Decrease in the number and apoptosis of alveolar bone osteoclasts in estrogen-treated rats

Bone is a mineralized tissue that is under the influence of several systemic, local and environmental factors. Among systemic factors, estrogen is a hormone well known for its inhibitory function on bone resorption. As alveolar bone of young rats undergoes continuous and intense remodeling to accommodate the growing and erupting tooth, it is a suitable in vivo model for using to study the possible action of estrogen on bone. Thus, in an attempt to investigate the possibility that estrogen may induce the death of osteoclasts, we examined the alveolar bone of estrogen-treated rats.Fifteen, 22-d-old female rats were divided into estrogen, sham and control groups. The estrogen group received estrogen and the sham group received corn oil used as the dilution vehicle. After 8 d, fragments containing alveolar bone were removed and processed for light microscopy and transmission electron microscopy. Sections were stained with hematoxylin and eosin and tartrate-resistant acid phosphatase (TRAP)-an osteoclast marker. Quantitative analysis of the number of TRAP-positive osteoclasts per mm of bone surface was carried out. For detecting apoptosis, sections were analyzed by the Terminal deoxynucleotidyl transferase-mediated dUTP Nick-End Labeling (TUNEL) method; TUNEL/TRAP combined methods were also used.The number of TRAP-positive osteoclasts per mm of bone surface was significantly reduced in the estrogen group compared with the sham and control groups. TRAP-positive osteoclasts exhibiting TUNEL-positive nuclei were observed only in the estrogen group. In addition, in the estrogen group the ultrastructural images revealed shrunken osteoclasts exhibiting nuclei with conspicuous and tortuous masses of condensed chromatin, typical of apoptosis.Our results reinforce the idea that estrogen inhibits bone resorption by promoting a reduction in the number of osteoclasts, thus indicating that this reduction may be, at least in part, a consequence of osteoclast apoptosis.

Cell death and quantitative reduction of rests of Malassez according to age

Background and Objective: Rests of Malassez are clusters of epithelial cells that remain in the periodontal ligament throughout life. However, it has been reported that the number of these structures decreases with age, and some epithelial cells undergo apoptosis in rests of Malassez of young and adult rats. Therefore, the purpose of the present study was to investigate the incidence of epithelial cell death and the quantitative changes in the rests of Malassez in rat molars of different ages.Material and Methods: Fragments containing the upper molars of rats aged 29, 45 and 120 d were fixed, decalcified and embedded for analysis by light microscopy. In the sections stained by hematoxylin and eosin, the number of rests of Malassez and the number of nuclei of these epithelial structures were obtained. Moreover, the nuclei exhibiting typical features of cell death were also counted in each rest of Malassez. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method for detection of cell death was also carried out.Results: In all groups examined, some rests of Malassez exhibited epithelial cell nuclei with typical features of apoptosis and some of them were also TUNEL positive. From 29 to 120 d of age in rats, the quantitative analysis showed a significant decrease in the total number of rests of Malassez in the cervical, middle and furcation regions of the periodontal ligament. Moreover, a significant decrease of epithelial cell nuclei was concomitant to an increase in the frequency of cell death in the oldest rats.Conclusion: These results suggest that epithelial cell death by apoptosis may be, at least in part, responsible for the reduction in the number of rests of Malassez according to age.

Area of Rests of Malassez in Young and Adult Rat Molars: Evidences in the Formation of Large Rests

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Morphometrical analysis of cleaning capacity of a hybrid instrumentation in mesial flattened root canals

The cleaning capacity of hybrid and rotary instrumentation techniques in mesial flattened canals of mandibular first molars was evaluated by morphometrical analysis in this study. Twenty human mandibular first molars were randomly assigned into two groups, according to instrumentation technique, as follows: group 1, instrumentation with ProTaper Starter Kit (Dentsply/Maillefer) rotary system; group 2, manual instrumentation using K files (Dentsply/Maillefer) by crown-down technique in middle and apical thirds, cervical preparation with Gates-Glidden #1 and #2 (Dentsply/Maillefer) burs, and to finalise the preparation, ProTaper F2 and F3 rotary files. Serial transverse cross-sections (5 mu m) of the apical third, stained with hematoxylin and eosin, were analysed at 100 x original magnification. The images were submitted to morphometrical analysis with an integration grid to determine the percentage of root canal area with debris. Statiscal analysis (t-Student, P < 0.05) showed significant difference between the techniques (P < 0.05), although neither completely cleaned the root canal.

