transforming-growth-factor-beta and Gingivitis

Topics: Alveolar Bone Loss; Animals; Biomarkers; Bone Resorption; Collagen; Collagen Type I; Endothelial Growth Factors; Epidermal Growth Factor; Extracellular Matrix Proteins; Gingival Crevicular Fluid; Gingivitis; Glycosaminoglycans; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Lymphokines; Osteocalcin; Peptides; Periodontal Diseases; Periodontal Index; Periodontitis; Platelet-Derived Growth Factor; Protein Isoforms; Proteoglycans; Transforming Growth Factor alpha; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Pregnancy is a special period marked with complicated changes in various immune responses. Although pregnant women are prone to developing gingival inflammation, its immunological mechanism remains to be clarified. In a modified ligature-induced periodontal disease murine model, pregnant mice developed more severe alveolar bone loss. Using this model, we investigated the Treg responses during exacerbated periodontal disease in pregnant mice. We tested Treg-associated molecules in gingival tissues by quantitative real-time PCR and found decreased gingival expression of Foxp3, TGFβ, CTLA-4, and CD28 in pregnant mice after periodontal disease induction. We further confirmed that lower number of Treg cells were present in the cervical lymph nodes of pregnant periodontitis mice. Treg cells from the cervical lymph nodes of ligated pregnant mice and non-pregnant mice were tested for their suppressive function in vitro. We manifested that Treg suppressive function was also down-regulated in the pregnant mice. Additionally, we demonstrated that more inflammatory Th17 cells were present in the cervical lymph nodes of ligated pregnant mice. Therefore, impaired Treg development and function, together with upregulated Th17 response, may contribute to the exacerbated periodontal disease during pregnancy.

Topics: Animals; Disease Models, Animal; Disease Progression; Down-Regulation; Female; Forkhead Transcription Factors; Gingivitis; Humans; Mice; Periodontal Diseases; Pregnancy; Pregnancy Complications; T-Lymphocytes, Regulatory; Th17 Cells; Transforming Growth Factor beta

Inhibitory signals mediated via molecules such as programmed death-1 (PD-1) play a critical role in downmodulating immune responses and maintaining peripheral tolerance. We investigated the involvement of cytokines and PD-1 engagement in mediating the T-cell unresponsiveness to bacterial and ubiquitous antigens in periodontal diseases.. Gingival and peripheral blood samples from healthy individuals and patients with chronic periodontitis were collected and used for the subsequent assays. Leukocytes in the lesion site and blood were evaluated using flow cytometry. The production of interferon-gamma, interleukin-10, and transforming growth factor-beta proteins was evaluated by enzyme-linked immunosorbent assay (ELISA), and the presence of PD-1+ cells in the inflamed gingiva was confirmed by immunofluorescence confocal microscopy for CD4 and PD-1 colocalization.. T cells from patients with chronic periodontitis proliferated poorly in response to Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) antigen. T-cell unresponsiveness was not associated with imbalanced cytokine production. However, T cells from patients with chronic periodontitis expressed significantly higher levels of PD-1 either upon isolation or after culture with antigens. Moreover, PD-1 blocking did not result in significant T-cell proliferation in cells cultured with phytohemagglutinin or bacterial antigens. The blockade of PD-1 resulted in the increased production of IFN-gamma. In addition, CD4+ and CD8+ T cells expressing PD-1 accumulated in lesions with chronic periodontitis.. These data show that PD-1 engagement could be involved in the modulation of IFN-gamma production by T cells in patients with chronic periodontitis.

Topics: Adult; Aged; Aged, 80 and over; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Antigens, Bacterial; Antigens, CD; Apoptosis Regulatory Proteins; Case-Control Studies; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Chronic Periodontitis; Down-Regulation; Female; Gingiva; Gingivitis; Humans; Interferon-gamma; Interleukin-10; Leukocytes; Lymphocyte Activation; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Pocket; Programmed Cell Death 1 Receptor; T-Lymphocyte Subsets; T-Lymphocytes; Transforming Growth Factor beta

The balance between inflammatory mediators and their counter-regulatory molecules may be crucial for determining the outcome of immune pathology of periodontal diseases. Based on clinical and immunological findings, the immune response in stable gingivitis lesion is supposed to be in balance, whereas the response is skewed towards the predominance of proinflammatory reactivity in progressive periodontitis lesion. However, this hypothesis has not been verified. Therefore, the aim of this study was to compare the gene expression profile of inflammatory mediators including proinflammatory cytokines and other inflammatory molecules, and anti-inflammatory cytokines by using quantitative real-time polymerase chain reaction in gingivitis and periodontitis lesions showing distinct clinical entities. For inflammatory mediators, interleukin (IL)-1beta, interferon (IFN)-gamma and receptor activator of nuclear factor (NF)-kappaB ligand tended to be higher in periodontitis, whereas tumour necrosis factor (TNF)-alpha and IL-12 p40 showed no difference. Heat-shock protein 60 (HSP60) expression was up-regulated significantly in periodontitis. For anti-inflammatory cytokines, transforming growth factor (TGF)-beta1 expression tended to be higher in periodontitis compared with gingivitis, whereas no difference was observed for IL-10 and IL-4. These findings support further our previous finding that autoimmune response to HSP60 may exert in periodontitis lesion, and suggest that perhaps subtle differences in the balance of cytokines may result in different disease expression.

