Orientadores: Getulio da Rocha Nogueira Filho, Francisco Humberto Nociti Junior / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-10T07:19:34Z (GMT). No. of bitstreams: 1 Peruzzo_DaianeCristina_D.pdf: 1082596 bytes, checksum: 5a19d83c292b5bded4f4f59bf5a943c6 (MD5) Previous issue date: 2008 / Resumo: Estudos em animais e epidemiológicos têm sugerido que o estresse pode alterar o estabelecimento e a progressão da doença periodontal (DP). Entretanto, dados relacionados ao efeito do estresse e seus mecanismos envolvidos na DP ainda são limitados. Os objetivos deste estudos foram: i) revisar sistematicamente a literatura sobre a influência do estresse crônico (EC) na DP; ii) analisar o impacto do EC induzido em ratos na progressão da DP e na regulação de genes relacionados à progressão da doença, bem como na alteração dos biomarcadores do estresse (catecolaminas e corticoesterona); iii) avaliar a viabilidade do uso da droga metirapone (MT), como um modelo experimental em ratos, para inibir a produção de glicocorticóides (GC), determinando, assim, o efeito do EC nos tecidos periodontais. Para a revisão sistemática, foi realizada uma busca na literatura e os dados dos estudos foram extraídos e avaliados por dois revisores independentes. Para os trabalhos em animais, foram realizados experimentos em ratos machos, Wistar, divididos em grupos com 20 animais cada: controle, DP induzida por ligadura, DP + EC (restrição de movimento e isolamento, 12h/dia) e DP + EC + administração de MT (3 doses/dia de 50mg/Kg). Após 30 dias todos os animais foram sacrificados. Amostras de sangue foram coletadas para mensurar os biomarcadores do EC, o tecido marginal, ao redor dos sítios com e sem ligaduras foi coletado para avaliar a expressão de genes por meio de PCRq (Reação de Polimerase em Cadeia quantitativa) e as mandíbulas foram removidas e fixadas para mensuração histométrica da perda óssea interradicular (POI). Análise dos dados demonstrou que: i) a maioria dos estudos analisados apresentaram um desfecho positivo entre EC e DP; ii) os biomarcadores do estresse, na presença de EC, podem localmente modular a DP por meio de um aumento local nas proporções dos genes pró-inflamatórios e pró-reabsorção, favorecendo, assim a destruição óssea periodontal; e, iii) a administração de MT resultou num importante efeito na redução dos níveis sistêmicos de GC, entretanto, pode-se observar que a administração da droga alterou a expressão de fatores importantes na modulação da DP e conseqüentemente refletiu nos níveis de POI. Dentro dos limites deste estudo, pode-se concluir que o EC significativamente apresenta uma relação com a DP e o aumento local de fatores pró-inflamatórios e pró-reabsorção pode ser o mecanismo envolvido na progressão da doença. Além disso, a administração de MT é capaz de reduzir os níveis sistêmicos de GC, entretanto, modula a expressão de fatores relacionados à progressão da DP, resultando em POI / Abstract: Animal and epidemiological studies have suggested that stress may modify the establishment and progression of periodontal disease (PD). However, data regarding the effect of stress and the mechanisms involved in PD are limited. The aim of this study was: i) to review systematically the literature about the influence of chronic stress (CS) on PD ii) to evaluate the impact of CS, induced in rats, in the progression of PD and regulation of genes related to the disease progression, as well as the variations of stress biomarkers (cathecolamines e corticoesterone); iii) to evaluate the feasibility of the use of metyrapone (MT) as an experimental model to inhibit glucocorticoid (GC) production and, therefore, as a method