Diferenciação in vitro de células-tronco mesenquimais da medula óssea de cães em precursores osteogênicos

O objetivo principal da nossa pesquisa foi avaliar o potencial de diferenciação osteogênica de células-tronco mesenquimais (MSC) obtidas da medula óssea do cão. As MSC foram separadas pelo método Ficoll e cultivadas sob duas condições distintas: DMEM baixa glicose ou DMEM/F12, ambos contendo L-glutamina, 20% de SFB e antibióticos. Marcadores de MSC foram testados, confirmando células CD44+ e CD34- através da citometria de fluxo. Para a diferenciação osteogênica, as células foram submetidas a quatro diferentes condições: Grupo 1, as mesmas condições utilizadas para a cultura de células primárias com os meios DMEM baixa glicose suplementado; Grupo 2, as mesmas condições do Grupo 1, mais os indutores de diferenciação dexametasona, ácido ascórbico e b-glicerolfosfato; Grupo 3, células cultivadas com meios DMEM/F12 suplementado; e Grupo 4, nas mesmas condições que no Grupo 3, mais indutores de diferenciação de dexametasona, ácido ascórbico e b-glicerolfosfato. A diferenciação celular foi confirmada através da coloração com alizarin red e da imunomarcação com o anticorpo SP7/Osterix. Nós observamos através da coloração com alizarin red que o depósito de cálcio foi mais evidente nas células cultivadas em DMEM/F12. Além disso, usando a imunomarcação com o anticorpo SP/7Osterix obtivemos positividade em 1:6 células para o Meio DMEM/F12 comparada com 1:12 para o meio DMEM-baixa glicose. Com base nos nossos resultados concluímos que o meio DMEM/F12 é mais eficiente para a indução da diferenciação de células-tronco mesenquimais caninas em promotores osteogênicos. Este efeito provavelmente ocorre em decorrência da maior quantidade de glicose neste meio, bem como da presença de diversos aminoácidos.

Concentração de imunoglobulinas G e M no soro sanguíneo de bezerros da raça Holandesa até os 90 dias de idade

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

Influence of low-level laser therapy on wound healing in nicotine-treated animals

Low-level laser therapy (LLLT) has been shown to have several biological effects that favor the healing process, and nicotine has been shown to delay the healing process. In this study we investigated the healing of open wounds created on the back of rats treated with nicotine with or without LLLT. of 115 animals, 59 received subcutaneous injections of saline solution, and the others received subcutaneous injections of nicotine (3 mg/kg body weight), twice a day throughout the study period. After 30 days, skin wounds were created on the back of the animals. The animals receiving saline injections were divided into two groups: group 1 (G1, n = 29), in which the wounds were left untreated, and group 2 (G2, n = 30), in which the wounds were treated with LLLT (GaAlAs, 660 nm, 30 mW, 5.57 J/cm(2) per point, 0.39 J, 13 s per point, 0.42 W/cm(2)). The animals receiving nicotine injections were also divided into two groups: group 3 (G3, n = 29), in which the wounds were left untreated, and group 4 (G4, n = 27), in which the wounds were treated with LLLT. The animals were killed 3, 7 or 14 days after surgery. Wound healing was evaluated histologically both qualitatively and semiquantitatively. Wounds of G2 showed a delay in epithelial migration and connective tissue organization compared to those of G1. Wounds of G2 showed faster healing than those of G1; similarly, wounds of G4 showed more advanced healing than those of G3. LLLT acted as a biostimulatory coadjuvant agent balancing the undesirable effects of nicotine on wound tissue healing.

Evaluation of the Effect of Retightening and Mechanical Cycling on Preload Maintenance of Retention Screws

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

Low-Fluoride Acidic Dentifrice: A Randomized Clinical Trial in a Fluoridated Area

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

The Effect of Different Fluoride Concentrations and pH of Dentifrices on Plaque and Nail Fluoride Levels in Young Children

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

Influence of the type of vehicle and limit of obturation on apical and periapical tissue response in dogs' teeth after root canal filling with mineral trioxide aggregate

