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 mu m, and the mantle dentin-enamel interface deviated +/- 15 mu m from the average plane over a distance of 520 mu m. In the highest resolution data (similar to 1.5 mu m 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. (C) 2007 Elsevier B.V. All rights reserved.

In situ and in vitro comparison of laser fluorescence with visual inspection in detecting occlusal caries lesions

The aim of this study was to compare the in situ and in vitro performances of a laser fluorescence (LF) device (DIAGNOdent 2095) with visual inspection for the detection of occlusal caries in permanent teeth. Sixty-four sites were selected, and visual inspection and LF assessments were carried out, in vitro, three times by two independent examiners, with a 1-week interval between evaluations. Afterwards, the occlusal surfaces were mounted on the palatal portion of removable acrylic orthodontic appliances and placed in six volunteers. Assessments were repeated and validated by histological analysis of the tooth sections under a stereomicroscope. For both examiners, the highest intra-examiner values were observed for the visual inspection when in vitro and in situ evaluations were compared. The inter-examiner reproducibility varied from 0.61 to 0.64, except for the in vitro assessment using LF, which presented a lower value (0.43). The methods showed high specificity at the D(1) threshold (considering enamel and dentin caries as disease). In vitro evaluations showed the highest values of sensitivity for both methods when compared to the in situ evaluations at D(1) and D(2) (considering only dentinal caries as the disease) thresholds. For both methods, the results of sensitivity (at D(1) and D(2)) and accuracy (at D(1)) showed significant differences between in vitro and in situ conditions. However, the sensitivity (at D(1) and D(2)), specificity and accuracy (both at D(1)) of the methods were not significantly different when the same condition was considered. It can be concluded that visual inspection and LF showed better performance in vitro than in situ.

Effect of low-level laser irradiation on odontoblast-like cells

Low-level laser therapy (LLLT), also referred to as therapeutic laser, has been recommended for a wide array of clinical procedures, among which the treatment of dentinal hypersensitivity. However, the mechanism that guides this process remains unknown. Therefore, the objective of this study was to evaluate in vitro the effects of LLL irradiation on cell metabolism (MTT assay), alkaline phosphatase (ALP) expression and total protein synthesis. The expression of genes that encode for collagen type-1 (Col-1) and fibronectin (FN) was analyzed by RT-PCR. For such purposes, oclontoblast-like cell line (MDPC-23) was previously cultured in Petri dishes (15000 cells/cm(2)) and submitted to stress conditions during 12 h. Thereafter, 6 applications with a monochromatic near infrared radiation (GaAlAs) set at predetermined parameters were performed at 12-h intervals. Non-irradiated cells served as a control group. Neither the MTT values nor the total protein levels of the irradiated group differed significantly from those of the control group (Mann-Whitney test; p > 0.05). on the other hand, the irradiated cells showed a decrease in ALP activity (Mann-Whitney test; p < 0.05). RT-PCR results demonstrated a trend to a specific reduction in gene expression after cell irradiation, though not significant statistically (Mann-Whitney test; p > 0.05). It may be concluded that, under the tested conditions, the LLLT parameters used in the present study did not influence cell metabolism, but reduced slightly the expression of some specific proteins.

Human pulp response to resin cements used to bond inlay restorations

Objective. The aim of this study was to evaluate the pulp response following cementation of inlays using two different resin cements.Methods. Deep Class V cavities were prepared on the buccal surface of 34 sound human premolars. impressions were taken and inlays were prepared which were cemented with the following luting materials-Group 1: Rely X(TM) Unicem. (3M ESPE); Group 2: Variolink(R) II (Ivoclar Vivadent). in Group 3 (control), after lining the cavity floor with Dycal(R) (Dentsply Caulk) the inlays were cemented with Rely X(TM) Unicem. Four additional teeth were used as an intact control group. For Variolink(R) II, the adhesive system Excite was used as part of the cementation procedure. After 7 or 60 days, the teeth were extracted and processed for histological assessment.Results. At 7 days, Rely X(TM) Unicern and Variolink(R) II system triggered in two samples a mild and moderate inflammatory response, respectively. At 60 days, the pulpal response decreased for both groups. A discrete persistent inflammatory response occurred in Group 2 in which displacement of resin components across the dentin tubules was observed. In the control group, normal histological characteristics were observed. The inflammatory response and tissue disorganization were related to the remaining dentin thickness between the cavity floor and the pulp tissue.Significance. Techniques for inlay cementation using distinct luting cements may cause specific pulpal damage. Variolink(R) II associated with the adhesive system Excite cause more aggressive effects to the pulp-dentin complex than Rely X(TM) Unicern cement when both are used to cement inlay restorations. (C) 2005 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Vitamin B-12 Supplement Exerts a Beneficial Effect on the Seminiferous Epithelium of Cimetidine-Treated Rats

