Impact of non-surgical periodontal treatment on salivary expression of cytokines related to bone metabolism.

We aimed to evaluate the impact of non-surgical periodontal treatment on the salivary expression of leptin, TNF-α, sclerostin, parathyroid hormone, osteoprotegerin, osteopontin, osteocalcin, IL-6, IL-1ß and fibroblast growth factor 23 in patients with chronic periodontitis after 1 year of follow-up. Fifteen patients with chronic periodontitis (56.0 ± SD 9.6 years) and 15 subjects with gingivitis (39.7 ± SD 4.4 years) were included in the study. Clinical periodontal parameters, such as probing pocket depth (PPD), clinical attachment level (CAL), % of plaque and bleeding on probing (BOP) were evaluated, and non-stimulated whole saliva was collected from all patients before periodontal treatment and after 1 year of follow-up. A bead-based multiplex assay measured cytokines. In the chronic periodontitis group, periodontal treatment significantly improved clinical parameters and reduced the salivary levels of IL-1ß, leptin and TNF-α (p = 0.002, 0.007 and 0.015, respectively). In the gingivitis group, there were also significant improvements in the mean patient %BOP, % Plaque, CAL and PPD. However, there were no significant changes in the cytokine's salivary levels. In conclusion, chronic periodontitis patients showed a significant reduction in the salivary levels of leptin, TNF-α and IL-1ß 1 year after periodontal treatment and a significant improvement in their clinical periodontal parameters suggesting that periodontal treatment alone can downregulate important cytokines associated with bone metabolism.

Anti-apoptotic traits in gingival tissue from patients with severe generalized chronic periodontitis.

AIM: In the present study, we aimed to evaluate the cytosolic and nuclear-mitochondrial expression of pro-apoptotic and anti-apoptotic markers in gingival tissue from patients with severe generalized chronic periodontitis (sGCP). METHODS: Twenty-four patients participated in the study: 15 (54.3 + 8.3 years) with sGCP and nine (38.2 + 5.4 years) with gingivitis alone. Gingival tissue was collected using a 1.5-mm diameter punch and homogenized using a cell disruptor. The supernatants were analyzed for the cytosolic and nuclear-mitochondrial fractions of caspase-3, Bax, Bak, Smac, lamin B, Bad, Bim, survivin, Bcl-xL, Mcl-1, and of the dimers Bcl-2/Bax, Bcl-xL/Bak and Mcl-1/Bak using a multiplex immunoassay. RESULTS: Significantly higher levels of cytosolic Bcl-xL/Bak, nuclear-mitochondrial Mcl-1/Bak, and cytosolic Bcl-xL were observed in gingival tissues from periodontitis patients compared to controls (P = 0.03, 0.03, and 0.05, respectively). The patients with gingivitis presented significantly increased levels of nuclear-mitochondrial Bad, cytosolic and nuclear-mitochondrial Bcl-2/Bax, and cytosolic Bim compared to the patients with periodontitis (P < 0.001, 0.03, 0.05, and 0.04, respectively). CONCLUSION: Significantly higher levels of anti-apoptotic markers, such as Bcl-xL/Bak, Mcl-1/Bak, and Bcl-xL, and lower levels of pro-apoptotic markers Bad and Bim in inflamed tissues indicate an anti-apoptotic trait in patients with sGCP.

Expressão de fatores apoptóticos em biópsias de tecido gengival de pacientes com periodontite crônica severa generalizada / Expression of apoptotics factors in gingival tissue biopsies in patients with severe generalized chronic periodontitis