The purpose of this study was to investigate the influence of the type of vehicle (distilled water or propyleneglycol) on the response of apical tissues of dogs' teeth after root canal filling with mineral trioxide aggregate (MTA) at two different limits. Forty roots of incisors and premolars of two adult dogs were used. After pulpectomy, the root canals were prepared biomechanically, and the roots had the apical cemental barrier penetrated with a #15 K-file and widened to a #25 K-file. The root canals were assigned to four groups according to the vehicle used for MTA (ProRoot-MTA; Tulsa Dental, Tulsa, OK) preparation and the limit of root canal filling: group 1, filling with MTA/distilled water to the limit of the cemental canal; group 2, overfilling with MTA/distilled water, group 3, filling with MTA/propyleneglycol to the limit of the cemental canal; and group 4, overfilling with MTAlpropyleneglycol. The animals were killed by anesthetic overdose 90 days after endodontic treatment and the anatomic pieces were prepared for histomorphological analysis. The sections were stained with hematoxylin and eosin and Brown and Brenn techniques. The results showed that MTA pastes prepared with either distilled water or propyleneglycol as vehicles had similar biological behavior (p > 0.05); root fillings placed at the cemental canal limit showed better results than the overfillings (p = 0.01), and MTA/propyleneglycol paste was more easily placed into the root canals than MTA/distilled water paste.

Antimicrobial potential of medicaments used in healing process in dogs' teeth with apical periodontitis

Estudou-se a influência de diferentes irrigantes no potencial antimicrobiano da pasta de hidróxido de cálcio em dentes de cães com periodontite apical. 48 pré-molares de cães adultos tiveram suas câmaras coronárias abertas e expostas à cavidade bucal por 6 meses. Os canais radiculares foram preparados, irrigados e medicados com diferentes substâncias, de acordo com os seguintes grupos: 1) 2,5% NaOCl + CHP; 2) 2% CHX + CHP; 3) vinagre + CHP; 4) vinagre + vinagre. No grupo 4, a solução irrigante e a medicação intracanal utilizada foi o vinagre. Neste grupo, a cada 7 dias, a solução era renovada. Cada amostra foi coletada, mantendo-se o cone de papel esterilizado em posição por 1 min, e a seguir transportado e imerso em 7 mL de Letheen broth, seguido de incubação a 37ºC por 48 h. O crescimento microbiano foi analisado por dois métodos, turbidade do meio de cultura e subcultura em meio nutritivo específico (brain heart infusion). Os resultados mostraram que em todos os grupos experimentais houve crescimento microbiano após 21 dias, em diferentes percentagens: grupo 1 - 30%; grupo 2 - 30%; grupo 3 - 40%; grupo 4 - 60%. Todos os materiais testados apresentaram potencial antimicrobiano. Entretanto, o processo de cura favorecido pela pasta de hidróxido de cálcio não pode ser esquecido, uma vez que muitos estudos já demonstraram sua ação antimicrobiana.

Influence of apical foramen widening and sealer on the healing of chronic periapical lesions induced in dogs' teeth

Objective. The aim of this study was to evaluate the influence of apical foramen widening on the healing of chronic periapical lesions in dogs' teeth after root canal filling with Sealer 26 or Endomethasone.Study design. Forty root canals of dogs' teeth were used. After pulp extirpation, the canals were exposed to the oral cavity for 180 days for induction of periapical lesions, and then instrumented up to a size 55 K-file at the apical cemental barrier. In 20 roots, the cemental canal was penetrated and widened up to a size 25 K-file; in the other 20 roots, the cemental canal was preserved (no apical foramen widening). All canals received a calcium hydroxide intracanal dressing for 21 days and were filled with gutta-percha and 1 of the 2 sealers: group 1: Sealer 26/apical foramen widening; group 2: Sealer 26/no apical foramen widening; group 3: Endomethasone/apical foramen widening; group 4: Endomethasone/no apical foramen widening. The animals were killed after 180 days, and serial histologic sections from the roots were prepared for histomorphologic analysis. Scores were assigned according to preestablished histomorphologic parameters and analyzed statistically by Kruskal-Wallis and Mann-Whitney U tests.Results. Regarding new cementum formation, repair of cementum and bone resorption areas, presence of microorganisms, inflammatory cell infiltrate and periodontal ligament conditions, significantly better periapical healing was obtained when foramen widening was done and Sealer 26 was used.Conclusion. Apical foramen widening and calcium hydroxide-containing sealer were more favorable to the healing of chronic periapical lesions. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;109:932-940)

Histologic evaluation of the use of membrane, bone graft, and MTA in apical surgery

