Effect of nonsteroidal anti-inflammatory drugs (NSAIDs) association on physicochemical and biological properties of tricalcium silicate-based cement

Abstract The aim of this study was to investigate the physicochemical and biological properties of an experimental tricalcium silicate-based repair cement containing diclofenac sodium (CERD). For the physicochemical test, MTA, Biodentine and CERD were mixed and cement disc were prepared to evaluate the setting time and radiopacity. Root-end cavity were performed in acrylic teeth and filled with cements to analyze the solubility up to 7 days. Polyethylene tubes containing cements were prepared and calcium ions and pH were measured at 3h, 24h, 72h and 15 days. For the biological test, SAOS-2 were cultivated, exposed to cements extracts and cell proliferation were investigated by MTT assay at 6h, 24h and 48h. Polyethylene tubes containing cements were implanted into Wistar rats. After 7 and 30 days, the tubes were removed and processed for histological analyses. Parametric and nonparametric data were performed. No difference was identified in relation to setting time, radiopacity and solubility. Biodentine released more calcium ion than MTA and CERD; however, no difference between MTA and CERD were detected. Alkaline pH was observed for all cements and Biodentine exhibited highest pH. All cements promoted a raise on cell proliferation at 24h and 48h, except CERD at 48h. Biodentine stimulated cell metabolism in relation to MTA and CERD while CERD was more cytotoxic than MTA at 48h. Besides, no difference on both inflammatory response and mineralization ability for all cement were found. CERD demonstrated similar proprieties to others endodontic cements available.

Resumo O objetivo deste estudo foi investigar as propriedades físico-químicas e biológicas de um cimento reparador experimental à base de silicato de tricálcio contendo diclofenaco de sódio (CERD). Para o teste físico-químico, MTA, Biodentine e CERD foram manipulados e discos de cimentos foram preparados para avaliar o tempo de presa e a radiopacidade. Retrocavidades foram feitas em dentes de acrílico e preenchidas com cimentos para análise de solubilidade por 7 dias. Tubos de polietileno contendo cimentos foram preparados e os íons cálcio e o pH foram mensurados às 3h, 24h, 72h e 15 dias. Para o teste biológico, SAOS-2 foram cultivadas, expostas aos extratos de cimentos e a proliferação celular foi investigada pelo ensaio de MTT às 6h, 24h e 48h. Tubos de polietileno contendo cimentos foram implantados em ratos Wistar. Após 7 e 30 dias, os tubos foram removidos e processados para análises histológicas. Dados paramétricos e não paramétricos foram realizados. Nenhuma diferença foi identificada em relação ao tempo de presa, radiopacidade e solubilidade. Biodentine liberou mais íons de cálcio do que MTA e CERD; no entanto, nenhuma diferença entre MTA e CERD foi detectada. O pH alcalino foi observado para todos os cimentos e o Biodentine exibiu o pH mais alto. Todos os cimentos promoveram aumento na proliferação celular às 24h e 48h, exceto o CERD às 48h. Biodentine estimulou o metabolismo celular em relação ao MTA e CERD, enquanto CERD foi mais citotóxico do que MTA em 48h. Além disso, nenhuma diferença foi encontrada na resposta inflamatória e na capacidade de mineralização para todos os cimentos. CERD demonstrou propriedades semelhantes a outros cimentos endodônticos disponíveis.

Cytotoxicity, biocompatibility and biomineralization of a new ready-for-use bioceramic repair material

Abstract New mineral trioxide aggregate (MTA) formulations are constantly introduced in the market, usually in a powder-and-liquid form. Bioceramic (Bio-C) Repair is a ready-for-use material suggested as substitute for MTA, but its properties need to be studied. This study evaluated the cytotoxicity, biocompatibility and biomineralization of Bio-C Repair compared to MTA Repair High-Plasticity (MTA-HP) and white MTA-Angelus (MTA-Ang). L929 fibroblasts were exposed to material-extracted (undiluted, ½ and » dilutions; 6, 24 and 48h). Polyethylene tubes with material or empty (control) were implanted in the subcutaneous tissue of rats. After 7 and 30 days (n=8), the specimens were removed for analysis (hematoxylin-eosin, von Kossa and polarized light). Cytotoxicity data were statistically analyzed by two-way ANOVA, and biocompatibility data by Kruskal-Wallis and Dunn tests (p<0.05). The cells exposed to the materials had greater viability at most of the periods compared with control (p<0.05). The undiluted and ½ dilutions of MTA-HP extract showed higher cytocompatibility than Bio-C Repair at 6 h and with the » dilution at 24 h (p<0.05); the white MTA-Ang showed higher cytocompatibility than Bio-C Repair at most of periods (p<0.05). The undiluted white MTA-Ang extract had higher cytocompatibility at 6 and 24h than MTA-HP, and with ½ dilution at 24h (p<0.05). The materials' cytocompatibility was similar at 48h for most dilutions (p>0.05). At 7 and 30 days, the groups had moderate and mild inflammation, respectively (p>0.05). All materials showed positive structures for von Kossa and polarized light. In conclusion, Bio-C Repair had similar cytocompatibility to MTA-based materials is biocompatible and induces biomineralization.

