O receptor tipo 2 ativado por protease (PAR2) é um receptor pro-inflamatório que pode ser ativado por tripsina, triptase, protease 3 produzida pelo neutrófilo e pela gingipaína (produzida por Porphyromonas gingivalis). Objetivo: o objetivo do presente estudo foi investigar a expressão do PAR2 na periodontite crônica em humanos, e avaliar se esta expressão está relacionada com a presença de atividade proteolítica no fluido crevicular. Metodologia: foram coletadas amostras de fluido crevicular gengival de indivíduos do grupo controle (sítios saudáveis com profundidade de sondagem &#8804; 3mm, ausência de sangramento à sondagem; n=40), grupo periodontite crônica com destruição de leve a moderada (n=40) e grupo periodontite crônica com destruição avançada (n=40). A expressão do PAR2 foi determinada por RT-PCR e a atividade proteolítica no fluido crevicular foi analisada utilizando-se o substrato BApNA. Resultados: observou-se um aumento significativo (p<0.001) da expressão do PAR2 no grupo periodontite em relação ao grupo controle, independentemente da severidade da doença. Além disso, no grupo periodontite houve um aumento significativo na atividade proteolítica (p<0.001) presente no fluido crevicular comparado com o grupo controle. Conclusões: concluiu-se que na periodontite crônica, há um aumento da expressão do PAR2 e um aumento da atividade proteolítica do fluido crevicular. Desta forma, sugere-se que o PAR2 pode estar envolvido com a inflamação periodontal em humanos. / Proteinase-activated receptor-2 (PAR2) is a pro-inflammatory receptor that can be activated by trypsin, tryptase, neutrophil-serine protease-3 (P3), and gingipain (produced by Porphyromonas gingivalis). Objectives: the objective of the present study was to investigate the expression of PAR2 in chronic periodontitis, and to evaluate whether its expression is related to the presence of proteolytic activity at the crevicular fluid. Methodology: gingival crevicular fluid (CF) samples were collected from subjects from control group (health sites with probing depth &#8804; 3mm, absence of bleeding on probing; n=40), chronic periodontitis group with slight to moderate destruction (n=40), and chronic periodontitis group with advanced destruction (n=40). PAR2 expression was determined by reverse transcriptase-PCR, and the proteolytic activity (PA) at the CF was quantified using the specific substrate BApNA. Results: PAR2 expression was significantly higher (p<0.001) in periodontitis compared to healthy individuals, independently of the diseases severity. In addition, in periodontitis group, there was a significantly increased proteolytic activity (p<0.001) compared with controls. Conclusions: we conclude that in chronic periodontitis there is an increased expression of PAR2 and an increased proteolytic activity at the crevicular fluid. Thus, it is suggested that PAR2 might be involved in human periodontal inflammation.

O presente estudo submeteu três cães ao afastamento gengival comparando a técnica de afastamento e moldagem com anel de cobre e godiva de baixa fusão com a técnica de afastamento gengival com cordão de algodão associado a cristais de cloreto de alumínio, e moldagem única com silicona de condensação. O objetivo foi comparar a biocompatibilidade periodontal entre as duas técnicas. Foram obtidas amostras nos períodos de 0 (zero), 3, 7, 14 e 21 dias, as quais foram avaliadas histologicamente, utilizando-se a técnica de coloração hematoxilina e eosina. Como resultado, para todos os métodos testados, as conseqüências histológicas para o periodonto foram consideradas reversíveis. / This study carried out three dogs to gingival displacement comparing the copper band and compound gingival displacement and impression technique with string impregnated in aluminium chloride with gingival displacement technique, and single impression by condensing silicone impression material. The objective was to compare the periodontal biocompability between the two techniques. Blocks specimens including the teeth and the soft tissue were taken immediately, 3, 7, 14 e 21 days, and had histological analysis with stain hematoxilin and eosin. The results showed that all groups had reversible histological consequences.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-03-10T18:03:44Z No. of bitstreams: 1 cristinadepaulanovaes.pdf: 2904372 bytes, checksum: 00165445711c1a78d289bd32e6a3e37a (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-03-13T19:22:11Z (GMT) No. of bitstreams: 1 cristinadepaulanovaes.pdf: 2904372 bytes, checksum: 00165445711c1a78d289bd32e6a3e37a (MD5) / Made available in DSpace on 2017-03-13T19:22:11Z (GMT). No. of bitstreams: 1 cristinadepaulanovaes.pdf: 2904372 bytes, checksum: 00165445711c1a78d289bd32e6a3e37a (MD5) Previous issue date: 2016-11-09 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Introdução: A doença periodontal (DP) e mucosite oral (MO) são condições imunoinflamatórias que podem ser caracterizadas pela presença constante de inflamação sistêmica e bacteremia. Objetivo: Avaliar o impacto da terapia periodontal na MO e investigar a ocorrência de patógenos periodontais em pacientes