transforming-growth-factor-beta and Dental-Caries

Trans-dentinal stimulation of tertiary dentinogenesis has long been recognized, and has traditionally been ascribed to diffusion of irritant substances arising during injury and restorative treatment. Identification of bio-active components, especially growth factors including TGF-beta s, sequestered within dentin matrix provides a new explanation for cellular signaling during tertiary dentinogenesis. Both isolated dentin matrix components and pure growth factors (TGF-beta s) have been shown to signal cellular events leading to reactionary and reparative tertiary dentinogenesis. Release of these bio-active components from dentin matrix may arise during carious attack and other injury to the tissue, and also during subsequent surgical intervention and restoration of the tooth. Both cavity-conditioning agents and leaching from restorative materials may contribute to release of these components. Distance of diffusion, as determined by cavity residual dentin thickness, and other restorative parameters may influence the signaling process after release of these components. Careful consideration of the interplay between tissue injury and surgical and restorative material factors is required for optimum exploitation of the exquisite regenerative capacity of dentin-pulp for more biological approaches to clinical treatment of dental disease.

Topics: Animals; Dental Caries; Dental Cavity Lining; Dental Materials; Dental Pulp; Dental Restoration, Permanent; Dentin; Dentin, Secondary; Dentinogenesis; Diffusion; Growth Substances; Humans; Irritants; Odontoblasts; Regeneration; Signal Transduction; Transforming Growth Factor beta

The purposes of this study were to evaluate a model of slow caries progression and to investigate the performance of a self-etch adhesive system for partial caries removal.. Rat molars were infected with Streptococcus sobrinus 6715 culture. Different time points were analyzed: days 78, 85, and 95 (± 2). After this, the samples were processed for morphological analysis. Additionally, the first molars were restored with zinc oxide and eugenol (IRM™; Dentsply; Brazil) or adhesive system (Clearfil SE Bond™; Kuraray Medical; Japan) 78 days after caries induction. After, 3 or 15 days post-treatment, the animals were euthanized, and their mandibles were processed for morphological analysis, classified by means of scores, and submitted to statistical analysis. Subsequently, immunohistochemical analysis was performed for osteonectin (OSN) and transforming growth factor-ß1 (TGF-ß1) expression.. According to the caries induction model used, on day 95 greater inflammatory infiltration (p < 0.001), and more extensive degradation of secondary/primary dentin were demonstrated than on day 78 (p < 0.05). Furthermore, the restorative materials presented similar performance (p > 0.05) and proved to be fundamental to control the carious lesion. The TGF-ß1 and OSN were shown to be active during the caries process.. The slow caries lesion model was feasible for morphological analysis of the dentin-pulp complex. The self-etch adhesive system triggered no acute inflammatory infiltration or pulp necrosis, instead it seemed to stimulate early pulp repair.. Clearfil SE Bond™ applied directly on caries-affected dentin did not predispose to pulp inflammation; instead, it appeared to provide early biological benefits.

Topics: Acid Etching, Dental; Animals; Dental Caries; Dental Cements; Disease Models, Animal; Disease Progression; Immunohistochemistry; Male; Mandible; Molar; Osteonectin; Rats; Rats, Wistar; Resin Cements; Streptococcus sobrinus; Surface Properties; Transforming Growth Factor beta; Zinc Oxide-Eugenol Cement

Topics: Dental Caries; Humans; Laser Therapy; Stem Cell Transplantation; Transforming Growth Factor beta

To investigate whether dental pulp fibroblasts express neuropeptide Y (NPY) and NPY-Y1 in vitro and to determine the effects of the cytokines including interleukin-1β (IL-1β), TGF- β(1) , substance P and NPY on the expression of NPY Y1.. Three primary fibroblast cell strains were obtained from freshly extracted human third molar teeth. RT-PCR was utilized to detect expression of NPY and mRNA expression. Membrane protein samples were isolated, and protein expression was determined by Western blotting. Radioimmunoassay was used to quantify NPY expression in healthy (n = 35) and carious (n = 39) whole pulp samples, and the student's t-test was used to test for statistical significance. In addition, the 3-(4,5-Dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to assay fibroblast cell growth.. mRNA transcripts were found in all three fibroblast cell populations with the cytokines having a stimulatory effect on its expression (P < 0.05). NPY mRNA was not detected in the cell strains. NPY-Y1 receptor protein expression was visualized by Western blotting, and there was no effect of IL-1β or TGF- β(1) on its expression. The mean concentration of NPY-Ir determined by radioimmunoassay in non-carious teeth was 19.40 ng x g(-1) (±17.03 SD) compared to 29.95 ng x g(-1) (±20.99 SD) in carious teeth (P < 0.05).. Human dental pulp fibroblasts express, but do not synthesize, NPY, demonstrating that the fibroblast is a target cell for NPY. The effect of proinflammatory cytokines suggests that fibroblasts play a neuroimmunomodulatory role in the pulpal response to dental caries and injury.