Response of mice connective tissue to intracanal dressings containing chlorhexidine

Substances containing chlorhexidine (CHX) have been studied as intracanal medicaments. The aim of the present study was to characterize the response of mouse subcutaneous connective tissue to CHX-containing medications by conventional optical microscopy. The tissue response was evaluated by implanting polyethylene tubes containing one of the substances evaluated: Calen paste + 0.5% CHX, Calen + 2% CHX, 2% CHX gel, and Calen paste (control). After experimental periods of 7, 21, and 63 days, the implants (n = 10) were removed along with the subcutaneous connective tissue. Tissue samples were subjected to histological processing, and sections were stained with hematoxylin and eosin. Qualitative and quantitative analyses of the number of inflammatory cells, blood vessels, and vascularized areas were performed. Results were analyzed by ANOVA and Tukey tests with the significance level set at 5%. We concluded that Calen + 0.5% CHX led to reparative tissue response in contrast with Calen + 2% CHX and 2% CHX gel, which induced persistent inflammatory response, pointing to the aggressive nature of this mixture. When Calen + 2% CHX and 2% CHX gel were compared, the latter induced more intense inflammatory response. Microsc. Res. Tech., 2012. (C) 2012 Wiley Periodicals, Inc.

Differential expression of myogenic regulatory factor MyoD in pacu skeletal muscle (Piaractus mesopotamicus Holmberg 1887: Serrasalminae, Characidae, Teleostei) during juvenile and adult growth phases

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Histochemical technique for the detection of chloride cells in fish

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Morphological changes in distant muscle fibers following thermal injury in Wistar rats

Purpose: Thermal injury causes catabolic processes as the body attempts to repair the damaged area. This study evaluated the effects of a scald injury on the morphology of muscle fibers belonging to a muscle distant from the lesion. Methods: Thirty Wistar rats were divided into control (C) and scalded (S) groups. Group S was scalded over 45% of the body surface, standardized by body weight. Rats in both groups were euthanized at four, seven and 14 days following the injury. The middle portions of the medial gastrocnemius muscles were sectioned, stained with hematoxylin and eosin and Picrosirius, and submitted to histological analysis. Results: Control group sections exhibited equidistantly distributed polygonal muscle fibers with peripheral nuclei, characteristic of normal muscle. The injured group sections did not consistently show these characteristics; many fibers in these sections exhibited a rounded contour, variable stain intensities, and greater interfiber distances. A substantially increased amount of connective tissue was also observed on the injured group sections. Conclusion: This experimental model found a morphological change in muscle distant from the site of thermal injury covering 45% of the body surface.

The Effects of a 785-nm AlGaInP Laser on the Regeneration of Rat Anterior Tibialis Muscle After Surgically-Induced Injury

