Elimination of intracanal infection in dogs' teeth with induced periapical lesions after rotary instrumentation: Influence of different calcium hydroxide pastes

The aim of this study was to evaluate the antiseptic efficacy of rotary instrumentation associated with calcium hydroxide-based pastes prepared with different vehicles and antiseptics. Chronic periapical lesions were experimentally induced in 72 premolar root canals of four dogs. Under controlled asepsis, after initial microbiological sampling (A1), the root canals were instrumented using the ProFile system in conjunction with 5.25% sodium hypochlorite and the intracanal medication was placed. Four experimental groups were formed according to the pastes used: group 1- Calen (n=18), group 2- Calen+CPMC (n=20), group 3- Ca(OH)2 p.a.+ anaesthetic solution (n=16) and group 4- Ca(OH)2 p.a.+ 2% chlorhexidine digluconate (n=18). After 21 days, the pastes were removed; the canals were emptied and 96 hours later a second microbiological sample was obtained (A2). The incidence of positive microbiological cultures and the number of cfus in stages A1 and A2 were compared statistically by the Wilcoxon test while the influence of the different treatments in intracanal infection was evaluated by Kruskal-Wallis test at 5% significance level (p<0.05). Large numbers of strict and facultative anaerobes, and viridans group streptococci were found in 100% of root canals of A1 samples. Among A2 samples, all treatments showed significant reduction of cfus and positive cultures (p<0.05), but only groups 3 and 4 showed 100% of root canals free of microorganisms. Rotary instrumentation plus NaOCl 5.25% associated with intracanal medication produced a drastic reduction or elimination of intracanal microbiota, whose performance was not influenced by the nature of the vehicle or the antiseptic added to the Ca(OH)2 p.a.

Genotoxicity in primary human peripheral lymphocytes after exposure to regular and white mineral trioxide aggregate

Objective: Taking into consideration that DNA damage plays an important role in carcinogenesis, the purpose of this study was to evaluate whether regular and white mineral trioxide aggregate (MTA) are able to induce genetic damage in primary human cells. Study design: Human peripheral lymphocytes obtained from 10 healthy volunteers were exposed to 2 presentation forms of MTA at final concentrations ranging from 1 to 1000 μg/mL for 1 hour at 37°C. The negative control group was treated with vehicle control (phosphate buffer solution, PBS) for 1 hour at 37°C and the positive control group was treated with hydrogen peroxide (at 100 μM) for 5 minutes on ice. Results were analyzed by the Friedman nonparametric test. Results: The results pointed out that either regular or white MTA in all concentrations tested did not induce DNA breakage in human peripheral lymphocytes as depicted by the mean tail moment. Conclusion: In summary, our results indicate that exposure to MTA may not be a factor that increases the level of DNA lesions in human peripheral lymphocytes as detected by single cell gel (comet) assay. © 2006 Mosby, Inc. All rights reserved.

In vitro antimicrobial activity of different gutta-percha points and calcium hydroxide pastes

The aim of this study was to evaluate the antimicrobial activity of different trademarks and compositions of gutta-percha points and calcium hydroxide pastes used in endodontic therapy. The evaluated material consisted of gutta-percha points containing calcium hydroxide (Roeko™), gutta-percha points containing chlorhexidine (Roeko™), two convencional gutta-percha points (Endo Points™ and Roeko™) and two calcium hydroxide pastes (Calen™ and Calen/PMCC™). Antimicrobial tests included five species of microorganisms: Escherichia coli (ATCC10538), Staphylococcus epidermidis (ATCC12228), Staphylococcus aureus (ATCC6538), Pseudomonas aeruginosa (ATCC27853), and Micrococcus luteus (ATCC9341). The Agar difusion method was employed. The plates were kept at room temperature for 2 h for prediffusion and then incubated at 37°C for 24 h. The triphenyltetrazolium chloride gel was added for optimization and the zones of inhibition were measured. Statistical evaluation was carried out using analysis of variance and Tukey Test. The obtained results showed that all microbial species used in the study were inhibited by the gutta-percha points containing chlorhexidine and by the calcium hydroxide pastes (Calen™ and Calen/ PMCC™), with similar results (p > 0.05). No antimicrobial activity was observed for the other groups. It was concluded that the gutta-percha points containing chlorhexidine presented antimicrobial activity, whereas the gutta-percha points containing calcium hydroxide did not.

