Influence of enamel matrix derivative (Emdogain((R))) and sodium fluoride on the healing process in delayed tooth replantation: histologic and histometric analysis in rats

Although it has already been shown that enamel matrix derivative (Emdogain((R))) promotes periodontal regeneration in the treatment of intrabony periodontal defects, there is little information concerning its regenerative capacity in cases of delayed tooth replantation. To evaluate the alterations in the periodontal healing of replanted teeth after use of Emdogain((R)), the central incisors of 24 Wistar rats (Rattus norvegicus albinus) were extracted and left on the bench for 6 h. Thereafter, the dental papilla and the enamel organ of each tooth were sectioned for pulp removal by a retrograde way and the canal was irrigated with 1% sodium hypochlorite. The teeth were assigned to two groups:in group I, root surface was treated with 1% sodium hypochlorite for 10 min (changing the solution every 5 min), rinsed with saline for 10 min and immersed in 2% acidulated-phosphate sodium fluoride for 10 min; in group II, root surfaces were treated in the same way as described above, except for the application of Emdogain((R)) instead of sodium fluoride. The teeth were filled with calcium hydroxide (in group II right before Emdogain((R)) was applied) and replanted. All animals received antibiotic therapy. The rats were killed by anesthetic overdose 10 and 60 days after replantation. The pieces containing the replanted teeth were removed, fixated, decalcified and paraffin-embedded. Semi-serial 6-mu m-thick sections were obtained and stained with hematoxylin and eosin for histologic and histometric analyses. The use of 2% acidulated-phosphate sodium fluoride provided more areas of replacement resorption. The use of Emdogain((R)) resulted in more areas of ankylosis and was therefore not able to avoid dentoalveolar ankylosis. It may be concluded that neither 2% acidulated-phosphate sodium fluoride nor Emdogain((R)) were able to prevent root resorption in delayed tooth replantation in rats.

Healing process of incisor teeth of diabetic rats replanted after storage in milk

Several local factors that influence the healing process of replanted teeth have been investigated. However, it remains unclear how systemic alterations, such as diabetes mellitus, affect the prognosis of these cases. The purpose of this study was to evaluate the healing process of incisors of non-controlled diabetic rats replanted after storage in bovine long shelf-life (UHT) whole milk. Thirty-two rats were randomly assigned to receive an endovenous injection of either citrate buffer solution (group I - control; n = 16) or streptozotocin dissolved in citrate buffer solution to induce diabetes (group II; n = 16). After confirmation of the diabetic status by analysis of the glycemic levels, the maxillary right incisor of each animal was extracted and immersed in milk for 60 min. The root canals of teeth were then instrumented, and were filled with a calcium hydroxide-based dressing and replanted into their sockets. All animals received systemic antibiotic and were killed by anesthetic overdose 10 and 60 days after replantation. The specimens containing the replanted teeth were removed, fixed, decalcified, and embedded in paraffin. Semi-serial 6-mu m-thick sections were obtained and stained with hematoxylin and eosin for histologic and histometric analyses. The results showed that the connective tissue adjacent to the root surface was less organized in the diabetic animals than in the control animals in both periods; the root dentin was less severely affected by root resorption in the diabetic rats; there were no significant differences between the control and diabetic groups regarding the occurrence of replacement resorption and inflammatory resorption.

Evaluation of alveolar socket response to Angelus MTA and experimental light-cure MTA

Objective. The aim of this study was to evaluate the rat alveolar socket response to implanted polyethylene tubes filled with a light-cured MTA or Angelus MTA.Study design. Fifty-seven Wistar rats received an implant filled with one of the materials in the dental socket of the right central incisor. After 30, 60, and 90 days, animals were killed, and the right hemimaxillas were removed and processed to be analyzed on light microscopy. After embedding in paraffin, 6-mu m-thick sections were obtained and stained with hematoxylin and eosin. Qualitative and quantitative analyses of the reaction were performed.Results. It was observed that the light-cured MTA presented a similar response compared with Angelus MTA, characterized by the reduction of the chronic inflammatory response and organization of the alveolar bone close to the materials, especially at the 90th day of observation.Conclusions. It was concluded that both materials were well accepted by the alveolar tissue of rats, with the formation of mineralized tissue close to the materials. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;110:e93-e97)

