Efeito citotóxico de agentes clareadores a base de peróxido de hidrogênio a 20% e 38% sobre células odontoblastóides

The aim of this in vitro study was to evaluate the trans-enamel and transdentinal cytotoxic effects of two in-office tooth bleaching techniques that employ bleaching gels containing 20% and 38% of H2 O2 on cultured odontoblast-like cell line (MDPC-23). Sixty enamel/dentin discs were obtained from bovine central incisors and placed individually in artificial pulp chambers. Six groups were formed according to the following enamel treatments: G1- 20% H2 O2 (1 application); G2- 20% H2 O2 (2 applications); G3- 38% H2 O2 (1 application); G4- 38% H2 O2 (2 applications); G5- 38% H2 O2 (3 applications); and G6- control (no treatment). In G1 and G2, the bleaching gel was left in contact with the enamel surface for 45 min in each application. However, in G3, G4, and G5 the bleaching gel was applied for only 10 min per application. After the last application, the extracts were collected and applied on previously cultured cells (30.000 cells/cm2 ) for 24 h. Cell metabolism was evaluated by the MTT assay and cell morphology was analysed by scanning electron microscopy. Cell metabolism decreased by 96.29%; 96.11%; 96.42%; 95.62%; and 97.18% in G1, G2, G3, G4, and G5, respectively. All treated groups differed significantly from non-treated control group (G6) (p < 0.05). However, the difference in cell metabolism among treated groups was not significant statistically. In addition, significant morphological cell alterations were observed in all treated groups. Under the tested experimental conditions, the extracts collected after both tooth bleaching techniques evaluated in this study caused severe toxic effects on cultured odontoblast-like cell MDPC-23.

Citotoxicidade transamelodentinária de uma fita clareadora com 10% de peróxido de hidrogênio sobre células de linhagem odontoblástica

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

Dose-Response and Time-Course of a-Tocoferol Mediating the Cytoprotection Of Dental Pulp Cells Against Hydrogen Peroxide

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Responses of Dental Pulp Cells to a Less Invasive Bleaching Technique Applied to Adhesively Restored Teeth

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

Transdentinal cytotoxicity of glutaraldehyde on odontoblast-like cells

This study investigated the transdentinal cytotoxicity of glutahaldehyde-containing solutions/materials on odontoblast-like cells. Dentin discs were adapted to artificial pulp chambers. MDPC-23 cells were seeded on the pulpal side of the discs and the occlusal surface was treated with the following solutions: water, 2% glutaraldehyde (GA), 5% GA, 10% GA, Gluma Comfort Bond+Desensitizer (GCB+De) or Gluma Desensitizer (GDe). Cell viability and morphology were assessed by the Alamar Blue assay and SEM. The eluates were collected and applied on cells seeded in 24-well plates. After 7 or 14 days the total protein (TP) production, alkaline phosphatase activity (ALP) and deposition of mineralized nodules (MN) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). GA solutions were not cytotoxic against MDPC-23. GCB+De (85.1%) and GDe (77.2%) reduced cell viability as well as TP production and ALP activity at both periods. After 14 days, GCB+De and GDe groups produced less MN. Affected MDPC-23 presented deformation of the cytoskeleton and reduction of cellular projections. The treatment with 2.5%, 5% and 10% GA was not harmful to odontoblast-like cells. Conversely, when GA was combined with other components like HEMA, the final material became cytotoxic. Glutaraldehyde has been used to decrease dentin hypersensitivity. This substance is also capable of preventing resin-dentin bond degradation by cross-linking collagen and MMPs. This study showed that GA might be safe when applied on acid etched dentin. However, when combined with HEMA the product becomes cytotoxic.

Responses of human dental pulp cells after application of a low-concentration bleaching gel to enamel

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

Evaluation of plasmatic MMP-8, MMP-9, TIMP-1 and MPO levels in obese and lean women

Objectives: To compare the plasma concentrations of matrix metalloproteinase (MMP)-9, tissue inhibitor of MMP (TIMP)-1, MMP-8, and myeloperoxidase (MPO) for obese and lean women. Design and methods: We recruited 30 lean and 36 obese women without comorbidities. The MMP-9, TIMP-1, and MMP-8 levels were measured using enzyme-linked immunosorbent assay (ELISA). MPO activity was assessed by a colorimetric assay. Results: Obese women had higher MMP-9 levels and MMP-9:TIMP-1 ratios than lean women. Conversely, the MMP-8 levels and MMP-8:TIMP-1 ratios in the obese women were significantly lower than those in the lean women despite neutrophil activation, which was assessed by MPO activity., Conclusion: We observed that MMP-9 and MMP-8 had distinct profiles, which suggested that these 2 enzymes play different roles in obesity. (C) 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Immunolocalization of podoplanin in benign odontogenic tumours with and without ectomesenchyme

Objectives: To investigate podoplanin expression in epithelial odontogenic tumours with and without ectomesenchyme and verify the association between its immunoexpression and proliferative activity in keratocystic odontogenic tumours (KCOTS) and orthokeratinized odontogenic cysts (OOCs). Design: Eight ameloblastomas, nine adenomatoid odontogenic tumours, twenty KCOTS, five OOC, one calcifying epithelial odontogenic tumour, two ameloblastic fibromas, four ameloblastic fibro-odontomas and five calcifying cystic odontogenic tumours were immunohistochemically analysed with anti-podoplanin antibody. For KCOTS and OOC, the cell proliferation index was determined with Ki-67 immunostaining and compared by Spearman correlation coefficient. Results: Podoplanin was expressed in the peripheral odontogenic epithelium of most tumours. Ectomesenchyme was negative, except for odontoblasts. KCOTS exhibited positive podoplanin expression while in OOC it was absent/weak. There was statistically significant correlation ( p = 0.006) between podoplanin expression and cellular proliferation index of KCOTS and OOC. Conclusion: Podoplanin seems to be related to the proliferative activity of KCOTS and may have a role in the process of local invasion of odontogenic tumours with and without ectomesenchyme