Bone healing in critical-size defects treated with bioactive glass/calcium sulfate: A histologic and histometric study in rat calvaria

Objective: The purpose of this study was to analyze histologically the influence of bioactive glass (BG) with or without a calcium sulfate (CS) barrier on bone healing in surgically created critical-size defects (CSD) in rat calvaria. Material and methods: A CSD was made in each calvarium of 48 rats. They were divided into three groups: C (control): blood clot only; BG: defect filled with BG; and BG/CS: defect filled with BG covered by a CS barrier. Animals were euthanized at 4 or 12 weeks. Formation of new bone was evaluated histomorphometrically. Results: No defect completely regenerated with bone. BG particles were observed in Groups BG and BG/CS at both periods of analysis. The thickness throughout the healing area in Groups BG and BG/CS was similar to the original calvarium, while Group C presented a thin connective tissue in the center of the defect in both periods of analysis. At 4 weeks, Groups C and BG/CS presented significantly more bone formation than Group BG. No significant differences were found between Groups C and BG/CS. At 12 weeks, no significant differences in the amount of bone formation were observed among the three groups. When comparing 4 and 12 weeks, there was a significant increase in new bone formation within groups BG and BG/CS, but not C. Conclusion: BG particles, used with or without a CS barrier, maintained the volume and contour of the area grafted in CSD. However, they did not lead to a significant difference in bone formation when compared with control at 12 weeks post-operative. © 2007 Blackwell Munksgaard.

Effects of hemostatic agents on the histomorphologic response of human dental pulp capped with calcium hydroxide

Objective: To evaluate the response of human pulps capped with a calcium hydroxide [Ca(OH)2] cement after bleeding control with 2 hemostatic agents. Method and Materials: Pulps were exposed on the occlusal floor, and the bleeding was controlled either with saline solution (SS) or 2.5% sodium hypochlorite (NaOCI) (SH). After that, the pulp was capped with Ca(OH) 2 cement and restored with resin composite. After 30 (groups SS30 and SH30) and 60 (groups SS60 and SH60) days, the teeth were extracted and processed with hematoxylin-eosin and categorized in a histologic score system. The data were subjected to Kruskal-Wallis and Mann-Whitney tests (α = .05). Results: Regarding dentin bridge formation, an inferior response of SH60 group was observed when compared to SS60 (P < .05). The response of the SH30 group generally was similar to that of the groups treated with saline solution. However, after 60 days, 2.5% NaOCl showed a trend toward having an inferior response. Conclusion: Using saline solution as a hemostatic agent before pulp capping with Ca(OH)2 resulted in a significantly better histomorphologic response than using 2.5% NaOCl as a hemostatic agent before capping with Ca(OH)2.

Residual antibacterial activity of chlorhexidine digluconate and camphorated p-monochlorophenol in calcium hydroxide-based root canal dressings

The purpose of this study was to evaluate the residual antibacterial activity of several calcium hydroxide [Ca(OH) 2]-based pastes, placed in root canals of dogs' teeth with induced chronic periapical lesions. Root canals were instrumented with the ProFile rotary system and filled with 4 pastes: G1 (n=16): Ca(OH) 2 paste + anesthetic solution; G2 (n=20): Calen® paste + camphorated pmonochlorophenol (CMCP); G3 (n=18): Calen®; and G4 (n=18): Ca(OH) 2 paste + 2% chlorhexidine digluconate. After 21 days, the pastes were removed with size 60 K-files and placed on Petri plates with agar inoculated with Micrococcus luteus ATCC 9341. Pastes that were not placed into root canals served as control. After pre-diffusion, incubation and optimization, the inhibition zones of bacterial growth were measured and analyzed by Mann-Whitney U test at 5% significance level. All pastes showed residual antibacterial activity. The control samples had larger halos (p<0.05). The mean residual antibacterial activity halos in G1, G2, G3 and G4 were 7.6; 10.4; 17.7 and 21.4 mm, respectively. The zones of bacterial growth of G4 were significantly larger than those of G1 and G2 (p<0.05). In conclusion, regardless of the vehicle and antiseptic, all Ca(OH) 2-based pastes showed different degrees of measurable residual antibacterial activity. Furthermore, unlike CMCP, chlorhexidine increased significantly the antibacterial activity of Ca(OH) 2.

