Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação

Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies

Calcium Hydroxide Mixed With Camphoric p-Monochlorophenol or Chlorhexidine in Delayed Tooth Replantation

This study evaluated the repair process after delayed replantation of rat teeth, using calcium hydroxide (Ca(OH)(2)) mixed with camphorated p-monochlorophenol (CMCP), chlorhexidine 2% (CHX), or saline as temporary root canal dressing to prevent and/or control inflammatory radicular resorption. Thirty Wistar rats (Rattus norvegicus albinos) had their right upper incisor extracted, which was bench-dried for 60 minutes. The dental papilla, the enamel organ, the dental pulp, and the periodontal ligament were removed. The teeth were immersed in 2% acidulated-phosphate sodium fluoride solution for 10 minutes. The root canals were dried with absorbent paper cones and divided into 3 groups of 10 animals according to root canal dressing used: group 1: Ca(OH)(2) + saline, group 2: Ca(OH)(2) + CMCP, and group 3: Ca(OH)(2) + CHX 2%. Before replanting, the teeth sockets were irrigated with saline. Histological analysis revealed the presence of inflammatory resorption, replacement resorption, and ankylosis in all 3 groups. Statistical analysis showed a significant difference between group 3 and the other groups. The use of Ca(OH)(2) mixed with CMCP or CHX did not show an advantage over the use of Ca(OH)(2) mixed with saline in preventing and/or controlling inflammatory resorption in delayed replantation of rat teeth.

Effects of light-curing time on the cytotoxicity of a restorative composite resin on odontoblast-like cells

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

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.

Effect of Er : YAG and diode laser irradiation on the root surface: Morphological and thermal analysis

Background: the aim of the present study was to compare the effects of Er:YAG and diode laser treatments of the root surface on intrapulpal temperature after scaling and root planing with hand instruments.Methods: Fifteen extracted single-rooted teeth were scaled and root planed with hand instruments. The teeth were divided into 3 groups of 5 each and irradiated on their buccal and lingual surfaces: group A: Er:YAG laser, 2.94 mum/100 mJ/10 Hz/ 30 seconds; group B: diode laser, 810 nm/1.0 W/0.05 ms/30 seconds; group C: diode laser, 810 nm/1.4 W/0.05 ms/30 seconds. The temperature was monitored by means of a type T thermocouple (copper-constantan) positioned in the pulp chamber to assess pulpal temperature during and before irradiation. Afterwards, the specimens were longitudinally sectioned, and the buccal and lingual surfaces of each root were analyzed by scanning electron microscopy.Results: In the Er:YAG laser group, the thermal analysis revealed an average temperature of -2.2 +/- 1.5degreesC, while in the diode laser groups, temperatures were 1.6 +/- 0.8degreesC at 1.0 W and 3.3 +/- 1.0degreesC at 1.4 W. Electronic micrographs revealed that there were no significant morphological changes, such as charring, melting, or fusion, in any group, although the specimens were found to be more irregular in the Er:YAG laser group.Conclusions: the application of Er:YAG and diode lasers at the utilized parameters did not induce high pulpal temperatures. Root surface irregularities were more pronounced after irradiation with an Er:YAG laser than with a diode laser.

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.

Single crowns versus conventional fillings for the restoration of root filled teeth

BackgroundEndodontic treatment, involves removal of the dental pulp and its replacement by a root canal filling. Restoration of root filled teeth can be challenging due to structural differences between vital and non-vital root filled teeth. Direct restoration involves placement of a restorative material e.g. amalgam or composite directly into the tooth. Indirect restorations consist of cast metal or ceramic (porcelain) crowns. The choice of restoration depends on the amount of remaining tooth which may influence long term survival and cost. The comparative in service clinical performance of crowns or conventional fillings used to restore root filled teeth is unclear.ObjectivesTo assess the effects of restoration of endodontically treated teeth (with or without post and core) by crowns versus conventional filling materials.Search methodsWe searched the following databases: the Cochrane Oral Health Group's Trials Register, CENTRAL, MEDLINE via OVID, EMBASE via OVID, CINAHL via EBSCO, LILACS via BIREME and the reference lists of articles as well as ongoing trials registries. There were no restrictions regarding language or date of publication. Date of last search was 13 February 2012.Selection criteriaRandomised controlled trials (RCTs) or quasi-randomised controlled trials in participants with permanent teeth which have undergone endodontic treatment. Single full coverage crowns compared with any type of filling materials for direct restoration, as well as indirect partial restorations (e.g. inlays and onlays). Comparisons considered the type of post and core used (cast or prefabricated post), if any.Data collection and analysisTwo review authors independently assessed trial quality and extracted data.Main resultsOne trial judged to be at high risk of bias due to missing outcome data, was included. 117 participants with a root filled premolar tooth restored with a carbon fibre post, were randomised to either a full coverage metal-ceramic crown or direct adhesive composite restoration. At 3 years there was no reported difference between the non-catastrophic failure rates in both groups. Decementation of the post and marginal gap formation occurred in a small number of teeth.Authors' conclusionsThere is insufficient evidence to support or refute the effectiveness of conventional fillings over crowns for the restoration of root filled teeth. Until more evidence becomes available clinicians should continue to base decisions on how to restore root filled teeth on their own clinical experience, whilst taking into consideration the individual circumstances and preferences of their patients.

