Root canal dressings for revascularization influence in vitro mineralization of apical papilla cells

Endodontic revascularization is based on cell recruitment into the necrotic root canal of immature teeth after chemical disinfection. The clinical outcome depends on the ability of surviving cells from the apical tissue to differentiate and promote hard tissue deposition inside the dentinal walls. Objective: To investigate the effect of calcium hydroxide (CH) and modified triple antibiotic paste (mTAP – ciprofloxacin, metronidazole and cefaclor) on the viability and mineralization potential of apical papilla cells (APC) in vitro. Material and Methods: APC cultures were kept in contact with CH or mTAP (250-1000 µg/mL) for 5 days, after which cell viability was assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Next, APCs were subjected to CH or mTAP at 250 µg/mL for 5 days before inducing the differentiation assay. After 14 and 21 days, calcium deposition was assessed by the Alizarin Red S staining method, followed by elution and quantification using spectrophotometry. Data were analyzed using ANOVA followed by Tukey post hoc test. Results: CH induced cell proliferation, whereas mTAP showed significant cytotoxicity at all concentrations tested. APC treated with CH demonstrated improved mineralization capacity at 14 days, while, for mTAP, significant reduction on the mineralization rate was observed for both experimental periods (14 and 21 days). Conclusion: Our findings showed that CH induces cell proliferation and improves early mineralization, whereas mTAP wa

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study

Objective: Obesity is a chronic disease that negatively affects an individual’s general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology: This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results: In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions: Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy

Effect of protease activated receptor type 1 (PAR1) activation on the osteogenic activity of periodontal ligament cells

O receptor ativado por protease do tipo 1 (PAR1) foi o primeiro membro clonado da família de receptores acoplados à proteína G. Sua ativação tem sido associada ao reparo tecidual e cicatrização óssea. O objetivo do presente estudo foi avaliar o efeito da ativação do PAR1 nas atividades osteogênica e cementogênica de células mesenquimais do ligamento periodontal (CMLP). CMLP obtidas de 3 indivíduos foram cultivadas e tratadas com meio clonogênico (MC) ou meio osteogênico (MO) por 2, 7 e 14 dias. Depósitos de cálcio, concentração de cálcio (sobrenadante), atividade de fosfatase alcalina (ALP), proliferação celular, expressão gênica (qPCR) e níveis proteicos (ELISA) de fatores osteogênicos e cementogênicos foram avaliados na presença de trombina, agonista do PAR1 ou antagonista do PAR1. A ativação do PAR1 levou ao aumento da formação de depósitos de cálcio (p<0,05), o que foi associado ao aumento da concentração de cálcio (p<0,05), atividade da ALP (p<0,05) e proliferação celular (p<0,05). Além disso, os ensaios qPCR e ELISA mostraram que a ativação do PAR1 pode aumentar a expressão gênica de Runx2, OPG e CEMP1 (p<0,05) e níveis proteicos de Runx2 e OPG (p<0,05). Em conclusão, nossos resultados demonstram que a ativação de PAR1 aumenta as atividades osteogênica e cementogênica de CMLP.Protease activated receptor 1 (PAR1) was the first cloned member of the G protein-coupled receptor family. Its activation has been associated to tissue repair and bone healing. The aim of the present study was to evaluate the effect of PAR1 activation on the osteogenic and cementogenic activities of human periodontal ligament stem cells (HPLSC). HPLSC obtained from 3 subjects and treated with a control medium or with an osteogenic medium for 2, 7 and 14 days. Calcium deposits, calcium concentration (supernatant), alkaline phosphatase activity (ALP), cell proliferation and gene (qPCR) and protein expression (ELISA assay) of osteogenic and cementogenic factors were assessed in the presence of thrombin, PAR1 specific agonist peptide or PAR1 antagonist peptide. The activation of PAR1 led to increased formation of calcium deposits (p<0.05), which was associated to increased calcium concentration (p<0.05), ALP activity (p<0.05) and cell proliferation (p<0.05). Further, qPCR and ELISA assay showed that activation of PAR1 may increase gene expression of Runx2, OPG and CEMP1 (p<0.05) and protein levels of Runx2 and OPG (p<0.05). In conclusion, our results demonstrate that PAR1 activation increases osteogenic and cementogenic activities of HPLSC

