Ultrastructure of early mineral deposition during hyaline layer formation in rat molars

There is no consensus on whether the first mineralized layer, the hyaline layer, that is juxtaposed to root dentine is a variety of dentine or cementum or even a tissue of epithelial origin. Some suggest that there is no intermediate tissue between the acellular extrinsic fibre cementum (AEFC) and the root dentine. Here, to study hyaline layer formation and mineralization we examined by transmission electron microscopy the early stages of root development in upper molars from 10 to 13 day old Wistar rats. In addition to conventionally processed material, undemineralized and unstained sections were examined, which showed the deposition of fine mineral crystals in contact with the mineralized surface of root dentine. Early mineralization of the hyaline layer occurred in the region of the inner basement membrane, which persisted between the inner cellular layer of Hertwig's epithelial root sheath and the outer mineralized root dentine. When the root sheath began its fragment, collagen fibrils From the developing periodontal ligament began to insert into the mineralising hyaline layer, which was 0.5-0.8 mum wide. As the fragmentation of the root sheath HERS increased, more collagen fibrils appeared intermingled with the mineralising hyaline layer. In more advanced stages, when the hyaline layer had become fully mineralized and the formation of the AEFC began, the hyaline layer could no longer be identified. Thus, the hyaline layer is clearly discernible at early stages of periodontal development. Subsequently, it is masked by intermingling of cementum and dentine and therefore it is not possible to detect it in the formed roots of rat molars. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Tissue response to an epoxy resin-based root canal sealer

The objective of the present study was to evaluate two different types of root canal sealers: AH Plus (an epoxy resin-based sealer) and Fill Canal (a zinc oxide-eugenol based sealer). A total of 34 root canals with vital pulp from dogs' premolars were used. After instrumentation, the root canals were filled with gutta-percha and AH Plus or gutta-percha and Fill Canal sealers using a classical technique of lateral condensation. After histological processing, the sections were stained with hematoxylineosin or Mallory's trichrome stain. Inflammatory cells or areas of necrosis were not associated with AH Plus. Hard tissue formation apically to the material was observed in 14 specimens. The Fill Canal sealer presented an inflammatory response of moderate intensity in the periapical region, mainly adjacent to the material.

Mechanism of micronuclei formation in polyploidizated cells of Allium cepa exposed to trifluralin herbicide

The trifluralin is an agent that promotes a cellular damage due to its direct action on the microtubules. This action leads to a decontrol in the cellular division, bringing about polyploid cells. In this work, we show the evidences that the exceeding genetical material of theses polyploidizated cells tends to be eliminated from the nucleus in the form of micronucleus. Our analyses prove this fact, both by the presence of a number of cells carrying micronucleus, and by the evidences of the elimination of the exceeding material itself, after exposition of the Allium cepa root tips tested with several concentration of trifluralin herbicide. It was noticed that the residual concentration induced a number of polyploid cells, micronuclei and mini cells. Inferences about the implications of the elimination of genetic material from micronuclei, such as cell viability and apoptosis, are also presented. (c) 2007 Elsevier B.V. All rights reserved.

In vitro evaluation of root canal preparation using oscillatory and rotary systems in flattened root canals

Objective: the aim of this study was to evaluate the biomechanical preparation of flattened root canals using the following systems: Endo-Eze AET stainless steel oscillatory instruments (Ultradent) and RaCe rotary NiTi instruments (FKG Dentaire). Materials and Methods: Twenty extracted human mandibular incisors were randomly assigned to two groups: Group I Instrumentation with oscillatory Endo-Eze AET files (oscillatory technique); Group 2 - Instrumentation with rotary NiTi RaCe files (rotary technique). The teeth were decoronated, had their apices and coronal openings sealed with sticky wax and were embedded in crystal-clear orthophtalic polyester resin. The roots were sectioned transversally with diamond discs at 10 mm (middle third) and 5 mm (apical third) from the apex and the segments were reassembled for instrumentation. The sections were photographed before and after root canal instrumentation and evaluated with respect to whether the original root canal shape was modified by instrumentation. To evaluate the differences in the root canal shape before and after biomechanical preparation, scores were given regarding the instruments touch on the intracanal walls. Results: In middle third of the root canals instrumented with the rotary system, there was a change in the original canal anatomy (p < 0.05), with formation of a protuberance in the mesiodistal direction. This protuberance did not occur when the oscillatory instrumentation was used. The oscillatory system had better results in the middle and apical thirds as evaluated by Dunn's multiple-comparison test (p > 0.05). Conclusion: Under the tested conditions, Endo-Eze oscillatory system yielded the instrumentation of all flattened oot canal walls, maintaining the canal original shape throughout the biomechanical preparation, and was more effective than RaCe rotary system.

