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.

What is the response profile of deciduous pulp fibroblasts stimulated with E. coli LPS and E. faecalis LTA?

BACKGROUND: Oral fibroblast immunological responses to bacterial stimuli are well known. However, there are few studies about pulp fibroblasts from deciduous teeth (HDPF) responses, which are important for the treatment of pulp infections in children. The aim of this study was to evaluate expression and production of inflammatory cytokines and chemokines by HDPF when challenged with bacterial antigens normally present in advanced caries lesions. METHODS: Triplicate HDPF from 4 children (n = 4; 2 boys and 2 girls) were cultured by explant technique and challenged or not with Escherichia coli lipopolysaccharide/1 µg/mL (EcLPS) or Enterococcus faecalis lipoteichoic acid/1 µg/mL (EfLTA) for 6 and 24 h. Most of published studies employed immortalized cells, i.e., without checking possible gender and genetic variables. mRNA expression and protein production were evaluated by RT-qPCR and ELISA MILLIPLEX®, respectively, for Interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-17, Chemokine C-C motif ligand 2/monocyte chemoattractant protein 1 (CCL2/MCP-1), Chemokine C-C motif ligand 3/macrophage inflammatory protein 1-alpha (CCL3/MIP1-α), Chemokine C-C motif ligand 5/ regulated on activation, normal T cell expressed and secreted (CCL5/RANTES), C-X-C motif chemokine 12/ stromal cell-derived factor 1 (CXCL12/SDF-1), Tumor Necrosis Factor-alpha (TNF-α), Interferon-gamma (IFN γ), Vascular Endothelial Growth Factor (VEGF), Colony stimulating factor 1 (CSF-1) and Macrophage colony-stimulating factor (M-CSF). RESULTS: EcLPS increased IL-1α, IL-1ß, IL-8, CCL2, CCL5, TNF-α and CSF-1 mRNA and protein levels while EfLTA was only able to positively regulate gene expression and protein production of IL-8. CONCLUSION: The results of the present study confirmed our hypothesis, since pulp fibroblasts from deciduous teeth are capable of increasing gene expression and protein production after being stimulated with EcLPS and EfLTA.

Endodontic infections increase leukocyte and lymphocyte levels in the blood.

OBJECTIVES: The aim of this study was to determine whether the presence of single or multiple apical periodontitis (AP) alters blood cell counts and cytokine production. MATERIAL AND METHODS: Thirty rats were divided into three groups: a control group comprising rats without AP, a group called 1AP comprising rats with AP in one tooth, and a group called 4AP comprising rats with AP in four teeth. Endodontic infection was induced by pulp exposure of the first right maxillary molar in the 1AP group or by exposing the first and second right maxillary and mandibular molars in the 4AP group. A blood count and cytokine levels were obtained 30 days after infection by collecting blood by cardiac puncture. The maxillae were dissected and stained with hematoxylin and eosin to evaluate the inflammatory infiltrate. The data were tabulated and subjected to statistical analysis (P < 0.05). RESULTS: Histological analysis showed a predominance of mononuclear inflammatory cells. In blood, significant increase of leukocytes, lymphocytes, and tumor necrosis factor alpha (TNF-α) in 4AP compared with the control and 1AP groups (P < 0.05) was observed. In addition, significant decrease of interleukin-4 (IL-4) in 1AP and 4AP groups compared with the control was observed (P < 0.05). CONCLUSIONS: In the rat model, the presence of multiple AP can affect health by increasing lymphocyte and TNF-α levels in the blood. CLINICAL RELEVANCE: The presence of endodontic infections can interfere with the blood profile, altering systemic health.

Effect of different methods of polymerizing ocular prosthesis acrylic resin on a human conjunctival cell line.

