Apoptotic bone cells may be engulfed by osteoclasts during alveolar bone resorption in young rats

The alveolar bone is a suitable in vivo physiological model for the study of apoptosis and interactions of bone cells because it undergoes continuous, rapid and intense resorption/remodelling, during a long period of time, to accommodate the growing tooth germs. The intensity of alveolar bone resorption greatly enhances the chances of observing images of the extremely rapid events of apoptosis of bone cells and also of images of interactions between osteoclasts and osteocytes/osteoblasts/bone lining cells. To find such images, we have therefore examined the alveolar bone of young rats using light microscopy, the TUNEL method for apoptosis, and electron microscopy. Fragments of alveolar bone from young rats were fixed in Bouin and formaldehyde for morphology and for the TUNEL method. Glutaraldehyde-formaldehyde fixed specimens were processed for transmission electron microscopy. Results showed TUNEL positive round/ovoid structures on the bone surface and inside osteocytic lacunae. These structures - also stained by hematoxylin - were therefore interpreted, respectively, as osteoblasts/lining cells and osteocytes undergoing apoptosis. Osteoclasts also exhibited TUNEL positive apoptotic bodies inside large vacuoles; the nuclei of osteoclasts, however, were always TUNEL negative. Ultrathin sections revealed typical apoptotic images - round/ovoid bodies with dense crescent-like chromatin - on the bone surface, corresponding therefore to apoptotic osteoblasts/lining cells. Osteocytes also showed images compatible with apoptosis. Large osteoclast vacuoles often contained fragmented cellular material. Our results provide further support for the idea that osteoclasts internalize dying bone cells; we were however, unable to find images of osteoclasts in apoptosis. (C) 2001 Harcourt Publishers Ltd.

Efeito da ciclofosfamida sobre o desenvolvimento de germes dentais de molares transplantados para a câmara anterior do olho de camundongos.

Mice foetus were killed between 13 and 16 days of the gestational period and their molar tooth germs grafted into the anterior chamber of the eye. The hosts were injected either with 6.25 mg/Kg or with 125 mg/Kg of cyclophosphamide and killed 15 days later. The cyclophosphamide irreversibly, interferes on tooth germ development causing either its total degeneration or an alteration on its morphology forming hypodeveloped tooth germs; its action is directly proportional to the foetus age and to the dose used; it does not induce the formation of osteodentin while some predentin partial cell inclusion may occur.

A hydrogel scaffold that maintains viability and supports differentiation of dental pulp stem cells

Objectives: The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation. Methods: DPSC cells were grown in 0.05-0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1. Results: DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation. Significance: Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics. © 2012 Academy of Dental Materials.

In vitro antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecalis in root canals

Objective: The aim of this study was to evaluate the antimicrobial activity of auxiliary chemical substances and natural extracts on Candida albicans and Enterococcus faecali's inoculated in root canals. Material and Methods: Seventy-two human tooth roots were contaminated with C. albicans and E. faecalis for 21 days. The groups were divided according to the auxiliary chemical substance into: G1) 2.5% sodium hypochlorite (NaOCl), G2) 2% chlorhexidine gel (CHX), G3) castor oil, G4) glycolic Aloe vera extract, g5) glycolic ginger extract, and G6) sterile saline (control). The samples of the root canal were collected at different intervals: confirmation collection, at 21 days after contamination; 1st collection, after instrumentation; and 2nd collection, seven days after instrumentation. Microbiological samples were grown in culture medium and incubated at 37°C for 48 hours. Results: The results were submitted to the Kruskal-Wallis and Dunn (5%) statistical tests. NaOCl and CHX completely eliminated the microorganisms of the root canals. Castor oil and ginger significantly reduced the number of CFU of the tested bacteria. Reduction of CFU/mL at the 1st and 2nd collections for groups G1, G2, G3 and G4 was greater in comparison to groups G5 and G6. Conclusion: It was concluded that 2.5% sodium hypochlorite and 2% chlorhexidine gel were more effective in eliminating C. albicans and E. faecalis, followed by the castor oil and glycolic ginger extract. The Aloe vera extract showed no antimicrobial activity.