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

Morphological evidences indicate that the interference of cimetidine on the peritubular components is responsible for detachment and apoptosis of Sertoli cells

Cimetidine, referred as antiandrogenic agent, has caused alterations in the seminiferous tubules, including alterations in the peritubular tissue and death of myoid cells by apoptosis. Regarding the structural and functional importance of the peritubular tissue for the maintenance of Sertoli cells (SC), we purpose to investigate the SC-basement membrane interface, focusing the morphological features of SC and their interaction with the basement membrane in the affected tubules by cimetidine. Ten animals were distributed into two groups, control (CG) and cimetidine (CmG) which received saline solution and 50 mg of cimetidine per kg of body weight, respectively, for 52 days. The testes were fixed, dehydrated and embedded for analyses under light and transmission electron microscopy. Paraffin sections were submitted to the TUNEL method; sections of testes embedded in glycol methacrylate were submitted to PAS method and stained by H&E for morphological and quantitative analyses of Sertoli Cells. In the CmG, the SC nuclei were positive to the TUNEL method and showed typical morphological alterations of cell death by apoptosis (from early to advanced stages). A significant reduction in the number of Sertoli Cells was probably due to death of these cells by apoptosis. A close relationship between SC nuclear alterations (including a high frequency of dislocated nuclei from the basal portion) and damage in the peritubular tissue was observed. The ultrastructural analysis showed a parallelism between the gradual advancement of apoptotic process in SC and detachment of the anchoring sites (hemidesmosomes) of SC plasma membrane from the lamina densa. The presence of portions of lamina densa underlying the detached hemidesmosomes indicates a continuous deposition of lamina densa, resulting in the thickening of the basal lamina. The results indicate a possible disarrangement of the SC cytoskeleton, including the focal adhesion structure. These alterations are related to SC apoptosis and probably result from disturbs induced by cimetidine on the peritubular tissue.

Factors affecting on bond strength of glass fiber post cemented with different resin cements to root canal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Penetration into dentin of sodium hypochlorite associated with acid solutions

Objective. The objective of this study was to evaluate the penetration of 2.5% NaOCl associated with 17.0% EDTA, 1.0% citric acid, and 1.0% peracetic acid into dentin tubules.Study design. The roots of 44 bovine incisors were cross-sectioned and 5-mm-long fragments were produced from their middle thirds. The specimens were instrumented with ProTaper hand files, stained in crystal violet, then sectioned mesiodistally. The buccal fragments were divided into 4 groups (n = 9) and subjected to 2 consecutive 10-minute immersion periods in one of the following acid solutions combined with 2.5% NaOCl: 17.0% EDTA (group 1), 1.0% citric acid (group 2), and 1.0% peracetic acid (group 3). Nine fragments were immersed in 2.5% NaOCl (group 4). The analysis of the penetration of NaOCl solutions into dentin was performed by measuring the depth of crystal violet stain that was bleached using a steromicroscope under x50 magnification. Statistical comparisons were carried out by Kruskal-Wallis and Dunn's tests at the 5% significance level.Results. Group 1 showed less penetration into dentin than group 4 (P < .05). No statistically significant differences were observed among groups 2, 3, and 4 (P > .05).Conclusions. Association of NaOCl with acid solutions did not increase its penetration depth into root dentin. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011;112:e155-e159)

Morphological studies on the heart ventricle of African catfish (Clarias gariepinus)

The histological and ultrastructural characteristics of the heart ventricle in Clarias gariepinus (African catfish) has been studied by light microscopy and transmission electron microscopy. The ventricle of the heart has a saccular shape and the myocardial wall consists of an outer thin compact myocardium and an inner well-developed spongy myocardium. The myocardial layer has small myocytes, interstitial spaces and blood vessels. The myocytes are the major constituents of the ventricular wall. They are long cells, with large nuclei, and predominantly euchromatin. The sarcoplasmic reticulum of the ventricular myocytes consists of a network of tubules and subsarcolemmal cisternae oriented mainly along the longitudinal axis of the myofibrils. In contrast to the ventricular structure of other fish species described in the literature (Greer-Walker et al., 1985 Santer, 1985 Sanchez-Quintana et al., 1995, 1996), the African catfish, a freshwater sedentary fish recently introduced in neotropical climatic environments, showed a saccular ventricle that consisted of two muscle layers, a thin compact layer with large vessels and a developed spongy layer. The ultrastructure of the ventricular myocardium of C.gariepinus is similar to that of other teleosts, inclusive that of fish with other swimming habits.

