Indiana University-Purdue University Indianapolis (IUPUI) / It has been proposed that jaw deformation during function may be a contributor to supraosseous tooth eruption. This could be through a transient pressure gradient in the flexed bone and/or tension in the PDL fibers produced by socket deformation. Thus, the purpose of this study was to determine if forces applied to a fresh frozen canine mandible can lead to occlusal displacement of a tooth. Seven hemisected canine mandibles (14 specimens) were used to test this hypothesis. A force cycle (10 N to 100 N to 300 N) was applied by a Bionix testing machine (MTS TM Systems Corp., Minneapolis, Minn.) to the mandible in three point bending. Digital radiographs were taken at each change in force level using computer digitized radiography (CDR, TM Schick Technologies, Inc., Long Island City, N.Y.). By measuring the relative movement of metal markers (1 mm steel balls) placed into the mandible and the crown of the 2nd premolar, the amount of tooth eruption was calculated. With cyanoacrylate cement (Archer® Instant Bonding Adhesive, Tandy Corp, Ft. Worth, Texas) injected into the PDL space of the same tooth, the test was repeated one week later. These "ankylosed" teeth served as controls. The eruption distances were compared between the experimental and the ankylosed teeth by means of repeated measures analysis of variance. The only significant occlusal displacement was noted as the force was initially raised to 100 N (21.7 ± 40.6 μm). Due to the possible breakdown of the cyanoacrylate cement, the ankylosed teeth also showed evidence of eruption with the continued application of force. Although significant eruption was noted, the displacements were very small. PDL degradation, viscoelastic behavior, measurement of small displacements, limited resolution radiography, and two dimensional analysis are contributing factors to the uncertainty of the results. Further investigation is required to determine if jaw deformation during function is a viable mechanism leading to the occlusal displacement of teeth.

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Objectives: Most bacterial species implicated as pathogens in human periodontitis are anaerobic in their metabolism. Systemic administration of metronidazole, an antibiotic specifically active against anaerobic bacteria, has been shown in multiple clinical trials to be beneficial in enhancing periodontal therapeutic outcomes beyond that attained by conventional mechanically-based forms of periodontal therapy alone, in large part by the drug inducing better reductions of major anaerobic pathogens in periodontal pockets. However, systemic metronidazole regimens in the treatment of periodontitis require multiple patient-administered drug doses per day, which may compromise treatment benefits in patients less compliant with prescribed oral drug consumption schedules. Tinidazole, a second-generation 2-methyl-5-nitroimidazole class antibiotic similar to metronidazole, also possesses marked antibacterial activity against anaerobic bacteria, and exhibits pharmacokinetic properties that enable its bioavailability with only a once-a-day oral drug dose, which may be an advantage for use in periodontitis patients unable to comply with more frequent drug dosing regimens. Little comparative data is available assessing the potential antimicrobial effects of tinidazole, as compared to metronidazole, against anaerobic periodontal pathogens, particularly “wild-type” clinical strains isolated from severely-diseased human periodontal pockets. As a result, this study tested fresh clinical subgingival isolates of selected anaerobic red and orange complex periodontal pathogens for their in vitro susceptibility to tinidazole, metronidazole, and three other antibiotics frequently employed in periodontal therapy. Methods: Paper point subgingival plaque biofilm specimens were removed from 31 adults with severe periodontitis, and transported in VMGA III medium from variousUnited States private periodontal practices to the Oral Microbiology Testing Service Laboratory at Temple University School of Dentistry. Within 24 hours, the samples were serial diluted and plated onto enriched Brucella blood agar plates with either no antimicrobials added, or supplemented with either tinidazole at 16 mg/L, metronidazole at 16 mg/L, doxycycline at 4 mg/L, amoxicillin at 8 mg/L, or clindamycin at 4 mg/L, which represent recognized non-susceptible drug breakpoint