Tissue reaction of the EndoREZ in root canal fillings short of or beyond an apical foramenlike communication

This study evaluated the response of periapical tissues to the endodontic sealer EndoREZ in root canal fillings short of or beyond the apical foramenlike communication. Twenty root canals of premolars and incisors of 2 mongrel dogs were used. After coronal access and pulp extirpation, the canals were instrumented up to a size 55 K-file and the apical cemental barrier was penetrated with a size 15 K-file to create an apical foramenlike communication, which was widened to a size 25 K-file. The canals were irrigated with saline at each change of file. The root canals were obturated either short of or beyond the apical foramenlike opening by the lateral condensation of gutta-percha and EndoREZ, originating 2 experimental groups: G1, EndoREZ/short of the apical foramenlike opening, and G2, EndoREZ/beyond the apical foramenlike opening. The animals were killed by anesthetic overdose 90 days after endodontic treatment. The individual roots were obtained and serial histological sections were prepared for histomorphological analysis (H&E and Brown and Brenn techniques) under light microscopy. The following parameters were examined: closure of the apical foramenlike communication and apical opening of accessory canals, apical cementum resorptions, intensity of the inflammatory infiltrate, presence of giant cells, and thickness and organization of the apical periodontal ligament. Each parameter was scored 1 to 4, 1 being the best result and 4 the worst. Data were analyzed statistically by the Wilcoxon nonparametric tests (P = .05). Comparing the 2 groups, the best result (P = .05) was obtained with root canal filling with EndoREZ short of the apical foramenlike opening. In conclusion, limiting the filling material to the root canal space apically was important to determine the best treatment outcome when EndoREZ was used as the sealer. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: e94-e99)

Comparison of the biocompatibility of different root canal irrigants

The purpose of this study was to compare the reaction of rat subcutaneous connective tissue to 0.9% sterile saline, 2.5% sodium hypochlorite (NaOCl), 5.25% NaOCl and 2% chlorhexidine gluconate solution or gel. Six circles were demarcated on the dorsal skin of 24 male Wistar rats, leaving 2 cm between each circle. Using a syringe, 0.1 mL of each root canal irrigant was injected subcutaneously into 5 circles. In the 6th circle, the needle of an empty syringe was introduced into the skin, but no irrigant was injected (control group). Evaluations were undertaken at 2 h, 48 h, 14 days and 30 days post-procedure. Tissue samples were excised, embedded in paraffin blocks and 3-mu m-thick sections were obtained and stained with hematoxylin and eosin. The areas of inflammatory reaction were evaluated and analyzed statistically by ANOVA and Tukey's test. The control group showed few or no inflammatory reaction areas in the subcutaneous tissue. 0.9% saline solution, 2.0% chlorhexidine solution and 2.5% NaOCl showed a good biocompatibility, as very mild inflammatory reaction was detected at 14 days and tissue repair occurred at 30 days. 5.25% NaOCl was the most toxic irrigant, as the number of inflammatory cells remained elevated at 14 and 30 days. The group treated with 2.0% chlorhexidine gluconate gel presented a moderate inflammatory response at 14 days, which decreased at 30 days, being considered similar to that of the control group, 0.9% saline solution, 2.0% chlorhexidine solution and 2.5% NaOCl at this experimental period.

Immunotherapy against experimental canine visceral leishmaniasis with the saponin enriched-Leishmune((R)) vaccine