Delayed replantation of rat teeth after use of reconstituted powdered milk as a storage medium

Minimal extraoral dry storage period and moist storage for the avulsed tooth are identified as key steps for the treatment protocol of tooth replantation. Among the possible moist storage media, bovine milk has stood out because of its capacity of preserving the integrity of the periodontal ligament (PDL) fibers. This condition has attracted the attention to investigate the use of powdered milk, which is one of the presentation forms of bovine milk, as a feasible storage medium in cases of delayed tooth replantation. The aim of this study was to evaluate the healing process after delayed replantation of rat teeth stored in reconstituted powdered milk and long shelf-life (ultra high temperature) whole milk. Forty maxillary right rat incisors were assigned to four groups (n = 10): group I - the teeth were extracted and immediately replanted into theirs sockets; group II - the teeth were stored for 60 min in 200 ml of freshly reconstituted powdered milk; group III - the teeth were stored for 60 min in 200 ml of long shelf-life whole milk; group IV - the teeth were kept dry for the same time. All procedures were performed at room temperature. Next, the root canals of teeth in groups II, III, and IV were instrumented, filled with a calcium hydroxide-based paste, and replanted into their sockets. All animals received systemic antibiotic therapy and were killed by anesthetic overdose 60 days after replantation. The pieces containing the replanted teeth were removed, fixed, decalcified, and paraffin-embedded. Semi-serial 6-mu m-thick sections were obtained and stained with hematoxylin and eosin for histomorphological analysis. There was statistically significant difference (P < 0.05) between groups I and IV regarding the presence of replacement resorption and PDL remnants on root surface. The powdered milk and long shelf-life whole milk presented similar results to each other and may be indicated as storage media for avulsed teeth.

Hybrid layer thickness and resin tag length of a self-etching adhesive bonded to sound dentin

Objectives: the purpose of this study is to employ optical microscopy to measure the thickness of the hybrid layer and the penetration (tags) of an aggressive self-etching adhesive system into sound dentin.Methods: occtusat cavities were prepared in 40 extracted human posterior teeth. The prepared teeth were randomly assigned to four experimental groups with 10 specimens each. The self-etching adhesive system Adper Prompt L-Pop was applied to the dentin surface as follows: Group 1: cavosurface enamel was etched for 60 s and dentin for 20 s with 35% phosphoric acid get, immediately followed by application of the self -etching adhesive with a brush to the entire cavity for 15 s; Groups 2, 3, and 4: no pre-etching was performed, and the self -etching adhesive was applied to both enamel and dentin for 15, 30 and 45 s, respectively. After curing, the cavities were fitted with composite resin Fittek Z250. Afterwards, the teeth were decalcified and the restorations were carefully removed for later embedding in paraffin. The specimens were serially sectioned at 6 mu m of thickness and sequentially mounted in glass slides. These sections were stained with Brown and Brenn staining for posterior analysis and measurement of the hybrid layer and resin tags on a tight microscope with a micrometric ocular 40/075. The results were submitted to analysis of variance at the 5% level.Results: whenever there was significance, the Tukey test was applied at the 5% level. The specimens receiving application of acid etching before the selfetching. adhesive displayed a larger thickness of the hybrid layer; on the other hand, specimens receiving only application of the self -etching adhesive on dentin for 15, 30 and 45 s exhibited similar thickness of the hybrid layer. As regards the resin tags, no statistically significant differences could be found between the study groups.Conclusions: it could be concluded that the increase in the time of application of the self-etching adhesive Adper Prompt L-Pop did not significantly influence the formation and thickness of hybrid layer, as well as its penetration into the sound dentin surface. (c) 2005 Elsevier Ltd. All rights reserved.

