RANKL Triggers Treg-Mediated Immunoregulation in Inflammatory Osteolysis.

The chronic inflammatory immune response triggered by the infection of the tooth root canal system results in the local upregulation of RANKL, resulting in periapical bone loss. While RANKL has a well-characterized role in the control of bone homeostasis/pathology, it can play important roles in the regulation of the immune system, although its possible immunoregulatory role in infectious inflammatory osteolytic conditions remains largely unknown. Here, we used a mouse model of infectious inflammatory periapical lesions subjected to continuous or transitory anti-RANKL inhibition, followed by the analysis of lesion outcome and multiple host response parameters. Anti-RANKL administration resulted in arrest of bone loss but interfered in the natural immunoregulation of the lesions observed in the untreated group. RANKL inhibition resulted in an unremitting proinflammatory response, persistent high proinflammatory and effector CD4 response, decreased regulatory T-cell (Treg) migration, and lower levels of Treg-related cytokines IL-10 and TGFb. Anti-RANKL blockade impaired the immunoregulatory process only in early disease stages, while the late administration of anti-RANKL did not interfere with the stablished immunoregulation. The impaired immunoregulation due to RANKL inhibition is characterized by increased delayed-type hypersensitivity in vivo and T-cell proliferation in vitro to the infecting bacteria, which mimic the effects of Treg inhibition, reinforcing a possible influence of RANKL on Treg-mediated suppressive response. The adoptive transfer of CD4+FOXp3+ Tregs to mice receiving anti-RANKL therapy restored the immunoregulatory capacity, attenuating the inflammatory response in the lesions, reestablishing normal T-cell response in vivo and in vitro, and preventing lesion relapse upon anti-RANKL therapy cessation. Therefore, while RANKL inhibition efficiently limited the periapical bone loss, it promoted an unremitting host inflammatory response by interfering with Treg activity, suggesting that this classic osteoclastogenic mediator plays a role in immunoregulation.

Analysis of anticaries potential of pit and fissures sealants containing amorphous calcium phosphate using synchrotron microtomography.

The aim of this study was to analyze the anticaries potential of pit and fissure sealants containing amorphous calcium phosphate (ACP) by synchrotron microtomography. Bovine enamel blocks (4×4 mm; n=50) were selected through surface hardness (Knoop) analysis. Slabs were obtained through cross-sections taken 1 mm from the border of the enamel. Five indentations, spaced 100 µm apart, were made 300 µm from the border. Ten specimens were prepared for each tested material (Ultraseal XT plus TM, Aegis, Embrace, Vitremer and Experimental Sealant). The materials were randomly attached to the sectioned surfaces of the enamel blocks and fixed with sticky wax. The specimens were submitted to pH cycling. After that, the surface hardness (SH1) was determined, and the blocks were submitted to synchrotron microcomputed tomography analysis to calculate the mineral concentration (ΔgHAp cm(-3)) at different areas of the enamel. The comparison between the SH1 and ΔgHAp cm(-3) showed a correlation for all groups (r=0.840; p<0.001). The fluoride groups presented positive values of ΔgHAp cm(-3), indicating a mineral gain that was observed mainly in the outer part of the enamel. The ACP showed mineral loss in the outer enamel compared with fluoride groups, although it inhibited the demineralization in the deeper areas of enamel. The combination of two remineralizing agents (fluoride and ACP) was highly effective in preventing demineralization.

Toxicological in vitro and subchronic evaluation of LASSBio-596.

LASSBio-596, 2-[4-(1,4-tiazinan-4-ylsulfonyl) phenylcarbamoyl] benzoic acid, is an achiral compound containing a subunit carboxylic amide, was capable of preventing induced mechanical and morphological changes in the lungs that commonly caused the onset of asthma. Previous studies to determine the acute toxicity of oral LASSBio-596 at dose of 2000mg/kg caused no deaths in any of the tested animals. To further evaluate the safety of LASSBio-596, in vitro and in vivo tests were carried out. Regarding to in vitro test were used renal, hepatic, pulmonary, cardiac, neurologic and intestinal cell lines. They were evaluated using neutral red (NR) and [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assays. Micronuclei also was performed. Concerning to in vivo was performed subchronic on Wistar rats at doses of 10, 50, and 250mg/kg and zebrafish test. The in vitro tests results showed the safety of LASSBio-596. However, subchronic toxicity study results revealed changes in the blood parameters of amylase, alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose and creatine kinase (CK) which is used for cardiotoxicity evaluation, although, did not identify any histopathological alterations. However, zebrafish test demonstrated cardiac damage. It was impossible to estimate the no-observed-adverse-effect-levels and lowest observed-adverse-effect level due to the presence of cardiotoxicity in all tested doses.