STATEMENT OF PROBLEM: Ocular prosthesis acrylic resins should be biocompatible regardless of the polymerization method. The authors are unaware of a study that evaluated the biocompatibility of ocular prostheses. PURPOSE: The purpose of this in vitro study was to evaluate the cytotoxicity of different methods of polymerizing ocular prosthesis acrylic resin. This was accomplished by analyzing the cell proliferation, production of proinflammatory cytokines, and expression of extracellular matrix proteins related to tissue remodeling and repair of a human conjunctival cell line. MATERIAL AND METHODS: Nine acrylic resin specimens were divided into 3 groups: polymerization in a water bath, by microwave, or by autopolymerization. Eluates (prepared for 72 hours) were exposed to cells for 72 hours. A medium without specimens served as negative control (nonstimulated group). The tetrazolium dye MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was performed to evaluate the cytotoxic effect, and an enzyme-linked immunosorbent assay was executed for analysis of interleukin 1 ß (IL1ß), IL6, tumor necrosis factor α (TNFα), and CCL3/MIP1α production. Also, real-time reverse transcriptase (RT)-PCR was performed for analysis of mRNA expression of type IV collagen (COL IV), TGFß, and MMP9, and data were tested using ANOVA with Bonferroni post hoc test (α=.05). RESULTS: Microwave-processed resin showed slight cytotoxicity due to a significant reduction in cell proliferation and an increase in IL6 quantity. Higher levels of mRNA expression of COL IV, MMP9, and TGFß were verified in water bath-processed resin, which were similar to those in the nonstimulated group. CONCLUSIONS: Microwave-processed resin showed a significant reduction in cell proliferation and an increase in IL6 quantity. Heat-polymerized resin exhibited a higher mRNA expression of COL IV, MMP9, and TGFß; this result was similar to that in the nonstimulated group.

Effect of processing methods on the cytotoxicity of methyl methacrylate-based ocular prostheses: An in vitro study.

The study evaluated the influence of cycles and methods of an ocular prosthesis resin on cytotoxicity toward human conjunctival cells. Resins were polymerized by water bath (WB, 74 °C or 100 °C for 30 min to 9 h), microwave (MW, 1200 W, 3 to 14 min and 30 s at 0 to 720 W), or autopolymerization (AP, room temperature for 20 min ± 60 °C for 30 min). Degree of conversion (DC), cytotoxicity, level of inflammatory mediators, gene expression of different markers, and apoptosis were evaluated. Data were submitted to ANOVA and Tukey test (p < 0.05). WB with longer processing time at higher temperature had highest DC (85.6%) and higher TGF ß1-gene expression (1.39); long cycle low power MW showed lowest DC (69.6%), lower cell proliferation (85.4%, MTT), and large IL-2 release (39,297 ng/mL). AP with additional processing time showed lower cell proliferation (75.3%, Alamar Blue), and AP polymerized at room temperature showed higher CASP 9-gene expression (1.21). AP methods showed higher IL-6 release (>277 pg/mL). Short cycle medium power MW had higher IL-23 release (534.2 pg/mL). MW (long and short cycles) and AP polymerizations have triggered a more intense inflammatory response. Among methods recommended by the manufacturer, WB showed high DC and less cytotoxicity.

Biocompatible silver nanoparticles incorporated in acrylic resin for dental application inhibit Candida albicans biofilm.

BACKGROUND: Although silver nanoparticles (SNP) have proven antimicrobial activity against different types of microorganisms, the effect of SNP incorporation into acrylic resin to control Candida albicans biofilm formation aiming at the prevention of Candida-associated denture stomatitis has not yet been fully elucidated. OBJECTIVES: This study aimed to evaluate the antimicrobial effect of an acrylic resin containing SNP on C. albicans biofilm growth, the flexural strength of this material and tissue reaction in the subcutaneous connective tissue of rats to SNP. METHOD: SNP were synthesized through silver nitrate reduction by sodium citrate. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to verify the size and colloidal stability. SNP were added to acrylic resin monomer (Lucitone 550) at 0.05, 0.5 and 5 vol%. The antimicrobial effect against C. albicans (ATCC 10231) was investigated by the enumeration of colony-forming units (CFUs) and SEM. The three-point bending test was performed to analyze the flexural strength. Tissue reaction was evaluated after 7 and 60 days of implantation in the connective tissue of Wistar rats. RESULTS: Spherical particles of 5 and 10 nm were obtained. SNP at 0.05 and 0.5% incorporated into acrylic resin was effective in reducing C. albicans biofilm growth (p < .001). SEM revealed that the material was able to disrupt C. albicans biofilm formation and did not reduce the flexural strength compared to control (p > .05). The inflammatory response observed 60 days after implantation SNP in the subcutaneous tissue was similar to control. CONCLUSION: It was concluded that SNP addition at 0.05 and 0.5% into acrylic resin exhibited antimicrobial effects against C. albicans biofilm, did not interfere in the flexural strength and may be considered biocompatible.