Identification of a fibronectin interaction site in the extracellular matrix protein ameloblastin

Mammalian teeth are composed of hydroxyapatite crystals that are embedded in a rich extracellular matrix. This matrix is produced by only two cell types, the mesenchymal odontoblasts and the ectodermal ameloblasts. Ameloblasts secrete the enamel proteins amelogenin, ameloblastin, enamelin and amelotin. Odontoblasts secrete collagen type I and several calcium-binding phosphoproteins including dentin sialophosphoprotein, dentin matrix protein, bone sialoprotein and osteopontin. The latter four proteins have recently been grouped in the family of the SIBLINGs (small integrin-binding ligand, N-linked glycoproteins) because they display similar gene structures and because they contain an RGD tripeptide sequence that binds to integrin receptors and thus mediates cell adhesion. We have prepared all the other tooth-specific proteins in recombinant form and examined whether they might also promote cell adhesion similar to the SIBLINGs. We found that only ameloblastin consistently mediated adhesion of osteoblastic and fibroblastic cells to plastic or titanium surfaces. The activity was dependent on the intact three-dimensional structure of ameloblastin and required de novo protein synthesis of the adhering cells. By deletion analysis and in vitro mutagenesis, the active site could be narrowed down to a sequence of 13 amino acid residues (VPIMDFADPQFPT) derived from exon 7 of the rat ameloblastin gene or exons 7-9 of the human gene. Kinetic studies and RNA interference experiments further demonstrated that this sequence does not directly bind to a cell surface receptor but that it interacts with cellular fibronectin, which in turn binds to integrin receptors. The identification of a fibronectin-binding domain in ameloblastin might permit interesting applications for dental implantology. Implants could be coated with peptides containing the active sequence, which in turn would recruit fibronectin from the patient's blood. The recruited fibronectin should then promote cell adhesion on the implant surface, thereby accelerating osseointegration of the implant.

Incremental lines of von Ebner in dinosaurs and the assessment of tooth replacement rates using growth line counts

Dinosaur dentine exhibits growth lines that are tens of micrometers in width. These laminations are homologous to incremental lines of von Ebner found in extant mammal and crocodilian teeth (i.e., those of amniotes). The lines likely reflect daily dentine formation, and they were used to infer tooth development and replacement rates. In general, dinosaur tooth formation rates negatively correlated with tooth size. Theropod tooth replacement rates negatively correlated with tooth size, which was due to limitations in the dentine formation rates of their odontoblasts. Derived ceratopsian and hadrosaurian dinosaurs retained relatively rapid tooth replacement rates through ontogeny. The evolution of dental batteries in hadrosaurs and ceratopsians can be explained by dentine formation constraints and rapid tooth wear. In combination with counts of shed dinosaur teeth, tooth replacement rate data can be used to assess population demographics of Mesozoic ecosystems. Finally, it is of historic importance to note that Richard Owen appears to have been the first to observe incremental lines of von Ebner in dinosaurs more than 150 years ago.

Differentiation of embryonic mesenchymal cells to odontoblast-like cells by overexpression of dentin matrix protein 1

Cells of the craniofacial skeleton are derived from a common mesenchymal progenitor. The regulatory factors that control their differentiation into various cell lineages are unknown. To investigate the biological function of dentin matrix protein 1 (DMP1), an extracellular matrix gene involved in calcified tissue formation, stable transgenic cell lines and adenovirally infected cells overexpressing DMP1 were generated. The findings in this paper demonstrate that overexpression of DMP1 in pluripotent and mesenchyme-derived cells such as C3H10T1/2, MC3T3-E1, and RPC-C2A can induce these cells to differentiate and form functional odontoblast-like cells. Functional differentiation of odontoblasts requires unique sets of genes being turned on and off in a growth- and differentiation-specific manner. The genes studied include transcription factors like core binding factor 1 (Cbfa1), bone morphogenetic protein 2 (BMP2), and BMP4; early markers for extracellular matrix deposition like alkaline phosphatase (ALP), osteopontin, osteonectin, and osteocalcin; and late markers like DMP2 and dentin sialoprotein (DSP) that are expressed by terminally differentiated odontoblasts and are responsible for the formation of tissue-specific dentin matrix. However, this differentiation pathway was limited to mesenchyme-derived cells only. Other cell lines tested by the adenoviral expression system failed to express odontoblast-phenotypic specific genes. An in vitro mineralized nodule formation assay demonstrated that overexpressed cells could differentiate and form a mineralized matrix. Furthermore, we also demonstrate that phosphorylation of Cbfa1 (osteoblast-specific transcription factor) was not required for the expression of odontoblast-specific genes, indicating the involvement of other unidentified odontoblast-specific transcription factors or coactivators. Cell lines that differentiate into odontoblast-like cells are useful tools for studying the mechanism involved in the terminal differentiation process of these postmitotic cells.