A histological study of the seminal vesicle of the armoured catfish Corydoras aeneus

Most species of Corydoras exhibited a reproductive behaviour called 'T-position', and exhibited an accessory gland in the male genital tract, called the seminal vesicle. It appeared that both the structure and the composition of the fluid varied considerably between the species investigated. Consequently, different opinions were proposed regarding the possible role of seminal vesicle on this particular reproductive behaviour. Male adults of Corydoras aeneus were collected, anaesthetized, and samples of seminal vesicle were fixed in Bouin's solution. The sections were stained with haematoxylin-eosin and periodic acid Schiff. The seminal vesicle showed a system of anastomosed secretory tubules, forming a vesicular collective network, which gave rise to the vesicular ducts. The latter fused with the testicular efferent ducts and formed the spermatic ducts. Considering this fusion, when the sperm cells reached the spermatic ducts, the fluid produced at the seminal vesicle covered them. Histochemical studies evidenced the presence of neutral and acid glycosaminoglycans in the seminal fluid. Considering the reproductive behaviour of C. aeneus, it is believed that the protection associated with the immobilization of the sperm cells assures the sperm integrity during the passage through female's intestine until fertilization.

Aquaporin 9 (AQP9) localization in the adult dog testis excurrent ducts by immunohistochemistry

Aquaporins (AQPs) are small, intrinsic membrane proteins that are present in many cell types involved in fluid transport. AQP9 is a major apical water channel that is expressed throughout the efferent ducts, epididymis, and vas deferens, as well as in other regions of the human and rodent male reproductive tract. The target of this study was to examine the expression of AQP9 in epithelial cells in the adult dog efferent ducts, epididymis, and vas deferens. Samples of dog male reproductive tract comprising fragments of the testis; initial segment, caput, corpus, and cauda of the epididymis; and vas deferens were obtained from eight adult mongrel dogs. Immunohistochemistry and Western blotting procedures were used to show AQP9 localization and distribution. AQP9 expression was not detected either in dog seminiferous tubules or rete testis. However, apical labeling for AQP9 was detected in the different regions of epididymis and vas deferens, with the reaction being less intense in the caput epididymis. Thus, AQP9 is abundantly expressed in dog male reproductive tract, in which it is an important apical pathway for transmembrane flow of water and neutral solutes.

Ultrastructural features of the glandular region of the stomach of Piaractus mesopotamicus (Holmberg, 1887) with emphasis on the oxyntopeptic cell

The characteristics of the lining and glandular epithelial types of the wall of the stomach of pacu (Piaractus mesopotamicus Holmberg, 1887) were studied by transmission electron microscopy. Columnar mucous cells were observed in the lining epithelial surface and the glandular epithelium that invaginated in the lamina propria showed secretory cells or oxyntopeptic cells as called by some authors in the literature. The columnar epithelial cells are narrow and show a basal nucleus. They are rich in organelles and contain secretory granules of round or rectangular shape heterogeneously electron dense in the apical portion. In the basal region of the glands there are secretory cells. The luminal half of these cells has abundant tubules (tubulo-vesicular system) that communicate with the external medium. Deeper basally in the cytoplasm there are the nucleus, mitochondria of various shapes and other scattered organelles.

Morphometric study of Rynchotus rufescens testis throughout the year

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

Morphological aspects of spermatogenic cells at different stages of sexual maturation in black isogenic C57BL/6/Uni mice

In order to study the morphological changes that occur in cells of the testes of isogenic black mouse C57BL/6/Uni into three periods during spermatogenetic used 15 mice divided into 3 groups of 5 animals with 40,50 and 60 days of age. The mice were sacrificed and weighed. Testicles were weighed and measured, and histologically processed and stained with HE, PAS and Masson Massom-H and evaluated under light microscopy. It was observed that group I with 40 days of age in the seminifcrous tubules had a lumen with sparse small amount of interstitial tubular cells. In the seminiferous epithelium type A spermatogonia, intermediate and B were identified, which occupied the compartment adbasal and intermingled with these cells in spermatocytes I in Pachytene and leptotene was observed, whereas in the adluminal compartment Golgi phase spermatids we observed the presence of acrosomal granule. In group II, the cells of the seminiferous epithelium were developed and it was observed in round spermatids cephalic hood phase plus many elongated spermatids in acrosome phase and Sertoli cells. In Group III, 60 days old, it was found that seminiferous epithelium which was of the tubules had elongated spermatids in acrosome phase and maturation, with elongated nuclei and acrosomal system typical of spermiation in the presence of sperm and residual bodies near the tubular lumen. Therefore morphological evolution of germ cell testicular spermatids can be checked and recognized in its four phases: Golgi, cap, acrosome and maturation over the age of the animal.

Structural evaluation of the effects of chronic ethanol ingestion on the testis of Calomys callosus

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