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.

High p16INK4a immunoexpression is not HPV dependent in oral leukoplakia.

OBJECTIVE: To evaluate the prevalence of HPV DNA detection in fresh tissue from oral leukoplakia by Linear Array assay, and its correlation with p16INK4a immunoexpression in the northwest region of the São Paulo state, Brazil. PATIENTS AND METHODS: Fifty patients diagnosed with oral leukoplakia were included in the study. Sociodemographic, clinicopathologic and lifestyle data, fresh tissue and formalin fixed paraffin embedded (FFPE) tissue samples were collected. The fresh tissue was stored at -80 °C and then submitted to further viral DNA detection by the Linear Array method. Immunohistochemical analysis for the p16INK4a expression was performed. RESULTS: Of the 50 patients included in the study, 62 % were men, and the age ranged from 25 to 82 years. Twenty-three (46 %) were elderly, 46 % were middle-aged adults, and only 12 % were young adults. Most patients were smokers (76 %), 14 % were former smokers, and 10 % were non-smokers. Most patients (56 %) were current drinkers, 22 % were ex-drinkers and 22 % were non-drinkers. Thirty-two percent of the lesions presented some degree of dysplasia. No lesions were positive to HPV by Linear Array detection. Thirty (60 %) OL had p16-low immunoexpression and 20 (40 %) had p16-high immunoexpression. CONCLUSION: HPV was not identified in the population studied. The high p16INK4a immunoexpression is not dependent on HPV in oral leukoplakia. Broader epidemiological studies are required to clarify the geographic variability in the prevalence of HPV in head and neck squamous cell carcinoma and oral potentially malignant lesions.

Cytotoxicity and inflammatory response of different types of provisional restorative materials.

AIM: To evaluate the cytotoxicity and inflammatory response of different types of provisional restorative materials to mice gingival fibroblasts. METHODS: Cytotoxicity of provisional material discs (thermal-polymerized acrylic resin; auto-polymerized acrylic resin; bisacrylic resin; nano-ceramic resin for CAD/CAM and prefabricated polymer block for CAD/CAM) to Mice (Balb/c) gingival cell were investigated under direct and indirect contact (extracts) at 24, 48 and 72 h, using the MTT and Alamar blue assays. Materials extracts (24 h) were applied to the cell culture (indirect contact) or cells were seeded on discs of provisional materials, and the cytotoxicity and production of IL-6, IL-1ß and TNF-α after 24, 48 and 72 h were analyzed through MTT, Alamar Blue® and ELISA. Culture medium was used as control for indirect contact assay (extract) and the surfaces of the wells without discs of provisional materials were used as control for direct contact assay. Results were analysed statistically by ANOVA followed by the Bonferroni-Test correction. Statistically significant differences were considered if P was < .05. RESULTS: Auto-polymerized and bisacrylic resins (direct contact) reduced cell viability after 24, 48 and 72 h compared to control (P < .05). Indirect contact (extract) was not cytotoxic to cells at all periods compared to control (P > .05). Auto-polymerized and bisacrylic resins increased IL-6, IL-1ß and TNF-α levels mainly at 24 h when compared to the other materials (P < .001). CONCLUSION: Auto-polymerized and bisacrylic were more cytotoxic to mice gingival fibroblasts. CAD/CAM nano ceramic resin and prefabricated polymer blocks are more predictable materials to preserve the periodontal soft tissues.

Cytotoxicity and antimicrobial effects of citronella oil (Cymbopogon nardus) and commercial mouthwashes on S. aureus and C. albicans biofilms in prosthetic materials.