to determine the effect of CS on periodontal tissues. A systematic literature search was performed and the data of the studies were independently extracted and evaluated by two reviewers. The animal studies were carried out on male Wistar rats assigned to 3 groups with 20 animals each: control, PD induced by ligature; PD associated with CS (restraint stress and isolation, 12 h/day) and PD + CS + MT administration (3 daily doses, 50mg/Kg). After 30 days, all animals were sacrificed. Blood samples were obtained and the concentrations of corticosterone and catecholamines measured as biomarkers of CS, marginal tissues around ligated and non-ligated teeth were harvested and gene expression assessed by qPCR (quantitative Polymerase Chain Reaction) and the jaws were removed and fixed to histometrically determine the interradicular bone loss (IBL). Data analysis demonstrated that: i) the majority of the studies showed a positive outcome between CS and PD; ii) the stress biomarkers may locally modulate PD by an increase of the local ratio of pro-inflammatory and pro-resorptive genes, thus favoring tissue destruction; and, iii) MT administration resulted in an important lowering effect of GC systemic levels, however, it could be observed that MT administration modified the expression of important factors which modulate PD, and consequently reflected the IBL. Within the limits of this study, it may be speculated that CS has a significant relationship with PD and the local increase in pro-inflammatory and pro-resorptive factors can be the mechanisms involved in disease progression. Moreover, MT administration is able to lower systemic levels of GC, however, it modulates the expression of factors related to periodontitis progression, resulting in IBL / Doutorado / Periodontia / Doutor em Clínica Odontológica

Orientadores: Marcio Zaffalon Casati, Sergio de Toledo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-08T14:46:02Z (GMT). No. of bitstreams: 1 Casarin_RenatoCorreaViana_M.pdf: 9678286 bytes, checksum: 95c6838ca5a99289bdbad65404d1cd7e (MD5) Previous issue date: 2007 / Resumo: O objetivo do presente estudo foi avaliar a resposta clínica das lesões de bifurcação proximais tratadas com proteínas derivadas da matriz do esmalte (EMD). Foram selecionados 24 pacientes com pelo menos uma lesão de bifurcação proximal classe II apresentando profundidade de sondagem (PS) = 5 mm e presença de sangramento a sondagem. As furcas foram divididas aleatoriamente em 2 grupos: Grupo controle (n=20) ¿ acesso para raspagem e alisamento radicular e aplicação de EDTA 24% (PrefGel®); Grupo Teste (n=20) ¿ acesso para raspagem e alisamento radicular + aplicação de EDTA 24% + aplicação das EMD (Emdogain®). Os pacientes foram avaliados quanto ao Índice de Placa (IP) e Sangramento à Sondagem (SS), Profundidade de Sondagem (PS), Posição da Margem Gengival (PMG), Nível Clínico de Inserção Vertical e Horizontal Relativo (NICVR e NICHR, respectivamente), Nível Ósseo Vertical e Horizontal (NOV e NOH) e fechamento da lesão. As avaliações foram realizadas antes do tratamento, 2, 4 e 6 meses após. Aos 6 meses de acompanhamento o ganho de NICVR dos grupos controle e teste foram de 0,63 e 0,74 mm, enquanto o ganho de NICHR foram 1,13 e 1,64 mm, ambos sem diferença entre os grupos. O ganho de NOV e NOH para os grupos controle e teste foram de 0,98 e 0,85 mm e 0,94 e 1,08 mm respectivamente, também sem diferença estatística entre os grupos. Na avaliação do número de bifurcações fechadas aos seis meses, no grupo teste houve fechamento de 4 lesões, enquanto no grupo controle não ocorreu nenhum fechamento (p<0,05). Pode-se concluir que, embora os defeitos de