Influence of prior 810-nm-diode intracanal laser irradiation on hydrophilic resin-based sealer obturation

Dentin wall structural changes caused by 810-nm-diode laser irradiation can influence the sealing ability of endodontic sealers. The objective of this study was to evaluate the apical leakage of AH Plus and RealSeal resin-based sealers with and without prior diode laser irradiation. Fifty-two single-rooted mandibular premolars were prepared and divided into 4 groups, according to the endodontic sealer used and the use or non-use of laser irradiation. The protocol for laser irradiation was 2.5W, continuous wave in scanning mode, with 4 exposures per tooth. After sample preparation, apical leakage of 50% ammoniacal silver nitrate impregnation was analyzed. When the teeth were not exposed to irradiation, the Real Seal sealer achieved the highest scores, showing the least leakage, with significant differences at the 5% level (Kruskal-Wallis test, p = 0.0004), compared with AH Plus. When the teeth were exposed to the 810-nm-diode laser irradiation, the sealing ability of AH Plus sealer was improved (p = 0282). In the Real Seal groups, the intracanal laser irradiation did not interfere with the leakage index, showing similar results in the GRS and GRSd groups (p = 0.1009).

Pulpal temperature increase with high-speed handpiece, ER : YAG laser and ultrasound tips

The aim of this study was to compare intrapulpal temperature increase produced by high-speed handpiece, Er:YAG laser and CVDentus ultrasound tips during cavity preparation. Thirty bovine mandibular incisors with an enamel/dentin thickness of 4 mm at buccal surface had their roots amputated and were allocated to the following groups (n=10): Group I- high-speed handpiece; Group II- noncontact Er:YAG laser (250 mJ/4Hz); and Group III- CVDentus ultrasouns tips. All devices were used with water cooling. Class V cavities were prepared to a depth of 3.5 mm, measured with a periodontal probe. A type T thermocouple was placed inside the pulp chamber to determine the temperature increase (degrees C), which was recorded by a data acquisition system ADS 2000 IP (Lynx Technology) linked to a notebook computer. Data were analyzed statistically by oneway ANOVA and Tukey's test (p=0.05). The mean temperature rises were: 1.10 degrees C ( 0.56) for Group 1, 0.84 degrees C (0.55) for Group II, and 3.00 degrees C (1.34) for Group III. There were no statistically significant differences (p > 0.05) between Groups I and II, but both of them differed significantly from Group III (p < 0.05). In conclusion, the use of Er:YAG laser and high-speed handpiece for cavity preparation resulted in similar temperature increase. Although ultrasound tips generated significantly higher intrapulpal temperature increase, it remained below the critical value of 5.5 degrees C and may be considered safe for use.

Estudo morfológico da polpa de molares de ratos Wistar frente a uma oclusão traumática experimental

As alterações morfológicas pulpares, decorrentes de um estímulo externo experimental (interferência oclusal), foram estudadas, em nível de microscopia de luz. Utilizaram-se restaurações de amálgama, em sobreoclusão nos primeiros molares superiores direitos de dez ratos Wistar, divididos em 3 grupos e sacrificados por perfusão transcardíaca com formol a 10%, aos 10, 20 e 30 dias. A avaliação foi feita nos molares inferiores direitos (lado experimental) e esquerdos (lado controle). As peças ósseas após descalcificação em solução de EDTA associada às microondas, seguiram técnica histológica de rotina e coloração por hematoxilina-eosina e tricrômico de Mallory. Verificou-se no lado controle uma reação intensa caracterizada por um posicionamento atípico dos odontoblastos, seguida pelo aparecimento de cálculos pulpares e posteriormente por uma aparente e uniforme acomodação tecidual em toda a polpa, com moderada incidência de fibras colágenas. No lado experimental, as alterações foram similares parecendo, porém, aumentar com o tempo, principalmente aos 30 dias, onde a imagem histológica era semelhante à do lado controle aos 10 dias. Os resultados obtidos permitiram concluir que a interferência oclusal provocou alterações no tecido conjuntivo pulpar tanto no lado experimental como no controle e que as mesmas foram proporcionais à direção dos movimentos mandibulares.