Although the effectiveness of some mouthwashes has been proven, phytotherapy is still a field to be explored as an alternative to commercial products. OBJECTIVE: To evaluate, in vitro, the cytotoxicity and efficacy of two solutions based on citronella oil (CN), on S. aureus and C. albicans biofilms (in formation-adhesion phase and 24 h-biofilm formation) on acrylic resin and nickel-chromium alloy samples (one trademark of each material), compared to two alcohol-free commercial mouthwashes. MATERIAL AND METHODS: Two solutions containing CN at concentrations of 5x and 10x the minimum bactericidal/fungicidal concentration (MBC/MFC) were prepared by microdilution. After contamination of the samples surfaces with these microorganisms, the mouthwashes (CN - 5x and 10x; CHX - 0,12% alcohol-free chlorhexidine and LT - alcohol-free essential oils) were evaluated. Mouthwash simulation was performed for 1 min at two moments, the first simulation after 4 h of microbial adhesion and 24 h-biofilm formation, and the second simulation, 6 h after the first simulation. For biofilm quantification, the number of cultured cells was evaluated by CFUs. The cytotoxicity assay was performed on HaCat epithelial cells and quantified by the MTT method. RESULTS: Tested solutions completely inhibited the growth of both microorganisms in the adhesion phase. All solutions showed inhibitory activity against 24 h-biofilm formation. However, CN led to greater microbial reduction, regardless of the surface of the sample. All solutions demonstrated a toxic effect. However, after serial dilution, CN presented the lowest cytotoxic effect. CONCLUSION: Citronella had a lower cytotoxic effect and a higher action compared to commercial solutions.

Influence of Preparation and Exposure Periods of Eluates from Ocular Prosthesis Acrylic Resin in Human Conjunctival Cell Line

Background: This study was undertaken to analyze if different preparation and exposure periods of eluates from ocular prosthesis acrylic resin influence the cytotoxicity for conjunctival cells. Methods: Twenty-four acrylic resin specimens were divided, according to the period of eluate exposure to Chang conjunctival cells (24 and 72 hours). Eluates were prepared in four different ways: 24, 48, and 72 hours of resin specimen immersion in medium and 24 hours of immersion in water, followed by 24 hours of immersion in medium. MTT assay was used to evaluate the cytotoxic effect. The production of IL-1ß, IL-6, TNF-α, and chemokine macrophage inflammatory protein 1α was evaluated by ELISA, while the mRNA expression of type IV collagen (COL IV), transforming growth factor ß (TGF-ß), and matrix metalloproteinase 9 (MMP9) were evaluated by real-time RT-PCR technique. The statistical analysis was carried out using ANOVA with Bonferroni post-hoc test and the student's t-test (p < 0.05). Results: Significant quantities of IL-6 (4.594 pg/mL) and mRNA expression of COL IV (1.58) were verified at 72 hours of eluate exposure to cells, as compared to 24 hours. After the 72-hour exposure of eluates to cells, lower cell proliferation (88.4%) and higher IL-6 quantities (12.374 pg/mL), as well as mRNA expression of COL IV (2.21), TGF-ß (2.02), and MMP9 (5.75) were observed, which corresponded to 72 hours of a specimen immersed in medium. Conclusion: Longer periods of eluate preparation and exposure from the acrylic resin to cells are related to higher production of proinflammatory cytokines and extracellular matrix proteins.

Effect of maternal periodontitis on GLUT4 and inflammatory pathway in adult offspring.

BACKGROUND: Maternal periodontal disease leads to low birth weight (LBW), insulin resistance (IR), increased TNF-α levels, and alterations in insulin signaling in adult offspring. TNF-α has been associated with the stimulation of IKKß/NF-κB, resulting in the decreased expression of GLUT4. Another mechanism that may be involved in decreasing GLUT4 expression is DNA methylation. This study aimed to evaluate in the adult offspring of rats with periodontal disease: IR, inflammatory pathways, DNA methylation, and expression of GLUT4. METHODS: Female Wistar rats were distributed into control and experimental periodontal disease groups. Seven days after induction of periodontal disease, both groups were mated with healthy male rats. After weaning, male offspring were distributed into control offspring (CN-o) and periodontal disease offspring (PED-o) groups. Body weights were measured from 0-75 days of age. At day 75, the following were measured in the offspring: IR (HOMA-IR index); TNF-α and NF-κBp65 content in the gastrocnemius muscle (GM) by western blotting; IKKα/ß, JNK, ERK 1/2, NF-κBp65, and NF-κBp50 phosphorylation status in the GM by western blotting; DNA methylation by restriction digest and real-time PCR(qAMP); and expression of GLUT4 mRNA in the GM by real-time PCR. RESULTS: LBW, IR, increases in TNF-α, IKKα/ß, ERK 1/2, NF-κBp65, and NF-κBp50 decreased expression of GLUT4 mRNA were observed in the PED-o rats. No differences were identified in JNK phosphorylation status and DNA methylation in the evaluated regions of the GLUT4-encoding gene Slc2a4. CONCLUSION: Maternal periodontal disease causes LBW, IR, activation of inflammatory pathways, and decreased GLUT4 expression in the GM of adult offspring.