bifurcação proximais tenham apresentado ganhos semelhantes de NICVR NICHR, NOV e NOH, o uso do EMD possibilitou um maior fechamento completo das bifurcações que o acesso para raspagem e alisamento radicular e biomodificação radicular com EDTA 24% / Abstract: The aim of the present study was to evaluate the clinical response of proximal furcations treated with enamel matrix derived proteins (EMD). Twenty four patients with at least one class II proximal furcation involvements presenting probing depth (PD) = 5 mm and bleeding on probing were selected. The patients were randomly assigned to: Control Group (n=20) ¿ Open Flap Debridement (OFD) + EDTA 24% conditioning (PrefGel®); Test Group (n=20) ¿ OFD + EDTA 24% conditioning + EMD application (Emdogain®). Plaque Index (PI), Bleeding on Probing (BOP), Probing Depth (PD), Gingival Margin Position (GMP), Relative Vertical and Horizontal Clinical Attachment Level (RVCAL and RHCAL, respectively), Vertical and Horizontal Bone Level (VBL and HBL) and Furcation Closure were evaluated immediately before and 2, 4 and 6 months after the surgeries. At 6th month the gain of RVCAL of control and test group were 0.63 e 0.74 mm, while the RHCAL gain were 1.13 e 1.64 mm, both without statistical difference between the groups. The VBL and HBL gain of control and test group were 0.98 and 0.85 mm and 0.94 and 1.08 mm respectively, also without statistical difference. Four closed furcations were observed at 6th month in the test group, while no closed furcations was observed at control group (p<0.05). It could be conclude that, although the proximal furcations showed similar gains of RVCAL, RHCAL, VBL and HBL at 6th month, the EMD application promote a superior, but unpredictable, frequency of closed furcations than that found with open flap debridement and EDTA 24% conditioning / Mestrado / Periodontia / Mestre em Clínica Odontológica

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Porphyromonas gingivalis à um patÃgeno extremamente associado com a etiologia da periodontite crÃnica e agressiva. O objetivo deste estudo foi avaliar atravÃs de reaÃÃo em cadeia da polimerase em tempo real (Real Time-PCR) a presenÃa de Porphyromonas gingivalis (Pg) e dos genÃtipos fimA II e IV em indivÃduos com periodontite agressiva generalizada (PAG). Quarenta indivÃduos com PAG (29,7  8,1 anos) foram analisados clinicamente - Ãndice de Placa (IP), Ãndice Gengival (IG), profundidade de sondagem (PS), nÃvel de inserÃÃo clÃnico (NIC) - e microbiologicamente, atravÃs de Real Time PCR, quanto à presenÃa de Pg e dos genÃtipos fimA II e IV. Amostras de biofilme subgengival foram colhidas do sÃtio proximal com maior PS e maior NIC. MÃdias de PS e NIC desses sÃtios foram respectivamente: 9,5  2,2 mm e 10,2  2,8 mm. P. gingivalis foi observado em 26 (65%) dos indivÃduos. O genÃtipo fimA II foi verificado em 16 (61,53%) enquanto o genÃtipo fimA IV em 7 (26,92%) dos que apresentaram P. gingivalis. Entretanto, nÃo foi observada diferenÃa estatÃstica entre os parÃmetros clÃnicos dos indivÃduos que apresentaram ou nÃo o microrganismo ou seus respectivos genÃtipos. TambÃm nÃo foi verificada associaÃÃo entre a presenÃa dos genÃtipos e idade ou gÃnero dos pacientes. Os dados sugerem uma associaÃÃo entre genÃtipos fimA II de Porphyromonas gingivalis quando da ocorrÃncia deste microrganismo em indivÃduos com periodontite agressiva generalizada. / Porphyromonas gingivalis is a pathogen strongly associated with the etiology of chronic and aggressive periodontitis. The purpose of this study was to evaluate by Real-Time polymerase chain reaction (Real Time-PCR) the presence of Porphyromonas gingivalis (Pg) and fimA genotypes