submetidos à quimioterapia para tumores sólidos e prévia ao transplante de medula óssea (TMO) Métodos: Avaliação dos dados sócio demográficos, dados da quimioterapia e comorbidades, além de avaliação sistemática orofacial. Estas ocorreram previamente e entre 7 a 14 dias após início da quimioterapia no grupo saúde periodontal (GSP). No grupo doença periodontal (GDP) somente após início da quimioterapia, 7 a 14 dias. A mucosa oral foi examinada para avaliar a presença e grau da MO induzida por quimioterapia. Ainda, testes clínicos para diagnóstico de disfagia, avaliação de xerostomia e qualidade de vida relacionada à saúde bucal (OHIP-14); e classe socioeconômica dos pacientes. Foram coletadas em ambos grupos, amostras subgengivais através da inserção de cones de papel estéreis, nos períodos descritos acima. Análise e quantificação da densidade bacteriana para cada espécie e número total de patógenos periodontais do “complexo vermelho” determinadas através do ensaio biomolecular de FISH (hibridação in situ fluorescente). Resultados: 51 pacientes com idade média de 51.1, sendo 38(74.5%) do sexo feminino e classe socioeconómico C 22(43.1%) predominante. Mama foi o local do tumor primário mais acometido 21(41.1%) e a doxorrubicina agente quimioterápico 22(43.1%) prevalente. A morbidade mais associada a hipertensão 20(39.2%). GSP incluiu 33 pacientes e GDP 18 pacientes. Análises estatísticas revelaram que as distribuições dos pacientes em relação à incidência de MO não diferiu entre estes grupos. Grau 1 de MO foi o mais incidente. Dezesseis(31%) pacientes desenvolveram MO, 9(56.2%) submetidos ao regime de condicionamento quimioterápico prévio ao transplante, 7(43.7%) em quimioterapia comum para tumores sólidos. Na qualidade de vida relacionada à saúde bucal, pacientes com complicações bucais mostraram média OHIP-14 de 1.36, enquanto aqueles sem complicações bucais média de 0.56, após quimioterapia para tumores sólidos. Conclusão: A saúde periodontal, pode afetar a incidência e grau da MO induzida por quimioterapia. Observando-se apenas pacientes com saúde periodontal submetidos ao regime de condicionamento quimioterápico e quimioterapia convencional, encontrou-se menor incidência de MO, comparado a evidências científicas. A necessidade de inclusão de pacientes com periodontite grave para avaliar essa relação dificultou tal julgamento. / Background: Periodontal disease (PD) and oral mucositis (OM) are immunoinflammatory conditions that can be characterized by the constant presence of systemic inflammation and bacteremia. Objective: Evaluate the impact of periodontal therapy in oral mucositis and investigate the occurrence of putative periodontal pathogens in patients undergoing chemotherapy for solid tumors and scheduled for hematopoietic stem cell transplantation (HSCT). Methods: This study included an evaluation of demographic data, data chemotherapy and comorbidities, and global systematic dental assessment. These were obtained prior and between 7- 14 days after initiated chemotherapy in periodontal health group (PH group). The Periodontal disease Group (PD Group) was evaluated during 7 to 14 days after of chemotherapy. The oral mucosa also was examined for scoring of chemotherapyinduced oral mucositis. Furthermore clinical tests for dysphagia diagnosis and evaluation of xerostomia and quality of life related to oral health; socio-economic class of patients was determined. Subgengival samples were collected by inserting sterile paper points for both groups, for same periods described above. Total number of bacterial cells and cells from each species determined by FISH (fluorescence in situ hybridization). Results: Fifty-one patients with solid tumors and scheduled for HSCT were assessed to evaluate the impact of periodontitis in OM. Mean age was 51.12, 38(74.5%) were female and socio-economic Class C 22(43.1%) was prevalent. The primary tumor site was most commonly breast cancer 21(41.1%) and prevalent chemotherapeutic agent was doxorubicin 22(43.1%). The prevalent morbidity associated was hypertension 20(39.2%). PH Group included 33 patients and PD Group enrolled 18 patients. Statistical analyses revealed that the distributions of patients with incidence of OM did not differ between both groups. The majority incidence of grade 1 chemotherapy-induced OM was observed. Sixteen(31%) patients developed oral mucositis, 9(56.2%) in patients undergoing conditioning regimen prior HSCT, and 7(43.7%) in patients undergoing common chemotherapy for solid tumors. Patients with oral complications showed a mean OHIP-14 index of 1.36, while those who did not develop oral complications had a mean OHIP-14 of 0.56, after chemotherapy for solid tumors. Conclusion: This study demonstrated that oral health, could affect experience and incidence of chemoinduced OM. When only patients periodontally healthy undergoing conditioning regimen of therapy and conventional chemotherapy were observed, was noted a lower incidence of oral mucositis, when compared with scientific evidences. The need of inclusion of patients with severe periodontitis to evaluate this relationship hampers such trial.

