transforming-growth-factor-beta and Pulpitis

To evaluate the Transforming Growth Factor-beta 1 (TGF-beta 1) expression in normal healthy pulps and in those with irreversible pulpitis.. Twenty-three normal, healthy pulps were removed from mandibular third molars, and 20 pulps were retrieved from teeth with irreversible pulpitis. TGF-beta 1 was evaluated in the odontoblastic and subodontoblastic layers, in the stromal cells (fibroblasts), and in the blood vessels. TGF-beta 1 expression was determined by evaluating 500 cells in the odontoblastic and subodontoblastic layers and 500 fibroblasts in the stroma for each specimen, and counting the number of positive cells. The number of the positive vessels was evaluated in 10 high power fields (HPF). In almost all cases, the cellular positivity was cytoplasmatic. Statistical analysis was performed using Mann-Whitney U- and Student's t-tests.. A higher expression of TGF-beta 1 was found in the odontoblastic-subodontoblastic layer of the irreversible pulpitis specimens; this difference was statistically significant (P = 0.0002). No statistically significant difference was observed between the two groups in TGF-beta 1 expression in the stromal cells (P = 0.54) or in the vascular component (P = 0.94).. The higher and statistically significant expression of TGF-beta 1 found in the odontoblastic-subodontoblastic layer of irreversible pulpitis specimens may indicate a role for TGF-beta 1 in the dentinal repair processes after pulp inflammation.

Topics: Adolescent; Adult; Blood Vessels; Dental Pulp; Humans; Odontoblasts; Pulpitis; Statistics, Nonparametric; Stromal Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1

Type IV matrix metalloproteinases (MMPs) are members of the family of MMPs and are thought to play an important role in degradation of extracellular components. Human pulp cells can secrete and produce these enzymes. Recent evidence shows that MMPs may play a role in pulpal inflammation. To date little is known regarding the regulation of MMPs in human pulp cell cultures. The purpose of this study was to determine the effects of cytokines (interleukin-1 and transforming growth factor-beta (TGF-beta), protein synthesis inhibitor cycloheximide (CD), and protein kinase C inhibitors (H7 and Go6976) on the secretion and production of MMPs by human pulp cell cultures using gelatin zymography. The main gelatinase secreted by human pulp cells migrated at 72 kDa and represented MMP-2. Minor gelatinolytic bands were also observed at 92 kDa regions that correspond to MMP-9. After an 8-day culture period TGF-beta, CD, H7, and Go6976 were found to depress MMP-2 production. The inhibition decreased in an order of CD > H7 > TGF-beta > Go6976. IL-1 was found to elevate MMP-2 production. Human pulp cells, however treated with either cytokines or pharmacological agents had no effect on the pattern of MMP-9 produced or secreted in either cell extracts or conditioned medium fractions. These observations suggest that the cytokines and pharmacological agents can regulate MMP-2 produced by human pulp cells. Inflammatory cytokines stimulate the production of elevated levels of MMP-2 and MMP-2 might play a role in pulpal inflammation. In addition agents that target protein synthesis or the protein kinase C pathway in human pulp cells inhibit MMP-2 production, and such inhibition may contribute to the pathogenesis of pulpal inflammation. Such inhibition might contribute to therapeutic efficacy.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Carbazoles; Cells, Cultured; Culture Media, Conditioned; Cycloheximide; Cytokines; Dental Pulp; Enzyme Inhibitors; Gelatinases; Humans; Indoles; Interleukin-1; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Protein Kinase C; Protein Synthesis Inhibitors; Pulpitis; Reproducibility of Results; Time Factors; Transforming Growth Factor beta

In vivo and ex vivo gene transfer are being developed for localized skeletal regeneration. These strategies for tissue regeneration were tested in an adult ferret model of vital pulp therapy. In this model a reversible pulpitis was induced first. Then after 3 d, the pulps were directly infected with recombinant virus or implanted with ex vivo transduced autologous dermal fibroblasts. The genome of the recombinant adenovirus contained a full-length cDNA encoding mouse bone morphogenetic protein (BMP)-7 (AdBMP7) or bacterial beta-galactosidase cDNA (AdlacZ). The BMP-7, but not lacZ, ex vivo transduced dermal fibroblasts induced reparative dentinogenesis with apparent regeneration of the dentin-pulp complex. In vivo infection with AdBMP-7 failed to produce reparative dentin in all cases. E. vivo gene transfer of BMP-7 may be an effective method for inducing dentin regeneration in teeth with reversible pulpitis.

Topics: Adenoviridae; Animals; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Dentin, Secondary; Ferrets; Fibroblasts; Gene Transfer Techniques; Male; Mice; Pulpitis; Regeneration; Transforming Growth Factor beta

Recombinant human BMP-7 (bone morphogenetic protein-7, osteogenic protein-1) is osteogenic, dentinogenic and cementogenic when implanted into the appropriate tissue in vivo. However, most studies characterizing the induction of these tissues have implanted BMP-7 into freshly surgerized, clinically healthy tissues. To determine if BMP-7 is dentinogenic in inflamed dental pulps, we applied BMP-7 to inflamed ferret pulps. A single application of 5 microg of a commercial preparation of lipopolysaccharide (LPS) from Salmonella typhimurium directly to the coronal pulp induced a reversible mixed inflammatory exudate of moderate intensity within 3 d. Treatment with a single application of 2.5, 7.5 or 25 microg recombinant human BMP-7/mg collagen (2 mg total mass/tooth) induced reparative dentinogenesis in controls but not LPS treated dental pulps. These data reveal that a single application of up to 50 microg/tooth of exogenous recombinant BMP-7 is insufficient to induce reparative dentinogenesis in ferret teeth with reversible pulpitis. Given that pulp cells in the inflamed tissues likely retain the capacity to respond to exogenous BMP-7, it is possible that insufficient active recombinant protein is available to induce tissue formation in experimentally inflamed dental pulps.

Topics: Animals; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Collagen; Dentin, Secondary; Dentinogenesis; Drug Carriers; Ferrets; Humans; Lipopolysaccharides; Pulpitis; Recombinant Proteins; Salmonella typhimurium; Transforming Growth Factor beta

Interstitial collagenase (matrix metalloprotease-1) is a member of a family of matrix metalloproteases and is thought to play a role in degradation of the extracellular components, such as collagens in normal extracellular matrix remodeling and in many disease processes. We examined interstitial collagenase mRNA expression in human dental pulp fibroblast cultures by Northern blot analysis. These cells did not express interstitial collagenase mRNA in an unstimulated condition. Inflammatory cytokines, such as interleukin-1 alpha, induced interstitial collagenase mRNA expression in these cells. The interstitial collagenase mRNA levels began to increase after 2-h exposure, reaching a maximum after 8 h, then dropping to the unstimulated level at 48 h. These effects were observed in a dose-dependent manner in a dose range of 0.1 to 10 ng/ml. Transforming growth factor-beta reduced the levels of interstitial collagenase mRNA expression that were induced by interleukin-1 alpha. These observations suggest that interstitial collagenase mRNA expression in human dental pulp fibroblasts is regulated by the inflammatory cytokines and that interstitial collagenase may play a role in tissue degradation in inflamed dental pulp.

Topics: Adolescent; Blotting, Northern; Cells, Cultured; Collagenases; Dental Pulp; DNA, Complementary; Dose-Response Relationship, Drug; Enzyme Induction; Fibroblasts; Humans; Interleukin-1; Male; Matrix Metalloproteinase 1; Pulpitis; Recombinant Proteins; RNA, Messenger; Stimulation, Chemical; Transforming Growth Factor beta