type II and type IV in patients with generalized aggressive periodontitis (GAgP). Forty individuals with aggressive periodontitis (AgP) (29.7  8.1 years) were clinical analyzed through plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL) and microbiologically, by Real Time-PCR for the presence of Pg and fimA genotypes type II and type IV. Subgingival biofilm samples were collected from the interproximal periodontal sites (> PD and > CAL). The PD and CAL average of this sites were respectively: 9,5  2,2 mm e 10,2  2,8 mm. P. gingivalis was observed in 26 (65%) of individuals. FimA genotypes type II was detected in 16 (61,53%) while fimA genotypes type IV in 7 (26,92%) of those with P. gingivalis. However, no differences were observed between the clinical parameters of patients who presented or not the organism or its genotypes. There was also no association between the presence of genotypes and age or gender of patients. The data suggest an association between P. gingivalis fimA genotypes upon the occurrence of this microorganism in patients with generalized aggressive periodontitis.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Periodontal disease is an infectious-inflammatory disease, and drugs have been studied as modulators of this inflammatory process. In this context, this thesis, constituted by 3 articles had by objective: (1) Perform a review about the effect of Bisphosphonates (BPs) on periodontal disease; (2) Investigate the effect of Alendronate (ALD) on Bone-specfic Alkaline Phosphatase (BALP) on alveolar bone loss (ABL) in rats; (3) Evaluate the effect of ALD and Atorvastatin (ATV) combination on ABL in rats. On study 1, we sought in data basis, using the keywords âBisphosphonatesâ and âPeriodontitisâ, pre-clinical and clinical studies, published in English and Portuguese, in the last 10 years. On study 2, 36 Wistar male rats, submitted to ligature-induced periodontitis, received 0.9% Saline (SAL) or ALD on the doses of 0.01; 0.05; 0.25 mg/kg-s.c., 30 min before ligature placement and daily during 11 days. It was evaluated: ABL (morphometry and histology) serum levels of Bone-specific Alkaline Phosphatase (BALP), transaminases, and Total Alkaline Phosphatase (TAP); and leukogram and corporal mass. On study 3, 78 Wistar male rats, submitted to ligature-induced periodontitis, received prophylactically (P): SAL or ALD (0.01; 0.25 mg/kg-s.c) or ATV (0.3; 27 mg/kg-v.o.) or the combination ALD+ATV (0.25+27; 0.01+0.3; 0.25+0.3; 0.01+27 mg/kg), 30 min before ligature and daily for 11 days; or the combination ALD+ATV (0.01+0.3 mg/kg) administered therapeutically (T), from the 5th day after ligature until the sacrifice. It was evaluated: ABL [morphometry, histology, histometry; immunohistochemistry for tartrate resistant acid phosphatase (TRAP); myeloperoxidase (MPO); BALP, transaminases; Leukogram and corporal mass]. The study 1 showed that BPs presented anti-resorptive and anti-inflammatory effects, reduced FAO and Telopeptide N-terminal of type I collagen (NTx) and improved periodontal clinical parameters. On article 2, ALD (0.25 mg/kg) prevented BALP and ABL reduction, and did not alter transaminases serum levels, but reduced TAP serum levels (p<0.05), it reduced neutrophilia and lymphomonocytosis (p<0.05), without causing important loss of weight. On the 3rd study, the isolated treatments in high doses, and all combinations controlled ABL (p<0.05). Low doses combination of ALD+ATV controlled ABL (P [38.96%] or T [53.53%]). The histological, histometric (p<0.05) and immunohistochemical analysis corroborated macroscopical findings. The low dose combination of ALD+ATV reduced MPO activity, prevented