concentrations for each of the antibiotics. After incubation at 37°C for 7 days in an 85% N2-10% H2-5% CO2 anaerobic atmosphere, all plates were examined with established phenotypic criteria for selected anaerobic red and orange complex periodontal pathogens, including Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia/nigrescens, Parvimonas micra, and Fusobacterium nucleatum group species. In vitro antibiotic resistance was noted when any of the test bacterial species displayed growth on one or more of the antibiotic-supplemented enriched Brucella blood agar plates. A paired t-test compared mean total subgingival proportions of the evaluated anaerobic red and orange complex periodontal pathogens per patient which were resistant in vitro to non- susceptible drug threshold concentrations of tinidazole as compared to metronidazole, as well as to doxycycline, amoxicillin, and clindamycin, with a P-value of < 0.05 required for statistical significance. Results: The study patients yielded an average 25.8% per patient of total subgingival proportions of the selected anaerobic red and orange complex periodontal pathogens. Among these species, P. micra was isolated from all (100%) study patients, and P. intermedia/nigrescens and F. nucleatum from 93.5% and 90.3% patients, respectively, with mean subgingival proportions of these species in positive patientsranging from 1.8% to 9.7%. T. forsythia at mean subgingival levels of 1.8% was recovered from 54.8% of the patients, whereas subgingival P. gingivalis averaged 9.1% in 5 (16.1%) patients. Tinidazole and metronidazole at 16 mg/L threshold concentrations inhibited in vitro growth of all test periodontal pathogens, except for a tinidazole-resistant strain of P. intermedia/nigrescens in one patient that was additionally resistant in vitro to doxycycline, amoxicillin and clindamycin. No statistically significant differences were found between tinidazole and metronidazole in mean total subgingival proportions of anaerobic red and orange complex periodontal pathogens per patient exhibiting in vitro resistance to a 16 mg/L drug concentration (P = 0.327, paired t-test). However, significantly greater total subgingival proportions of anaerobic red and orange complex periodontal pathogens per patient were resistant in vitro to breakpoint concentrations of either doxycycline, amoxicillin, or clindamycin, as compared to tinidazole or metronidazole (all P-values < 0.006, paired t-test). Conclusions: Tinidazole performed in vitro similar to metronidazole, but significantly better than doxycycline, amoxicillin, or clindamycin, in antimicrobial activity against freshly-isolated clinical strains of human subgingival anaerobic red and orange complex periodontal pathogens. As a result of its similar spectrum of antimicrobial inhibition against anaerobic bacteria, and its more convenient once-a-day oral drug dosing properties, tinidazole may be prescribed for clinical systemic use in place of metronidazole in severe human periodontitis treatment regimens where patient compliance with multiple dose per day systemic drug consumption is anticipated to be poor or difficult to attain. / Oral Biology

Resumo: O prognóstico de um dente reimplantado está relacionado a integridade do ligamento periodontal cementário e da camada de cemento. Esta, por sua vez, e influenciada por vários fatores, sendo os principais, o periodo extra-alveolar e o meio de conservação. Existem dúvidas, no entanto, se alterações sistêmicas, como o diabetes, também podem interferir no prognóstico do reimplante dentário. Portanto, o propósito deste trabalho foi avaliar o processo de reparo do reimplante de incisivos de ratos diabéticos nao controlados após a manutenção dos dentes em leite bovino pasteurizado. Para tanto, 32 ratos foram divididos em dois grupos de 16 animais cada, sendo que no Grupo I foi administrada, por via endovenosa, injeção de tampão citrato e no Grupo II inoculada estreptozotocina dissolvida em tampão citrato pela mesma via, para a indução do diabetes. Após controle glicêmico e a comprovação das altas taxas de glicemia, o incisivo superior direito de cada animal foi extraído e mantido em leite durante sessenta minutos. Na sequência, realizou-se o tratamento endodontico com a colocação de curativo de hidróxido de cálcio e o posterior reimplante dos dentes em seus respectivos alveolos. A antibioticoterapia sistêmica foi instituida no pós-operatório imediato. Decorridos 10 e 60 dias após o reimplante, realizou-se