MTA-induced neutrophil recruitment: a mechanism dependent on IL-1β, 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-1β (IL-1β) 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-1β 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-1β antibodies. In the exudates, IL-1β and MIP-2 were detected. Conclusions: This study suggests that MTA induces NM via a mechanism dependent on MAST and MO mediated by IL-1β, MIP-2, and LTB4. © 2008 Mosby, Inc. All rights reserved.

Evidence for the role of calcineurin in morphogenesis and calcium homeostasis during mycelium-to-yeast dimorphism of Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is a dimorphic fungus that causes paracoccidioidomycosis, the most prevalent human deep mycosis in Latin America. The dimorphic transition from mycelium to yeast (M-Y) is triggered by a temperature shift from 25°C to 37°C and is critical for pathogenicity. Intracellular Ca 2+ levels increased in hyphae immediately after temperature-induced dimorphism. The chelation of Ca 2+ with extracellular (EGTA) or intracellular (BAPTA) calcium chelators inhibited temperature-induced dimorphism, whereas the addition of extracellular Ca 2+ accelerated dimorphism. The calcineurin inhibitor cyclosporine A (CsA), but not tacrolimus (FK506), effectively decreased cell growth, halted the M-Y transition that is associated with virulence, and caused aberrant growth morphologies for all forms of P. brasiliensis. The difference between CsA and FK506 was ascribed by the higher levels of cyclophilins contrasted to FKBPs, the intracellular drug targets required for calcineurin suppression. Chronic exposure to CsA abolished intracellular Ca 2+ homeostasis and decreased mRNA transcription of the CCH1 gene for the plasma membrane Ca 2+ channel in yeast-form cells. CsA had no detectable effect on multidrug resistance efflux pumps, while the effect of FK506 on rhodamine excretion was not correlated with the transition to yeast form. In this study, we present evidence that Ca 2+/calmodulin-dependent phosphatase calcineurin controls hyphal and yeast morphology, M-Y dimorphism, growth, and Ca 2+ homeostasis in P. brasiliensis and that CsA is an effective chemical block for thermodimorphism in this organism. The effects of calcineurin inhibitors on P. brasiliensis reinforce the therapeutic potential of these drugs in a combinatory approach with antifungal drugs to treat endemic paracoccidioidomycosis. Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Determination of calcium and magnesium in hydroethanolic extracts of propolis by atomic absorption flame spectrophotometry

Propolis is a natural product collected by honeybees and has a large range of pharmacological activity, including antimicrobial, antitumoral, antioxidant and anti-inflammatory. Its use as a popular medicine is increasing all over the world, creating a need for quality control of the commercial products. In this study the levels of calcium and magnesium in commercial hydroalcoholic propolis extracts from varies states of Brazil were determined by atomic absorption flame spectrophotometry and different values were obtained for northern and southern states. This study can be extended to the analysis of metals that are harmful to health. The results showed that the calibration curves were linear over a wide concentration range (0.5-4.0 μg.mL -1 for calcium and 0.05-0.4 μg.mL -1 for magnesium) with good correlation coefficients (0.999 and 0.988, respectively). Good analytical recovery (94%) was obtained. The proposed method showed adequate precision and relative standard deviation lower than 2 %. The method is accurate and precise as well as having advantages such as simplicity and speed.

