Microbial Profile and Endotoxin Levels in Primary Periodontal Lesions with Secondary Endodontic Involvement.

This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

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 was found cytotoxic and reduced the mineralization potential in vitro of APCs.

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.

Efeito da ativação do receptor ativado por protease do tipo 1 (PAR1) sobre as atividades osteogênica e cementogênica de células mesenquimais do ligamento periodontal / 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.

Effects of periodontal treatment on primary sjȫgren's syndrome symptoms.

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-1ß, 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.

Effects of periodontal treatment on primary sjogren'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.

Aspectos relevantes da síndrome de Sjõgren para o cirurgião-dentista / Sjogren’s syndrome’s relevant aspects for the dental surgeon

A síndrome de Sjõgren (SS), conhecida como síndrome sicca, é uma doença autoimune caracterizada pela hipofunção das glândulas salivares e lacrimais, cuja prevalência na população mundial é de aproximadamente 0,5% a 1%. Por ser uma doença autoimune complexa e de difícil diagnóstico, é sub-diagnosticada e sub-tratada segundo o consenso realizado em 2012 pelo Colégio Americano de Reumatologia (ACR). O Cirurgião-Dentista pode desempenhar papel importante na detecção de possíveis alterações compatíveis com a síndrome, além de auxiliar no tratamento de diversas patologias orais decorrentes da síndrome. Este trabalho tem como objetivo explanar aspectos importantes referentes ao diagnóstico e tratamento da síndrome aqui discutida. A SS apesar de ser considerada uma doença de evolução lenta, em estágios avançados pode ser fatal, principalmente por aumentar as chances dos pacientes virem a desenvolver linfoma não Hodking. O tratamento odontológico dos pacientes com SS deve principalmente ser profilático, com a recomendação do uso de repositores de saliva e controle rígido da higiene bucal.

Sjogren’s syndrome (SS), known as the sicca syndrome, is an autoimmune disease characterized by salivary and lacrimal glands hypofunction which prevalence in the world population is approximatel y around 0,5% to 1%. For being a complex autoimmune disease and with difficult diagnosis, it is sub diagnosed and miss treated according to the consensus occurred in 2012 by the American College of Rheumatology (ACR). The surgeon-dentist (SD) may play a important role on the detection of possible changes compatible to the syndrome, besides can help in the treatment of many oral pathologies caused by the syndrome. This work has the main purpose to explain the important aspects regards to the correct diagnosis and treatment of this syndrome.The SS besides been considered a slow evolution disease, in advanced stages it can be fatal,mainly for increasing the patient’s chances of developing non-Hodking lymphoma. The dental treatment of patients with SS must be prophylactic, with the recomedations of the use of salivary replenishing and careful control of the oral hyigiene.

Antibioticoterapia como coadjuvante no tratamento periodontal não cirúrgico de pacientes diabéticos / Antibiotic therapy as an adjunct to non-surgical periodontal treatment in diabetic patients ­ a literature review

Objetivo: verificar se o uso de antibióticos sistêmicos, adjunto à terapia periodontal não cirúrgica, em pacientes diabéticos pode trazer benefícios adicionais no controle glicêmico e parâmetros clínicos periodontais. Material e métodos: após a busca nos bancos de dados PubMed e Lilacs, foram incluídos nove ensaios clínicos randomizados para essa revisão da literatura. Resultados: nos desfechos periodontais, seis dos nove ensaios clínicos encontraram algum benefício a favor da antibioticoterapia. Quanto ao controle glicêmico, a maioria dos estudos falhou ao tentar relacionar o uso de antibióticos a uma maior redução da hemoglobina glicada. Conclusão: com base nesta revisão, concluiu-se que o uso de antibióticos sistêmicos em conjunto com o tratamento periodontal não cirúrgico pode oferecer benefícios adicionais, quando comparado ao tratamento periodontal sozinho, no tratamento da periodontite em pacientes com diabetes. Não há evidências de que a antibioticoterapia possa contribuir no controle glicêmico desses pacientes, sendo necessários mais estudos clínicos.