Effect of protease activated receptor type 1 (PAR1) activation on the osteogenic activity of periodontal ligament cells

O receptor ativado por protease do tipo 1 (PAR1) foi o primeiro membro clonado da família de receptores acoplados à proteína G. Sua ativação tem sido associada ao reparo tecidual e cicatrização óssea. O objetivo do presente estudo foi avaliar o efeito da ativação do PAR1 nas atividades osteogênica e cementogênica de células mesenquimais do ligamento periodontal (CMLP). CMLP obtidas de 3 indivíduos foram cultivadas e tratadas com meio clonogênico (MC) ou meio osteogênico (MO) por 2, 7 e 14 dias. Depósitos de cálcio, concentração de cálcio (sobrenadante), atividade de fosfatase alcalina (ALP), proliferação celular, expressão gênica (qPCR) e níveis proteicos (ELISA) de fatores osteogênicos e cementogênicos foram avaliados na presença de trombina, agonista do PAR1 ou antagonista do PAR1. A ativação do PAR1 levou ao aumento da formação de depósitos de cálcio (p<0,05), o que foi associado ao aumento da concentração de cálcio (p<0,05), atividade da ALP (p<0,05) e proliferação celular (p<0,05). Além disso, os ensaios qPCR e ELISA mostraram que a ativação do PAR1 pode aumentar a expressão gênica de Runx2, OPG e CEMP1 (p<0,05) e níveis proteicos de Runx2 e OPG (p<0,05). Em conclusão, nossos resultados demonstram que a ativação de PAR1 aumenta as atividades osteogênica e cementogênica de CMLP.Protease activated receptor 1 (PAR1) was the first cloned member of the G protein-coupled receptor family. Its activation has been associated to tissue repair and bone healing. The aim of the present study was to evaluate the effect of PAR1 activation on the osteogenic and cementogenic activities of human periodontal ligament stem cells (HPLSC). HPLSC obtained from 3 subjects and treated with a control medium or with an osteogenic medium for 2, 7 and 14 days. Calcium deposits, calcium concentration (supernatant), alkaline phosphatase activity (ALP), cell proliferation and gene (qPCR) and protein expression (ELISA assay) of osteogenic and cementogenic factors were assessed in the presence of thrombin, PAR1 specific agonist peptide or PAR1 antagonist peptide. The activation of PAR1 led to increased formation of calcium deposits (p<0.05), which was associated to increased calcium concentration (p<0.05), ALP activity (p<0.05) and cell proliferation (p<0.05). Further, qPCR and ELISA assay showed that activation of PAR1 may increase gene expression of Runx2, OPG and CEMP1 (p<0.05) and protein levels of Runx2 and OPG (p<0.05). In conclusion, our results demonstrate that PAR1 activation increases osteogenic and cementogenic activities of HPLSC

Estudo da diversidade microbiana e níveis de endotoxinas em lesões endo-periodontais