Leukotriene B4 is essential for selective eosinophil recruitment following allergen challenge of CD4+ cells in a model of chronic eosinophilic inflammation.

Subcutaneous heat-coagulated egg white implants (EWI) induce chronic, intense local eosinophilia in mice, followed by asthma-like responses to airway ovalbumin challenge. Our goal was to define the mechanisms of selective eosinophil accumulation in the EWI model. EWI carriers were challenged i.p. with ovalbumin and the contributions of cellular immunity and inflammatory mediators to the resulting leukocyte accumulation were defined through cell transfer and pharmacological inhibition protocols. Eosinophil recruitment required Major Histocompatibility Complex Class II expression, and was abolished by the leukotriene B4 (LTB4) receptor antagonist CP 105.696, the 5-lipoxygenase inhibitor BWA4C and the 5-lipoxygenase activating protein inhibitor MK886. Eosinophil recruitment in EWI carriers followed transfer of: a) CD4+ (but not CD4-) cells, harvested from EWI donors and restimulated ex vivo; b) their cell-free supernatants, containing LTB4. Restimulation in the presence of MK886 was ineffective. CC chemokine receptor ligand (CCL)5 and CCL2 were induced by ovalbumin challenge in vivo. mRNA for CCL17 and CCL11 was induced in ovalbumin-restimulated CD4+ cells ex vivo. MK886 blocked induction of CCL17. Pretreatment of EWI carriers with MK886 eliminated the effectiveness of exogenously administered CCL11, CCL2 and CCL5. In conclusion, chemokine-producing, ovalbumin-restimulated CD4+ cells initiate eosinophil recruitment which is strictly dependent on LTB4 production.

Stress hormones concentrations in the normal microenvironment predict risk for chemically induced cancer in rats.

Evidence show that stress hormones can influence cancer progression, but its role in carcinogenesis is poorly understood. In this study, we used a new method based on oral carcinogenesis model in rats to test the hypothesis that physiological levels of stress hormones in the normal tissue microenvironment would have significant predictive value for chemically induced cancer occurrence. Male Wistar rats were submitted to a tongue biopsy for measuring not-stress induced levels of norepinephrine, corticosterone, adrenocorticotropic hormone (ACTH) and brain-derived neurotrophic factor (BDNF) in the tissue before carcinogenic induction. Rats were treated with the 4-nitroquinoline-1-oxide (4NQO) chemical carcinogen for twenty weeks and then euthanized for microscopic evaluation of the tongue lesions. Increased pre-carcinogen norepinephrine concentrations and reduced basal corticosterone levels in the normal tissue microenvironment were predictive for oral squamous cell carcinoma (OSCC) occurrence. Likewise, increased pre-carcinogen norepinephrine levels in the normal microenvironment were associated a lower expression of pCDKN2a-p16 in OSCCs. Post-carcinogen levels of corticosterone and BDNF in oral leukoplakia tissues (precursor lesion of OSCC) and post-carcinogen corticosterone concentrations in OSCCs were higher than basal levels in the normal mucosa. Increased norepinephrine concentrations in OSCCs were associated to a greater tumor volume and thickness. Furthermore, higher levels of norepinephrine, ACTH and BDNF in OSCCs were associated to a lesser intensity of the lymphoplasmocytic infiltrate. This study shows that pre-carcinogen stress hormones levels in the normal microenvironment may be predictive for chemically induced cancer in rats. Moreover, chemical carcinogenesis can promote stressor-like effects with hormonal changes in the tissue microenvironment, which may be associated to tumor progression.

Evaluation of the Cytotoxicity and Biocompatibility of New Resin Epoxy-based Endodontic Sealer Containing Calcium Hydroxide.