Comparison of the biocompatibility of different root canal irrigants

The purpose of this study was to compare the reaction of rat subcutaneous connective tissue to 0.9% sterile saline, 2.5% sodium hypochlorite (NaOCl), 5.25% NaOCl and 2% chlorhexidine gluconate solution or gel. Six circles were demarcated on the dorsal skin of 24 male Wistar rats, leaving 2 cm between each circle. Using a syringe, 0.1 mL of each root canal irrigant was injected subcutaneously into 5 circles. In the 6th circle, the needle of an empty syringe was introduced into the skin, but no irrigant was injected (control group). Evaluations were undertaken at 2 h, 48 h, 14 days and 30 days post-procedure. Tissue samples were excised, embedded in paraffin blocks and 3-mu m-thick sections were obtained and stained with hematoxylin and eosin. The areas of inflammatory reaction were evaluated and analyzed statistically by ANOVA and Tukey's test. The control group showed few or no inflammatory reaction areas in the subcutaneous tissue. 0.9% saline solution, 2.0% chlorhexidine solution and 2.5% NaOCl showed a good biocompatibility, as very mild inflammatory reaction was detected at 14 days and tissue repair occurred at 30 days. 5.25% NaOCl was the most toxic irrigant, as the number of inflammatory cells remained elevated at 14 and 30 days. The group treated with 2.0% chlorhexidine gluconate gel presented a moderate inflammatory response at 14 days, which decreased at 30 days, being considered similar to that of the control group, 0.9% saline solution, 2.0% chlorhexidine solution and 2.5% NaOCl at this experimental period.

Bone healing in critical-size defects treated with either bone graft, membrane, or a combination of both materials: a histological and histometric study in rat tibiae

Objectives: The aim of the present investigation was to histologically analyze the effect of using lyophilized bovine bone (GenOx (R) organic matrix) with (or without) guided tissue regeneration (using a decalcified cortical osseous membrane [GenDerm (R)]) on bone healing in surgically created critical-size defects created in rat tibia.Material and methods: Surgical critical-size bone defects were created in 64 animals that were randomly divided into four groups: group I (control); group II (defect filled with GenOx (R)); group III (defect covered by GenDerm (R)); group IV (defect filled with GenOx (R) and covered by GenDerm (R)). Animals were killed at 30 or 90 days post-surgery. The specimens were embedded in paraffin, serially cut, and stained with hematoxylin and eosin for analysis under light microscopy. The formation of new bone in the cortical area of the defect was histomorphometrically evaluated.Results: All experimental groups demonstrated superior bone healing compared with the control group. However, group IV samples showed evidence of more advanced healing at both 30 and 90 days post-surgery as compared with the other experimental groups.Conclusions: The bovine organic bone graft GenOx (R) associated with GenDerm (R) this produced the best treatment results in the case of critical-size defects in rat tibia.

Correlation between koilocytes and human papillomavirus detection by PCR in oral and oropharynx squamous cell carcinoma biopsies

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

Immunosuppressant Prograf (R) (Tacrolimus) Induces Histopathological Disorders in the Peritubular Tissue of Rat Testes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Structural alterations in the seminiferous tubules of rats treated with immunosuppressor tacrolimus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Presença do papilomavirus humano em lesões malignas de mucosa oral

O objetivo deste estudo foi investigar a prevalência do papilomavírus humano 6/11 e 16/18 em pacientes, com lesões orais clínicamente diagnosticadas como leucoplasias, atendidas na Faculdade de Odontologia de Araraquara, UNESP, Brasil. Após a inclusão em parafina, os cortes corados com H&E, foram selecionadas 30 biópsias e separadas em 3 grupos: lesões sem displasia (n=10), lesões com diferentes graus de displasia (n=10) e carcinoma espinocelular invasivo(n=10). As lesões que apresentaram displasia epitelial foram classificadas de acordo com os critérios histopatológicos propostos por Van Der Waal. As lesões foram investigadas para a presença de HPV por hibridização in situ com sondas biotiniladas de amplo espectro, 6/11 e 16/18. HPV 16/18 foi detectado em 20% (n=2) das biópsias com displasia severa. A presença de HPV 16/18 em lesões malignas sugere sua importância como fator de risco na carcinogênese oral.