Evaluation of two mineral trioxide aggregate compounds as pulp-capping agents in human teeth

Aim: The present randomized, controlled prospective study evaluated the histomorphological response of human dental pulps capped with two grey mineral trioxide aggregate (MTA) compounds. Methodology: Pulp exposures were performed on the occlusal floor of 40 human permanent pre-molars. The pulp was capped either with ProRoot (Dentsply) or MTA-Angelus (Angelus) and restored with zinc oxide eugenol cement. After 30 and 60 days, teeth were extracted and processed for histological examination and the effects on the pulp were scored. The data were subjected to Kruskal-Wallis and Conover tests (α = 0.05). Results: In five out of the 40 teeth bacteria were present in pulp tissue. No significant difference was observed between the two materials (P > 0.05) in terms of overall histological features (hard tissue bridge, inflammatory response, giant cells and particles of capping materials). Overall, 94% and 88% of the specimens capped with MTA-Angelus and ProRoot, respectively, showed either total or partial hard tissue bridge formation (P > 0.05). Conclusions: Both commercial materials ProRoot (Dentsply) and MTA-Angelus (Angelus) produced similar responses in the pulp when used for pulp capping in intact, caries-free teeth. © 2009 International Endodontic Journal.

In vivo biological behavior of calcium phosphate cements with different Ca/P ratio

Bioceramics with different Ca/P ratio were prepared from a mechanical mixture of NaPO3, CaCO3, Ca(OH)2 and phosphate buffer solution and implanted in rats subcutaneous tissues. The cements were characterized by Thermo gravimetric analysis (TG-TDA), X-ray diffraction and 31P-NMR. The implant sites were excised after 1, 4 and 16 weeks, fixed, dehydrated, included in paraffin wax for serial cutting and examined under the light transmitted microscope. They were biocompatible and biodegradable when implanted in rat subcutaneous. None of the materials induced ectopic osteogenesis. According to the results, the studied materials seem to be able for manufacturing reabsorbable bone implants.

Biological response to wollastonite doped α-tricalcium phosphate implants in hard and soft tissues in rats

The biological response following subcutaneous and bone implantation of β-wollastonite(β-W)-doped α-tricalcium phosphate bioceramics in rats was evaluated. Tested materials were: tricalcium phosphate (TCP), consisting of a mixture of α- and β-polymorphs; TCP doped with 5 wt. % of β-W (TCP5W), composed of α-TCP as only crystalline phase; and TCP doped with 15 wt. % of β-W (TCP 15), containing crystalline α-TCP and β-W. Cylinders of 2×1 mm were implanted in tibiae and backs of adult male Rattus norvegicus, Holtzman rats. After 7, 30 and 120 days, animals were sacrificed and the tissue blocks containing the implants were excised, fixed and processed for histological examination. TCP, TCP5W and TCP15W implants were biocompatible but neither bioactive nor biodegradable in rat subcutaneous tissue. They were not osteoinductive in connective tissue either. However, in rat bone tissue β-W-doped α-TCP implants (TCP5W and TCP 15W) were bioactive, biodegradable and osteoconductive. The rates of biodegradation and new bone formation observed for TCP5W and TCP15W implants in rat bone tissue were greater than for non-doped TCP.

Mitigating stress effects during transportation of matrinxã(Brycon amazonicus Günther, 1869; Characidae) through the application of calcium sulfate