In vitro evaluation of the effect of mouth rinse with trimetaphosphate on enamel demineralization.

OBJECTIVE: The aim of this study was to investigate the effectiveness of sodium trimetaphosphate (TMP) addition to mouth rinses to inhibit enamel demineralization. DESIGN: Bovine enamel blocks (n = 88) were selected by surface hardness and divided into eight treatment groups (n = 11 per group): placebo, 100 or 225 µg F/ml; the rinses with 100 µg F/ml had differing TMP concentrations (range 0-0.6%). The blocks were subjected to pH cycling for 5 days and treated twice a day with mouth rinses. After that, surface and cross-sectional hardness as well as fluoride in enamel were measured. RESULTS: The groups containing both 100 µg F/ml and 0.4% TMP inhibited demineralization most effectively (p < 0.001). This formulation yielded lower values of lesion areas than the formulations containing 100 or 225 µg F/ml but no TMP. The addition of 0.4% TMP increased the fluoride in enamel. CONCLUSION: It is possible to improve the effectiveness of a mouth rinse with 100 µg F/ml by addition of TMP, this being superior in inhibiting enamel demineralization compared with mouth rinses containing 225 µg F/ml.

Toxicological, toxicokinetic and gastroprotective evaluation of the benzaldehyde semicarbazone.

Benzaldehyde semicarbazone (BS) has presented positive results in several pharmacological models, including anticonvulsivant and anti-inflammatory models. The present study evaluated the preclinical toxicity (acute and subchronic), as well as the toxicokinetic and gastroprotective effects of BS against ethanol lesions. Oral doses of 300 and 2000mg/kg were used in the preclinical acute toxicity study; 100, 200, and 300mg/kg were used in both the subchronic toxicity evaluation and the gastric study; and 300mg/kg was used in the toxicokinetic study. No impact from the dose of 300mg/kg could be identified; while, one animal died at 2000mg/kg in the acute toxicity test. In the subchronic toxicity test, changes in the biochemical parameters of the liver, as well as in the histopatological evaluation, demonstrated that BS is a hepatotoxic drug. BS proved to be effective for moderate and severe gastric lesions. In the toxicokinetics study, BS presented a low concentration and rapid plasma disappearance. Several results also indicate that BS is likely to be mostly eliminated from the liver and may well undergo a first-pass effect after oral absorption. It was impossible to estimate the noobserved-adverse-effect-levels (NOAEL) and lowest-observed-adverse-effect-levels (LOAEL) due to the presence of hepatotoxicity in all tested doses.

Campomanesia lineatifolia Ruiz & Pav. as a gastroprotective agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia lineatifolia Ruiz and Pav. (Myrtaceae) is a native edible species found in the Amazon Rainforest, commonly known as gabiroba. In Brazil, Campomanesia species are frequently used in traditional medicine for gastrointestinal disorders. MATERIALS AND METHODS: The present study performed phytochemical analyses and determined both the in vitro antioxidant activity of the ethanolic extract of Campomanesia lineatifolia leaves (EEC) as well as its ethyl acetate fraction (EAFC). In this analysis, quercetin was used as a positive control. Gastroprotective activity was also investigated at different oral doses in two experimental models in rats - gastric lesion induced by ethanol and gastric lesion induced by indomethacin. In this analysis, cimetidine and sucralfate were used as positive controls. The area of gastric lesion underwent macroscopic and histomorphometric evaluations, while the mucus content was estimated by applying the periodic acid-Schiff stain. Oral acute toxicity was also assessed. RESULTS: Phytochemical studies revealed the presence of flavonoids and tannins. Catechin and quercitrin were isolated by bioguided chromatographic fractionation of EAFC. EEC and EAFC presented in vitro antioxidant activity. The oral administration of EEC and EAFC at doses 100-400 mg/kg (ethanol model) and at doses of 400-1200 mg/kg (indomethacin model) proved to be effective in preventing gastric ulcerations in rats. Pretreatment with EAFC (400mg/kg) significantly increased the gastric mucus content in the ethanol model. No animals died during the acute oral toxicology test. CONCLUSIONS: Results confirm the Brazilian ethnopharmacological use of Campomanesia lineatifolia as a gastroprotective agent and indicate that the anti-ulcer effect is most likely mediated by scavenging free radicals due to the polyphenol content and, at least in part, by increasing the mucus secretion and the mucosal defense. In addition, EEC and EAFC were found to be safe when applied to a 2000 mg/kg single oral dose.

Effect of mouth rinses with fluoride and trimetaphosphate on enamel erosion: an in vitro study.