a eutanasia dos animais e as peças obtidas foram submetidas ao processamento laboratorial para a obtenção dos cortes, os quais foram corados com hematoxilina e eosina e Tricromicro de Masson para análise histologica e histométrica. Os resultados demonstraram que o tecido conjuntivo adjacente a superfície radicular se apresentou desorganizado nos animais diabéticos. Ìndices menores de reabsorção radicular externa e anquilose também foram encontrados neste grupo, porém, sem diferenças estatísticas significantes. / Abstract: The prognosis of a replanted tooth is related to the integrity of the cemental periodontal ligament and the cementum layer, which, in turn, is influenced by several factors. Among these factors, the extra-alveolar time and the storage medium are the most important. However, it remains unclear whether systemic alterations, such as diabetes, may also interfere with the prognosis of tooth replantation. Therefore, the purpose of this investigation was therefore to evaluate the healing process of non-controlled diabetic rats' incisors replanted after storage of the teeth in bovine pasteurized milk. A total of 32 rats were assigned to two groups of 16 animals each. In group I, a citrate buffer injection was administered intravenously to rats while the animals in group II received an intravenous injection of streptozotocin dissolved in citrate buffer to induce diabetes. After confirmation of high glycemic levels, the maxillary right incisor of each animal was extracted and stored in milk during 60 minutes. Thereafter, the teeth were endodontically treated, with placement of a calcium hydroxide-based intracanal dressing, and the teeth replanted into their sockets. Systemic antibiotic therapy started in the immediate postoperative period. The animals were euthanized 10 and 60 days postoperatively. The anatomical pieces were removed, submitted to routine laboratorial processing and serially sectioned to obtain cuts that were stained by hematoxylin and eosin and Masson's Trichrome for histologic and histometric analyses. The results showed that the connective tissue adjacent to the root surface was disorganized in the diabetic rats. Lower indices of external root resorption and ankylosis were also observed in the diabetic group, though with no statistically significant difference. / Orientador: Celso Koogi Sonoda / Coorientador: Alessandra Marcondes Aranega / Banca: Luis Eduardo Marques Padovan / Banca: Wilson Roberto Poi / Mestre

Este estudo avaliou os efeitos da administração tópica de Bdellovibrio bacteriovorus HD100 na periodontite experimental em ratos. 32 ratos foram alocados nos grupos CT, DPT, CT-HD100 e DPT-HD100. No dia 0 do experimento, os animais dos grupos DPT e DPT-HD100 receberam ligaduras de seda ao redor dos primeiros molares inferiores (PMIs). Nos grupos CT-HD100 e DPT-HD100, suspensões de 1 mL contendo B. bacteriovorus HD100 foram administradas topicamente na região subgengival de PMIs nos dias 0, 3 e 7. Nos grupos CT e DPT, administrações tópicas foram realizadas com uma suspensão não contendo B. bacteriovorus HD100. Todos os animais foram submetidos à eutanásia no dia 14 do experimento. O tecido gengival, hemi-mandíbulas e biofilme bucal foram coletados para avaliação dos seguintes parâmetros: i) microarquitetura óssea, volume ósseo e nível ósseo alveolar (microtomografia computadorizada por transmissão de raios X micro-CT); ii) níveis de inserção conjuntiva (análise histomorfométrica); iii) microbiota bacteriana (checkerboard DNA-DNA hybridization); iv) expressão de citocinas inflamatórias e fatores de transcrição (análise imunoenzimática - Multiplex e reação em cadeia da polimerase por transcriptase reversa em tempo real); iii) padrão de imunomarcação para beta defensinas (BD), receptores do tipo Toll (TLR) e grupamentos de diferenciação (CD) (reações imunohistoquímicas). Testes in vitro foram também realizados para avaliar o potencial antimicrobiano de B. bacteriovorus HD100 contra periodontopatógenos. Os dados foram analisados estatisticamente (p < 0,05). O grupo DPT-HD100 apresentou menores porosidade óssea, separação trabecular, nível ósseo alveolar e nível de inserção conjuntiva, bem como maiores volume ósseo e número de trabéculas ósseas quando comparado ao Grupo DPT (p < 0,05). O grupo DPT-HD100 apresentou maiores proporções de espécies semelhantes à Actinomyces e Streptococcus e menores proporções de espécies semelhantes à Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Fusobacterium