In order to assess the immunotherapeutic potential on canine visceral leishmaniasis of the Leishmune (R) vaccine, formulated with an increased adjuvant concentration (1 mg of saponin rather than 0.5 mg), 24 mongrel dogs were infected with Leishmania (L.) chagasi. The enriched-Leishmune (R) vaccine was injected on month 6, 7 and 8 after infection, when animals were seropositive and symptomatic. The control group were injected with a saline solution. Leishmune (R)-treated dogs showed significantly higher levels of anti-FML IgG antibodies (ANOVA; p < 0.0001), a higher and stable IgG2 and a decreasing IgG I response, pointing to a TH1 T cell mediated response. The vaccine had the following effects: it led to more positive delayed type hypersensitivity reactions against Leishmania lysate in vaccinated dogs (75%) than in controls (50%), to a decreased average of CD4+ Leishmania-specific lymphocytes in saline controls (32.13%) that fell outside the 95% confidence interval of the vaccinees (41.62%, CI95% 43.93-49.80) and an increased average of the clinical scores from the saline controls (17.83) that falls outside the 95% confidence interval for the Leishmune (R) immumotherapy-treated dogs (15.75, CI95% 13.97-17.53). All dogs that received the vaccine were clustered, and showed lower clinical scores and normal CD4+ counts, whereas 42% of the untreated dogs showed very diminished CD4+ and higher clinical score. The increase in clinical signs of the saline treated group was correlated with an increase in anti-FML antibodies (p < 0.0001), the parasitological evidence (p = 0.038) and a decrease in Leishinania-specific CD4+ lymphocyte proportions (p = 0.035). These results confirm the immunotherapeutic potential of the enriched-Leishmune (R) vaccine. The vaccine reduced the clinical symptoms and evidence of parasite, modulating the outcome of the infection and the dog's potential infectiosity to phlebotomines. The enriched-Leishmune (R) vaccine was subjected to a safety analysis and found to be well tolerated and safe. (c) 2007 Elsevier Ltd. All rights reserved.

In vitro cytotoxicity of five glass-ionomer cements

To evaluate the cytotoxic effects of five glass-ionomer cements (GICs) on an odontoblast cell line (MDPC-23), disks of every material were prepared and divided into Group 1: Vitrebond, Group 2: Vitremer, Group 3: Fuji IILC, Group 4: Fuji IX GP, Group 5: Ketac-Molar, Group 6: Z-100 (positive control). In Group 7, phosphate-buffered saline solution (negative control) was applied on filter paper. After placing the samples in the bottom of wells, the cells (30,000 cells/cm(2)) were plated and incubated for 72 h. The cell number was counted, the cell morphology was assessed by scanning electron microscopy and the cell metabolism was evaluated using methyltetrazolium assay. The statistical analysis of Kruskal-Wallis was used to determine if the scores obtained for the cell metabolism and number of cells were different at the 95% confidence level. In groups 1, 2, 3, 4, 5, and 6 the materials decreased the cell number by 74.5% 75.5%, 45.5%, 29.5%, 32.5%, and 88.5%, respectively. In groups 1, 2, 3, 4, and 5, the experimental GICs reduced the cell metabolism by 79%, 84%, 54%, 40%, and 42.5%, respectively. Despite the fact that all experimental materials were cytotoxic to the MDPC-23 cells, the GICs were the least cytotoxic. on the other hand, the RMGICs caused the highest cytophatic effects. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Biocompatibility of resin-based materials used as pulp-capping agents

Aim To evaluate and compare the response of pulps of rats capped with resin-modified glass-ionomer cement (RMGIC) or self-etching adhesive system.Methodology Class I cavities were prepared on the occlusal surface of 54 maxillary first molars of 27 rats. Pulp exposure was performed on the cavity floor. The following resin-based materials were applied as pulp-capping agents: G1, Clearfil Liner Bond 2V (CLB 2V; Kuraray Co., Japan); G2, Vitrebond (VIT; 3M/ESPE, USA). In group 3 (control group), a calcium hydroxide/saline paste (CH; Labsynth, Brazil) was used. The cavities were restored with amalgam. After 7, 30 and 60 days, the animals were sacrificed and the jaws were processed for microscopic evaluation.Results Despite the inflammatory response caused by the experimental and the control materials at 7 days, pulpal healing associated with calcified barrier formation was observed at 60 days following the pulp therapy. Both resin-based materials promoted a large zone of cell-rich fibrodentine matrix deposition on the pulp horn related to the pulp exposure site, which was larger to VIT than to CLB 2V specimens. Tertiary dentine underneath the fibrodentine matrix was deposited by a layer of elongated pulpal cells. The remaining pulpal tissue exhibited normal histological characteristics. In the control group, healing and dentine-bridge formation was observed at 30 days. Pulpal breakdown occurred only when bacterial infection occurred.Conclusion Both experimental pulp-capping agents allowed pulpal healing characterized by cell-rich fibrodentine and tertiary dentine deposition as well as calcified barrier formation.