BALP reduction, reduced neutrophilia and lymphomonocytosis (p<0.05), without altering transaminases serum levels and without causing loss of weight. In this way, we can conclude that BPs presented anti-resorptive and anti-inflammatory effects reduced levels of biochemical markers of bone metabolism and improved periodontal parameters. ALD, administered isolated prevented BALP and ABL reduction, without causing systemic problems, and the combination of ALD+ATV, in low doses, reduced ABL and periodontal inflammation, without causing important systemic alterations as well. / A doenÃa periodontal à uma desordem infecto-inflamatÃria, e fÃrmacos tÃm sido estudados como moduladores deste processo inflamatÃrio. Neste contexto, esta tese, constituÃda por 3 artigos, teve por objetivo: (1) Realizar uma revisÃo sobre o efeito de Bisfosfonatos (BFs) na doenÃa periodontal; (2) Investigar o efeito do Alendronato (ALD) nos nÃveis de Fosfatase Alcalina Ãssea (FAO) na perda Ãssea alveolar (POA) em ratos; (3) Avaliar o efeito da combinaÃÃo entre ALD e Atorvastatina (ATV) na POA em ratos. No estudo 1 buscou-se, em bases de dados, utilizando as palavras chave: âBisphosphonatesâ e âPeriodontitisâ, estudos prÃ-clÃnicos e clÃnicos, publicados em lÃngua Inglesa ou Portuguesa, nos Ãltimos 10 anos. No estudo 2, 36 ratos Wistar machos, submetidos à periodontite induzida por ligadura, receberam soluÃÃo Salina (SAL) 0,9% ou ALD nas doses de 0,01; 0,05; 0,25 mg/kg-s.c, 30 min antes da colocaÃÃo do fio e diariamente por 11 dias. Avaliou-se: POA (morfometria e histologia); nÃveis sÃricos de FAO, transaminases e Fosfatase Alcalina Total (FAT); Leucograma e Peso. No estudo 3, 78 ratos Wistar machos, submetidos à periodontite induzida por ligadura, receberam de forma profilÃtica (P): SAL ou ALD (0,01; 0,25 mg/kg-s.c) ou ATV (0,3; 27 mg/kg-v.o.) ou a combinaÃÃo ALD+ATV (0,25+27; 0,01+0,3; 0,25+0,3; 0,01+27 mg/kg), 30 min antes da ligadura e diariamente por 11 dias; ou ainda a combinaÃÃo ALD+ATV (0,01+0,3 mg/kg) na forma terapÃutica (T), ou seja administrada a partir do 5 dia apÃs ligadura, atà o sacrifÃcio. Avaliou-se: POA [morfometria, histologia, histometria; imunohistoquÃmica para fosfatase Ãcido tÃrtaro resistente (TRAP); mieloperoxidase (MPO); FAO, transaminases; Leucograma e Peso]. O artigo 1 mostrou que BFs apresentaram efeitos antirreabsortivo e anti-inflamatÃrio, reduziram FAO e TelopeptÃdeo N-terminal de colÃgeno tipo I (NTx) e melhoraram os parÃmetros clÃnicos periodontais. No artigo 2, o ALD (0,25 mg/kg) preveniu a reduÃÃo de FAO e POA, nÃo alterou nÃveis de transaminases, mas nÃo preveniu reduÃÃo dos nÃveis de FAT (p<0,05), preveniu neutrofilia e linfomonocitose (p<0,05), sem causar perda de peso importante. No 3 estudo, os tratamentos isolados, em altas doses, e todas as combinaÃÃes avaliadas controlaram POA (p<0,05). A combinaÃÃo de ALD+ATV em baixas doses controlou POA (P [38,96%] ou T [53,53%]). As anÃlises histolÃgicas, histomÃtricas (p<0,05) e imunohistoquÃmicas corroboraram os achados macroscÃpicos. A combinaÃÃo de ALD+ATV em baixas doses reduziu a atividade de MPO, preveniu reduÃÃo de FAO, reduziu neutrofilia e linfomonocitose (p<0,05), sem alterar os nÃveis de transaminases e causar perda de peso. Desta forma conclui-se que os BFs apresentaram efeitos antirreabsortivo e anti-inflamatÃrio, reduziram nÃveis de marcadores bioquÃmicos do metabolismo Ãsseo e melhoraram os parÃmetros clÃnicos periodontais. O ALD, administrado isoladamente, preveniu reduÃÃo de FAO, POA, sem repercussÃes sistÃmicas e a combinaÃÃo de ALD+ATV, em baixas doses, reduziu POA e inflamaÃÃo periodontal, tambÃm sem causar alteraÃÃes sistÃmicas importantes.