Este estudo transversal teve por objetivo avaliar e relacionar entre si densidade mineral óssea (DMO), composição corporal e doença periodontal em 22 pacientes do gênero masculino, de 22 a 48 anos, portadores de obesidade, com IMC40kg/m2, candidatos à cirurgia bariátrica, atendidos no Hospital Amaral Carvalho (HAC) pelo Sistema Único de Saúde (SUS). Foram realizadas as avaliações antropométrica, do DXA e da condição periodontal. As variáveis qualitativas e quantitativas foram analisadas por meio da correlação linear pelo índice de correlação de Pearson, considerando o intervalo de confiança de 95% e nível de significância de 5% (p<0,05). O peso médio foi de 148kg (122-198); IMC médio de 48,8 (40,5-62,5); CMO, em média, de 3,31kg (2,7-4,1); DMO de corpo total média de 1,3 g/cm2 (1,1-1,5); e Z-score médio de 1,2 (0,0-2,9). A média de gordura corporal total foi de 43,3% (37,4-52,1), e a porcentagem de massa livre de gordura total foi de 56,6 (47,9-82,3). O IMG (Índice de massa gorda) médio foi de 20,6 (14,4-27,0), relação A/G de 1,2 (1.0-1,6) e o peso de tronco médio, 85,8kg (68,3-116,2). Dos pacientes, 29% apresentaram doença periodontal severa. Na casuística, ocorreu uma diferença a menor de 4,5kg (1,0 a 7,8) no peso de DXA comparado ao peso antropométrico, o que corresponde a uma diferença média de 3,06% (0,05-5,27). Na análise de correlação da avaliação odontológica vs resultado DXA, obteve-se apenas correlação significativa entre o índice de sangramento gengival e o IMG (p=0,031, R= -0,525). Pode-se concluir que o índice de massa corporal esteve inversamente relacionado ao sangramento gengival. Estudos futuros deverão ser conduzidos para esclarecer melhor estes achados. / The present cross-sectional study was aimed at evaluating and establishing the relation between bone mineral density (BMD), body composition and periodontal disease in 22 male patients, aged 22-48 years, with obesity (BMI of 40kg / m2), eligible for bariatric surgery, at Hospital Amaral Carvalho Hospital (HAC) by the Brazilian Unified Health System (SUS). Patients underwent anthropometric evaluations, whole-body DXA and periodontal assessment. Qualitative and quantitative variables were analyzed by means of linear correlation analyzes using the Pearsons correlation coefficient with 95% confidence interval and significance level of 5% (p <0.05). The data analyzes were performed with the aid of the SPSS v.22 program. Mean patients weight was 148kg (122-198); mean BMI, 48.8 (40.5-62.5); mean BMC, 3.31kg (2.7-4.1); mean whole-body BMD, 1.3 g / cm2 (1.1-1.5); and mean Z-score, 1.2 (0.0-2.9). Mean total body fat was 43.3% (37.4-52.1), and the percentage of total lean mass was 56.6 (47.9-82.3). Mean FMI (fat mass index) was 20.6 (14.4- 27.0); A/G ratio 1.2 (1.0-1.6) and mean trunk weight, 85.8 kg (68.3-116, 2). In the present study, 29% of the patients presented with severe periodontal disease. In the sample, there was a shortfall of 4.5 kg (1.0 to 7.8) in the DXA weight compared to anthropometric weight, which corresponds to an average difference of 3.06% (0.05- 5.27). In the correlation analysis between Dentistry assessment and DXA score, only a significant correlation was found between gingival bleeding index and FMI (p = 0.031, R = -0.525). It could be concluded that the body mass index was inversely related to de gingival bleeding. Future studies are needed to better clarify these findings.