O objetivo deste estudo foi avaliar a expressão dos marcadores de apoptose caspase-3, survivina e dos dímeros Bcl-xL/Bak e Mcl-1/Bak, em biópsias de tecido gengival de pacientes com Periodontite Crônica Generalizada (PCG) severa. No grupo teste foram avaliados 15 pacientes portadores de PCG severa com média de idade de 54,7 + 8,4 anos. O grupo controle foi composto por 10 pacientes com gengivite com média de idade de 38,9 + 5,4 anos. Os pacientes foram avaliados em todos os elementos dentários, exceto os terceiros molares, para Profundidade de Bolsa (PB), Nível de Inserção (NI), Índice de Placa Visível (IPV) e Índice de Sangramento Gengival (ISG). Nos dentes onde as biópsias foram coletadas, também foram avaliados os critérios de Índice de Placa (IP) e Índice Gengival (IG). Foram coletadas para cada paciente, duas biópsias no grupo teste e uma no grupo controle. As biópsias foram coletadas com um punch de 1,5 mm de diâmetro, acima do fundo da bolsa ou sulco e foram armazenadas a -70° Celsius (C). Posteriormente, as amostras de tecido gengival foram pesadas, maceradas em um disruptor de células e cada biópsia foi dividida em fração citosólica e nuclear/mitocondrial. Essas frações foram analisadas em um imunoensaio com microesferas do tipo multiplex e a expressão dos marcadores de apoptose caspase-3, survivina e dos dímeros Bcl-xL/Bak e Mcl-1/Bak foi apresentada em nanograma por miligrama de tecido (ng/mg). Os dados foram apresentados em mediana e intervalo inter-quartil e avaliados pelo teste U de Mann-Whitney com p < 0,05. A correlação de Spearman foi considerada relevante sendo r > 0,6 com p < 0,01. Os valores de PB, NI e ISG foram significativamente maiores no grupo teste, comparados ao grupo controle (p < 0,001, p < 0,001, p = 0,035, respectivamente). O grupo de pacientes com periodontite apresentou, na fração nuclear/mitocondrial, concentrações de Mcl-1/Bak e survivina significativamente maiores, em comparação ao grupo de pacientes com gengivite (p = 0,05 em ambos). A correlação entre a caspase-3 e o Bcl-xL/Bak foi significantemente positiva, tanto para a fração citosólica quanto para a fração nuclear/mitocondrial (p < 0,001, p = 0,003, respectivamente). Concluímos que os pacientes com periodontite apresentaram níveis significantemente maiores dos marcadores antiapoptóticos, survivina e do dímero Mcl-1/Bak, na fração nuclear/mitocondrial, sugerindo um possível efeito de retardo no processo de apoptose.

The aim of this study was to evaluate the expression of the apoptotic markers caspase-3, survivin and the dimers Bcl-xL/Bak and Mcl-1/Bak, in gingival tissue biopsies in subjects having severe Generalized Chronic Periodontitis (GCP). In the test group were assessed 15 subjects affected by severe GCP with mean age of 54,7 + 8,4 years. The control group was formed with 10 subjects with gingivitis with mean age of 38,9 + 5,4 years. All teeth in the patients were evaluated (except the third molars) for Pocket Depth (PD), Attachment Level (AL), Visible Plaque Index (VPI) and Gingival Bleeding Index (GBI). In the teeth where the biopsies were collected, Plaque Index (PI) and Gingival Index (GI) were also measured. Two biopsies in the test group and one biopsy in the control group were collected using a 1,5 mm of diameter punch and stored at -70º Celsius (C). Just before analysis, the samples were weighted and macerated with a cell disrupter. Each biopsy was divided into cytosolic and nuclear/mitochondrial fractions. These fractions were analyzed in a multiplex microsphere immunoassay and the expression of the apoptotic markers caspase-3, survivin and the dimers Bcl-xL/Bak and Mcl-1/Bak were shown in nanogram per milligram of tissue (ng/mg). The data were presented in median and interquartile range and measured using the Mann-Whitney U Test with p < 0,05. Spearman's correlation was deemed relevant when r > 0,6 with p < 0,01. The values of PD, AL and GBI were significantly higher in the test group, when compared to the control (p < 0,001, p < 0,001, p = 0,035, respectively). The group of pacients suffering with periodontitis presented, in the nuclear/mitochondrial fraction, concentrations of Mcl-1/Bak and survivin significantly higher when compared to the control (p = 0,05 for both). The correlation between the caspase-3 and the Bcl-xL/Bak was significantly positive, from both the cytosolic and the nuclear/mitochondrial fractions (p < 0,001, p = 0,003, respectively). We conclude that the subjects having periodontitis presented significantly higher levels of the antiapoptotic markers survivin and the dimer Mcl-1/Bak, in the nuclear/mitochondrial fraction, suggesting a possible retarding effect on the process of apoptosis.

Efeito da temperatura de armazenamento na resistência coesiva de um compósito / Effect of storage temperature on the composite cohesive strength

O objetivo deste estudo foi avaliar a resistência à tração diametral de um compósito fotopolimerizável em diferentes temperaturas de armazenamento. Foram confeccionados 12 espécimes cilíndricos (n = 6),que foram divididos em dois grupos: G1- resina à temperatura ambiente (25ºC) e G2- resina utilizada imediatamente após remoção do refrigerador (5ºC). Cada espécime foi confeccionado em três incrementos, cada incremento polimerizado por 40 segundos. Realizouse o teste a uma velocidade de 1.0 mm/min até a falha. Após cada teste, a resistência à tração diametral foi calculada (MPa) e os resultados submetidos à análise estatística, pelo teste t-student (p > 0,05). Não houve efeito da temperatura de armazenamento na resistência coesiva da resina utilizada neste estudo.