented rods) and B. forsythus, a microorganism with an important role on pulpoperiapical pathosis. The results showed a decrease of bacterial strains population indirectly, based on bands presence (amplicons) where the calcium hydroxide dressing pastes were applied. Concerning the intrinsic efficacy of each material, the Groups I, II and III didn't show significant statistical differences among them. / Doutor

Considerando-se a necessidade da obturação tridimensional e o adequado selamento marginal do sistema de canais radiculares, como fatores decisivos para o êxito da terapia endodôntica, este estudo objetivou avaliar as respostas inflamatórias periapicais resultantes da infiltração coronária em dentes de cães obturados com diferentes cimentos resinosos (AH Plus®, RealSeal® e RealSeal SE®) e técnicas obturadoras (condensação lateral e termoplástica). Para tanto, pré-molares e incisivos centrais e laterais de 5 cães tiveram o tratamento endodôntico realizado empregando-se limas de NiTi e creme de Endo PTC associado ao hipoclorito de sódio a 1%, estabelecendo-se como limite de instrumentação à distância de 1mm a 1.5mm aquém do vértice radiográfico. A seguir os condutos foram obturados de acordo com os grupos experimentais (n=12) e grupos controles (n=10): GI- AH Plus/condensação lateral, GII- AH Plus/termoplástificação, GIIIRealSeal/ condensação lateral, GIV- RealSeal/ termoplástificação, GV- RealSeal SE/condensação lateral, GVI- RealSeal SE/ termoplástificação, Grupo controle positivo e Grupo controle negativo. As cavidades coronárias ficaram expostas ao meio bucal por um período de 75 dias e, passado este período experimental, os cães foram eutanásiados por superdosagem de anestésico. Foi realizada a remoção cirúrgica das partes correspondentes aos pré-molares e incisivos da maxila e da mandíbula, fixação das mesmas em solução de formol a 10% por 72h e desmineralização em solução de ácido fórmico a 20% + citrato de sódio a 10% pelo período mínimo de 90 dias. Posteriormente, realizou-se a rotina histológica com cortes seriados com espessura de 6m e estes foram corados com hematoxilina e eosina (HE). Cada corte histológico foi digitalizado e analisado em relação à existência de patologia periapical por três avaliadores independentes. Os dados obtidos foram confrontados estatisticamente utilizando o teste de Kruskal-Wallis seguido do teste comparativo de Dunn, observando-se que em relação às técnicas, a termoplástificação por ondas contínuas de condensação associada à técnica de injeção mostrou melhores condições periapicais quando comparada à técnica de condensação lateral (p=0.0055), sendo que a utilização desta técnica resultou em reações inflamatórias mais intensas para o cimento RealSeal SE (p=0.002). Em relação aos cimentos avaliados, o RealSeal SE mostrou-se menos eficaz em evitar a percolação, produzindo maiores níveis de inflamação crônica independentemente do sistema de obturação (p=0.0088). / Considering the need for three-dimensional obturations and adequate marginal sealing of the radicular canal system as being decisive factors for successful endodontic therapy, this study had as its objective the evaluation of periapical inflammatory responses resulting from coronary infiltration in dog\'s teeth obturated using different resin sealers (AH Plus, RealSeal and RealSeal SE) and different obturation techniques (lateral condensation and thermoplastic). For this study, premolars, central and lateral incisors from five dogs had endodontic treatment employing NiTi files and Endo PTC cream associated with 1% sodium hypochlorite. The working length was established at a distance of 1.0 to 1.5mm short of the radiographic apex. After this, the teeth were obturated according to their experimental groups (n=12) and control groups (n=10). The groups were defined as follows: GI - AH Plus/lateral condensation, GII - AH Plus/thermoplastic, GIII - RealSeal/lateral condensation, GIV - RealSeal/thermoplastic, GV - RealSeal SE/lateral condensation, GVI - RealSeal SE/ thermoplastic, a positive control group and a negative control group. The coronal access were left exposed to the oral environment for a period of seventy-five days, and after this experimental period, the dogs were euthanized by anesthetic overdose. The parts corresponding to the premolars and the maxilar incisors were surgically removed. These were then submitted to a solution of 10% formol for 72 hours and then demineralized in a solution of 20% formic acid and 10% sodium citrate for a minimum period of ninety days. Before hand, a histological routine of serial cuts having a width of 6m was undertaken and these were colored with hematoxilin and eosin (HE). Each histological cut was digitalized and analyzed with respect to the existence of periapical inflammation by three independent evaluators. The data obtained was statistically compared, firstly using the Kruskal-Wallis test and then the comparative Dunn test, where it was noted that in relation to the techniques, thermoplastification by continuous wave of condensation associated with the injection technique showed better periapical conditions than what was seen for the lateral condensation technique (p=0.0055). On the other hand, for the sealers evaluated, the condensation technique resulted in more intense inflammatory reactions when the sealer RealSeal SE was used (p=0.002). In relation to the sealers evaluated, RealSeal SE was seen to be less effective against percolation, thus producing greater levels of chronic inflammation irrespective of the obturation system employed (p=0.0088).

Indiana University-Purdue University Indianapolis (IUPUI) / Introduction: Elimination of bacteria and active infection within an infected root canal system is one of the primary objectives of nonsurgical root canal treatment. One of the measures of successful root canal treatment is subsequent bone healing of periapical lesions caused by previous infection. A previous study by Yumoto et al. showed that electromagnetic wave stimulation can increase proliferation of osteoblastic cells with no cytotoxicity, and it can also up-regulate growth factors such as vascular endothelial growth factor and platelet-derived growth factor.18 They also showed increased proliferation of an immortalized osteoblastic MC3T3-E1 cell line 3 days following electromagnetic stimulation (EMS).18 Previously, Pauly et al. found increased alkaline phosphatase (ALP) activity with 10 mA EMS application to primary murine calvaria-derived osteoblastic cells with 5 pulses at 1 second per pulse, but no significant differences were found for MTS proliferation nor mineral deposition compared to a negative control group.82 Optimization of the different variables including post-treatment incubation time, current delivery, and number of pulses per treatment may be necessary to improve osteogenic activity. The use of mesenchymal stem cells from murine bone marrow may also offer a physiologically relevant model for osteoblastic regeneration of periapical lesions. Objectives: The goal of this study was to investigate and optimize the effects of electromagnetic wave stimulation (EMS) on murine bone marrow mesenchymal stem cells (MSCs) by evaluating the proliferation and differentiation of the cells after exposure to different EMS treatment regimens. Materials and Methods: 5 x104 stromal osteoblasts (SOBs) were cultured in 24-well plates in α-MEM containing 10% fetal bovine serum. Cells were then subjected to pulsed EMS treatments of 1 mA, 10 mA, and 50 mA. EMS was generated using an electromagnetic apical treatment (EMAT) device created by J. Morita MFG Corp. Proliferation was assessed via MTS assay 1 days after treatment. For osteogenic differentiation, ascorbic acid and β-glycerol phosphate were added to the culture media, and SOBs were cultured for 14 days. Afterwards, alkaline phosphatase (ALP) activity and Alizarin-red S mineral deposition were quantified as measures of osteoblast activity. Cells grown in osteogenic media without EMS treatment served as the negative control. Results: Although MSC proliferation was unaffected by different EMS treatment regimens, 50 mA EMS resulted in a decrease in ALP activity and mineral deposition by osteoblasts. Conclusions: Our findings suggest bone healing by EMS may involve a different cellular mechanism, that is not reproduced in vitro in our studies. Utilizing different amperage and EMS regimens may improve osteogenic differentiation.