polymorphum, Eikenella corrodens, Eubacterium nodatum, Campylobacter gracilis, Capnocytophaga sputigena e Veillonella parvula quando comparado ao Grupo DPT. Nas análises de parâmetros imunoinflamatórios, o Grupo DPT-HD100 apresentou maiores níveis de Proteina quimioatrativa de monócito1 (MCP-1), Células T normais expressas e secretadas, reguladas por ativação (RANTES), Osteoprotegerina (OPG), Fator de Crescimento Transformador (TGF)-&alpha; e Interleucina (IL)-10 e menores níveis de Fator de Necrose Tumoral (TNF)-&beta;, bem como maior padrão de imunomarcação para BD-1, BD-2 e BD-3 quando comparado ao Grupo DPT (p < 0,05). Para os níveis de IL-1, IL-6, Fator Estimulador de Colônias de Macrófagos (M-CSF), Ligante do Receptor Ativador de Fator Nuclear kappa-B (RANK-L) e padrões de imunomarcação para TLR-2, TLR-4, CD-4, CD-8 e CD-57, não foram observadas diferenças entre os grupos DPT e DPT-HD100. Na análise de expressão gênica, o Grupo DPT-HD100 apresentou maior expressão de IL-17, IL-10 e Forkhead box P3 (FOXP3) quando comparado ao Grupo DPT (p < 0,05). Nos testes in vitro, as co-culturas de periodontopatógenos (F. nucleatum, P. intermedia e A. ctinomycetemcomitans) e B. bacteriovorus HD100 apresentaram menor densidade óptica do que as culturas isoladas de periodontopatógenos em 48 horas. O uso tópico de B. bacteriovorus HD100 modifica os parâmetros imunoinflamatórios e microbiológicos, promovendo um efeito protetor contra a perda óssea alveolar e perda de inserção do tecido conjuntivo em ratos com periodontite experimental / This study evaluated the effects of topical administration of Bdellovibrio bacteriovorus HD100 on experimental periodontitis in rats. Thirty-two rats were allocated in the CT, DPT, CT-HD100 and DPT-HD100 groups. At day 0 of the experiment, animals of the DPT and DPT-HD100 groups received silk ligatures around the mandibular first molars (MFMs). In the CT-HD100 and DPT-HD100 groups, 1 ml suspensions containing B. bacteriovorus HD100 were topically administered in the subgingival region of MFMs on days 0, 3 and 7. In the CT and DPT groups, topical administrations were performed with a suspension without B. bacteriovorus HD100. All animals were submitted to euthanasia on day 14 of the experiment. The gingival tissue, hemi-mandibles and oral biofilm were collected to evaluate the following parameters: i) bone microarchiteture, bone volume and alveolar bone level (X-ray micro-computed tomography - micro-CT); ii) levels of conjunctive insertion (histomorphometric analysis); iii) microbiological profile (checkerboard DNA-DNA hybridization); iv) expression of inflammatory cytokines and transcription factors (immunoenzymatic analysis - Multiplex and Real-time reverse transcription polymerase chain reaction); iii) immunostaining pattern for Beta defensins (BD), Toll-like receptors (TLR) and Cluster of differentiation (CD) (immunohistochemical reactions). In vitro tests were also performed to evaluate the antimicrobial potential of B. bacteriovorus HD100 against periodontopathogens. The data were analyzed statistically (p <0.05). The DPT-HD100 Group presented lower bone porosity, trabecular separation, alveolar bone level and connective tissue attachment level, as well as higher bone volume and number of bone trabeculae when compared to the DPT Group (p <0.05). The DPT-HD100 Group presented higher proportions of Actinomyces and Streptococcus-like species and smaller proportions of Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Fusobacterium polymorphum, Eikenella corrodens, Eubacterium nodatum, Campylobacter gracilis, Capnocytophaga sputigena and Veillonella parvula-like species when compared to the DPT Group. In the analysis of immunoinflammatory parameters, the DPT-HD100 Group presented higher levels of Monocyte-1 chemoattractant protein (MCP-1), Regulated on activation, normal T cell expressed and secreted (RANTES), Osteoprotegerin (OPG), Transforming Growth Factor (TGF)-&alpha; and Interleukin (IL) -10 and lower levels of Tumor Necrosis Factor (TNF)-&beta;, as well as a higher immunolabeling pattern for BD-1, BD-2 and BD-3 when compared to the DPT Group (p <0.05). For levels of IL-1, IL-6, Macrophage Colony Stimulating Factor (M-CSF), Receptor activator of nuclear factor kappa-B ligand (RANK-L) and immunolabeling patterns for TLR-2 TLR- 4, CD-4, CD-8 and CD-57, no differences were observed between the DPT and DPT-HD100 groups. In the analysis of gene expression, the DPT-HD100 group presented higher expression of IL-17, IL-10 and Forkhead box P3 (FOXP3) when compared to the DPT Group (p <0.05). In the in vitro tests, co-cultures of periodontopathogens (F. nucleatum, P. intermedia and A. ctinomycetemcomitans) and B. bacteriovorus HD100 showed lower optical density than isolated cultures of periodontopathogens at 48 hours. Topical use of B. bacteriovorus HD100 modifies immunoinflammatory and microbiological parameters, promoting a protective effect against alveolar bone loss and loss of connective tissue attachment in rats with experimental periodontitis