Tesis Magister en Ciencias Odontológicas con Mención en Periodontologia / Introducción: Estudios epidemiológicos han establecido una asociación entre la periodontitis apical asintomática (PAA) y enfermedades cardiovasculares, particularmente aterogénesis, sin embargo no existe evidencia mecanística que sustente dicha asociación. El objetivo de este estudio fue determinar los niveles de marcadores de inflamación sistémica, endotoxinas y anticuerpos IgG anti-P. endodontalis en suero de voluntarios sanos, y pacientes con PAA con y sin infección endodóntica por P. endodontalis, en línea base y post terapia endodóntica. Materiales y métodos: Se incluyeron 23 pacientes con PAA y 25 voluntarios sin PAA, todos sistémicamente sanos, entre 18-40 años de edad que acudieron a la clínica odontológica de la Facultad de Odontología, Universidad de Chile. Se excluyeron individuos con medicación sistémica, periodontitis crónica (marginal). Se determinaron los niveles de MPO, IL-6, CRP, ICAM-1, VCAM-1s, y Selectina-Es como marcadores de inflamación sistémica mediante ensayo multiplex, niveles de endotoxinas por ensayo LAL y anticuerpos IgG anti-P. endodontalis por ensayo de ELISA. Se determinó la presencia de P. endodontalis mediante PCR y cultivo bacteriano. Se determinó significancia estadística si p<0,05. Resultados: Se encontró una tendencia hacia niveles mayores de marcadores de inflamación sistémica entre PAA e individuos sanos (p>0,05), particularmente para Selectina-Es (p=0,06), mientras que hsCPR alcanzó niveles de riesgo CV medio (1,5mg/L). Los pacientes con PAA infectados con P. endodontalis mostraron un aumento significativo en la presión diastólica (p=0,004) y niveles séricos de VCAM-1s (p=0,02), mientras que IL-6 demostró tendencia al aumento (p=0,06), comparados con los no infectados. La endotoxemia fue significativamente mayor en pacientes con PAA no infectados versus infectados (p=0,02). Se encontró una reducción significativa de los niveles séricos de VCAM-1s tras una semana (p=0,0008) y un mes (p=0,026) post tratamiento respecto de los niveles basales. Asimismo se observó una reducción significativa de los niveles séricos de anticuerpos IgG anti-P. endodontalis una semana post tratamiento en comparación con los niveles basales (p=0,02). Conclusiones: Pacientes con PAA presentaron riesgo CV medio en relación con los controles que presentaron riesgo CV bajo, determinados por los niveles séricos de hsCRP. Pacientes con PAA infectados con P. endodontalis mostraron un aumento significativo en la presión diastólica y en los niveles séricos de VCAM-1s. El tratamiento endodóntico redujo los niveles séricos de VCAM-1s y los niveles séricos de anticuerpos IgG anti-P. endodontalis en pacientes con PAA. El tratamiento endodóntico conservador contribuiría a reducir la inflamación sistémica y el potencial riesgo de disfunción endotelial asociado en pacientes con PAA. / Adscrito Proyecto FONDECYT 1120138 y 1160741

Memoria para optar al título de Bioquímico / La periodontitis es una enfermedad inflamatoria crónica de las estructuras que soportan las piezas dentales, la cual está asociada a un proceso infeccioso causado por un cambio en la ecología local de la biopelícula bacteriana oral. Un 90% de la población chilena presenta signos clínicos de esta patología. Porphyromonas gingivalis (P. gingivalis) se ha propuesto como un agente etiológico clave en la periodontitis. En nuestro laboratorio, se obtuvieron aislados clínicos de P. gingivalis provenientes de individuos sanos, en los que se vio ausencia de la región del antígeno O (AgO) del lipopolisacárido (LPS). En cambio, en aislados clínicos provenientes de pacientes con periodontitis, el LPS está completo. Además, se ha observado anteriormente que cuando células de epitelio gingival (GECs, por sus siglas en inglés) eran infectadas con una cepa silvestre de P. gingivalis (W50) había un aumento del ARNm del Toll-like receptor 4 (TLR4), a diferencia de lo que ocurre cuando son infectadas con una mutante isogénica carente de AgO polimérico (cepa ΔPG1051), sugiriendo que el AgO sería importante para el reconocimiento de la bacteria. Una de las consecuencias de la activación de TLRs es el incremento en los niveles de citoquinas pro-inflamatorias. Esto se ha relacionado con un aumento en la capacidad migratoria en varios tipos celulares, incluyendo a GECs, donde la migración favorece la formación de saco periodontal / Periodontitis is a highly prevalent chronic inflammatory disease in Chile and worldwide that mainly affects the tissues supporting de teeth. This disease is associated with an infectious process caused by a change in the local ecology of the subgingival biofilm. It has been proposed that the bacterium Porphyromonas gingivalis (P. gingivalis) plays a key role in disease onset and progression because this bacterium induces dysbiosis that contributes