Cleaning capacity promoted by motor-driven or manual instrumentation using ProTaper Universal system: Histological analysis

The aim of this study was to assess the cleaning capacity of the Protaper system using motor-driven or manual instrumentation. Materials and Methods: Ten mandibular molars were randomly separated into 2 groups (n = 5) according to the type of instrumentation performed, as follows: Group 1 - instrumentation with rotary nickel-titanium (Ni-Ti) files using ProTaper Universal System (Dentsply/Maillefer); and, Group 2 - instrumentation with Ni-Ti hand files using ProTaper Universal (Dentsply-Maillefer). Afterwards, the teeth were sectioned transversely and submitted to histotechnical processing to obtain histological sections for microscopic evaluation. The images were analyzed by the Corel Photo-Paint X5 program (Corel Corporation) using an integration grid superimposed on the image. Results: Statistical analysis (U-Mann-Whitney - P < 0.05) demonstrated that G1 presented higher cleaning capacity when compared to G2. Conclusions: The rotary technique presented better cleaning results in the apical third of the root canal system when compared to the manual technique.

Cutaneous healing after topical application of nebacetin and gingilone to infected wounds. Clinical and histological study in rats

It was performed a study to examine the cutaneous cicatrization of infected wound of skin after daily topic application of Nebacetin and Gingilone. Following clinical observations the animals were sacrificed after 4, 10 and 17 post-operative days. The pieces were treated, analyzed, in an optical microscope. The topic application of Nebacetin showed to be more efficient in the epithelial reparation comparatively to the Gingilone and control group.

Histological analysis of effects of 24% EDTA gel for nonsurgical treatment of periodontal tissues

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diabetes triggers the loss of tooth structure associated to radiographical and histological dental changes and its evolution to progressive pulp and periapical lesions in rats

The aim of this study was to evaluate the putative influence of diabetes without metabolic control in the loss of tooth structure as well as histological changes in dentin and pulp tissue in rats. Diabetes was induced in Wistar rats (n=25) by intravenous administration of alloxan (42mg/kg). Diabetic and non-diabetic control rats were evaluated at 1, 3, 6, 9 and 12 months of follow-up. In order to evaluate the presence and progression of dental caries and periapical lesions, hemimandibles were removed and submitted to radiographical, histological, and morphometrical procedures. Dental caries were detected after radiographical and histological evaluations in diabetic group from the third month of diabetes onset, increasing gradually in frequency and severity in periods. Diabetic rats dental pulps also presented significant reduction in volume density of collagen fibers and fibroblasts at third month, parallel with a trend towards the increase in inflammatory cells volume density. Diabetic rats presented a generalized pulp tissue necrosis after 6 months of diabetes induction. Moreover, periapical lesions were not detected in control group, while these lesions were observed in all rats after 3, 6, 9, and 12 months of diabetes induction. Uncontrolled diabetes seems to trigger the loss of tooth structure, associated to histological dental changes and mediates its evolution to progressive severe pulp and periapical lesions in rats. Therefore, diabetes may be considered a very important risk factor regarding alterations in dental pulp, development of dental caries, and periapical lesions.

Does spinal block through tattooed skin cause histological changes in nervous tissue and meninges?: an experimental model in rabbits