Staining methods applied to glycol methacrylate embedded tissue sections

The use of glycol methacrylate (GMA) avoids some technical artifacts, which are usually observed in paraffin-embedded sections, providing good morphological resolution. on the other hand, weak staining have been mentioned during the use of different methods in plastic sections. In the present study, changes in the histological staining procedures have been assayed during the use of staining and histochemical methods in different GMA-embedded tissues.Samples of tongue, submandibular and sublingual glands, cartilage, portions of respiratory tract and nervous ganglion were fixed in 4% formaldehyde and embedded in glycol methacrylate. The sections of tongue and nervous ganglion were stained by H&E. Picrosirius, Toluidine Blue and Sudan Black B methods were applied, respectively, for identification of collagen fibers in submandibular gland, sulfated glycosaminoglycans in cartilage (metachromasia) and myelin lipids in nervous ganglion. Periodic Acid-Schiff (PAS) method was used for detection of glycoconjugates in submandibular gland and cartilage while AB/PAS combined methods were applied for detection of mucins in the respiratory tract. In addition, a combination of Alcian Blue (AB) and Picrosirius methods was also assayed in the sublingual gland sections.The GMA-embedded tissue sections showed an optimal morphological integrity and were favorable to the staining methods employed in the present study. In the sections of tongue and nervous ganglion, a good contrast of basophilic and acidophilic structures was obtained by H&E. An intense eosinophilia was observed either in the striated muscle fibers or in the myelin sheaths in which the lipids were preserved and revealed by Sudan Black B. In the cartilage matrix, a strong metachromasia was revealed by Toluidine Blue in the negatively-charged glycosaminoglycans. In the chondrocytes, glycogen granules were intensely positive to PAS method. Extracellular glycoproteins were also PAS positive in the basal membrane and in the region occupied by the lamina externa and reticular fibers surrounding each smooth muscle cells of the blood vessels. In the epithelial cells of the respiratory tract, acid and neutral mucins were histochemically detected by AB and PAS methods, respectively. Moreover, granules containing acid and neutral mucins were revealed in purple by AB and PAS concomitantly. In the sublingual gland sections, a distinct affinity of acid mucins by AB (in turquoise-blue) and collagen fibers by Picrosirius (in red) was obtained when these methods were combined. Although some routine dyes used in paraffin sections have showed a weak stain in historesin sections, our results showed that different dyes could be applied in GMA sections if modified staining procedures were assayed. Therefore, appropriate staining contrast and, thus, detection of one or different substances in a same section can be acquired in association to the good morphological resolution provided by GMA. (C) 2003 Elsevier Ltd. All rights reserved.

Processo de reparo em feridas de extração dentária em camundongos tratados com o complexo Symphytum officinale e Calendula officinallis

Medicamentos homeopáticos como o Symphytum officinalle e a Calendula officinallis são dotados de propriedades anti-sépticas, antiinflamatória, cicatrizantes e também agem como promotores da consolidação de fraturas ósseas. Neste trabalho, uniram-se esses dois medicamentos similares em um complexo para verificar o seu efeito no reparo em feridas de extração dentária em camundongos. O complexo Symphytum officinalle e Calendula officinallis nas potências de 6CH e 3CH, respectivamente, foi ministrado por via oral ao grupo tratado durante 5 dias antes e após a extração do incisivo superior direito. No grupo controle, administraram-se 5ml de álcool etílico a 70% diluídos em 30 ml de soro fisiológico. Após a proservação, os animais foram sacrificados, a maxila direita separada da esquerda, fixada e processada para inclusão em parafina. Após a microtomia, os cortes obtidos foram corados pela H/E. A análise histológica mostrou que, tanto no grupo controle como no tratado, o alvéolo dentário estava preenchido por tecido de granulação e tecido ósseo neoformado, com graus variáveis de maturação, rico em osteócitos. No entanto, nos animais tratados, o processo de reparo em feridas após extração dentária do incisivo superior direito mostrou um avanço progressivo de neoformação óssea mais acentuado quando comparado ao grupo controle, em tempos equivalentes. Estes resultados enfatizam as propriedades biológicas do complexo Symphytum officinalle e Calendula officinallis e sua possível utilização como recurso terapêutico na Odontologia.

Immunohistochemical detection of estrogen receptor beta in alveolar bone cells of estradiol-treated female rats: possible direct action of estrogen on osteoclast life span

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mast cells and MMP-9 in the lamina propria during eruption of rat molars: quantitative and immunohistochemical evaluation

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)