Philosophiae Doctor - PhD / More than 20 million infants in the world (15.5 % of all births) are born with low birth weight. Ninety-five % of them are in developing countries. Oral colonization of Gramnegative anaerobes has been implicated as a risk factor for preterm delivery of low birth weight (PLBW) infants. The objective of this study was to investigate the association between periodontal pathogens and pre-term delivery of low birth weight (PLBW) infants. The study sample included 200 randomly selected women admitted to the department of obstetrics-gynecology of the teaching hospital of Butare in Rwanda. Mothers were asked to complete a questionnaire in order to identify factors which might pose a health risk to them and their infants. Gingival crevicular fluid (GCF) was collected from each quadrant of the mother’s month (using paper points) within 24 hours of delivery. Ten ml of foetal cord serum samples were collected at delivery and 10 ml of maternal serum samples were collected within 48 of delivery. GCF was examined by PCR for the presence of 5 periodontopathogens and ELISA was used for the evaluation of cytokines (IL-6 and IL-10) and immunoglobulins (IgM, IgG) in foetal cord and maternal blood against the periodontopathogens. P. intermedia showed significant associations either on its own or in combinations with most indicators of periodontal disease used in this study, while Aa and members of the red complex were significantly associated with gum bleeding and reduced frequency of tooth brushing. A strong association between PLBW and maternal and foetal cord serum sample levels of IL-10 was observed. Also, a good association was observed between PLBW and FCB sample levels of IL-6. Significant associations were observed between PLBW and maternal IgG against the different peridontopathogens. The findings of this study may suggest that the levels of maternal IgG and foetal IgM against the different periodontopathogens are associated with dissemination of maternal periodontopathogens to the foetus thereby illiciting an inflammatory response which contributes to PLBW. / South Africa

Philosophiae Doctor - PhD / More than 20 million infants in the world (15.5 % of all births) are born with low birth weight. Ninety-five % of them are in developing countries. Oral colonization of Gramnegative anaerobes has been implicated as a risk factor for preterm delivery of low birth weight (PLBW) infants. The objective of this study was to investigate the association between periodontal pathogens and pre-term delivery of low birth weight (PLBW) infants. The study sample included 200 randomly selected women admitted to the department of obstetrics-gynecology of the teaching hospital of Butare in Rwanda. Mothers were asked to complete a questionnaire in order to identify factors which might pose a health risk to them and their infants. Gingival crevicular fluid (GCF) was collected from each quadrant of the mother’s month (using paper points) within 24 hours of delivery. Ten ml of foetal cord serum samples were collected at delivery and 10 ml of maternal serum samples were collected within 48 of delivery. GCF was examined by PCR for the presence of 5 periodontopathogens and ELISA was used for the evaluation of cytokines (IL-6 and IL-10) and immunoglobulins (IgM, IgG) in foetal cord and maternal blood against the periodontopathogens. P. intermedia showed significant associations either on its own or in combinations with most indicators of periodontal disease used in this study, while Aa and members of the red complex were significantly associated with gum bleeding and reduced frequency of tooth brushing. A strong association between PLBW and maternal and foetal cord serum sample levels of IL-10 was observed. Also, a good association was observed between PLBW and FCB sample levels of IL-6. Significant associations were observed between PLBW and maternal IgG against the different peridontopathogens. The findings of this study may suggest that the levels of maternal IgG and foetal IgM against the different periodontopathogens are associated with dissemination of maternal periodontopathogens to the foetus thereby illiciting an inflammatory response which contributes to PLBW.