Objective: This study aims to investigate the effects of low-level laser therapy (LLLT) on muscle regeneration. For this purpose, the anterior tibialis muscle of 48 male Wistar rats received AlGaInP laser treatment (785 nm) after surgically-induced injury.Background Data: Few studies have been conducted on the effects of LLLT on muscle regeneration at different irradiation doses.Materials and Methods: The animals were randomized into four groups: uninjured rats (UN); uninjured and laser-irradiated rats (ULI); injured rats (IN); and injured and laser-irradiated rats (ILI). The direct contact laser treatment was started 24 h after surgery. An AlGaInP diode laser emitting 75 mW of continuous power at 785 nm was used for irradiation. The laser probe was placed at three treatment points to deliver 0.9 J per point, for a total dose of 2.7 J per treatment session. The animals were euthanized after treatment sessions 1, 2, and 4. Mounted sections were stained with hematoxylin and eosin and used for quantitative morphological analysis, in which the number of leukocytes and fibroblasts were counted over an area of 4480 mu m(2). The data were statistically analyzed by analysis of variance (ANOVA) and the Bonferroni t-test.Results: Quantitative data showed that the number of both polymorphonuclear and mononuclear leukocytes in the inflammatory infiltrate at the injury site was smaller in the ILI(1), ILI(2), and ILI(4) subgroups compared with their respective control subgroups (IN(1), IN(2), and IN(4)) for sessions 1, 2, and 4, respectively (p < 0.05). on the other hand, the number of fibroblasts increased after the fourth treatment session (p < 0.05). With regard to the regeneration of muscle fibers following injury, only after the fourth treatment session was it possible to find muscle precursor cells such as myoblasts and some myotubes in the ILI(4) subgroup.Conclusion: During the acute inflammatory phase, the AlGaInP laser treatment was found to have anti-inflammatory effects, reducing the number of leukocytes at the injury site and accelerating the regeneration of connective tissue.

Fructose-rich diet leads to reduced aerobic capacity and to liver injury in rats

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Clinical and histomorphometric characteristics of three different families with hereditary gingival fibromatosis

Objective. To examine the histomorphologic and histomorphometric features of tissue from 3 unrelated families with hereditary gingival fibromatosis (HGF).Study design. Twelve affected individuals from 3 HGF families and 3 control subjects were evaluated. Gingival samples were fixed in formalin and embedded in paraffin for hematoxylin and eosin stain to count the number of fibroblast and inflammatory cells. Sirius red staining was performed to quantitate the amount of collagen present.Results. Histomorphologic analysis of HGF showed extension of epithelial rete ridges into the underlying lamina propria and the presence of collagen bundles in the connective tissue. Analysis of the mean area fraction of collagen showed that there were significant increases in the collagen fraction for all HGF types compared with control subjects (P < .05). There were significant increases in the number of fibroblasts for HGFa and HGFb compared with control subjects (P < .05). The number of fibroblasts for HGFc were similar to that for control subjects.Conclusions. The collagen fraction was significantly greater in all HGF types compared with controls. The number of fibroblasts was significantly increased in 2 of the 3 HGF types compared with controls. These data indicate that different mechanisms may be responsible for tissue enlargement in different forms of HGF.

Scattering of positronium by H, He, Ne, and Ar

The low-energy scattering of the ortho-positronium (Ps) by H, He, Ne, and Ar atoms has been investigated in the coupled-channel framework by using a recently proposed time-reversal symmetric non-local electron-exchange model potential with a single parameter C. For H and He, we use a three-Ps-state coupled-channel model and, for Ar and Ne, we use a static-exchange model. The sensitivity of the results is studied with respect to the parameter C. Present low-energy cross-sections for He, Ne and Ar are in good agreement with experiment. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Ps-He scattering using the three-state positronium close-coupling approximation

A three-state target elastic positronium close-coupling approximation (CCA) is employed to investigate Ps-He scattering in the energy range 0-200 eV with and without electron exchange. Low-lying phase shifts below the first excitation threshold and the total integrated cross sections using both the models are reported. Estimation of integrated excitation cross sections for Ps(1s --> 2s) and Ps(1s --> 2p) using CCA are presented for the first time. The present total cross sections are in good agreement with the measured data in the incident Ps energy range 20-30 eV.

Prediction of a weakly bound excited state in the He-4(2)-Li-7 molecule

A scale-independent approach, valid for weakly bound three-body systems, is used to analyze the existence of excited Thomas-Efimov states in molecular systems with three atoms: a helium dimer together with isotopes of lithium (Li-6 and Li-7) and sodium (Na-23). With the present study and the available data, we can clearly predict that the He-4(2)-Li-7 system supports an excited state with binding energy close to 2.31 mK. (C) 2000 American Institute of Physics. [S0021-9606(00)30442-1].

Nonmesonic weak decay spectra of He-4(Lambda)

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