Blood clot stabilization on root dentin conditioned by the combination of tetracycline and EDTA

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

Controvérsias no tratamento das fraturas mandibulares em crianças: relato de caso clínico

Introduction: The treatment of mandibular fractures has undergone many changes, especially after the introduction of rigid internal fixation techniques. Currently, the use of resorbable materials presents some advantages over metallic devices, particularly when applied in children. Objective: The authors discuss the controversies involved in the treatment of mandibular fractures in children, mainly related to fixation of fractures. Case Report: In this paper, we present a case of a 5 years treatment patient with mandibular angle fracture treated by open reduction and internal fixation with plates and screws. Final Comments: Aiming to not interfere in the development of tooth germs and mandibular bone growth a second surgery for removal of the fixation devices is indicated.

Human pulp responses to in-office tooth bleaching

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

Response of human pulps after professionally applied vital tooth bleaching

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Correlation between pH and surface structural changes on human enamel submitted to different tooth whitening agents

The intense valuation of an esthetic pleasant smile guided the dentistry to bleached tooth due the popularity of whitening treatments. The consequence of it is an increasing interest in searching the effect of peroxides in hard dental tissues. The aim of this work was to analyze qualitatively in vitro the human enamel after three different bleaching treatments: Opalescence PF 10%, White Class 7.5% and Opalescence Xtra Boost 38%, correlating the structural changes in the surface of the enamel with its respective pH. A total of 40 sound human pre-molars were randomly divided into four groups of 10 elements, which had been immersed in artificial saliva during all the experiment. Bleaching protocols followed the recommendations of the respective manufacturers. Each bleached sample and control group were submitted to a scanning electronic microscopy analysis and compared with one another. Bleaching agents used in this experiment had modified the morphologic aspect of the surface of the dental enamel; however, it did not have correlation between the degrees of severity of the alterations and pH. There is a correlation between hydrogen peroxide concentration and changes in the enamel, where G4 showed more severe alterations, followed for G3 and G2.

Evaluation of 2 endodontic techniques used to treat human primary molars with furcation radiolucency area: A 48-month radiographic study

Objective: To evaluate 2 techniques for the treatment of human primary molars with necrotic Pulp and bifurcation bone loss by means of radiographic examination for 48 months. Method and Materials: Fifty-one mandibular primary molars were evaluated in children ranging from 4.5 to 6.5 years of age. The teeth with necrotic pulp and bifurcation bone loss were diagnosed by radiographic examination. The teeth were divided into 2 groups: group 1 (28 teeth)-pulpotomy technique using formocresol as a temporary dressing between sessions and coronal chamber obturation with zinc oxide-eugenol cement; and group 2 (23 teeth)-pulpectomy technique with calcium hydroxide paste as a temporary dressing between sessions and root canal obturation with a dense Calcium hydroxide paste. Standardized radiographs were taken immediately after the fillings were completed and after 12, 24, 36, and 48 months. The radiographs were digitized and analyzed with software that outlined and measured the bifurcation radiolucency. Results: Bifurcation radiolucency reduced significantly or repaired completely for both treatnients in the first 12 months. Minor radiographic reduction of the lesion was observed from 12 to 24 months, and no significant reduction of the remaining radioulcent area was observed from 24 to 48 months after treatment. Conclusion: The 2 endodontic techniques evaluated showed similar results. The main effect of treatment on the lesion repair was obtained in the first year after treatment.