Although there is no documented evidence that tattoo pigments can cause neurological complications, the implications of performing neuraxial anesthesia through tattooed skin are unknown. In this study, we aimed to assess whether spinal puncture performed through tattooed skin of rabbits determines changes over the spinal cord and meninges. In addition, we sought to evaluate the presence of ink fragments entrapped in spinal needles. Thirty-six young male adult rabbits, each weighing between 3400 and 3900 g and having a spine length between 38.5 and 39 cm, were divided by lot into 3 groups as follows: GI, spinal puncture through tattooed skin; GII, spinal puncture through tattooed skin and saline injection; and GIII, spinal puncture through skin free of tattoo and saline injection. After intravenous anesthesia with ketamine and xylazine, the subarachnoid space was punctured at S1-S2 under ultrasound guidance with a 22-gauge 2½ Quincke needle. Animals in GII and GIII received 5 μL/cm of spinal length (0.2 mL) of saline intrathecally. In GI, the needle tip was placed into the yellow ligament, and no solution was injected into the intrathecal space; after tattooed skin puncture, 1 mL of saline was injected through the needle over a histological slide to prepare a smear that was dyed by the Giemsa method to enable tissue identification if present. All animals remained in captivity for 21 days under medical observation and were killed by decapitation. The lumbosacral spinal cord portion was removed for histological analysis using hematoxylin-eosin stain. None of the animals had impaired motor function or decreased nociception during the period of clinical observation. None of the animals from the control group (GIII) showed signs of injuries to meninges. In GII, however, 4 animals presented with signs of meningeal injury. The main histological changes observed were focal areas of perivascular lymphoplasmacyte infiltration in the pia mater and arachnoid. There was no signal of injury in neural tissue in any animal of both groups. Tissue coring containing ink pigments was noted in all GI smears from the spinal needles used to puncture the tattooed skin. On the basis of the present results, intrathecal injection of saline through a needle inserted through tattooed skin is capable of producing histological changes over the meninges of rabbits. Ink fragments were entrapped inside the spinal needles, despite the presence of a stylet.

Homogenous demineralized dentin matrix and platelet-rich plasma for bone tissue engineering in cranioplasty of diabetic rabbits: biochemical, radiographic, and histological analysis

This study evaluated the effects of homogenous demineralized dentin matrix (HDDM) slices and platelet-rich plasma (PRP) in surgical defects created in the parietal bones of alloxan-induced diabetic rabbits, treated with a guided bone regeneration technique. Biochemical, radiographic, and histological analyses were performed. Sixty adult New Zealand rabbits were divided into five groups of 12: normoglycaemic (control, C), diabetic (D), diabetic with a PTFE membrane (DM), diabetic with a PTFE membrane and HDDM slices (DM-HDDM), and diabetic with PTFE membrane and PRP (DM-PRP). The quantity and quality of bone mass was greatest in the DM-HDDM group (respective radiographic and histological analyses: at 15 days, 71.70±16.50 and 50.80±1.52; 30 days, 62.73±16.51 and 54.20±1.23; 60 days, 63.03±11.04 and 59.91±3.32; 90 days, 103.60±24.86 and 78.99±1.34), followed by the DM-PRP group (respective radiographic and histological analyses: at 15 days 23.00±2.74 and 20.66±7.45; 30 days 31.92±6.06 and 25.31±5.59; 60 days 25.29±16.30 and 46.73±2.07; 90 days 38.10±14.04 and 53.38±9.20). PRP greatly enhanced vascularization during the bone repair process. Abnormal calcium metabolism was statistically significant in the DM-PRP group (P<0.001) for all four time intervals studied, especially when compared to the DM-HDDM group. Alkaline phosphatase activity was significantly higher in the DM-HDDM group (P<0.001) in comparison to the C, D, and DM-PRP groups, confirming the findings of intense osteoblastic activity and increased bone mineralization. Thus, HDDM promoted superior bone architectural microstructure in bone defects in diabetic rabbits due to its effective osteoinductive and osteoconductive activity, whereas PRP stimulated angiogenesis and red bone marrow formation.

The Fresh Frozen Bone Allograft (FFBA) to reconstruc the atrophic maxilla: evaluation of the clinical and histological viability at six and nine months in the maxillary sinus

This work aimed to analyze clinically and histologically the allogen bone graft behavior at 6 and 9 months. A leukoderm, female, 55 years old patient sought dental care for oral rehabilitation with dental implants and implant supported fixed prosthesis in the maxilla. Bilateral sinus lifting procedure were performed in a same patient, the analysis were made after 6 and 9 months, respectively. At 6 months, there was lack of a better bone remodeling in the region, associated to the presence of fibrous connective tissue within the collected tissue, showing us that this is not the best stage of tissue maturation to place dental implants. The 9 months period showed an improvement, with a more organized bone tissue surrounding particles of homogenous bone, what possibly had increased implant stability at the time of surgery. There is no doubt about the increase of clinical applications of FFBA, but there are no studies available regarding the standardization of time to load implants inserted in allografts. So papers with long-term monitoring are necessary to eliminate questions that still remain to be answered.