Viability of human fibroblasts in coconut water as a storage medium

P>AimTo evaluate the effectiveness of a new storage medium for avulsed teeth, coconut water, in maintaining the viability of human fibroblasts.MethodologyCell viability after different time periods was evaluated in the following storage media: coconut water, coconut water with sodium bicarbonate, milk, saline and still mineral water. Human fibroblasts were seeded in Eagle's minimal essential medium (EMEM) supplemented with 7.5% foetal calf serum. After trypsinisation, 100 mu L of culture medium containing approximately 10(4) cells mL(-1) were collected and pipetted into the wells of 96-well plates, which were incubated overnight in 5% CO(2) and 95% air mixture at 37 degrees C. EMEM was then replaced by the storage media and the plates were incubated at 37 degrees C for 1, 2 and 4 h. Cell viability was determined using the neutral red assay. The proportions of viable cells after exposure to the storage media were analysed statistically by anova and the least significant difference (LSD) test (alpha = 5%).ResultsMilk had the greatest capacity to maintain cell viability (P < 0.05), followed by coconut water with sodium bicarbonate and saline. Coconut water was significantly worse at maintaining cell viability compared to milk, coconut water with sodium bicarbonate and saline. The smallest number of viable cells was observed for mineral water (P < 0.05).ConclusionCoconut water was worse than milk in maintaining human fibroblast cell viability.

Long-term treatment with alendronate increases the surgical difficulty during simple exodontias - an in vivo observation in Holtzman rats

Background: Atraumatic teeth extractions protocols are highly encouraged in patients taking bisphosphonates (Bps) to reduce surgical trauma and, consequently, the risk of jaws osteonecrosis development. In this way, this paper aims to report the findings of increased surgical difficulty during simple exodontias in animals treated with bisphosphonates.Methods: Sixty male Holtzman rats were randomly distributed into three groups of 20 animals and received daily subcutaneous administration of 1 mg/kg (AL1) or 3 mg/kg (AL3) of alendronate or saline solution (CTL). After 60 days of drug therapy all animals were submitted to first lower molars extractions under general anesthesia. Operatory surgical time and the frequency of teeth fractures were measured as principal outcomes and indicators of surgical difficulty degree.Results: Animals treated with alendronate (AL1 and AL3) were associated to higher operatory times and increased frequency of teeth fractures compared to match controls.Conclusions: The bisphosphonate therapy may be associated with an increased surgical difficulty and trauma following simple exodontias protocols, which is considered a critical issue when it comes to osteonecrosis development.

Effect of Drilling Technique on the Early Integration of Plateau Root Form Endosteal Implants: An Experimental Study in Dogs

Purpose: This study tested the hypothesis that early integration of plateau root form endosseous implants is significantly affected by surgical drilling technique.Materials and Methods: Sixty-four implants were bilaterally placed in the diaphysial radius of 8 beagles and remained 2 and 4 weeks in vivo. Half the implants had an alumina-blasted/acid-etched surface and the other half a surface coated with calcium phosphate. Half the implants with the 2 surface types were drilled at 50 rpm without saline irrigation and the other half were drilled at 900 rpm under abundant irrigation. After euthanasia, the implants in bone were nondecalcified and referred for histologic analysis. Bone-to-implant contact, bone area fraction occupancy, and the distance from the tip of the plateau to pristine cortical bone were measured. Statistical analyses were performed by analysis of variance at a 95% level of significance considering implant surface, time in vivo, and drilling speed as independent variables and bone-to-implant contact, bone area fraction occupancy, and distance from the tip of the plateau to pristine cortical bone as dependent variables.Results: The results showed that both techniques led to implant integration and intimate contact between bone and the 2 implant surfaces. A significant increase in bone-to-implant contact and bone area fraction occupancy was observed as time elapsed at 2 and 4 weeks and for the calcium phosphate-coated implant surface compared with the alumina-blasted/acid-etched surface.Conclusions: Because the surgical drilling technique did not affect the early integration of plateau root form implants, the hypothesis was refuted. (C) 2011 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 69: 2158-2163, 2011

Effect of diabetes mellitus and insulin therapy on bone density around osseointegrated dental implants: a digital subtraction radiography study in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of root-surface conditioning with acid and chelating agents on clot stabilization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Treatment of experimental periodontal disease with antimicrobial photodynamic therapy in nicotine-modified rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Commissural nucleus of the solitary tract is important for cardiovascular responses to caudal pressor area activation