INTRODUCTION: Many endodontic sealers are available, but the search for the ideal sealer continues. This study evaluated the cytotoxicity and biocompatibility of Sealer Plus, a new resin epoxy-based endodontic sealer containing calcium hydroxide. AH Plus, Endofill, and SimpliSeal endodontics sealers were used for comparison. METHODS: L929 fibroblasts were cultured, and an MTT assay was used to determine the cytotoxicity of the sealer extracts at 6, 24, 48, and 72 hours. Tubes containing materials or empty tubes for control were inserted into the subcutaneous tissues of 20 rats. After 7 and 30 days, the rats were killed, and the tubes were removed with the surrounding tissues for histologic analysis. The data were submitted to statistical tests (P < .05). RESULTS: Undiluted Sealer Plus exhibited less cytotoxicity compared with other undiluted extracts at 6 hours (P < .05), and cell viability was higher for all Sealer Plus extracts after 24 hours (P < .05). At 48 hours, the undiluted and ½ Sealer Plus dilution were the extracts with less cytotoxicity (P < .05). At 72 hours, cell viability was higher for the undiluted and ½ Sealer Plus dilution compared with the other sealers (P < .05). At 7 days, Endofill and SimpliSeal had higher inflammation compared with the control and Sealer Plus (P < .05); AH Plus had moderate inflammation (P > .05). At 30 days, control, Sealer Plus, and AH Plus had less inflammation (P < .05). The fibrous capsule was thick at 7 days and thin at 30 days, except for SimpliSeal. CONCLUSIONS: In general, Sealer Plus promoted greater cell viability and was more biocompatible compared with the other sealers.

Periapical Lesions Increase Macrophage Infiltration and Inflammatory Signaling in Muscle Tissue of Rats.

INTRODUCTION: Our previous studies have shown that periapical lesions (PLs) in rats cause systemic disorders such as increased tumor necrosis factor-α plasma levels, insulin resistance, and impairment in insulin signal transduction in muscle tissue. However, the mechanisms involved in these alterations are not fully understood. Under chronic inflammatory conditions such as obesity, it has been shown that the skeletal muscle is affected by inflammation, and the number of resident macrophages that are associated with impairments of insulin action and sensitivity is increased. This study aimed to investigate the presence of macrophages, activation of inflammatory pathways in muscle tissue, glycemia, and insulinemia of rats with PLs. METHODS: Sixty Wistar rats were distributed into a control group; a group with 1 PL (1PL), which was induced in the right maxillary first molar; and a group with 4 PLs (4PL), which were induced in the right upper and lower first and second molars. We quantified macrophage content by immunohistochemistry for the F4/80 protein. We evaluated Jun N-terminal kinase and IKKα/ß phosphorylation status in the muscle tissue by Western blotting. Serum levels of lipopolysaccharide (LPS) and HSP70 and plasma levels of glucose and insulin were assessed by using commercial kits. RESULTS: The 1PL and 4PL groups showed increase in macrophage content, IKKα/ß, and Jun N-terminal kinase phosphorylation status, serum LPS and HSP70 levels, and insulin resistance and no changes in glycemia and insulinemia compared with the control group. There was no difference in these parameters between the 1PL and 4PL groups. CONCLUSIONS: PLs promoted an increase in macrophage infiltration, activation of inflammatory pathways in muscle tissue, and serum concentrations of HSP70 and LPS in rats. The present study improves the knowledge on the impact of oral inflammations on the development of systemic alteration, which can induce insulin resistance.

Evaluation of photodynamic therapy on fibroblast viability and cytokine production.

BACKGROUND: The aim of this study was to evaluate the effects of photodynamic therapy with curcumin (PDT) comparatively to 5% sodium hypochlorite (NaOCl) and saline solution on cell viability and cytokine (IL-1ß and IL-6) production by mouse fibroblasts. METHODS: Sixty seconds of pre-irradiation time with curcumin 500mg/L and Led wavelength (λ) 480nm, 72Jcm(2), for 300s was used for PDT. Solutions were diluted in culture medium DMEM (1×10(4) cells) and placed into 24-well cell culture plates with mouse fibroblasts L-929. Culture medium was used as control. After 6, 24 and 48h, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT) was used to evaluate the cell viability and the supernatant was collected for cytokine evaluation using enzyme-linked immunosorbent assay (ELISA). The results were statistically analyzed by ANOVA and BonFerroni correction (p<0.05) for MTT and Kruskal-Wallis test and Dunn (p<0.05) for ELISA. RESULTS: PDT and saline solution presented low cytotoxic effect similar to the control group (p>0.05) while 5% NaOCl was more cytotoxic than PDT (p<0.05) in all periods of time. All materials similarly expressed IL-1ß and IL-6 regardless to the experimental period (p<0.05). CONCLUSIONS: PDT with curcumin was not cytotoxic to L929 fibroblasts differently from 5% NaOCl. In all groups occurred similar expression of IL-1ß and IL-6.