Existem poucos estudos na área de periodontia envolvendo indivíduos portadores de fissura labiopalatina; pesquisas que avaliem as condições periodontais destes pacientes durante tratamento ortodôntico são ainda mais escassas. O objetivo deste trabalho foi fazer um estudo da condição periodontal, durante o tratamento ortodôntico, dos pacientes com fissura labiopalatina com comprometimento de rebordo alveolar e avaliar se a região da fissura apresenta maior incidência e severidade de alterações periodontais. O grupo estudado foi selecionado no Ambulatório da Disciplina de Prótese Bucomaxilofacial da Faculdade de Odontologia da Universidade de São Paulo (FO-USP), sendo constituído por 43 pacientes na faixa etária de 11 a 35 anos, portadores de fissuras labiopalatinas, não sindrômicos, já submetidos às cirurgias reparadoras primárias e em tratamento com ortodontia fixa há mais de 6 meses. Durante o exame clínico periodontal da maxila realizamos medidas de profundidade clínica de sondagem, nível clínico de inserção, sangramento à sondagem e índice de placa. Todos os exames foram feitos por um único pesquisador, especialista em periodontia, com calibração prévia. A avaliação periodontal mostrou que a região da maxila afetada pela fissura apresentou maior índice de sangramento à sondagem o que está associado a maior ocorrência de gengivite. Os dentes da região da fissura apresentaram menores profundidades clínicas de sondagem que a média da maxila, indicando que não há maior prevalência de bolsas periodontais na região fissurada. Não encontramos diferenças estatisticamente significantes no nível clínico de inserção dos dentes adjacentes à fissura, sendo assim a presença da fissura não esteve associada a perda óssea. Os pacientes da amostra apresentaram uma profundidade de sondagem rasa e pouca perda de inserção. A doença periodontal em indivíduos portadores de fissura, neste estudo, demonstrou não ter maior incidência ou severidade na região da fissura. / There is a shortage of studies in the periodontics field about individuals with cleft lip and palate, studies that evaluate the periodontal conditions of these patients during orthodontic treatment are even scarcer. The aim of this research was to analyze the periodontal condition during orthodontic treatment, of patients with cleft lip and palate with compromised alveolar bone and evaluate if the cleft region on maxilla presented a higher incidence and severity of periodontal disease. The study group was selected from the Clinical Division of Bucomaxilofacial Prosthodontics subject, from School of Dentistry of University of São Paulo, which consisted of 43 patients between 11 and 35 years old with cleft lip and palate, non-syndromic, who had already undergone primary reconstructive surgeries, and who have been in treatment with fixed orthodontics for more than 6 months. During a periodontal clinical exam of the maxilla, the measurements of depth probing, clinical level of insertion, bleeding on probing and plaque index were carried out. The exams were performed by an experienced periodontal examiner who was calibrated previously. The periodontal evaluation showed that the region of the maxilla affected by the cleft had a higher index of bleeding on probing, which is associated with a higher occurrence of gingivitis. The teeth in the cleft region showed smaller clinical depths of probing than the maxilla average, indicating that there is no greater prevalence of periodontal pockets in the region of the cleft. We did not find any significant statistical differences in the clinical level of insertion of the teeth adjacent to the cleft, so the presence of cleft was not associated with a higher bone loss. Patients in the sample had a shallow probing depth and low insertion loss. This study showed that periodontal disease in individuals with cleft lip and palate occurs in a similar way at all the maxilla.