Reconstruction of the anterior wall of the frontal sinus using bovine bone matrix: histological study with monkeys

The purpose of this study was to evaluate the repair process in the reconstruction of the anterior wall of the frontal sinus of monkeys with bovine bone matrix. Four adult Cebus apella monkeys underwent an ostectomy of the anterior wall of the frontal sinus. The frontal sinus mucosa and the nasofrontal duct were not manipulated. Reconstruction occurred with implants of bovine bone matrix laminae measuring 2.0 x 2.5 cm and 0.4 mm thick, stabilized under pressure in the lateral wall of the frontal sinus. The monkeys were sacrificed over a period of 150 days and routine laboratory procedures were followed for hematoxylin-eosin staining and histologic evaluation of the specimens. Neoformed bone tissue was observed in contact with the frontal sinus mucosa and the bovine bone matrix. The frontal sinus mucosa remained whole without fibrous tissue or cystic formations. There was no occurrence of cellularization as well as revascularization of the bovine bone matrix, though it has permitted bone conduction on this surface. It was possible to conclude that the demineralized bovine bone matrix was biotolerable, being incorporated into the bone without the presence of inflammatory cells with characteristics of inertness and antigenicity and behaved as an osteoconductive material.

Immunohistochemical, tomographic, and histological study on onlay bone graft remodeling. Part III: allografts

Objective In the last decades aroused the interest for bone tissue bank as an alternative to autogenous grafting, avoiding donor sites morbidity, surgical time, and costs reduction. The purpose of the study was to compare allografts (ALg) with autografts (AUg) using histology, immunochemistry, and tomographic analysis. Material and methods Fifty-six New Zealand White rabbits were submitted to surgical procedures. Twenty animals were donors and 36 were actually submitted to onlay grafting with ALg (experimental group) and AUg (control group) randomly placed bilaterally in the mandible. Six animals of each group were sacrificed at 3, 5, 7, 10, 20, and 60 postoperative days. Immunolabeling was accomplished with osteoprotegerin (OPG); receptor activator of nuclear factor-k ligand (RANKL); alkaline phosphatase (ALP); osteopontin (OPN); vascular endothelial growth factor (VEGF); tartrate-resistant acid phosphatase (TRAP); collagen type I (COL I); and osteocalcin (OC). Density and volume of the grafts was evaluated on tomography obtained at the surgery and sacrifice. Results The ALg and AUg exhibited similar patterns of density and volume throughout the experiments. The intra-group data showed statistical differences at days 7 and 60 in comparison with other time points (P = 0.001), in both groups. A slight graft expansion from fixation until day 20 (P = 0.532) was observed in the AUg group and then resorbed significantly at the day 60 (P = 0.015). ALg volume remained stable until day 7 and decreased at day 10 (P = 0.045). The light microscopy analysis showed more efficient incorporation of AUg onto the recipient bed if compared with the ALg group. The immunohistochemical labeling picked: at days 10 and 20 with OPG in the AUg group and at day 7 with TRAP in the ALg group (P = 0.001 and P = 0.002, respectively). Conclusions ALg and AUg were not differing in patterns of volume and density during entire experiment. Histological data exhibit more efficient AUg incorporation into recipient bed compared with the ALg group. Immunohistochemistry outcomes demonstrated similar pattern for both ALg and AUg groups, except for an increasing resorption activity in the ALg group mediated by TRAP and in the AUg group by higher OPG labeling. However, this latter observation does not seem to influence clinical outcomes.