Response of human pulps following acid conditioning and application of a bonding agent in deep cavities

Objectives. The aim of this in vivo study was to evaluate the human dental pulp response when a one-bottle adhesive system was applied on etched or unetched deep dentine.Methods. Eighteen class V deep cavity preparations were divided in three groups: group 1-total etching + two coats of single bond (SB) + composite resin (Z-100); group 2-enamel etching + two coats of SB + Z-100, group 3-cavity floor lined with a calcium hydroxide liner (Dycal) + acid-etching of enamel and lateral walls + two coats of SB + Z-100. Two teeth were used as intact control group. After 30 days the teeth were extracted and processed through H and E, Masson's trichrome and Brown and Brenn staining techniques.Results. Moderate inflammatory response, disorganization of pulp tissue, as well as, deposition of thin layer of reactionary dentin were observed in group 1 teeth in which the remaining dentin thickness (RDT) was less than 300 mum. These histological findings appear to be related to long resin tags formation and bonding agent diffusion through dentinal tubules. In group 2, slight inflammatory response was observed only in one tooth in which the RDT was 162 mum. In group 3, all the teeth showed normal histological characteristics which were similar to the intact control group. Presence of bacteria was not correlated with the intensity of pulpal response. The patients reported no symptoms during the experiment. Radiographic evaluation showed no periapical pathology for any of the teeth,Significance and conclusions. Acid-etched deep dentin (RDT less than 300 mum) lined with SB causes more intense pulpal response than unetched deep dentin. Based on the results observed in the present study and the conditions in which it was carried out, we recommend the application of a biocompatible liner before etching deep dentin and applying SB. (C) 2002 Academy of Dental Materials. Published by Elsevier B.V. Ltd. All rights reserved.

Effects of mechanical cycling on the bonding of zirconia and fiber posts to human root dentin

Purpose: To evaluate the effect of cyclical mechanical loading on the bond strength of a fiber and a zirconia post bonded to root dentin.Materials and Methods: Forty single-rooted human teeth (maxillary incisors and canines) were sectioned, and the root canals were prepared at 12 mm. Twenty randomly seleced specimens received a quartz fiber post (FRC) (D.T. Light-Post) and 20 others received a zirconia post (ZR) (Cosmopost). The posts were resin luted (All Bond 2 + resin cement Duo-link) and each specimen was embedded in epoxy resin inside a PVC cylinder. Ten specimens with FRC post and 10 specimens with ZR post were submitted to fatigue testing (2,000,000 cycles; load: 50 N; angle of 45 degrees; frequency: 8 Hz), while the other 20 specimens were not fatigued. Thus, 4 groups were formed: G1: FRC+O cycles; G2: FRC+2,000,000 cycles; G3: ZR+O cycles; G4: ZR+2,000,000 cycles. Later, the specimens were cut perpendicular to their long axis to form 2-mm-thick disk-shaped samples (4 sections/specimen), which were submitted to the push-out test (1 mm/min). The mean bond strength values (MPa) were calculated for each tooth (n = 10) and data were submitted to statistical analysis (alpha = 0.05).Results: Two-way ANOVA revealed that the bond strength was significantly affected by mechanical cycling (p = 0.0014) and root post (p = 0.0325). The interaction was also statistically significant (p = 0.0010). Tukey's test showed that the mechanical cycling did not affect the bonding of FRC to root dentin, while fatigue impaired the bonding of zirconium to root dentin.Conclusion: (1) the bond strength of the FRC post to root dentin was not reduced after fatigue testing, whereas the bonding of the zirconia post was significantly affected by the fatigue. (2) Cyclical mechanical loading appears to damage the bond strength of the rigid post only.