The nucleus of the solitary tract (NTS) is the site of the first synapse of cardiovascular afferent fibers in the central nervous system. Important mechanisms for cardiovascular regulation are also present in the caudal pressor area (CPA) localized at the caudal end of the ventrolateral medulla. In the present study we sought to investigate the role of the commissural subnucleus of the NTS (commNTS) on pressor and tachycardic responses induced by L-glutamate injected into the CPA. Male Holtzman rats (n=8 rats/group) anesthetized with urethane (1.2 g/kg of body weight, iv) received injections of the GABAA receptor agonist muscimol into the commNTS. Unilateral injection of L-glutamate (10 nmol/ 100 nL) into the CPA increased mean arterial pressure (MAP, 31 4 mm Hg, vs. saline: 3 +/- 2 mm Hg) and heart rate (HR, 44 8 bpm, vs. saline: 10 7 bpm). inhibition of commNTS neurons with muscimol (120 pmol/60 nL) abolished the increase in MAP (9 4 mm Hg) and HR (17 7 bpm) produced by L-glutamate into the CPA. The present results suggest that the pressor and tachycardic responses to CPA activation are dependent on commNTS mechanisms.

Involvement of L-glutamate and ATP in the neurotransmission of the sympathoexcitatory component of the chemoreflex in the commissural nucleus tractus solitarii of awake rats and in the working heart-brainstem preparation

Peripheral chemoreflex activation with potassium cyanide (KCN) in awake rats or in the working heart-brainstem preparation (WHBP) produces: (a) a sympathoexcitatory/pressor response; (b) bradycardia; and (c) an increase in the frequency of breathing. Our main aim was to evaluate neurotransmitters involved in mediating the sympathoexcitatory component of the chemoreflex within the nucleus tractus solitarii (NTS). In previous studies in conscious rats, the reflex bradycardia, but not the pressor response, was reduced by antagonism of either ionotropic glutamate or purinergic P2 receptors within the NTS. In the present study we evaluated a possible dual role of both P2 and NMDA receptors in the NTS for processing the sympathoexcitatory component (pressor response) of the chemoreflex in awake rats as well as in the WHBP. Simultaneous blockade of ionotropic glutamate receptors and P2 receptors by sequential microinjections of kynurenic acid (KYN, 2 nmol (50 nl)(-1)) and pyridoxalphosphate-6-azophenyl-2',4'-disulphonate (PPADS, 0.25 nmol (50 nl)(-1)) into the commissural NTS in awake rats produced a significant reduction in both the pressor (+38 +/- 3 versus +8 +/- 3 mmHg) and bradycardic responses (-172 +/- 18 versus -16 +/- 13 beats min(-1); n = 13), but no significant changes in the tachypnoea measured using plethysmography (270 +/- 30 versus 240 +/- 21 cycles min(-1), n = 7) following chemoreflex activation in awake rats. Control microinjections of saline produced no significant changes in these reflex responses. In WHBP, microinjection of KYN (2 nmol (20 nl)(-1)) and PPADS (1.6 nmol (20 nl)(-1)) into the commissural NTS attenuated significantly both the increase in thoracic sympathetic activity (+52 +/- 2% versus +17 +/- 1%) and the bradycardic response (-151 +/- 17 versus -21 +/- 3 beats min(-1)) but produced no significant changes in the increase of the frequency of phrenic nerve discharge (+0.24 +/- 0.02+0.20 +/- 0.02 Hz). The data indicate that combined microinjections of PPADS and KYN into the commissural NTS in both awake rats and the WHBP are required to produce a significant reduction in the sympathoexcitatory response (pressor response) to peripheral chemoreflex activation. We conclude that glutamatergic and purinergic mechanisms are part of the complex neurotransmission system of the sympathoexcitatory component of the chemoreflex at the level of the commissural NTS.