Histological evaluation of MTA as a root-end filling material

Aim To assess the histological response associated with grey mineral trioxide aggregate (GMTA) and zinc oxide eugenol (ZOE) as root-end filling materials in teeth where the root canals were not filled and the coronal access cavities were not restored.Methodology Periapical lesions were developed in 24 premolar teeth in three dogs. The root canals were prepared and half of them were dried, filled and the coronal access restored (closed). The remaining teeth were not root filled and no coronal restoration was placed (open). Apical root-end resections were performed 3 mm from the apex, and root-end cavities were prepared with ultrasonic tips. These were randomly filled with either ZOE or GMTA in the same number of specimens using MAPSYSTEM device. After 180 days the animals were killed and blocks of tissues removed and processed for histological examination. Periradicular tissue reaction was evaluated, including severity of inflammation and cementum formation. Statistical analysis was performed using ANOVA analysis and Tukey's test.Results A significant difference was found between the levels of inflammation in the periradicular tissues of the GMTA/closed group, compared with the ZOE/open and ZOE/closed groups (P < 0.05) but not between GMTA/closed and GMTA/open groups. Cementum formation was not found over any ZOE specimens but over MTA in all specimens. No microorganisms were found in the interface between the material and the dentinal walls.Conclusions GMTA was associated with less periapical inflammation and tissue response when used as a root-end filling material, even when no root filling or coronal restoration was present.

Healing of dehiscence defects following root surface demineralization with tetracycline: A histologic study in monkeys

Background: the purpose of this study was to histologically evaluate the healing of experimental dehiscence defects after surface demineralization with tetracycline hydrochloride.Methods: Six adult male monkeys (Cebus apella) were used in this study. Dehiscence defects were surgically created on the buccal aspect of the mandibular lateral incisors in all animals. The root surfaces were debrided and planed. In a split-mouth design, a 10% tetracycline hydrochloride solution was applied to one tooth for 4 minutes (T group), followed by irrigation with saline. The contralateral tooth served as a control (C group). The flaps were repositioned and sutured. The animals were sacrificed at 6 months postoperatively and histological sections were processed. Computer-assisted histomorphometric analysis was used to evaluate the formation of new cementum, new bone, new connective tissue attachment, and length of the epithelium (junctional and sulcular).Results: Bone regeneration was similar in both groups (1.5 +/- 0.3 mm for the T group and 1.5 +/- 0.6 mm for the C group). The C group showed more new cementum than the T group (2.3 +/- 0.3 mm versus 2.2 +/- 0.3 mm) as well as a longer epithelium (1.0 +/- 0.3 mm versus 0.9 +/- 0.2 mm). The T group presented more new connective tissue attachment (3.1 +/- 0.2 mm) than the C group (2.9 +/- 0.6 mm). However, no statistically significant differences were detected between the two groups.Conclusions: the amount of new attachment was similar in both groups. Root conditioning with 10% tetracycline solution did not produce any additional new attachment in comparison to the controls.

Hard tissue formation adjacent to implants of various size and configuration immediately placed into extraction sockets: an experimental study in dogs

ObjectivesTo evaluate the influence of implant size and configuration on osseointegration in implants immediately placed into extraction sockets.Material and methodsImplants were installed immediately into extraction sockets in the mandibles of six Labrador dogs. In the control sites, cylindrical transmucosal implants (3.3 mm diameter) were installed, while in the test sites, larger and conical (root formed, 5 mm diameter) implants were installed. After 4 months of healing, the resorptive patterns of the alveolar crest were evaluated histomorphometrically.ResultsWith one exception, all implants were integrated in mineralized bone, mainly composed of mature lamellar bone. The alveolar crest underwent resorption at the control as well as at the test implants. This resorption was more pronounced at the buccal aspects and significantly greater at the test (2.7 +/- 0.4 mm) than at the control implants (1.5 +/- 0.6 mm). However, the control implants were associated with residual defects that were deeper at the lingual than at the buccal aspects, while these defects were virtually absent at test implants.ConclusionsThe installment of root formed wide implants immediately into extraction sockets will not prevent the resorption of the alveolar crest. In contrast, this resorption is more marked both at the buccal and lingual aspects of root formed wide than at standard cylindrical implants.To cite this article:Caneva M, Salata LA, de Souza SS, Bressan E, Botticelli D, Lang NP. Hard tissue formation adjacent to implants of various size and configuration immediately placed into extraction sockets: an experimental study in dogs.Clin. Oral Impl. Res. 21, 2010; 885-895.doi: 10.1111/j.1600-0501.2010.01931.x.