Histological analysis of healing process after rat teeth replantation maintained in Resveratrol dissolved in dimethyl sufoxide (DMSO)

Tissue repair after replantation of avulsed teeth is directly related to the extent of damage to the cells of the periodontal ligament. Thus, immediate replantation is the treatment of choice for avulsed permanent teeth. To achieve more favorable prognostics, adequate storage media must be used to preserve periodontal ligament cells. A series of storage media are studied and show good results, such as saliva, milk, Hank's balanced solution (HBSS) and ViaSpan. However, recent studies were performed using news and promising storage media. Resveratrol has been extensively studied because of its antioxidant properties and its ability to prolong life of many organisms from yeast to mammals. One of its limitations is its poor solubility in aqueous vehicles. For this reason, the aim of this study was to evaluate the healing repair process after replantation of teeth of rats kept in Resveratrol using dimethyl sulfoxide (DMSO) as a vehicle. This study was approved by the Ethics Committee on Research with Animals, of the School of Dentistry of Araçatuba, Univ. Estadual Paulista, UNESP, Araçatuba, SP, Brazil. Were used 40 male rats, under general anesthesia upper right incisor were extracted and replanted. Treatments were done, dividing in four groups, of 10 animals each. In group I, the teeth were be extracted and immediately replanted into their sockets of origin (positive control). In group II, the teeth were immersed in 50 mL of resveratrol in DMSO (0.0512 g / ml) for 60 minutes. In group III teeth were kept for 60 minutes in 50 ml of DMSO. In group IV, the teeth were kept in dry for the same period (negative control). Then the teeth of animals in Groups II, III and IV were replanted in their sockets. Systemic antibiotics were administrated in all groups, and 60 days post-operative the animals were euthanized. The specimens were processed and stained in HE for histomorphological analysis. The results showed that resveratrol as storage media, was not able to improve the rep

A Field-Test of Rhodamine B as a Biomarker in Raccoons

Rhodamine B is a dye that when ingested results in fluorescent bands in growing hair and whiskers of many mammals. Previous research at Wildlife Services’ (WS) National Wildlife Research Center (NWRC) found that rhodamine B is a successful biomarker in raccoon whiskers and that raccoons do not have a taste aversion to the dye when it comprises ≤ 3% of a bait. Our study assessed the ease of bait distribution, whisker collection, and evaluation of the biomarker for potential use in the Oral Rabies Vaccination (ORV) program administered by the WS National Rabies Management Program (NRMP). In collaboration with WS operations personnel from Ohio and Michigan, 750 fishmeal polymer baits each containing 150 mg of rhodamine B were hand distributed at NASA’s Plum Brook Station, Sandusky, Ohio in the summer of 2008. Four weeks after baits were distributed whiskers from 162 raccoons were collected. Wildlife Services biologists and technicians evaluated the whiskers for fluorescence using a handheld UV magnifying lamp. Biologists then sent the whiskers to the NWRC, Ft. Collins, Colorado for confirmation of fluorescence under a UV microscope. Results suggest a high level of agreement between the two methods of evaluation. Surveys completed by biologists confirmed that the ease of use, less invasive sampling techniques and promptness of results obtained through the use of rhodamine B are advantageous to the tetracycline biomarker presently used by the ORV program. All participants recommended further evaluation of rhodamine B for its inclusion in future efforts requiring biomarker evaluation.

Integrated biomarker responses as environmental status descriptors of a coastal zone (Sao Paulo, Brazil)

Sao Paulo state (Brazil) has one of the most overpopulated coastal zones in South America, where previous studies have already detected sediment and water contamination. However, biological-based monitoring considering signals of xenobiotic exposure and effects are scarce. The present study employed a battery of biomarkers under field conditions to assess the environmental quality of this coastal zone. For this purpose, the activity of CYP 450, antioxidant enzymes, DNA damage, lipid peroxidation and lysosomal membrane were analysed in caged mussels and integrated using Factorial Analysis. A representation of estimated factor scores was performed in order to confirm the factor descriptions characterizing the studied areas. Biomarker responses indicated signals of mussels` impaired health during the monitoring, which pointed to the impact of different sources of contaminants in the water quality and identified critical areas. This integrated approach produced a rapid, sensitive and cost-effective assessment, which could be incorporated as a descriptor of environmental status in future coastal zones biomonitoring. (C) 2011 Elsevier Inc. All rights reserved.