This study verified the effects of CaSO4 on physiological responses of the tropical fish matrinxãBrycon amazonicus(200.2 ± 51.1 g) in water containing CaSO4 after a 4-h transportation at concentrations of: 0, 75, 150, and 300 mg L-1. Blood samples were collected prior to transportation (initial levels), immediately after packaging, at arrival, and 24 h and 96 h after transportation (recovery). Cortisol levels increased after ackaging (118.2 ± 14.2 ng ml-1), and decreased slightly after transportation in water containing CaSO4 (106.8 ± 14.1), but remained higher than initial levels (21.0 ± 2.6 ng ml)1). Fish kept at 150 mg L-1 CaSO4 reached the pre-transportation levels at 24 h of recovery. Blood glucose increased after transportation in all treatments (8.2 ± 0.2 mmol L-1) and declined after full recovery to values below initial levels (4.8 ± 0.1 mmol L-1). Chloride levels did not change in CaSO4 treatments; serum sodium concentrations decreased after packaging and after transportation. Serum calcium levels did not differ among treatments, but decreased after packaging and increased at 96 h of recovery. Hematocrit and the number of red blood cells were higher in all treatments after packaging and arrival, except in fish exposed to 300 mg L-1 CaSO4. Mean corpuscular volume increased in 75 mg L-1 CaSO4, which reached the higher VCM after transportation. Hemoglobin levels increased only after transportation, regardless of calcium sulfate levels. Handling before transportation and transportation itself were both stressful to fish; calcium sulfate at concentrations tested in the present work had a moderate influence in the reduction of stress responses. © 2009 Blackwell Verlag, Berlin.

Evaluation of the diffusion capacity of calcium hydroxide pastes through the dentinal tubules

This study aimed to evaluate the diffusion capacity of calcium hydroxide pastes with different vehicles through dentinal tubules. The study was conducted on 60 extracted single-rooted human teeth whose crowns had been removed. The root canals were instrumented and divided into 4 groups according to the vehicle of the calcium hydroxide paste: Group I - distilled water; Group II - propylene glycol; Group III - 0.2% chlorhexidine; Group IV - 2% chlorhexidine. After placement of the root canal dressings, the teeth were sealed and placed in flasks containing deionized water. After 1, 2, 7, 15, 30, 45 and 60 days, the pH of the water was measured to determine the diffusion of calcium hydroxide through the dentinal tubules. The data were recorded and statistically compared by the Tukey test. The results showed that all pastes presented a similar diffusion capacity through dentin. Group IV did not present difference compared to group I. Group II presented difference compared to the other groups, as did Group III. In conclusion, groups I and IV presented a better diffusion capacity through dentin than groups II and III; 2% chlorhexidine can be used as a vehicle in calcium hydroxide pastes. © 2009 Sociedade Brasileira de Pesquisa Odontológica.

Cytotoxic effects of white-MTA and MTA-Bio cements on odontoblast-like cells (MDPC-23)

This study evaluated the cytotoxic effects of 2 mineral trioxide aggregate (MTA) cements - White-MTA-Angelus and a new formulation, MTA-Bio - on odontoblast-like cell (MDPC-23) cultures. Twenty-four disc-shaped (2 mm diameter x 2 mm thick) specimens were fabricated from each material and immersed individually in wells containing 1 mL of DMEM culture medium for either 24 h or 7 days to obtain extracts, giving rise to 4 groups of 12 specimens each: G1 - White-MTA/24 h; G2 - White-MTA/7 days; G3 - MTA-Bio/24 h; and G4 - MTA-Bio/7 days. Plain culture medium (DMEM) was used as a negative control (G5). Cells at 30,000 cells/cm 2 concentration were seeded in the wells of 24-well plates and incubated in a humidified incubator with 5% CO 2 and 95% air at 37°C for 72 h. After this period, the culture medium of each well was replaced by 1 mL of extract (or plain DMEM in the control group) and the cells were incubated for additional 2 h. Cell metabolism was evaluated by the MTT assay and the data were analyzed statistically by ANOVA and Tukey's test (α=0.05). Cell morphology and the surface of representative MTA specimens of each group were examined by scanning electron microscopy. There was no statistically significant difference (p>0.05) between G1 and G2 or between G3 and G4. No significant difference (p>0.05) was found between the experimental and control groups either. Similar cell organization and morphology were observed in all groups, regardless of the storage periods. However, the number of cells observed in the experimental groups decreased compared to the control group. MTA-Bio presented irregular surface with more porosities than White-MTA. In conclusion, White-MTA and MTA-Bio presented low cytotoxic effects on odontoblast-like cell (MDPC-23) cultures.