The induction of cytotoxicity, oxidative stress, and genotoxicity by root canal sealers in mammalian cells

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

Direct and indirect connections between cochlear root neurons and facial motor neurons: Pathways underlying the acoustic pinna reflex in the albino rat

Cochlear root neurons (CRNs) are involved in the acoustic startle reflex, which is widely used in behavioral models of sensorimotor integration. A short-latency component of this reflex, the auricular reflex, promotes pinna movements in response to unexpected loud sounds. However, the pathway involved in the auricular component of the startle reflex is not well understood. We hypothesized that the auricular reflex is mediated by direct and indirect inputs from CRNs to the motoneurons responsible for pinna movement, which are located in the medial subnucleus of the facial motor nucleus (Mot7). To assess whether there is a direct connection between CRNs and auricular motoneurons in the rat, two neuronal tracers were used in conjunction: biotinylated dextran amine, which was injected into the cochlear nerve root, and Fluoro-Gold, which was injected into the levator auris longus muscle. Under light microscopy, close appositions were observed between axon terminals of CRNs and auricular motoneurons. The presence of direct synaptic contact was confirmed at the ultrastructural level. To confirm the indirect connection, biotinylated dextran amine was injected into the auditory-responsive portion of the caudal pontine reticular nucleus, which receives direct input from CRNs. The results confirm that the caudal pontine reticular nucleus also targets the Mot7 and that its terminals are concentrated in the medial subnucleus. Therefore, it is likely that CRNs innervate auricular motoneurons both directly and indirectly, suggesting that these connections participate in the rapid auricular reflex that accompanies the acoustic startle reflex. © 2008 Wiley-Liss, Inc.

Blood cells attachment after root conditioning and PRP application: An in vitro study

Aim: Root conditioning is aimed at smear layer removal and at dental matrix collagen exposure, which may promote periodontal regeneration. This in vitro study assessed smear layer removal, collagen fiber exposure and the influence of PRP (platelet-rich plasma) application on adhesion of blood cells to the root surface using scanning electron microscopy (SEM). Materials and methods: Scaled root samples (n = 160) were set in five groups and conditioned with: group I - control group (saline solution); group II (EDTA 24%); group III (citric acid 25%); group IV (tetracycline hydrochloride 50 mg/ml); group V (sodium citrate 30%). Eighty samples were assessed using the root surface modification index (RSMI). The other eighty samples were set in two groups. The first group (n = 40) received PRP gel application with a soft brush and the second group (n = 40) received PRP application and then a blood drop. The fibrin clot formation was assessed in the first group and the blood cells adhesion was assessed in the second group using the BEAI (blood elements adhesion index). A previously trained, calibrated, and blind examiner evaluated photomicrographs. Statistical analysis was performed using the Kruskal-Wallis's and Dunn's tests. Results: Group III attained the best results for RSMI and BEAI. Moreover, it was the only group showing fibrin clot formation. Conclusion: Citric acid was the most efficient conditioner for smear layer removal, collagen fiber exposure and blood cell adhesion. Moreover, it was the only group showing fibrin clot formation after PRP application. Clinical significance: This study demonstrated that root conditioning followed by PRP application may favor blood cell adhesion on root surface which may optimize periodontal healing.