Mutagenicity of Blue Rayon Extracts of Fish Bile as a Biomarker in a Field Study

Blue rayon (BR) in combination with the Salmonella/microsome assay was used to evaluate the mutagenicity of fish bile samples. Specimens of Mugil curema from two sites were collected over a 1-year period. Piacaguera channel contains high concentrations of total polycyclic aromatic hydrocarbons (PAHs) and other contaminants, while Bertioga channel was considered the reference sites in this study. Bile was extracted with BR and tested with TA98, TA100, and YG1041 strains with and without S9 in dose response experiments. PAH metabolite equivalents were analyzed using reverse-phase high performance liquid chromatography /fluorescence. Higher mutagenic responses were observed for the contaminated site; YG1041 with S9 was the most sensitive strain/condition. Mutagenicity ranged from 3,900 to 14,000 rev./mg at the contaminated site and from 1,200 to 2,500 rev./mg of BR at the reference site. The responses of YG1041 were much higher in comparison with the TA98 indicating the presence of polycyclic compounds from the aromatic amine class that cause frameshift mutation. TA100 showed a positive mutagenic response that was enhanced following S9 treatment at both sites suggesting the presence of polycyclic compounds that require metabolic activation. benzo(a)pyrene, naphthalene, and phenanthrene metabolite equivalents were also higher in the bile of fish collected at the contaminated site. It was not possible to correlate the PAH metabolite quantities with the mutagenic potency. Thus, a combination of the Salmonella/microsome assay with YG1041 with S9 from BR bile extract seems to be an acceptable biomarker for monitoring the exposure of fish to mutagenic polycyclic compounds. Environ. Mal. Mutagen. 51:173-179, 2010. (C) 2009 Wiley-Liss, Inc.

Biomarker responses as indication of contaminant effects in Oreochromis niloticus

The current study investigated oxidative stress parameters (enzymes activities, metallothionein content and lipid peroxidation) in freshwater fish, Oreochromis niloticus, tilapia exposure to Monjolinho River (in 4 months of year: January, April, July and November). One critical site in Monjolinho River (site B) was assessed in comparison to a reference site (site A). Water pH and oxygen concentration was lower than that recommended by CONAMA (Brazilian National Environmental Committee), resolution 357/2005 for protection of aquatic communities, and ammonium and the metals Cu, Zn, Mn and Fe (on all months) concentrations were higher than the maximum concentration recommended. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significantly decreased in liver and muscle in tila. pia from Monjolinho River, throughout the year, in relation to reference except in gills that SOD activity increased. Glutathione S-transferase (GST) activity was significantly increased in liver of the tilapia from Monjolinho River in all sites, in relation to reference except in gills that GST activity increased in July and decreased in November, suggesting that GST activity could be induced to neutralize the pollutants toxicity. On the other hand, GST activity was significantly decreased in white muscle indicating a toxic effect of pollutants, resulting in a decreased ability of tilapia to perform defense reactions associated to GSTs. The decrease of catalase (CAT) activity in gills of the O. niloticus together with the increase of SOD activity, could explain the increased lipid peroxidation (LPO) level in this organ. Metallothionein levels in liver and gills were significantly high in all sites. Results indicate that the exposure to metals caused severe damage to tissues; despite the consensually assumed antioxidant induction as a sign of exposure to contaminants the effects seem in part to be mediated by suppression of antioxidant system with SOD, CAT and GPx as potential candidates for tissues toxicity biomarkers of pollutants. (c) 2012 Elsevier Ltd. All rights reserved.