The endo-periodontal lesion is characterized by the association of endodontic and periodontal disease in the same tooth. The evolution of this lesion occurs by the presence of microorganisms with vast majority gram-negative anaerobes. Therefore this study aims to identify the microorganisms in endo-periodontal lesions by DNA-DNA checkerboard hybridization method, evaluate and compare the levels of endotoxins with clinical signs and symptoms. 10 teeth with endo-periodontal lesions in agree with the inclusion criteria were selected from patients in the ICT-UNESP, 10 endodontic and 10 periodontal samples were collected, stored and subsequently performed the laboratory assays. The results showed higher levels of endotoxin in periodontal pockets with mean and standard deviation of 39096 ± 39312. The mean and standard deviation values of endotoxins in root canals were 3240 ± 4001 (significantly lower p = 0.002). The Pearson correlation test showed moderate correlation, with p = 0.084. P. nigrescens species was associated with periodontal microflora (p = 0.0198) whereas in endodontic microbiota the species E. faecium (p = 0.0031), P. acnes (p = 0.0325), G. morbillorum (p = 0.0108) C. sputigena (p = 0.0031) and L. buccalis (p = 0.0055) were strongly correlated. The species P. intermedia, V parvula and P. endodontalis showed low correlation in endo-periodontal microflora. The study concluded that the periodontal pocket has higher levels of endotoxins in relation to the root canal. The microbiota of endo-periodontal lesions seems to be almost the same in their infections separated. The study suggests that the orange complex must play an important role in this type of infectionA lesão endo-periodontal é caracterizada pela associação da doença endodôntica e periodontal no mesmo elemento dentário. A evolução dessa lesão ocorre pela infecção por microrganismos, na sua grande maioria gram-negativas anaeróbias. O presente estudo teve como objetivo, identificar os microrganismos presentes em lesões endo-periodontais pelo método do DNA-DNA checkerboard hybridization, avaliar os níveis de endotoxinas pelo ensaio LAL e correlacionar com sinais e sintomas clínicos de pacientes com lesões endo-periodontais. Dez pacientes com lesões endo-periodontais foram selecionados do ICT-UNESP, foram coletadas 10 amostras endodônticas e 10 periodontais, as amostras foram armazenadas e posteriormente realizado os ensaios laboratoriais(LAL para endotoxinas e Checkerboard para microbiológico). Os resultados mostraram maiores níveis de endotoxinas na bolsa periodontal com média e desvio padrão de 39096 ± 39312. A média e desvio padrão dos valores de endotoxinas no canal radicular foram 3240 ± 4001 (significativamente menores p = 0,002). O teste de correlação de Pearson mostrou correlação moderada, com p = 0,084. A espécie P. nigrescens foi associada com a microbiota periodontal (p = 0,0198) enquanto na microbiota endodôntica as espécies E. faecium(p =0,0031), P. acnes(p= 0,0325), G. morbillorum(p=0,0108), C. sputigena(p = 0,0031) e L. buccalis(p = 0,0055), foram fortemente relacionadas. Os microrganismos P. intermedia, V parvula e P. endodalis tiveram pequena correlação quando associadas as microbiotas endodônticas e periodontais. O estudo concluiu que a bolsa periodontal apresenta maiores níveis de endotoxinas em relação ao canal radicular. O estudo sugere que a microbiota das lesões endo-periodontais parece a mesma presente em lesões endodônticas e periodontais separadamentes. No entanto,devido a sua grande prevalência, o complexo laranja parece exercer um papel importante neste tipo de lesã

Root canal dressings for revascularization influence in vitro mineralization of apical papilla cells

Abstract Endodontic revascularization is based on cell recruitment into the necrotic root canal of immature teeth after chemical disinfection. The clinical outcome depends on the ability of surviving cells from the apical tissue to differentiate and promote hard tissue deposition inside the dentinal walls. Objective To investigate the effect of calcium hydroxide (CH) and modified triple antibiotic paste (mTAP – ciprofloxacin, metronidazole and cefaclor) on the viability and mineralization potential of apical papilla cells (APC) in vitro . Material and Methods APC cultures were kept in contact with CH or mTAP (250-1000 µg/mL) for 5 days, after which cell viability was assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Next, APCs were subjected to CH or mTAP at 250 µg/mL for 5 days before inducing the differentiation assay. After 14 and 21 days, calcium deposition was assessed by the Alizarin Red S staining method, followed by elution and quantification using spectrophotometry. Data were analyzed using ANOVA followed by Tukey post hoc test. Results CH induced cell proliferation, whereas mTAP showed significant cytotoxicity at all concentrations tested. APC treated with CH demonstrated improved mineralization capacity at 14 days, while, for mTAP, significant reduction on the mineralization rate was observed for both experimental periods (14 and 21 days). Conclusion Our findings showed that CH induces cell proliferation and improves early mineralization, whereas mTAP was found cytotoxic and reduced the mineralization potential in vitro of APCs