Imidazoline receptors of the paraventricular nucleus on the pressor response induced by stimulation of the subfornical organ

In the present experiments we investigated a possible involvement of imidazoline receptors of the paraventricular nucleus (PVN) of the hypothalamus on the presser effects of the angiotensin LI (ANG II) injected into the subfornical organ (SFO), in male Holtzman rats (250-300 g) with a cannula implanted into the third ventricle (3rdV), PVN and SFO. At first we tested the participation of alpha(2) and imidazoline agonist and antagonist compounds on the presser effect of ANG II injected into the 3rdV. Based on the results we may conclude that clonidine associated with rilmenidine was able to block the hypertensive response to ANG IT. The ANG II (20 pmol) injected into SFO induced a robust increase in blood pressure (37 +/- 2 mmHg). Isotonic saline (0.15 M) NaCl did not produce any change in blood pressure (5 +/- 2 mmHg). The injection of rilmenidine (30 mu g/kg/l mu L), an imidazoline agonist agent injected into PVN before ANG II injection into SFO, blocked the presser effect of ANG II (5 +/- 2 mmHg). Also, the injection of idazoxan (60 mu g/kg/mu L) before rilmenidine blocked the inhibitory effect of rilmenidine on blood pressure (39 +/- 4 mmHg). The injection of clonidine (20 nmol/mu L) prior to ANG II into the 3rdV produced a decreased in arterial blood pressure (37 +/- 2 mmHg) to (15 +/- 4 mmHg). The injection of yohimbine (80 nmol/mu L) prior to clonidine blocked the effect of clonidine on the effect of ANG II (27 +/- 2 mmHg). The injection of rilmenidine prior to ANG TI also induced a decrease in arterial blood pressure (10 +/- 3 mmHg). The injection of idazoxan prior to rilmenidine also blocked the inhibitory effect of rilmenidine (24 +/- 3 mmHg). In summary, the present study demonstrated that rilmenidine decreases the hypertensive effect of ANG II, with more potency than clonidine, even when injected into 3rdV or PVN. This study established that the PVN interacts with SFO by imidazoline receptors in order to control the arterial blood pressure. (C) Elsevier, Paris.

5-HT2 and 5-HT3 receptors in the lateral parabrachial nucleus mediate opposite effects on sodium intake

The present study investigated the role of several 5-HT receptor subtypes in the lateral parabrachial nucleus (LPBN) in the control of sodium appetite (i.e. NaCl consumption). Male Holtzman rats had cannulas implanted bilaterally into the LPBN for the injection of 5-HT receptor agonists and antagonists in conjunction with either acute fluid depletion or 24-h sodium depletion. Following these treatments, access to 0.3 M NaCl was provided and the intakes of saline and water were measured for the next 2 h. Bilateral injections of the 5-HT2A receptor antagonist, ketanserin or the 5-HT2C receptor antagonist, mianserin into the LPBN increased 0.3 M NaCl intake without affecting water intake induced by acute fluid-depletion. Bilateral injections of the 5-HT2B receptor agonist, BW723C86 hydrochloride, had no effect on 0.3 M NaCl or water intake under these conditions. Treatment of the LPBN with the 5-HT2B/2C receptor agonist, 2-(2-methyl-4-clorophenoxy) propanoic acid (mCPP) caused dose-related reductions in 0.3 M NaCl intake after 24 h sodium depletion. The effects of mCPP were prevented by pretreating the LPBN with the 5-HT2B/2C receptor antagonist, SDZSER082. Activation of 5-HT3 receptors by the receptor agonist, 1-phenylbiguanicle (PBG) caused dose-related increases in 0.3 M NaCl intake. Pretreatment of the LPBN with the 5-HT3 receptor antagonist, 1-methyl-N-[8-methyl-8-azabicyclo (3.2.1)-oct-3-yl]-1H-indazole-3-carboxamide (LY-278,584) abolished the effects of PBG, but LY-278,584 had no effects on sodium or water intake when injected by itself. PBG injected into the LPBN did not alter intake of palatable 0.06 M sucrose in fluid replete rats. The results suggest that activation of the 5-HT2A and 5-HT2C receptor subtypes inhibits sodium ingestion. In contrast, activation of the 5-HT3 receptor subtype increases sodium ingestion. Therefore, multiple serotonergic receptor subtypes in the LPBN are implicated in the control of sodium intake, sometimes by mediating opposite effects of 5-HT. The results provide new information concerning the control of sodium intake by LPBN mechanisms. (C) 2007 IBRO. Published by Elsevier Ltd. All rights reserved.