The effect of mouth rinses containing fluoride (100 µg/ml) and sodium trimetaphosphate (TMP) on enamel erosion was evaluated in vitro. Bovine enamel blocks were subjected to erosive challenges 4 times per day for 5 min, followed by treatment with placebo, 225 µg F/ml, 100 µg F/ml, 100 µg F/ml and TMP (0.2, 0.4 and 0.6%) solutions (30 s) and storage in artificial saliva, over a duration of 5 days. TMP groups showed lower enamel wear than fluoride-only and placebo groups (p < 0.05). Addition of TMP at a TMP:NaF molar proportion between 1.24:1 and 3.72:1 to a solution containing 100 µg F/ml presented a greater protective effect under erosive conditions than a solution containing 225 µg F/ml, in the absence of TMP.

Effect of rinsing with water immediately after neutral gel and foam fluoride topical application on enamel remineralization: An in situ study.

OBJECTIVE: The aim of this study was to evaluate, in situ, the effect of rinsing with water immediately after neutral fluoride foam application (Foam) or fluoride gel application (Gel). DESIGN: Ten volunteers wore acrylic palatal appliances containing 4 enamel blocks selected by surface hardness with artificial caries. Five experimental regimes of 3 days each were set according to treatment: placebo; Gel and Foam followed by no rinsing or consuming of liquids or solids for the next 30min; Gel and Foam followed by immediately washing with water jet. After each phase, surface hardness was again measured for analysis of mineral gain, evaluated through percentage of surface hardness recovery (%SHR) and integrated loss of subsurface hardness (ΔKHN). The concentration of loosely bound fluoride (CaF2) and firmly bound fluoride (FA-like) formed and retained were also determined. RESULTS: Fluoride treatments produced greater remineralization (%SHR and ΔKHN) compared to placebo group (p<0.05). There was no difference in the ability to promote remineralization and in the concentration of fluoride formed and retained, in each analysis, between Gel and Foam (p>0.05). CONCLUSION: The data suggest that rinsing with water immediately after topical application does not seem to have an influence on the ability of fluoride to promote remineralization.

Comparison of methods for evaluating mineral loss: hardness versus synchrotron microcomputed tomography.

This study analyzed degrees of demineralization in bovine enamel using synchrotron microcomputed tomography (SMCT) and hardness measurements (Knoop hardness number, KHN). For 5 days, 40 bovine enamel blocks were individually subjected to a pH cycling model and treatment with fluoride dentifrices (placebo, 275, 550 and 1,100 microg F/g) diluted in deionized water twice a day. Surface hardness number and cross-sectional profiles of hardness and mineral concentration (by SMCT) were determined. Integrated hardness (KHN x microm) for sound and demineralized specimens was calculated and subtracted to give the integrated loss of hardness (DeltaKHN) for the lesions. Increasing fluoride concentration in the dentifrices led to higher values for surface hardness after pH cycling and mineral concentration (g(HAp) cm(-3)), and lower values for DeltaKHN (p < 0.05). From the present results, it may be concluded that hardness measurements revealed demineralization in all groups, which was lower in groups treated with dentifrice with a higher F concentration. SMCT and hardness measurements gave similar results in areas with higher demineralization, but diverged in areas with lower demineralization.

Synchrotron microComputed Tomography of the mature bovine dentinoenamel junction.

The mature dentinoenamel junction (DEJ) is viewed by some investigators and the current authors, not as a fossilized, sharp transition between enamel and dentin, but as a relatively broad structural transition zone including the mantle dentin and the inner aprismatic enamel. In this study, the DEJ structure in bovine incisors was studied with synchrotron microComputed Tomography (microCT) using small cubes cut parallel to the tooth surface. The reconstructions revealed a zone of highly variable punctate contrast between bulk dentin and enamel; the mean linear attenuation coefficients and their standard deviations demonstrated that this zone averaged less mineral than dentin or enamel but had more highly variable structure than either. The region with the punctuate contrast is, therefore, the mantle dentin. The thickness of the mantle dentin seen in a typical data set was about 30 microm, and the mantle dentin-enamel interface deviated +/-15 microm from the average plane over a distance of 520 microm. In the highest resolution data ( approximately 1.5 microm isotropic voxels, volume elements), tubules in the dentin could be discerned in the vicinity of the DEJ. Contrast sensitivity was high enough to detect differences in mineral content between near-surface and near-DEJ volumes of the enamel. Reconstructions before and after two cubes were compressed to failure revealed cracks formed only in the enamel and did not propagate across the mantle dentin, regardless of whether loading was parallel to or perpendicular to the DEJ.