The aim of this study was to evaluate the periapical healing after the use of membrane, bone graft, and mineral trioxide aggregate (MTA) in apical surgery of dogs' teeth. Apical lesions were induced in 48 roots of 6 dogs after coronal access and pulpal removal. Apical surgery consisted of osteotomy with trephine bur for the standardization of the critical surgical cavities, followed by apicoectomy, curettage, preparation of the root-end cavities with the aid of the ultrasonic device, and retrofilling with MTA. The surgical sites were divided into: group 1-filled with blood; group 2-filled with blood and recovered with membrane; group 3-filled with bone graft; and group 4-filled with bone graft and recovered with membrane. The results showed that the inflammatory infiltrate, the periapical healing process, and the behavior of MTA was the same in all groups, including the mineralization stimulation. It was concluded that the use of membranes and bone graft materials isolated or associated in apical surgery did not alter the periapical healing process after the root-end filling with MTA. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109: 309-314)

In vivo evaluation of the biocompatibility of three current bonding agents

The aim of this in vivo study was to evaluate the biocompatibility of three current bonding agents and calcium hydroxide cement. Sixty polyethylene tubes filled with the following materials: Group 1: Prime & Bond NT (PB - Dentsply, US; Group 2: Bond 1 (BO - Jeneric/Pentron, US); Group 3: Optibond Solo (OP - Kerr, US); and Group 4 (control): calcium hydroxide cement - Dycal (CH - Dentsply, US) were implanted into the connective tissue of 30 rats. After 15, 30 and 60 days, the implants were excised and the animals sacrificed. The biopsies were immersed in Karnovsky (pH, 7.2) fixative solution for 48 hours, and processed using routine histological technique. Six-micron-thick sections were cut and stained with hematoxilin and eosin and Masson's trichome technique. Microscopic evaluation was used to compare the connective tissue reactions caused by the experimental and control materials adjacent to the tube opening. At 15 days, the experimental and control materials triggered a moderate to intense inflammatory response which gave rise to a thick capsule adjacent to the tube opening. With time, the inflammatory reaction decreased. At 60 days, the connective tissue adjacent to the bonding agents exhibited a persistent inflammatory response mediated by macrophages and giant cells which were engulfing displaced resin components. on the other hand, for the control group (calcium hydroxide) no inflammatory response associated with a thin capsule adjacent to the material was observed even at the 30-day period. The hard-setting calcium hydroxide cement allowed complete healing and was considered more biocompatible than the bonding agents.

In vitro cytotoxicity of five glass-ionomer cements

To evaluate the cytotoxic effects of five glass-ionomer cements (GICs) on an odontoblast cell line (MDPC-23), disks of every material were prepared and divided into Group 1: Vitrebond, Group 2: Vitremer, Group 3: Fuji IILC, Group 4: Fuji IX GP, Group 5: Ketac-Molar, Group 6: Z-100 (positive control). In Group 7, phosphate-buffered saline solution (negative control) was applied on filter paper. After placing the samples in the bottom of wells, the cells (30,000 cells/cm(2)) were plated and incubated for 72 h. The cell number was counted, the cell morphology was assessed by scanning electron microscopy and the cell metabolism was evaluated using methyltetrazolium assay. The statistical analysis of Kruskal-Wallis was used to determine if the scores obtained for the cell metabolism and number of cells were different at the 95% confidence level. In groups 1, 2, 3, 4, 5, and 6 the materials decreased the cell number by 74.5% 75.5%, 45.5%, 29.5%, 32.5%, and 88.5%, respectively. In groups 1, 2, 3, 4, and 5, the experimental GICs reduced the cell metabolism by 79%, 84%, 54%, 40%, and 42.5%, respectively. Despite the fact that all experimental materials were cytotoxic to the MDPC-23 cells, the GICs were the least cytotoxic. on the other hand, the RMGICs caused the highest cytophatic effects. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

A discrepância de tamanho dentário, de Bolton, na oclusão normal e nos diferentes tipos de más oclusões, bem como sua relação com a forma de arco e o posicionamento dentário

Esta pesquisa teve como objetivo geral avaliar a discrepância de tamanho dentário, na oclusão normal e nos diferentes tipos de más oclusões e a sua relação com as medidas que determinam a forma de arco e o posicionamento dentário na região anterior. Para tanto, foram estudados 185 pares de modelos de gesso, divididos em 4 grupos: Grupo 1 (composto por 41 pares com Oclusão Normal, sendo 20 do gênero masculino e 21 do gênero feminino); Grupo 2 (composto por 44 pares com má oclusão de Classe I, divisão 1, sendo 22 do gênero masculino e 22 do gênero feminino); Grupo 3 (composto por 54 pares com má oclusão de Classe II, sendo 28 do gênero masculino e 26 do gênero feminino) e Grupo 4 (composto por 46 pares com Classe III, sendo 23 do gênero masculino e 23 do gênero feminino). Observou-se que não ocorreu dimorfismo sexual entre as discrepâncias de tamanho dentário e os diferentes tipos de oclusão dentária; as proporções estabelecidas por Bolton não se aplicaram ao grupo com Oclusão Normal; na Oclusão Normal, Classe I, Classe II e Classe III, houve um predomínio de excesso dentário total (RAZ12) no arco inferior; na Classe I houve uma igualdade na distribuição de excesso dentário anterior (RAZ6) nos arcos superior e inferior; na Oclusão Normal, Classe II e Classe III, ocorreu um predomínio de excesso dentário anterior (RAZ6) no arco inferior, em relação ao arco superior; os excessos dentários não contribuíram na ocorrência das más oclusões e as discrepâncias total e anterior (RAZ12 e RAZ6) não interferiram diretamente nas larguras e comprimentos dos arcos, bem como no posicionamento dos dentes anteriores.