Probióticos do gênero Lactobacillus estão sendo amplamente investigados no tratamento da periodontite. Contudo, os efeitos de microrganismos do gênero Bifidobacterium ainda são pouco conhecidos. Este estudo avaliou os efeitos do probiótico Bifidobacterium animalis subsp. lactis (B. lactis) HN019 como adjuvante à raspagem e alisamento radicular (RAR) no tratamento da periodontite experimental (PE) em ratos. No dia 0 do experimento, 32 ratos foram alocados em 4 grupos: C (controle), PROB (probiótico), PE-RAR e PE-RARPROB. Nos grupos PE-RAR e PE-RAR-PROB, a PE foi induzida pela colocação de ligaduras de seda ao redor dos primeiros molares inferiores dos animais. No 14&deg; dia, as ligaduras foram removidas e realizou-se a RAR. Nos animais dos grupos PROB e PE-RARPROB, o probiótico B. lactis HN019 foi administrado diariamente (10 mL/dia de 109 unidades formadoras de colônia) por 15 dias tendo seu início no 14&deg; dia do experimento. Os animais de todos os grupos foram submetidos à eutanásia 29 dias após o início do experimento. As hemimandíbulas e amostras de intestino delgado foram coletadas. Foram realizadas análises histomorfométricas, microtomográficas e imunohistoquímicas. Foram investigados, também, os efeitos microbiológicos de B. lactis HN019 no biofilme associado às ligaduras durante o desenvolvimento da PE. Todos os dados foram analisados estatisticamente. O Grupo PE-RAR-PROB apresentou menores reabsorção óssea alveolar e perda de inserção conjuntiva quando comparado ao Grupo PE-RAR, bem como menor número de osteoclastos, maior expressão de citocinas anti-inflamatórias e menor expressão de citocinas pró-inflamatórias (p <0,05). No grupo PE-RAR-PROB, os valores médios da profundidade da cripta do jejuno e duodeno foram significativamente maiores que aqueles do grupo PE-RAR. A proporção de bactérias aeróbias/anaeróbias foi maior nas amostras de biofilme de animais tratados com B. lactis HN019 em relação àquelas de animais não tratados (p <0,05). Dentro dos limites deste estudo, pode-se concluir que a utilização de B. lactis HN019 como adjuvante à RAR promove benefícios histológicos, microtomográficos e imunológicos adicionais no tratamento da PE em ratos, bem como melhora a morfologia intestinal. / Lactobacillus probiotics have been investigated in periodontitis. However, the effects of the genus Bifidobacterium on periodontitis are hardly known. This study evaluated the effects of the probiotic Bifidobacterium animalis subsp. lactis (B. lactis) HN019 as an adjunct to scaling and root planing (SRP) in rats with experimental periodontitis (EP). At baseline, 32 rats were assigned to 4 groups: C (control), PROB, EP-SRP and EPSRP- PROB. In groups EP-SRP and EP-SRP-PROB, the mandibular first molars of the animals received a ligature. At day 14, the ligatures were removed and SRP was performed. Animals of groups PROB and EP-SRP-PROB were orally administered with 10 mL/day of 109 colony forming units of B. lactis HN019 for 15 days, starting at day 14. Animals were euthanized at day 29. The jaws and samples of the duodenum, jejunum, and ileum were resected. Histomorphometric, microtomographic and immunohistochemical analyses were performed. Microbiological effects of B. lactis on biofilm were also evaluated. Data were statistically analyzed. Group EP-SRP-PROB presented reduced alveolar bone resorption and attachment loss when compared with Group EP-SRP (p<0.05). Group EP-SRP-PROB showed significantly fewer osteoclasts, increased expression of anti-inflammatory cytokines and reduced expression of proinflammatory cytokines compared with Group EP-SRP (p<0.05). In group EP-SRPPROB, the mean values of crypt depth of the jejunum and dudoenum were significantly higher than the ones from group EP-SRP. B. lactis promoted a higher ratio between aerobic and anaerobic bacteria in biofilm samples (p<0.05). Within the limits of this study it can be concluded that the use of B. lactis HN019 as an adjunct to SRP promotes additional histologic, microtomographic and immunologic benefits in the treatment of EP in rats and improves the intestinal morphology.