Os tecidos periodontais estão em confronto continuo com microorganismos capazes de disparar mecanismos da resposta imune inata, dando origem ao infiltrado inflamatório. Estudos recentes mostraram a importancia dos receptores do tipo Toll (TLRs) na fase inicial de reconhecimento de diferentes patogenos. A participação de receptores tipo Toll (TLRs) na resposta de neutrófilos e macrófagos frente a periodontopatógenos precisa ser determinada. Nesse estudo procuramos caracterizar o infiltrado inflamatório presente no peritônio de animais deficientes de TLR4-/-, avaliar a atividade fagocítica, bem como a produção de óxido nítrico (NO) e a atividade de mieloperoxidase (MPO) no curso da infecção por Aggregatibacter actinomycetemcomitans. A ausência de TLR4 não influenciou a quimiotaxia de neutrófilos e macrófagos para o local da infecção, a produção de óxido nítrico, a atividade de MPO e a viabilidade celular. No entanto, neutrófilos e macrófagos de animais TLR4-/- apresentaram menor atividade fagocítica quando comparado ao grupo controle (camundongos WT). Em relação a doença periodontal induzida experimentalmente com Aggregatibacter actinomycetemcomitans em camundongos deficientes de TLR4, os resultados mostraram que 100% dos animais deficientes de TLR4 sobreviveram a infecção durante o período de observação. Em relação a análise de perda óssea, os dados revelaram uma menor perda progressiva de osso alveolar na região dos molares de animais deficientes de TLR4. A ausência do receptor interferiu na disseminação da bactéria, uma vez que se observou um grande número de bacilos no linfonodo e baco dos animais que não expressaram TLR4, diferente do observado para os animais selvagens (WT). Os resultados indicam a importância da sinalização via TLR4 durante a resposta imune contra Aggregatibacter actinomycetemcomitans. / Aggregatibacter actinomycetemcomitans is an oral gram negative bacteria that contributes to periodontitis progression. Isolated antigens from A. actinomycetemcomitans could be activating innate immune cells through Toll-like receptors (TLRs), molecules that recognize structural components conserved among microorganisms. In this study, we evaluated the role of TLR4 in the recognition of Aggregatibacter actinomycetemcomitans. Neutrophils and macrophage from TLR4 deficient mice and WT mice were collected and used for the subsequent assays. The phagocytosis of leukocytes against A. actinomycetemcomitans and the presence of apoptotic cells were determined by flow cytometry. The in vivo and in vitro production of NO and MPO was evaluated 24h after A. actinomycetemcomitans challenge. In addition, we examined the mechanisms that modulate the outcome of A. actinomycetemcomitans-induced periodontal disease in TLR4-/- mice. The results show that inflammatory cells influx in peritoneal cavity of TLR4-/- mice was similar to that observed into their littermate controls. The phagocytic activity was diminished by cells from TLR4-/- mice. In addition, we did not observe difference in NO and MPO production and the frequency of apoptotic cells between cells from TLR4-/- and WT mice. The results showed that TLR4-deficient mice developed less severe periodontitis after A. actinomycetemcomitans infection, characterized by significantly lower bone loss and inflammatory cell migration to periodontal tissues. Together, these data demonstrate the role TLR4 signals for neutrophils activation after A. actinomycetemcomitans infection and development of periodontal disease.

Hipótese: A principal hipótese deste estudo foi que o uso de enxaguatório placebo associado ao tratamento periodontal não cirúrgico é tão eficaz quanto o uso de clorexidina para bochecho. Objetivo: O objetivo deste ensaio clínico controlado randomizado foi avaliar a resposta placebo em dois modelos terapêuticos periodontais; terapia periodontal mecânica convencional por quadrante e `one-stage full-mouth disinfection. Metodologia: Para responder esta hipótese foram incluídos indivíduos com periodontite crônica distribuídos aleatoriamente em seis grupos, Full mouth (Full Mouth disinfection (FMD), Full Mouth placebo (FMP) e Full Mouth Controle (FMC) e quadrante (Raspagem convencional Clorexidina (RCC), raspagem convencional placebo (RCP) e raspagem convencional (RC), sendo cada grupo com 10 indivíduos. Mensurações tanto no momento inicial (baseline), 90 e 180 dias após os procedimentos de raspagem foram considerados na avaliação geral da resposta placebo em toda a população. Nesta análise foram consideradas medidas clínicas de profundidade de sondagem, nível de inserção clínico, índice de placa e gengival. Em adição, o impacto do efeito placebo foi também investigado com relação à avaliação das analises salivares de proteína total, atividade argininolítica e nitrito. A avaliação de dor/ansiedade em função da intervenção, qualidade de vida e impacto sobre a saúde bucal foram também objeto de análise durante todo o ensaio. O experimento foi conduzido no delineamento casualizado com 10 repetições. Os dados foram submetidos à análise de variância, os modelos