Evaluation of the systemic effect of bone formation marker released by endodontic calcium silicate-based sealers in local tissues, the bloodstream, and body organs.

Calcium silicate-based sealers are bioactive materials that release ions when in contact with body fluids. Therefore, this study aims mapping/trace bone formation markers released by MTA Fillapex, BioRoot RCS, and experimental tricalcium silicate-based sealer (CEO) into subcutaneous tissues, bloodstream and body organs. Toward, polyethylene tubes filled with sealers were implanted into connective tissue of Wistar rats. On days 7, 15, 30, and 45 after implantation, blood samples were collected to measure calcium (Ca2+), phosphorus (P), and alkaline phosphatase (ALP) levels. Thereafter, the animals were killed, and the brain, liver, kidneys, and subcutaneous tissue were removed and processed to determine the concentrations of Ca2+ and P by ICP-OES. Similar Ca2+ levels were observed in subcutaneous tissue for all groups, although, at 45 days, it was identified a reduction in Ca2+ serum levels of CEO compared to those two other sealers and an increase in Ca2+ levels in the liver compared to those released by MTA Fillapex. In contrast, no trace of P was detected in any tissue; moreover, plasma P and ALP serum levels of MTA Fillapex were higher at day 30. Our findings showed that Ca2+ were identified in local tissues, bloodstream, and organs from all sealers. The up-regulation of bone marker levels promoted by sealers can modify body homeostasis and induce tissue damage. Besides, MTA Fillapex was associated with a raise of bone marker levels, suggesting a possible systemic effect. The sealer composition can affect not only the local repair process but also the systemic health.

Biological and antimicrobial properties of the association Ambroxol and a water-soluble viscous liquid as a vehicle for a tricalcium silicate-based sealer.

This study aimed to investigate the antimicrobial and biological properties of Ambroxol associated with glycerin (GLI), propylene glycol (PG), and polyethylene glycol (PEG) as a possible vehicle for an experimental tricalcium silicate sealer, with the intention of developing a new biomaterial. Mouse undifferentiated dental pulp cells (OD-21) were cultured, and the effects of different association on cell proliferation and inflammatory cytokine production were investigated. Antimicrobial adhesion of Enterococcus faecalis to setting sealers at 2 h was evaluated. Polyethylene tubes containing experimental sealers and empty tubes were implanted into dorsal connective tissues of 12 male 3- to 4-months-old Wistar rats (250-280 g). After 7 and 30 days, the tubes were removed and processed for histological and immunohistochemical analyses. ANOVA followed by Bonferroni correction and ANOVA followed by Tukey test was used for parametric data and Kruskal-Wallis followed by Dunn for nonparametric (p < 0.05). Cell proliferation was dose-dependent, since all association were cytotoxic at higher concentrations; however, Ambroxol-PEG showed significantly higher cytotoxicity than other association (p < 0.05). In addition, irrespective of the association, no cytokine production was observed in vitro. Ambroxol-GLI reduced bacterial viability, whereas Ambroxol-PEG increased (p < 0.05). Histological examination showed no significant difference in the inflammatory response (p > 0.05) and mineralization ability in all association. Additionally, IL-1ß and TNF-α were upregulated on Ambroxol-PEG in relation to Control at 07 days (p < 0.05). Ambroxol-GLI was the best vehicle for experimental tricalcium silicate sealer, as it promoted an increase in antimicrobial activity without altering the inflammatory response or mineralization ability.

Biological assessment of a new ready-to-use hydraulic sealer.

OBJECTIVES: This study compared the cytotoxicity, biocompatibility, and tenascin immunolabeling of a new ready-to-use hydraulic sealer (Bio-C Sealer) with MTA-Fillapex and white MTA-Angelus. MATERIALS AND METHODS: L929 fibroblasts were cultivated and exposed to undiluted and diluted material extracts. Polyethylene tubes with or without (the control) the materials were implanted into the dorsa of rats. At 7 days and 30 days, the rats were euthanized, and the specimens were prepared for analysis; inflammation and immunolabeling were measured, and statistical analysis was performed (p < 0.05). RESULTS: MTA-Fillapex exhibited greater cytotoxicity than the other materials at all time points (p < 0.05). The undiluted Bio-C Sealer exhibited greater cytocompatibility at 6 and 48 hours than white MTA-Angelus, with higher cell viability than in the control (p < 0.05). White MTA-Angelus displayed higher cell viability than the control at 24 hours, and the one-half dilution displayed similar results at both 6 and 48 hours (p < 0.05). At 7 days and 30 days, the groups exhibited moderate inflammation with thick fibrous capsules and mild inflammation with thin fibrous capsules, respectively (p > 0.05). At 7 days, moderate to strong immunolabeling was observed (p > 0.05). After 30 days, the control and MTA-Fillapex groups exhibited strong immunolabeling, the white MTA-Angelus group exhibited moderate immunolabeling (p > 0.05), and the Bio-C Sealer group exhibited low-to-moderate immunolabeling, differing significantly from the control (p < 0.05). CONCLUSIONS: Bio-C Sealer and white MTA-Angelus exhibited greater cytocompatibility than MTA-Fillapex; all materials displayed adequate biocompatibility and induced tenascin immunolabeling.

Cytotoxicity and biocompatibility of a new bioceramic endodontic sealer containing calcium hydroxide.