Wound healing of dehiscence defects following different root conditioning modalities: An experimental study in dogs

Objective: The purpose of this study was to investigate the periodontal healing pattern of dehiscence-type defects following different chemical root conditioning modalities. Materials and methods: Buccal osseous dehiscence defects were created on six teeth of seven dogs. After dental plaque accumulation, defects were treated with sterile saline solution (control group) or one chemical conditioning modality: citric acid (CA group), ethylenediaminetetraacetic acid (EDTA group), tetracycline (TTC group), citric acid + tetracycline (CA + TTC group), or tetracycline + citric acid (TTC + CA group). After 3 months of healing, clinical parameters were evaluated, and the animals were killed. Histological sections were processed, and a computer-assisted histometric analysis was used to evaluate the formation of new cementum, new bone, and epithelial apical migration. Results: All treatments yielded significant improvements in terms of probing depth decrease and clinical attachment level gain compared to baseline values; however, without significant differences among the groups (p > 0.05; one-way ANOVA). The highest amount of new cementum was noted in the EDTA group (3.72 ± 0.83 mm, 77.6 %), while the lowest amount of new bone was observed in the TTC group (0.7 ± 0.94 mm, 14.3 %). However, no statistically significant differences could be observed among the groups regarding epithelial apical migration, new cementum, and alveolar bone formation (p > 0.05). Conclusion: Chemical root surface conditioning did not promote any significant improvement in periodontal healing pattern of dehiscence-type defects in dogs. Clinical Relevance: Chemical root surface conditioning after surgical debridement did not promote positive or negative effects on periodontal healing pattern of dehiscence-type defects. © 2012 Springer-Verlag Berlin Heidelberg.

Araçatuba formation: palustrine deposits from the initial sedimentation phase of the Bauru Basin

A Bacia Bauru (Cretáceo Superior), acumulou uma seqüência sedimentar continental essencialmente arenosa. Numa fase inicial desértica, o seu substrato basáltico foi soterrado por extensa e monótona cobertura de areias eólicas com intercalações subordinadas de depósitos de loesse. O relevo original do substrato favoreceu a formação de uma drenagem regional endorrêica, sob clima semi-árido, propiciando assim condições de formação do Paleopantanal Araçatuba. Os depósitos paludiais (Formação Araçatuba) constituem estratos tabulares de siltitos e arenitos de cor cinza claro esverdeado típica, eventualmente cimentados por carbonato de cálcio. Moldes e pseudomorfos de cristais de gipsita e dolomita foram identificados na unidade. Aparentemente, estão associados com gretas de ressecação, marcas de raízes e intervalos com laminação tipo climbing ripple, que indicam ambiente de águas salinas rasas e relativamente calmas, submetidas a fases de exposição subaérea e ressecação. Nos limites da área de ocorrência da Formação Araçatuba, as unidades arenosas podem exibir feições sigmóides e estratificação contorcida, comuns em depósitos deltaicos marginais. A Formação Araçatuba é contornada e posteriormente encoberta por depósitos eólicos da Formação Vale do Rio do Peixe.

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)

Platelet-Rich Plasma Derived From Bone Marrow Aspirate Promotes New Cementum Formation

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of the association between citric acid and EDTA on root surface etching

Objective: This study aims to compare the clot stabilization on root surfaces conditioned with citric acid and ethylenediaminetetraacetic acid (EDTA). Materials and methods: Scaled root samples (n = 100) were set in five groups: group I?control group (saline solution); group II (24% EDTA); group III (25% citric acid); group IV (EDTA + citric acid); group V (citric acid + EDTA). Fifty samples were assessed using the root surface modification index (RSMI). The other 50 received a blood drop after conditioning. Clot formation was assessed using blood elements adhesion index (BEAI). A blind examiner evaluated photomicrographs. Statistical analysis considered p < 0.05. Results: Groups-III and G-V attained the best results for RSMI and BEAI in comparison to control. The worst results for clot stabilization were seen in group-II. EDTA employment before citric acid (group-IV) reduced clot formation in comparison to citric acid use alone (group-III). Conclusion: Root conditioning with citric acid alone and before EDTA had the best results for smear layer removal and clot stabilization. EDTA inhibited clot stabilization on root surface and must have a residual activity once it has diminished clot adhesion to root even after citric acid conditioning. Thus, EDTA can be used to neutralize citric acid effects on periodontal cells without affecting clot stabilization. Clinical significance: To demonstrate that citric acid use on root surfaces previously affected by periodontal disease may favor clot stabilization and may have a beneficial effect on surgical outcomes. Also, EDTA can be used to neutralize citric acid effects on periodontal cells.