Topics: Blotting, Western; Cell Culture Techniques; Cell Proliferation; Cells, Cultured; Coloring Agents; Dental Caries; Dental Pulp; Fibroblasts; Humans; Interleukin-1beta; Membrane Proteins; Neuroimmunomodulation; Neuropeptide Y; Receptors, Neuropeptide Y; Reverse Transcriptase Polymerase Chain Reaction; Substance P; Tetrazolium Salts; Thiazoles; Transforming Growth Factor beta

Transforming growth factor (TGF)-beta s in dentin matrix provide a pool of bioactive molecules, but association with latency-associated peptides (LAPs) may influence their activity. We investigated TGF-beta 1, -beta 2, and -beta 3 LAP expression in sound and carious human teeth. Teeth were fixed and processed immediately following extraction prior to staining with rabbit polyclonal antibodies to the TGF-beta LAPs. A soluble dentin matrix fraction was prepared from dissected human dentin and sequential extraction of pulpal ECM was performed prior to purification. Fractions were Western blotted and probed with the LAP antibodies. All three LAPs were present in odontoblasts, cells of the pulp, and predentin; however, no staining of mineralized dentin matrix was seen. Similar patterns of expression were seen in carious tissue. Expression of TGF-beta LAPs in cells and pulpal matrix of healthy and carious teeth will be important in regulation of TGF-beta activity and may modulate the tissue response to injury.

Topics: Dental Caries; Dental Pulp; Dentin; Extracellular Matrix; Humans; Immunohistochemistry; Molar; Peptide Fragments; Peptides; Protein Precursors; Reference Values; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta3

Members of the TGF-beta family of growth factors are important in modulation of odontoblast secretory activity during dental tissue repair. Odontoblast expression of TGF-beta isoforms during development leads to their sequestration within the dentin matrix, from where they may be released during carious injury and participate in reparative processes. Two receptors, implicated in TGF-beta-mediated cell signaling, have been identified immunohistochemically in both odontoblasts and pulpal cells of healthy and carious human molar teeth. This study aimed to characterize the expression of the TGF-beta receptors I and II in sound and carious teeth by means of in situ hybridization, to help our understanding of the response of these cells to TGF-beta stimulation. Sound and carious human third molar teeth were routinely processed immediately following extraction, and 10-microns paraffin-embedded sections prepared. These sections were hybridized with 32P-labeled probes to TGF-beta receptors I and II, and the subsequent signal was detected by autoradiography. mRNA for both receptors I and II was mainly detected within the odontoblasts and nerve-associated cells of healthy tissues, with expression at lower levels seen within the subodontoblast and pulp core cells. The expression in odontoblasts was higher for TGF-beta receptor I than for receptor II. Expression of both receptors was more homogenous in all pulp cells within carious teeth, because of an increase of signal within the underlying pulp cell population, including blood-vessel-associated cells. We conclude that the TGF-beta receptors I and II were expressed in odontoblasts and pulp cells, and that subtle variations in the levels of their expression could be involved in the tissue response to injury.

Topics: Activin Receptors, Type I; Autoradiography; Dental Caries; Dental Pulp; Dentin; Gene Expression; Humans; In Situ Hybridization; Molar, Third; Odontoblasts; Phosphorus Radioisotopes; Protein Isoforms; Protein Serine-Threonine Kinases; Radiopharmaceuticals; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; RNA, Messenger; Signal Transduction; Transforming Growth Factor beta; Wound Healing

Several growth factors and extracellular matrix molecules, which are expressed during embryonic tooth development, are re-expressed in dental tissues under pathological conditions. Pathological conditions such as caries lesions and dental injuries are often lethal to the odontoblasts, which are then replaced by other pulp cells. These cells are able to differentiate into odontoblast-like cells and produce a reparative dentin. Here we demonstrate the in vivo distribution of several molecules in human permanent teeth under normal and pathological conditions. The intermediate filament protein nestin, which is a marker of young odontoblasts, is absent from old permanent teeth. Similarly, the Notch protein, which is involved in cell fate specification and is localized in the sub-odontoblastic cell layer during odontogenesis, is not detected in adult dental tissues. In carious and injured teeth, nestin is expressed in a selective manner in odontoblasts surrounding the injury site, while Notch is expressed in the sub-odontoblastic layer of cells. We reproduced this physiological event in an in vitro culture system. Pulp cells cultured in the presence of beta-glycerophosphate formed mineralization nodules. As odontoblasts, pulp cells contributing to the nodule formation express type I collagen, osteonectin, dentin sialophosphoprotein, and nestin. In this in vitro assay system, nestin is up-regulated after local application of Bone Morphogenetic Protein 2 and 4. Fourier transform infrared microspectroscopy showed that both the organic and the mineral compositions of the nodules have the characteristics of human dentin and differ from those of enamel and bone. These findings show that both the molecular and the mineral characteristics of the human dentin matrix are respected in the in vitro culture conditions.

Topics: Adult; Bone Morphogenetic Protein 2; Bone Morphogenetic Protein 4; Bone Morphogenetic Proteins; Cell Differentiation; Collagen Type I; Culture Techniques; Dental Caries; Dental Pulp; Dentin; Dentin, Secondary; Dentinogenesis; Extracellular Matrix Proteins; Fetus; Humans; Intermediate Filament Proteins; Membrane Proteins; Nerve Tissue Proteins; Nestin; Odontoblasts; Odontogenesis; Osteonectin; Phosphoproteins; Protein Precursors; Receptors, Notch; Sialoglycoproteins; Spectroscopy, Fourier Transform Infrared; Tooth Calcification; Tooth Diseases; Tooth Injuries; Trans-Activators; Transforming Growth Factor beta; Up-Regulation