This study evaluated the cytotoxicity and biocompatibility of a new bioceramic endodontic sealer (i.e., Sealer Plus BC) in comparison with those of MTA Fillapex and AH Plus. L929 fibroblasts were cultured and Alamar Blue was used to evaluate cell viability of diluted extracts (1:50, 1:100, and 1:200) from each sealer at 24 h. Polyethylene tubes that were filled with material or empty (as a control) were implanted in the subcutaneous tissue of rats. The rats were killed after 7 and 30 d (n = 8), and the tubes were removed for histological analysis. Parametric data was analyzed using a one-way ANOVA test, and nonparametric data was analyzed via the Kruskal-Wallis test followed by the Dunn test (p < 0.05). A reduction in cell viability was observed in the extracts that were more diluted for Sealer Plus BC when compared to that of Control and AH Plus (p < 0.05). However, the 1:50 dilution of the Sealer Plus BC was similar to that of the Control (p > 0.05). Conversely, more diluted extracts of MTA Fillapex (1:200) and AH Plus (1:100 and 1:200) were similar to the Control (p > 0.05). Histological analysis performed at 7 d did not indicate any significant difference between tissue response for all materials, and the fibrous capsule was thick (p > 0.05). At 30 d, Sealer Plus BC was similar to the Control (p > 0.05) and MTA Fillapex and AH Plus exhibited greater inflammation than the Control (p < 0.05). The fibrous capsule was thin for the Control and for most specimens of Sealer Plus BC and AH Plus. Thus, Sealer Plus BC is biocompatible when compared to MTA Fillapex and AH Plus, and it is less cytotoxic when less-diluted extracts are used.

Th1/Th2/Th17/Treg Balance in Apical Periodontitis of Normoglycemic and Diabetic Rats.

INTRODUCTION: The aim of this study was to evaluate the inflammatory profile of T helper (Th) cells in normoglycemic (N) and diabetic rats with apical periodontitis (AP). METHODS: Twenty male Wistar rats were divided in 2 groups: N rats and rats with diabetes mellitus (DM). DM was induced using streptozotocin, and AP was induced by dental pulp exposure of the first mandibular molar to the oral environment. After 30 days, the mandibles were removed and processed for histologic analysis, bacterial analysis, and immunochemical assays for interleukin (IL)-6, tumor necrosis factor alpha, IL-17, IL-23, interferon gamma, and IL-10. The Mann-Whitney U test and Student t test were used for statistical analysis (P < .05). RESULTS: The DM group showed more intense inflammatory infiltrate with larger sizes of bone reabsorption and a greater presence of bacteria than the N group (P < .05). Proinflammatory cytokine levels in the DM group were also greater than those in the N group (P < .05). However, interferon gamma was more intense in the N group than in the DM group (P < .05). CONCLUSIONS: The inflammatory profile of AP in DM is different from that in the N group, suggesting that Th1 is a secondary strain and the Th17 strain is predominant in DM.

Hypertension Undermines Mineralization-inducing Capacity of and Tissue Response to Mineral Trioxide Aggregate Endodontic Cement.

INTRODUCTION: This study evaluated the effect of hypertension on tissue response to and mineralization capacity of white and gray mineral trioxide aggregate (MTA) (Angelus Industry Ontological Products, Londrina, Brazil), an endodontic reparative cement. METHODS: Polyethylene tubes containing gray MTA, white MTA, or intermediate restorative material (positive control) or an empty tube (negative control) were implanted into the dorsal connective tissue of spontaneous hypertensive and Wistar rats (n = 12 each). Six rats in each group were sacrificed after 7 days, and the remainder after 30 days. Tubes with surrounding tissue were removed, and a histologic analysis was performed using hematoxylin-eosin and von Kossa staining and examination by polarized light microscopy. RESULTS: The inflammatory response to all materials was greater in hypertensive compared with normotensive rats (P < .05). Positive von Kossa staining and birefringent structures in polarized light were observed for both gray and white MTA (P > .05), but these were more pronounced in normotensive rats (P < .05). Necrotic areas with positive von Kossa staining were observed for intermediate restorative material. CONCLUSIONS: Hypertension undermines tissue repair and mineralization, which can negatively affect treatment outcome. Nonetheless, mineralization in response to MTA was observed even under hypertensive conditions.

Influence of diabetes mellitus on tissue response to MTA and its ability to stimulate mineralization.

AIM: To evaluate the influence of diabetes mellitus on the tissue response to mineral trioxide aggregate (MTA) and its ability to stimulate mineralization. METHODS: Twenty-four male Wistar rats were divided into a non-diabetic control group and another with Alloxan-induced diabetes. Two polyethylene tubes, one kept empty as a control and the other containing Angelus MTA(®) , were implanted into the dorsal connective tissue of all rats for 30 days. Animals in each group received injections of calcein, alizarin, and oxytetracycline on day 7, 14, and 21, respectively. Animals were killed after 30 days; specimens were prepared by staining with hematoxylin and eosin (HE) and von Kossa technique as well as for examination of unstained sections with polarized light and fluorescence microscopy. RESULTS: The inflammatory reaction to the implanted tubes was equally mild in both groups. Structures staining with von Kossa were seen in response to Angelus MTA(®) , as were birefringent structures visualized on polarized light analysis; these had no relation to diabetic condition (P < 0.05). Fluorescence intensity was not changed in diabetic rats either (P < 0.05). CONCLUSION: Diabetes mellitus did not influence the tissue response to Angelus MTA(®) or the mineralization stimulated by it.