Resumo Novas formulações de agregado de trióxido mineral (MTA) são constantemente introduzidas no mercado, geralmente em forma de pó e líquido. O Biocerâmico (Bio-C) Reparador (Repair) é um material pronto para uso sugerido como substituto do MTA, mas suas propriedades precisam ser estudadas. Este estudo avaliou a citotoxicidade, biocompatibilidade e biomineralização do Bio-C Repair comparado ao MTA-High Plasticity (MTA-HP) e MTA branco da Angelus (MTA-Ang). Fibroblastos L929 foram expostos a extratos dos materiais (não diluído, ½ e » diluições; 6, 24 e 48 h). Tubos de polietileno contendo os materiais ou vazios (controle) foram implantados no tecido subcutâneo de ratos. Após 7 e 30 dias (n=8), os espécimes foram removidos para análises (hematoxilina-eosina, von Kossa e luz polarizada). Os dados da citotoxicidade foram analisados estatisticamente pelo teste de two-way ANOVA, e os dados da biocompatibilidade pelos testes de Kruskal-Wallis e Dunn (p<0,05). As células expostas aos materiais apresentaram maior viabilidade celular na maior parte dos períodos, comparados com o controle (p<0,05). O extrato não diluído e ½ diluição do MTA-HP apresentaram maior citocompatibilidade do que Bio-C Repair às 6h, e com » diluição às 24h (p<0,05); o MTA-Ang branco apresentou maior citocompatibilidade do que o Bio-C Repair na maior parte dos períodos (p<0,05). O extrato não diluído do MTA-Ang branco apresentou maior citocompatibilidade às 6 e 24 h comparado ao MTA-HP, e com ½ diluição às 24h (p<0,05). A citocompatibilidade dos materiais foi semelhante às 48 h para a maior parte das diluições (p>0,05). Aos 7 e 30 dias, os grupos apresentaram inflamação moderada e leve, respectivamente (p>0,05). Todos os materiais mostraram estruturas positivas para von Kossa e luz polarizada. Em conclusão, o Bio-C Repair teve citocompatibilidade semelhante aos materiais à base de MTA, é biocompatível e induz à biomineralização.

CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS

ABSTRACT Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts from permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by increasing concentrations of PgLPS (0 – 10 µg/mL) at 1, 6 and 24 h. The cells were tested for viability through MTT assay, and production of the chemokines CCL3 and CXCL12 was determined through ELISA. Comparisons among samples were performed using One-way ANOVA for MTT assay and Two-way ANOVA for ELISA results. Results: Cell viability was not affected by the antigen after 24 h of stimulation. PgLPS induced the production of CCL3 by dental pulp fibroblasts at similar levels for both permanent and deciduous pulp fibroblasts. Production of CXCL12, however, was significantly higher for PDPF than DDPF at 1 and 6 h. PgLPS, in turn, downregulated the production of CXCL12 by PDPF but not by DDPF. Conclusion: These data suggest that dental pulp fibroblasts from permanent and deciduous teeth may present a differential behavior under PgLPS stimulation. .

Functional activity of neutrophils and systemic inflammatory response of Down’s syndrome patients with periodontal disease

Periodontal disease (PD) is characterized as an inflammatory process that compromises the support and protection of the periodontium. Patients with Down’s syndrome (DS) are prone to develop PD. Neutrophils (NE) are the first line of defense against infection and their absence sets the stage for disease. Aim: To compare the activity and function of NE in the peripheral blood from DS patients with and without PD, assisted at the Center for Dental Assistance to Patients with Special Needs affiliated with the School of Dentistry of Araçatuba, Brazil. Methods: Purified NE were collected from peripheral blood of 22 DS patients. NE were used to detect the 5-lypoxigenase (5-LO) expression by RT-PCR. Plasma from peripheral blood was collected to measure tumor necrosis factor-a (TNF-a) and interleukin-8 (IL-8) by ELISA and nitrite (NO3) using a Griess assay. Results: Data analysis demonstrated that DS patients with PD present high levels of TNF-a and IL-8 when compared with DS patients without PD. However, there was no statistically significant difference in the levels of NO3 production between the groups. The levels of the inflammatory mediator 5-LO expression increased in DS patients with PD. Conclusions: According with these results, it was concluded that TNF-a and IL-8 are produced by DS patients with PD. Furthermore, DS patients with PD presented high levels of 5-LO expression, suggesting the presence of leukotriene B4 (LTB4) in PD, thus demonstrating that the changes in NE function due to the elevation of inflammatory mediators contribute to PD.

Evaluation of tissue reaction, cell viability and cytokine production induced by Sealapex Plus

OBJECTIVE: The aim of this study was to investigate the effects of mineral trioxide aggregate (MTA), Sealapex, and a combination of Sealapex and MTA (Sealapex Plus) on the reaction of subcutaneous connective tissue of rats, and on cell viability and cytokine production in mouse fibroblasts. MATERIAL AND METHODS: The tissue reaction was carried out with dentin tubes containing the materials implanted in the dorsal connective tissue of rats. The histological analysis was performed after 7 and 30 days. Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts to evaluate the cell viability by MTT assay. ELISA assays were also performed after 24 h of exposure of the mouse fibroblasts to set material disks. RESULTS: Histopathologic examination showed Von Kossa-positive granules that were birefringent to polarized light for all the studied materials at the tube openings. No material inhibited the cell viability in the in vitro test. It was detected IL-6 production in all root-end filling materials. MTA and Sealapex Plus induced a slight raise of mean levels of IL-1β. CONCLUSIONS: The results suggest that Sealapex Plus is biocompatible and stimulates the mineralization of the tissue.