to the inflammatory process. In a previous study from our group, we observed that the O antigen (OAg) region of the lipopolysaccharide contributes to the inhibition of apoptosis induced by P. gingivalis in epithelial cells, which correlates with an increase in the expression of TLR4. One of the consequences of the activation of TLRs is the increase in the levels of pro-inflammatory cytokines. This has been related to an increase in the migratory capacity of several cell types, including human oral epithelial cells, where enhanced migration is associated with formation of periodontal pockets. Considering this background, the objective was to determine if TLR4 activation mediated by AgO can increase the levels of pro-inflammatory cytokines and to promote migration an oral epithelial cell line OKF6 / TERT2. For this, the level of TLR4 (flow cytometry), TNF-α, IL-8 and IL-1β (multiplex) and migration (Boyden's chamber) were measured after infecting with the previously mentioned strains. Our results show that infection with P. gingivalis does not change the TLR4 levels on the surface of OKF6 / TERT2 cells, independent of the presence or absence of AgO. On the other hand, levels of IL-8 decreased only with the virulent strain of P. gingivalis W50, unlike the isogenic mutant in LPS that does not produce AgO, which does not change them. These changes are independent of TLR4. Finally, the strain with a complete LPS (W50) produces an increase in migration. However, in the absence of AgO there are not significant changes with respect to uninfected cells. In conclusion, in this work we found that the AgO of P. gingivalis participates in the modulation of pro-inflammatory marker levels (IL-8) and cell migration, which could contribute to the development of periodontitis / CONICYT-FONDAP 15130011; Fondecyt 1170925; FIOUCH 17/020

A DNA probe was developed for the identification of the periodontal pathogen, Actinobacillus actinomycetemcomitans. Chromosomal DNA was extracted from A. actinomycetemcomitans, digested with a restriction enzyme, Sau3A, ligated to plasmid DNA (pUC18) and transformed into JM109 cells to give a partial A. actinomycetemcomitans library. The library was screened using Southern blot analysis. Out of the nine inserts tested, one was found to be species specific as it did not cross-hybridise to Haemophilus aphrophilus, a closely related organism which occurs in the normal oral microflora, nor did it cross-hybridise with 7 species of Bacteroides tested. A level of detection of 104 cells or 50ng of A. actinomycetemcomitans was obtained. The probe has a length of 779bp and out of 30 restriction enzymes tested, only SspI was found to have a restriction site in the insert. The probe was tested on clinical specimens obtained from five different periodontitis patient groups and was shown to correlate with culture results in eighteen out of twenty-two cases in detecting A. actinomycetemcomitans.

Objectives: Earlier reports suggested that aggressive periodontitis is common in certain African populations and is associated with the JP2 clone of Aggregatibacter actinomycetemcomitans (Aa). There are few studies that investigated the type of microorganisms that colonize the subgingival sites in young subjects inflicted with a subcategory of aggressive periodontitis that is associated with the Aa-JP2 clone. Hence, the objective of this study was to characterize the subgingival microbiome of JP2 clone-associated aggressive periodontitis. Methods: The study subjects were drawn from a large survey among 14-18 years old schoolchildren in Morocco. The sample included 7 JP2-positive aggressive periodontitis subjects and 14 JP2-negative controls. The controls were selected to be either JP2-positive, JP2-negative (but Aa positive), or Aa-negative. Subgingival samples from these subjects were sequenced for the V1-V3 region (16S rRNA gene) on a Miseq platform. High-quality, non-chimeric merged reads were classified with our previously reported BLASTn-algorithm. Downstream analysis was performed with QIIME and LEfSe. Results: There were no significant differences between the groups in species richness. However, aggressive periodontitis subjects showed significantly lower alpha diversity. The microbiomes of aggressive periodontitis clustered distinctively from the controls. However, there was no significant separation between the subgroups of the control group. Species associated with health included Streptococcus spp., Haemophilus spp., Neisseria spp., Gemella spp., Rothia spp., Fusobacterium nucleatum subsp. polymorphum, Porphyromonas oral taxon 279, Veillonella parvula, Granulicatella adiacens and Lautropia mirabilis. Important periodontal pathogens, including Treponema spp., Fretibacterium spp. P. gingivalis and Tannerella forsythia were significantly enriched in aggressive periodontitis subjects. However, the taxa detected in high abundance and showed strongest association with aggressive periodontitis but not the controls were Pseudomonas oral taxon C61 and Enterobacter cloacae. Conclusions: The results suggest that several periodontal pathogens involved in chronic periodontitis also play a role in aggressive periodontitis. Future studies should investigate the role of Pseudomonas and Enterobacter spp. in the pathogenesis of aggressive periodontitis. / Oral Biology