Objective: to verify whether systemic antibiotic therapy along with non-surgical periodontal management in diabetic patients can bring additional benefi ts on the glycemic control and periodontal parameters. Material and methods: after an electronic search on Medline and Lilacs databases, 9 RCTs were included. Results: six articles found favorable outcomes for periodontal clinical parameters. Regarding the glycemic control, most studies failed to prove a relationship between antibiotic use and the reduction on glycated hemoglobin levels. Conclusion: within the limits of this review, it can be concluded that systemic antibioticotherapy added to non-surgical periodontal treatment can bring additional benefits when compared to non-surgical treatment alone for diabetes-affected periodontal patients. However, there is no evidence that antibiotics can contribute to the glycemic control and more studies are necessary on this subject.

O efeito da sinvastatina na osseointegração / The effect of simvastatin on osseointegration ­ a literature review

O crescente aumento de instalações de implantes dentais com carga imediata e de suas fixações em tecido ósseo de baixa densidade tem gerado a necessidade real de aceleração e aprimoramento do processo de osseointegração, e muitos métodos e técnicas têm sido propostos para solucionar este problema. Neste contexto, a presente revisão teve por objetivo discutir os principais aspectos do papel da sinvastatina como intervenção para a melhoria no processo de osseointegração. Os resultados indicaram que a sinvastatina apresentou efeitos positivos em estudos experimentais, sendo necessários ensaios clínicos randomizados.

With increasing applications of dental implants with immediate loading and their installation in low quality bone, there is a real need to speed up and improve the osseointegration process. This review aimed to discuss the use of simvastatin in improving the osseointegration. The results indicate that simvastatin showed positive results in experimental studies, although randomized clinical trials are necessary.

Titanium scaffold osteogenesis in healthy and osteoporotic rats is improved by the use of low-level laser therapy (GaAlAs).

The present study aimed to assess the effects of low-level laser therapy (GaAlAs) on the bone repair process within titanium scaffolds in the femurs of healthy and osteoporotic rats. Fifty-six rats were divided into four groups: group Sh: SHAM animals that received scaffolds; group LSh: SHAM animals that received scaffolds and were subjected to laser therapy; group OV: ovarietomized (OVX) animals that received scaffolds; and group LOV: OVX animals that received scaffolds and were subjected to laser therapy. Thirty days following ovariectomy or sham surgery, scaffolds were implanted in the left femurs of all animals in the study. Immediately after opening the surgical site, the inner part of the surgical cavity was stimulated with low-level laser (GaAlAs). In addition to this procedure, the laser group was also subjected to sessions of low-level laser therapy (LLLT) at 48-h intervals, with the first session performed immediately after surgery. The rats were sacrificed at 2 and 6 weeks, time in which femur fragments were submitted for histological and histomorphometric examination, and skin tissue above the scaffold was submitted to histological analysis. At the end of the study, greater bone formation was observed in the animals submitted to LLLT. At 2 and 6 weeks, statistically significant differences were observed between LSh and Sh groups (p = 0.009 and 0.0001) and LOV and OV (p = 0.0001 and 0.0001), respectively. No statistical difference was observed when assessing the estrogen variable. On the basis of our methodology and results, we conclude that LLLT improves and accelerates bone repair within titanium scaffolds in both ovariectomized and healthy rats, when compared to animals not subjected to radiation.

Clinical and Microbiological Evaluation of Povidone-Iodine 10% as an Adjunct to Nonsurgical Periodontal Therapy in Chronic Periodontitis: A Randomized Clinical Trial.