Effect of adhesive system type and tooth region on the bond strength to dentin

Purpose: This study evaluated the bond strength of two etch-and-rinse adhesive systems (two- and three-step) and a self-etching system to Coronal and root canal dentin.Materials and Methods: The root canals of 30 human incisors and canines were instrumented and prepared with burs. The posts used for luting were duplicated with dual resin cement (Duo-link) inside Aestheti Plus #2 molds. Thus, three groups were formed (n = 10) according to the adhesive system employed: All-Bond 2 (TE3) + resin cement post (rcp) + Duo-link (DI); One-Step Plus (TE2) + rcp + DI; Tyrian/One-Step Plus (SE) + rcp + DI. Afterwards, 8 transverse sections (1.5 mm) were cut from 4 mm above the CEJ up to 4 mm short of the root canal apex, comprising coronal and root canal dentin. The sections were submitted to push-out testing in a universal testing machine EMIC (1 mm/min). Bond strength data were analyzed with two-way repeated measures ANOVA and Tukey's test (p < 0.05).Results: The relationship between the adhesives was not the same in the different regions (p < 0.05). Comparison of the means achieved with the adhesives in each region (Tukey; p < 0.05) revealed that TE3 (mean standard deviation: 5.22 +/- 1.70) was higher than TE2 (2.60 +/- 1.74) and SE (1.68 +/- 1.85).Conclusion: Under the experimental conditions, better bonding to dentin was achieved using the three-step etch-and-rinse system, especially in the coronal region. Therefore, the traditional etch-and-rinse three-step adhesive system seems to be the best choice for teeth needing adhesive endodontic restorations.

Penetration of 38% hydrogen peroxide into the pulp chamber in bovine and human teeth submitted to office bleach technique

This study evaluated the pulp chamber penetration of peroxide bleaching agent in human and bovine teeth after office bleach technique. All the teeth were sectioned 3 mm apical of the cement-enamel junction and were divided into 2 groups, A (70 third human molars) and B (70 bovine lateral incisors), that were subdivided into A1 and B1 restored by using composite resin, A2 and B2 by using glass ionomer cement, and A3 and B3 by using resin-modified glass ionomer cement; A4, A5, B4, and B5 were not restored. Acetate buffer was placed in the pulp chamber, and the bleaching agent was applied for 40 minutes as follows: A1-A4 and B1-B4, 38% hydrogen peroxide exposure and A5 and B5, immersion into distilled water. The buffer solution was transferred to a glass tube in which leuco crystal violet and horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined by spectrophotometer and converted into microgram equivalents of hydrogen peroxide. Data were submitted to analysis of variance and Dunnett, Kruskal-Wallis, and Tukey tests (5%). A higher level of hydrogen peroxide penetrated into the pulp chamber in resin-modified glass ionomer cements in bovine (0.79 +/- 0.61 mu g) and human (2.27 +/- 0.41 mu g) groups. The bleaching agent penetration into the pulp chamber was higher in human teeth for any experimental situation. The penetration of the hydrogen peroxide depends on restorative materials, and under the conditions of this study human teeth are more susceptible to penetration of bleaching agent into the pulp chamber than bovine teeth.

Influence of fluoride dentifrice on brushing abrasion of eroded human enamel: An in situ/ex vivo study

This in situ/ex vivo study assessed the effect of fluoride dentifrice on eroded enamel subjected to brushing abrasion. In a crossover study performed in 2 phases, 10 volunteers wore acrylic palatal appliances, each containing 3 human enamel blocks. Dentifrice was used to brush the volunteers' teeth and the specimens subjected to abrasion. In phases A and B the dentifrices used had the same formulation, except for the absence or presence of fluoride, respectively. The blocks were subjected to erosion by immersion of the appliances in a cola drink for 5 min, 4 times a day. Then the blocks were brushed, and the appliance was replaced into the mouth. Enamel alterations were determined using profilometry and percentage change in surface microhardness (%SMHC) tests. The data were tested using the paired t test. The mean wear values (+/- SD, mu m) were: group A 6.84 +/- 1.72 and group B 5.38 +/- 1.21 (p = 0.04). The mean %SMHC values (+/- SD) were: group A 54.6 +/- 16.2 and group B 45.7 +/- 6.8 (p = 0.04). Fluoride dentifrice had a protective effect on eroded enamel subjected to brushing abrasion. Copyright (c) 2007 S. Karger AG, Basel.