Salivary alpha amylase and cortisol levels in children with global developmental delay and their relation with the expectation of dental care and behavior during the intervention.

The purpose of this study was to analyze the alpha-amylase (sAA) and cortisol levels in children with Global developmental delay (GDD) before and after dental treatment and its association with the children's behavior during treatment. The morning salivary cortisol levels and activity of sAA of 33 children with GDD were evaluated before and after dental treatment and were compared to 19 healthy children. The behavior of children with GDD during dental care was assessed by the Frankl scale. Children with GDD showed lower levels of sAA activity than healthy children, but this result was not significant. The salivary cortisol levels were similar between GDD and healthy children. GDD children showed increased levels of sAA (but not cortisol) prior to the dental treatment as compared to the post-treatment phase. GDD children who showed less favorable behavior during dental care had higher levels of sAA and salivary cortisol than GDD children with more favorable behavior, but only the sAA results were significant. In conclusion, GDD children show hyperactivity of the SNS-axis in anticipation of dental treatment which indicates the need for strategies to reduce their anxiety levels before and during dental care.

MTA-induced neutrophil recruitment: a mechanism dependent on IL-1beta, MIP-2, and LTB4.

OBJECTIVE: The objective of this study was to investigate the mediators and the resident peritoneal cells involved in the neutrophil migration (NM) induced by mineral trioxide aggregate (MTA) in mice. STUDY DESIGN: MTA (25 mg/cavity) was injected into normal and pretreated peritoneal cavities (PC) with indomethacin (IND), dexamethasone (DEX), BWA4C, U75302, antimacrophage inflammatory protein-2 (MIP-2), and anti-interleukin-1beta (IL-1beta) antibodies and the NM was determined. The role of macrophage (MO) and mast cells (MAST) was determined by administration of thioglycollate 3% or 48/80 compound, respectively. The concentration of IL-1beta and MIP-2 exudates was measured by ELISA. RESULTS: MTA induced dose- and time-dependent NM into mice PC, with the participation of MO and MAST. NM was inhibited by DEX, BWA4C, and U75302, as well as anti-MIP-2 and anti-IL-1beta antibodies. In the exudates, IL-1beta and MIP-2 were detected. CONCLUSIONS: This study suggests that MTA induces NM via a mechanism dependent on MAST and MO mediated by IL-1beta, MIP-2, and LTB(4).

Mechanism of calcium hydroxide-induced neutrophil migration into air-pouch cavity.

The aim of this study was to investigate cellular migration induced by calcium hydroxide to air-pouch cavities in mice. The migration was more specific to neutrophil and was dose and time dependent (peaking 96 h after stimulation). This migration was inhibited by pretreatment with thalidomide, indomethacin, MK886, meloxicam, dexamethasone, MK886 associated with indomethacin, and MK886 associated with indomethacin and dexamethasone. The air-pouch exudate from animals stimulated with calcium hydroxide showed an increase of leukotriene-B(4) (LTB(4)), interleukin-1beta, tumor necrosis factor alpha (TNF-alpha), cytokine-induced neutrophil chemoattractant (KC), and macrophage inflammatory protein 2 (MIP-2) release. Pretreatment with 3% thioglycollate increased the macrophage population in the air pouch but did not change neutrophil migration. Depleting the resident mast cells through chronic pretreatment with compound 48/80 did not alter neutrophil migration in response to calcium hydroxide. It was possible to conclude that calcium hydroxide-induced neutrophil migration to the air-pouch cavity in mice is mediated by LTB(4), TNF-alpha, KC, MIP-2, and prostaglandins, but it was not dependent on macrophages or mast cells.