AIM: To evaluate the clinical and microbiological effects of irrigation with 10% povidone-iodine (PVP-I) as an adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis (ChP). METHODS: Thirty-six subjects with ChP were randomly assigned to receive SRP + irrigation with saline solution (control group) or SRP + irrigation with 10% PVP-I (test group). Subjects received clinical and microbiological evaluation at baseline, 30 and 90 days post-treatment. Six subgingival plaque samples/subject were collected at each time point and individually analyzed for 40 bacterial species by checkerboard DNA-DNA hybridization. RESULTS: No statistically significant differences were observed between the two groups at 90 days post-treatment. At 30 days, the test group presented with lower levels of Tannerella forsythia and lower proportions of red complex pathogens in comparison to the control group. However, these benefits were not maintained at 90 days. CONCLUSION: The results of this study were unable to show a significant clinical benefit from the adjunctive use of 10% PVP-I in the treatment of chronic periodontitis. However, the microbiological benefit observed with the use of this substance at 30 days might indicate a role for PVP-I in periodontal care, possibly through its repeated application during supportive therapy.

Estudo da diversidade microbiana e níveis de endotoxinas em lesões endo-periodontais / Study of microbiology diversity and endotoxin levels in endoperiodontal lesions

A 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 lesão

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 infection

Evaluation of dental staining using a dentifrice containing chlorhexidine and zinc acetate. A doble blind randomizedclinical trial / Avaliação de dentifrício contendo clorexidina e acetato de zinco. Estudo clínico controlado randomizado duplo cego

Objective: The aim of this study was to evaluate whether the association of chlorhexidine with zinc acetate in dentifrices formulations could reduce the emergence of extrinsic tooth stain. As second outcome check the clinical gingival parameters. Methods: 30 volunteers were randomly divided into three groups: CHX+Z, consisting of 10 participants who used a dentifrice with 0.8% chlorhexidine gluconate and zinc acetate 1.16%; CHX group, with 10 participants who used a similar formulation dentifrice without zinc acetate, and the Placebo group also with 10 participants who used a dentifrice formulation similar but without the chlorhexidine and zinc acetate. Patients were assessed at baseline and 60 days when the indexes of plaque, gingival bleeding and staining were collected. Results: The results showed that by day 60, there was a decrease of both plaque index(PI) and the gingival index(GI) for all groups. Additionally, it was observed that both groups using dentifrice containing chlorhexidine, showed more stain than placebo. The CHX+Z group showed less stain compared to the CHX group, but the difference was not statistically significant. The difference between Placebo and Chlorhexidine Groups was statistically significant (p < 0.05) when considered the stain intensity and area plus intensity scores. The CHX+Z group was as efficient in PI and GI reduction as the CHX group. Conclusions: The association of chlorhexidine with zinc acetate showed no additional benefits regarding reducing the staining. The dentifrices containing chlorhexidine presented higher reduction of GI and PI when compared to Placebo group.

Objetivo: O objetivo deste estudo foi avaliar se a associação de clorexidina com acetato de zinco em dentifrícios poderia reduzir o aparecimento de manchas dentárias extrínsecas. Como segundo objetivo verificar os parâmetros clínicos gengivais. Materiais e métodos: 30 voluntários foram divididos aleatoriamente em três grupos: CHX + Z, composta por 10 participantes que usaram um creme dental com 0,8% de gluconato de clorexidina e acetato de zinco 1,16%; grupo CHX, com 10 participantes que usaram uma formulação de dentifrício semelhante, porém sem acetato de zinco, e o grupo do placebo também com 10 participantes que usaram uma formulação dentífrica semelhante, mas sem a clorexidina e sem acetato de zinco. Os pacientes foram avaliados no início do estudo e após 60 dias, quando foram coletados os índices de placa, sangramento gengival e manchas. Resultados: Os resultados revelaram que ao dia 60, havia uma redução em ambos os índices, de placa (IP) e do índice gengival (IG) em todos os grupos. Além disso, observou-se que em ambos os grupos que utilizou dentifrício contendo clorexidina, mostrou um maior índice de manchas maior do que o placebo. O grupo CHX + Z apresentou um índice de manchas menor em relação ao grupo