de regressão foram testados pelo teste de Fischer, os coeficientes angulares dos modelos de regressão foram testados pelo teste t, as médias foram comparadas pelo teste Duncan a 5% de probabilidade e a análise de correlação teve a significância testada pelo teste t. Resultados: Os grupos FMP e RCP tiveram uma tendência à aumentar a atividade da arginase aos 90 dias e diminuir aos 180 dias, porém na concentração de nitrito foi observada uma diminuição em todos os tempos. Nos parâmetros clínicos observou-se uma diminuição de suas médias em todos os grupos avaliados. Com relação aos questionários observou-se uma tendência às respostas em escores que indicavam pouca dor pós-tratamento e a maioria respondeu concordar que a saúde bucal está relacionada com a qualidade de vida. Conclusão: Todos os grupos apresentaram melhora nas médias, relacionadas aos parâmetros clínicos, após o tratamento periodontal, porém não foi estatisticamente significante. Os grupos placebo não apresentaram melhoras significantes que se possa afirmar sobre a influência da resposta placebo, observando-se uma resposta negativa à nossa hipótese. / Hypothesis: The main hypothesis of this study was that the use of mouthwash placebo combined with non-surgical periodontal treatment is as effective as the use of chlorhexidine mouthwash. Objective: The objective of this randomized controlled clinical trial was to evaluate the placebo response in two periodontal therapeutic models; conventional mechanical periodontal therapy quadrant and `one-stage full-mouth disinfection. Methodology: To answer this hypothesis, subjects with chronic periodontitis were randomly divided into 6 groups, Full mouth (FMD, and FMC FMP) and quadrant (RCC, RC and RCP), each group had 10 subjects in. Measurements were taken at baseline (baseline), 90 and 180 days after scraping procedures were considered in the overall evaluation of the placebo response in the whole population. This analysis considered clinical measures of probing depth, clinical attachment level, plaque index, gingival. In addition, the impact of placebo effect was also investigated in the analysis of the salivary total protein and activity argininolítica nitrite assessment. The assessment of pain / anxiety due to the intervention, quality of life and impact on oral health were also analyzed throughout the test. The experiment was conducted in randomized design with 10 replications. The data was subjected to analysis of variance, regression models were tested by Fischers test, the slopes of the regression models were tested by t test, means were compared by Duncan test at 5% probability and correlation analysis had its significance tested by t test. Results: The FMP and RCP groups had a tendency to increase the activity of arginase at 90 days and decrease at 180 days, but the nitrite concentration decrease was observed at all times. In clinical parameters a decrease in their average in all groups evaluated was observed. Regarding the questionnaires, they revealed a trend in the responses that indicated little pain scores after treatment and most responses agreed that oral health is related to quality of life. Conclusion: All groups showed improvement in average, related to clinical parameters after periodontal treatment, however it was not statistically significant. The placebo group showed no significant improvements that can be said about the influence of the placebo response, observing a negative answer to our hypothesis.

Indiana University-Purdue University Indianapolis (IUPUI) / Periodontal disease is the leading cause of tooth loss in adults. Bacterial biofilm on tooth surfaces is the primary initiator of periodontal disease. Various factors contribute to the severity of periodontal disease including the different virulence factors of the bacteria within the biofilm. In the progression of periodontal disease, the microflora evolves from a predominantly Gram positive microbial population to a mainly Gram negative population. Specific gram negative bacteria with pronounced virulence factors have been implicated in the etiology and pathogenesis of periodontal disease, namely Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola which form the red complex of bacteria. The orange complex bacteria become more dominant in the maturation process of dental plaque and act to bridge the early colonizers of plaque with the later more dominant red complex bacterial and consists of such bacteria as Campylobacter showae, Campylobacter rectus, Fusobacterium nucleatum and Prevotella intermedia. Perhaps the most investigated contributing factor is the relationship between smoking and periodontal disease. When examining the association between cigarette smoking and interproximal bone loss, greater bone loss is associated with higher cigarette consumption, longer duration (i.e., pack year history) and higher lifetime exposure. The presence of various virulence factors such as the production of a capsular material, as well as the proteolytic activity of the various periopathodontic bacteria has been associated