[Erosion effect of a newly developed soft drink]. / De erosiepotentie van een nieuwe frisdrank.

In this article, the erosion effect of a newly developed soft drink available in 3 different flavours is compared with the erosion effect of some common soft drinks. Several basic parameters of the soft drinks were determined: pH, titratable acidity, and the calcium as well as phosphate concentration. Three methods were used to assess in vitro the erosion depth of the soft drinks in enamel specimens. Erosion depth is defined as the mean height of lost enamel. The methods used were atomic absorption spectroscopy for calcium loss, a colorimetric ammonium molybdate method for phosphate loss and non-contact profilometry for enamel volume loss. The newly developed soft drink realized statistically significantly less erosion depth when compared with cola and apple juice and statistically non-significantly less erosion depth when compared with orange soda.

Conservative treatment of a radicular cyst in a 5-year-old child: a case report.

The present report describes the management of a radicular cyst in a 5-year-old child. The treatment comprised extraction of the primary teeth involved followed by marsupialization. A removable appliance with a resin extension penetrating into the cystic cavity was used to help decompress the lesion. This treatment allowed rapid healing of the lesion and eruption of the permanent incisors without the need for orthodontic treatment.

Molecular and biochemical screening for the diagnosis and management of medullary thyroid carcinoma in multiple endocrine neoplasia type 2A.

Patients with Multiple Endocrine Neoplasia (MEN) type 2A are at risk for early medullary thyroid carcinoma (MTC). We performed different screening tests for MTC--a recently reported biochemical screening test using omeprazole-induced calcitonin (CT) stimulation and DNA analysis--in fifteen members of two non-consanguineous Brazilian families with MEN 2A. RET proto-oncogene analysis was carried out by direct DNA sequencing of PCR-amplified products for exons 10 and 11. Family 1 showed a germline mutation (C634Y) in three individuals; a sister and a brother with symptomatic MTC; the former also presented with pheochromocytoma and hyperparathyroidism, and her son was a nine-year-old boy of previously unknown status. Family 2 showed the C634R mutation only in the index case, who presented with cutaneous lichen amyloidosis in addition to MTC, pheochromocytoma and hyperparathyroidism. Neither her parents nor her four brothers showed this genetic abnormality, suggesting a de novo RET proto-oncogene mutation in this patient. The controls and patients presented normal basal gastrin levels and a significant increase after omeprazole. Basal CT levels were elevated in patients with MTC and undetectable in control and asymptomatic family members. No subject showed any increase in CT levels after omeprazole treatment. In conclusion, the two most frequent RET proto-oncogene mutations in MEN 2A are present in Brazilian families. In addition, the specificity of basal and omeprazole-stimulated calcitonin is rather limited, and the efficacy of the omeprazole test still needs to be systematically examined. Therefore, RET proto-oncogene analysis must be the first choice for a screening procedure to identify gene carriers in MEN 2A family members and to permit early prophylactic treatment of MTC.

Oral trauma in Brazilian patients aged 0-3 years.

The aim of the present investigation was to gather data pertaining to dental trauma in 1654 patients aged 0-3 years, attended at the Baby Clinic of the School of Dentistry at Araçatuba-UNESP, Brazil. The prevalence of traumatic injuries was 16.3%. There was greater involvement of boys (62.6%), of children aged 1-2 years (39.9%) and of the maxillary central incisors (86%). Falls were more often the etiology for dental injuries (58.3%). There was a predominance of uncomplicated crown fractures (48.4%).

Use of dental implants in children: a literature review.

The aim of this literature review is to discuss the use of dental implants in growing patients and the influence of maxillary and mandibular skeletal and dental growth on the stability of those implants. It is recommended to wait for the completion of dental and skeletal growth, except for severe cases of ectodermal dysplasia.

Treatment of mucus retention phenomena in children by the micro-marsupialization technique: case reports.

The purpose of the present study was to emphasize the technique of micro-marsupialization as an alternative for the treatment of mucus retention phenomena. Out of 41 patients, 14 were selected for treatment by the micro-marsupialization technique on the basis of clinical criteria. Patient age ranged from 5-9 years. The technique was performed as follows: the area was disinfected with 0.1% iodine; a topical anesthetic was applied to cover the entire lesion for approximately 3 min; a 4.0 silk suture was passed through the internal part of the lesion along its widest diameter; and a surgical knot was made. Of the original 14 patients treated by the micro-marsupialization technique, 12 presented full regression one week after treatment. Recurrence occurred in two cases. It was possible to conclude that the micro-marsupialization technique is an alternative to be considered, especially in pediatric dentistry.