Periodontal disease is initiated by bacterial plaque that induces a chronic inflammatory condition with subsequent leukocyte infiltration, osteoclast activation and alveolar bone resorption. The ideal treatment for periodontal disease is the complete regeneration of the lost periodontium. Resolvin E1 (RvE1) is an endogenous anti-inflammatory lipid mediator derived from omega-3 fatty acids. Animal studies showed that topical treatment of periodontitis with RvE1 significantly decreased osteoclast counts, prevented alveolar bone loss, and restored lost periodontal tissues including bone. It is not known if RvE1 directly impacts osteoblast functions and bone formation. The objective of this study was to determine RvE1 mechanism of action on osteoblasts. Results showed that topical RvE1 treatment prevented the progression of ligature-induced periodontitis in mice compared to the vehicle. RvE1 receptor, chemR23, was expressed in murine calvaria osteoblasts and the expression was not changed in the inflammatory environment with or without RvE1 treatment. RvE1 treatment resulted in a significant increase in the ALP activity after 2 and 7 days of treatment compared to the control as well as in the inflammatory milieu. Similarly, RvE1 treatment enhanced murine calvaria osteoblasts mineralization in vitro compared to the inflammatory environment. The proliferation of mouse calvaria osteoblasts was significantly increased with RvE1 treatment after 2 and 10 days of treatment compared to the control. Results for evaluating the impact of RvE1 on OPG/RANKL axis showed that RvE1 treatment markedly elevated OPG production compared to the IL-6/IL-6R treatment and decreased RANKL. Overall, RvE1 increased the OPG/RANKL ratio to favor bone formation. RvE1 treatment resulted in a significant increase in the phosphorylation of AKT, ERK1/2 and ribosomal S6 (rS6) kinase. Those pathways were further confirmed via using specific pharmacological inhibitors which showed a significant reduction in OPG production and in the osteoblast proliferation as well. In conclusion, RvE1 has a positive anabolic impact in ligature-induced periodontitis model via direct actions on osteoblasts. RvE1 increases the OPG/RANKL production ratio favoring the bone formation through a pathway that includes phosphorylation of AKT, ERK1/2 and rS6 kinase. The data suggest that RvE1 stimulates bone formation in inflammatory conditions by directly modulating both the osteoclast and osteoblast functions.

Periodontal infection can manifest itself in many different clinical presentations. The aggressive form of this disease is frequently seen in the younger patient population referred for treatment to the Temple University Kornberg School of Dentistry (TUKSD). This study was done to assess the demographics of aggressive periodontitis cases and the types of periodontal treatment methods provided to these patients, antibiotic prescription patterns and compliance with treatment. A chart review was conducted to identify cases of aggressive periodontitis in patients &lt;30 years of age referred for treatment at the Graduate Periodontology and Oral Implantology Clinic, TUKSD. The diagnosis of aggressive periodontitis was validated by presence of characteristic radiographic bone loss at permanent incisors and molars. Exclusion criteria were deficient radiographs, and a medical history of systemic diseases that compromise the immune response. Twenty-two aggressive periodontitis cases were identified among 300 charts surveyed. All patients were 12-26 years old. The patient sample was comprised mainly of African American race-ethnicity, with no predominance of a sex group. Initial treatment with scaling and root planing, was done in 64% of cases with 36% dropout before treatment. Microbial plaque testing was done in 46% of cases, and 59% received systemic antibiotics. A combination antibiotic therapy regimen was often used in combination with nonsurgical periodontal therapy. Most patients did not present for treatment beyond the non-surgical phase, and some even before the treatment started. Because periodontal non-surgical treatment of aggressive periodontitis cases in the pre-doctoral clinic takes relatively long time, it is recommended that the treatment of these cases be expedited by referring the patients to the graduate clinic for all periodontal treatment including the initial phase. / Biology