Projeto ONCO: preparo odontológico para pacientes portadores de neoplasias malignas

Introdução: É esperado que pacientes com câncer desenvolvam algum tipo de complicação na região oral, que pode estar relacionada às condições de saúde bucal do paciente e ao tipo de terapia utilizada. É de grande importância que o profissional da odontologia conheça as modalidades de tratamento do câncer de boca para o preparo da cavidade bucal antes, durante e após a terapia. Objetivos: O objetivo do projeto é possibilitar a formação de profissionais qualificados e competentes para atender adequadamente a pacientes portadores de neoplasias malignas, no pré e pós-operatório, no pré ou pós- tratamento de radioterapia e/ou quimioterapia, bem como, pesquisar, inovar e desenvolver novos protocolos, técnicas e metodologias, correlacionando as alterações clínicas com as características sistêmicas, dentro de um enfoque multidisciplinar e multiprofissional. Métodos: O projeto Onco atendeu mais de 200 pacientes oncológicos, realizando mais de 600 procedimentos, entre exodontias, dentística restauradora, endodontia, biopsias e cirurgias oncológicas em lábio, língua e mucosa oral com expectativas de cura. Acompanhamos e controlamos pacientes que desenvolveram osteoradionecrose e osteoradiomielite de mandíbula, pós- radioterapia, e proservação dos cânceres de cirurgia de cabeça e pescoço, operados e diagnosticados neste serviço. Resultados: O Projeto Onco vem se desenvolvendo de maneira excepcional, tendo sido procurado, por Hospitais Oncológicos, tais como, o Instituto de Oncologia e Radioterapia do Vale do Paraíba e pelo serviço de Transplante de Medula Óssea do Hospital Pio XII, para ser sua referência nos tratamentos odontológicos. Já hoje, desenvolvemos atendimento odontológico de alto nível para pacientes oncológicos e transplantados, cujo campo de atuação é extremamente carente em todo o país

Effects of periodontal treatment on primary sjȫgren’s syndrome symptoms

Abstract The aim of this longitudinal prospective study was to evaluate the effects of periodontal treatment on the clinical, microbiological and immunological periodontal parameters, and on the systemic activity (ESSDAI) and subjective (ESSPRI) indexes in patients with primary Sjögren’s Syndrome (pSS). Twenty-eight female patients were divided into four groups: pSS patients with or without chronic periodontitis (SCP, SC, respectively), and systemically healthy patients with or without chronic periodontitis (CP, C, respectively). Periodontal clinical examination and immunological and microbiological sample collection were performed at baseline, 30 and 90 days after nonsurgical periodontal treatment (NSPT). Levels of interleukin IL-1β, IL-8 and IL-10 in saliva and gingival crevicular fluid (GCF) were evaluated by ELISA, as well as the expression of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans, (Aa) Tannerella forsythia (Tf), and Treponema denticola (Td), by qPCR. Systemic activity and pSS symptoms were evaluated by ESSDAI and ESSPRI. NSPT resulted in improved periodontal clinical parameters in both SCP and CP groups (p>0.05). Pg, Aa, and Tf levels decreased after NSPT only in CP patients (p<0.05). Significantly greater levels of IL-10 in GCF were verified in both SCP and CP groups (p<0.05). SCP patients showed increased salivary flow rates and decreased ESSPRI scores after NSPT. In conclusion, NSPT in pSS patients resulted in improved clinical and immunological parameters, with no significant effects on microbiological status. pSS patients also showed increased salivary flow and lower ESSPRI scores after therapy. Therefore, it can be suggested that NSPT may improve the quality of life of pSS patients