Utilization of autogenous bone, bioactive glasses, and calcium phosphate cement in surgical mandibular bone defects in Cebus apella monkeys

Purpose: the purpose of the present study was to evaluate the histologic results of bone cavities that were surgically created in the mandibles of Cebus apella monkeys and filled with autogenous bone, PerioGlas, FillerBone, or Bone Source. Materials and Methods: Surgical cavities 5 mm in diameter were prepared through both mandibular cortices in the mandibular angle region. The cavities were randomly filled, and the animals were divided into groups according to the material employed: Group 1 cavities were filled with autogenous corticocancellous bone; group 2 cavities were filled with calcium phosphate cement (BoneSource); and group 3 and group 4 cavities were filled with bioactive glass (FillerBone and PerioGlas, respectively). After 180 days the animals were sacrificed, and specimens were prepared following routine laboratory procedures for hematoxylin/eosin staining and histologic evaluation. Results: the histologic analysis showed that autogenous bone allowed total repair of the bone defects; bioactive glasses (FillerBone and PerioGlas) allowed total repair of the defects with intimate contact of the remaining granules and newly formed bone; and the cavities filled with calcium phosphate cement (BoneSource) were generally filled by connective fibrous tissue, and the material was almost totally resorbed. Discussion: the autogenous bone, FillerBone, and PerioGlas provided results similar to those in the current literature, showing that autogenous bone is the best Choice for filling critical-size defects. Synthetic implanted materials demonstrated biocompatibility, but the bioglasses demonstrated osteoconductive activity that did not occur with calcium phosphate (BoneSource). Conclusion: According to the methodology used in this study, it can be concluded that the utilization of autogenous bone and bioactive glasses permitted the repair of surgically created critical-size defects by newly formed bone; the synthetic implanted materials demonstrated biocompatibility, and the bioactive glasses demonstrated osteoconductive activity. The PerioGlas was mostly resorbed and replaced by bone and the remaining granules were in close contact with bone; the FillerBone showed many granules in contact with the newly formed bone; BoneSource did not permit repair of the critical-size defects, and the defects were generally filled by connective fibrous tissue.

Comparison of Biomaterial Implants in the Dental Socket: Histological Analysis in Dogs

Background:Bone graft procedures have been used commonly in buco-maxillo-facial surgery. For this reason, many researchers have evaluated the bone substitutes.Purpose:The present study evaluated soft and hard tissue reactions to two different hydroxyapatites HAs (synthetic HA and natural HA) and bioactive glass implanted into the sockets immediately after extraction.Materials and Methods:First and third upper and lower premolars, on both sides, were extracted from six female dogs. The alveolar sockets were randomly assigned to four groups: Group 1 - control (unfilled), Group 2 - filled with synthetic hydroxyapatite, Group 3 - filled with bovine bone mineral (natural HA), and Group 4 - filled with bioactive glass. The animals were euthanized at 4 weeks (n = 2), 8 weeks (n = 2), and 28 weeks (n = 2) after extraction. The mandible and maxilla of each animal were removed for histological analysis to determine soft tissue reactions, newly formed bone, bone characteristics, and presence or absence of implanted materials.Results:Most particles of synthetic hydroxyapatite had bone formation on their surface, although some particles showed a layer of fibrous connective tissue. The bovine bone mineral group exhibited particles partially replaced with bone formation. The bioactive glass group showed particles with a thin layer of calcified tissue, but was absent in some specimens, suggesting complete resorption.Conclusion:All biomaterials had similar behavior. Bovine bone mineral, compared to synthetic hydroxyapatite and bioactive glass, showed a larger number of particles covered with osseous tissue. All biomaterials interfered with the socket repair process.