with the pathogenesis of periodontitis. Even though many different enzymes are produced in large quantities by these periodontal bacteria, trypsin-like enzymes, chymotrypsin-like enzymes and elastase-like enzymes, as well as dipeptidyl peptidase-like enzymes, have been thought to increase the destructive potential of the bacterium and mediate destruction of the periodontal apparatus. More specifically, it is hypothesized that the proteolytic activity of other clinically important periodontal pathogens, such as Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas assacharolyticus, is increased in the presence of nicotine. The purpose of this study was to determine the effects of nicotine on F. nucleatum, P. intermedia and P. assacharolyticus proteolytic activity. Cultures were maintained on anaerobic blood agar plates containing 3% sheep blood. Bacterial cells were harvested from the plates and washed. Washed F. nucleatum, P. intermedia and P. assacharolyticus cells were incubated with 1 mg/ml of nicotine. Bacterial cells not incubated with nicotine were used as positive controls. Secreted enzymatic activity was measured using the synthetic chromogenic substrates glycyl-L-proline-p-nitroanilide (GPPNA), N-succinyl-L-alanyl-L-alanyl-L-alanyl-p-nitroanilide (SAAAPNA), N-succinyl-alanine-alanine-proline-phenylalanine-p-nitroanilide (SAAPPPNA) and N-α-benzoyl-L-arginine-p-nitroanilide (L-BAPNA) (Sigma-Aldrich Products, St. Louis, MO, USA). Appropriate means and standard deviations were determined for each of the enzymatic activities measured and analysis of variance (ANOVA) was used to compare the groups utilizing a 5% significance level for all comparisons. Results demonstrated that after 60 minutes of incubation of F. nucleatum, P. intermedia and P. assacharolyticus cells with 1 mg/ml of nicotine and the various synthetic substrates, had the following proteolytic activity for GPPNA: 0.83 ± 0.14, 0.72 ± 0.03 and 0.67 ± 0.10, respectively; SAAAPNA: 0.82 ± 0.06, 0.76 ± 0.05 and 0.68 ± 0.08, respectively; SAAPPPNA: 0.90 ± 0.13, 0.85 ± 0.17 and 0.72 ± 0.03, respectively; and BAPNA: 0.81 ± 0.15, 0.74 ± 0.13 and 0.74 ± 0.16, respectively. In conclusion, the results indicate that in the presence of 1 mg/ml of nicotine, the proteolytic activity of F. nucleatum and P. assacharolyticus was increased with all of the synthetic substrates (with statistical significance seen only in the increases with F. nucleatum and GPPNA, SAAAPNA and BAPNA). The proteolytic activity exhibited an increasing trend in activity for P. intermedia with SAAPPPNA and BAPNA but a decreasing trend in activity with GPPNA and SAAAPNA when incubated with 1 mg/ml of nicotine, once again demonstrating no statistical significance for any of the substrates. Therefore, it could be concluded that based on these results nicotine at a concentration of 1 mg/ml may increase the proteolytic activity of periodontal pathogens and thus may increase periodontal disease activity and subsequent periodontal breakdown. Further studies are needed to validate these results utilizing different concentrations of nicotine.

Background: Periodontal disease has been associated with gestational complications and both conditions have a high prevalence in rural populations from developing regions. A cross-sectional study was carried out to explore the relationship between periodontal inflamed surface area (PISA), blood pressure (BP), and, serum uric acid levels (UA) in a group of rural North Chinese pregnant women in the third trimester of pregnancy. Methods: Three hundred and thirty-five rural women aged 20–34 years, with normal body mass index (BMI) were examined in a cross-sectional study during their third trimester of gestation. Exclusion criteria were history of pregnancy complications,multiple pregnancy, smoking habits, diabetes, hypertension or any known infectious disease. Socio-demographic variables, including age and socioeconomic status (SES), systolic blood pressure (SBP) and diastolic blood pressure (DBP) readings, serum UA levels, and PISA values were recorded. A structural equation model was implemented with two constructed latent variables including “Dem” (comprising of age and SES category to represent unobserved demographic variables) and, “BP” (comprising of SBP and DBP to account for measurement error and lack of multiple BP readings). The model accounted for co-variance of BP and UA, and implemented simultaneous regressions for BP and UA as outcomes, upon Dem and PISA values as exogenous variables. Results: The median PISA score was 1,081.7 (IQR = 835.01), reflecting high levels of periodontal inflammation in the sample. SEM showed a significant association of PISA with BP (estimate = 0.011, 95%CI = 0.009–0.012 p < 0.001) and UA (estimate = 0.001, 95% CI = 0.001–0.001, p < 0.001). Conclusion: Higher PISA values were significantly associated with higher blood pressure and uric acid levels among rural pregnant women in a cross-sectional sample from a center in North China after accounting for a latent demographic construct derived from age and SES.