Prevalence of different types of accessory canals in the furcation area of third molars

Background: Although there are several studies that show the prevalence and diameter of accessory root canals in the furcation area, there is a scarceness of studies that observe the trajectory and different types of cavo-interradicular canals. The aim of this study was to verify the prevalence of the different morphologic types of accessory canals in the furcation region in an attempt to show their trajectories.Methods: Forty submerged mandibular third molars were used, which were extracted and decalcified so that the microtomy procedure in the mesio-distal axial plane could be performed, obtaining semiserial sections with thicknesses of 5 mu m. The sections were stained with hematoxylin and eosin and observed under optical microscopy.Results: All of the morphologic types were found, whereas the proper accessory canals, type-A canals, were present in 10% of the specimens. The most prevalent canals were the sealed ones (type D), with a prevalence of 87.5%, followed by the blind ones (type B), with a prevalence of 75%. The loop accessory canals (type C), observed in only 5% of the teeth, were the least prevalent ones.Conclusions: Different morphologic types of accessory canals were found in the furcation area of submerged mandibular molars. The histologic method was effective to show the canal trajectories.

Prevalence of taurodontism in premolars

Taurodontism does not involve molar teeth exclusively. To call attention to the occurrence of this phenomenon in premolars, 4,459 extracted mandibular and maxillary premolars were examined. Eleven showed evidence of taurodontism. Roentgenograms taken from a buccolingual direction confirmed the diagnosis of taurodont teeth.

Radiographic and microbiologic evaluation of posttreatment apical and periapical repair of root canals of dogs' teeth with experimentally induced chronic lesion

The objective of the present study was to evaluate radiographically and bacteriologically apical and periapical repair in dogs' teeth with induced chronic periapical lesions with the use of two different operative techniques (techniques 1 and 2). The study was conducted on 40 root canals of upper and lower premolars from two dogs aged approximately 12 months. Periapical lesions were induced by leaving the root canals exposed to the oral environment for 5 days and then sealing them with zinc oxide-eugenol for 45 days. After this period, radiographic examination revealed the occurrence of a radiolucent lesion and endodontic treatment was started. The two techniques did not differ in terms of chemomechanical preparation, final filling, or type of cement, but differed in terms of irrigating solution and the presence of an antibacterial dressing. Thus 4% to 6% hypochlorite and hydrogen peroxide (10 volumes) were used in technique 1 during chemomechanical preparation and an antibacterial dressing based on calcium hydroxide was applied between sessions, whereas Dakin's fluid (0.5% sodium hypochlorite solution) and a final filling with no antibacterial dressing were used in technique 2. After chemomechanical preparation, the root canals were filled with gutta-percha cones and Sealapex (Sealapex-Sybron, Kerr, Sao Paulo, Brazil), and the animals were killed 270 days after the final filling. Blocks were cut into 6-μm sections and stained by the Brown and Brenn method. Radiographic, histomicrobiologic and statistical analysis permitted us to conclude the following: radiographically there was a marked reduction or even the disappearance of the radiolucent area present before treatment with greater success in the group treated with technique 1 (group I) than in the group treated with technique 2 (group II); the extent of bacterial invasion of dentinal tubules was greater and more intense in group II than in group I; and the amount of microorganisms detected in the ramifications of the apical delta and in the lumen of the root canal was intense in group II and mild or absent in group I. © 1994.

Pulpal and periodontal reactions of immature permanent teeth in the dog to intrusive trauma.

Pulpal and periodontal tissues of immature incisors of 10 dogs were radiographically and histopathologically evaluated immediately, 7, 15, 30 and 60 days after experimental intrusion induced by mechanical blows. Forty upper central and mesial lateral incisors showing incomplete root formation on radiographs were submitted to intrusive force. After the observation periods, the dogs were killed, two at a time. The hemi-maxilas were removed and processed for histopathologic examination. The traumatized teeth showed accelerated apical formation with reduced radicular length. Pulpal vitality was maintained and the subjacent tissues did not present irreversible changes. All traumatized teeth re-erupted spontaneously.

Pulpal lesions in normal and cyclosporin a treated rats

The objective of this study was to examine the development of pulpal lesions in the lower molar of control and cyclosporin A (CyA) treated rats. The pulps of the first lower molars of 20 normal and 20 CyA treated rats were exposed and left open into the oral cavity. Five animals of each group were killed at 7, 14, 21, and 28 days after the pulp exposure. The specimens were sectioned sagittally at a thickness of 7 μm and stained with hematoxylin and eosin. The pulpal lesions were similar for both normal and CyA treated rats in all studied periods and the differences between both groups were not statistically significant by the Student t test at the 5% (0.05) level of significance, indicating that the immunosuppression did not alter the evolution of the inflammatory process. Copyright © 1997 by The American Association of Endodontists.