Topics: Adult; Carrier Proteins; Chaperonin 60; Chronic Disease; Cytokines; Female; Gene Expression; Gingivitis; Humans; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-4; Male; Membrane Glycoproteins; Middle Aged; NF-kappa B; Periodontitis; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha; Up-Regulation

CD4+CD25+ regulatory T (Tr) cells are critical in regulating the immune response and thereby play an important role in the defense against infection and control of autoimmune diseases. Our previous studies demonstrated the involvement of autoimmune responses in periodontitis. The aim of this study was to identify CD4+CD25+ Tr cells in periodontitis tissues and compare them with those in gingivitis tissues. Immunohistological analysis of CD4, CD25, and CTLA-4 and the gene expression analysis of FOXP3, TGF-beta1, and IL-10 on gingival biopsies revealed the presence of CD4+CD25+ Tr cells in all tissues. In periodontitis, the percentage of CD4+CD25+ Tr cells increased with increasing proportions of B-cells relative to T-cells. FOXP3, a characteristic marker for CD4+CD25+ Tr cells, TGF-beta1 and IL-10 were expressed more highly in periodontitis compared with gingivitis. These findings suggest that CD4+CD25+ Tr cells and possibly other regulatory T-cell populations do exist and may play regulatory roles in periodontal diseases.

Topics: Aged; CD4 Antigens; Chemotaxis, Leukocyte; DNA-Binding Proteins; Forkhead Transcription Factors; Gene Expression Profiling; Gingivitis; Humans; Immunohistochemistry; Interleukin-10; Middle Aged; Periodontitis; Receptors, Interleukin-2; Reference Values; T-Lymphocyte Subsets; T-Lymphocytes; Transforming Growth Factor beta; Transforming Growth Factor beta1

In this study we examine the properties of a vegetable extract from seeds of Lupinus albus (LU 105). In previous works we demonstrated that LU 105 reduced the expression, by gingival fibroblasts, of both matrix metalloproteinase (MMP)-2 and MMP-9. We decided to study the impact of LU 105 on cell proliferation and morphology. Using organ culture media we also studied the MMP and tissue inhibitors of metalloproteinases (timp) expression AND THE cytokines secretion.. Healthy and inflamed gingival biopsies were placed in appendage culture with or without LU 105. The organ culture media were analyzed using Western blottings (MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MMP-13, TIMP-1, and TIMP-2) and gelatine zymography. A reverse transcription polymerase chain reaction (RT-PCR) was also performed on healthy and inflamed gingival biopsies, which were maintained in culture with or without LU 105 0.1%. Then, we decided to determine the amount of cytokines present in the organ culture media such as interleukin (IL)-1 beta, IL-4, IL-6, transforming growth factor (TGF)-beta, and tumor necrosis factor (TNF)-alpha.. When gingival biopsies derived from inflamed tissues were cultured with LU 105 0.1% in the culture media, the MMP and TIMP expression and activity decreased significantly when compared to cultures without LU 105. Moreover, we did not note any statistical difference in the cell proliferation compared with human gingival fibroblast cultures without LU 105. Furthermore, IL-1 beta, IL-6, TGF-beta, and TNF-alpha amounts in the culture media decreased significantly, whereas IL-4 increased significantly when LU 105 0.1% was added to the culture media.. LU 105, a novel metalloproteinase inhibitor with few consequences on cell proliferation and morphology, is a vegetable extract with potential clinical capacity.

Topics: Analysis of Variance; Cell Proliferation; Cell Shape; Cells, Cultured; Cytoskeleton; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gingiva; Gingivitis; Humans; Interleukins; Lupinus; Metalloproteases; Oligopeptides; Plant Extracts; Protease Inhibitors; Reverse Transcriptase Polymerase Chain Reaction; Seeds; Tissue Inhibitor of Metalloproteinases; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Both the virulence factors of periodontopathic bacteria and the immune response against them have been involved in tissue destruction observed in periodontal disease. Considering the regulatory role of cytokines produced by T cells, the purpose of this study was to compare the CD3+, CD4+, and CD8+ subpopulations of T cells, and to characterize the mRNA of cytokines involved in the adaptive immune response in a group of healthy/gingivitis 1 (HI/G1) individuals and aggressive periodontitis (AgP) patients.. The percentages of T-cell subpopulations were analyzed in 10 gingival samples of HI/G1 individuals and 10 gingival samples of AgP patients by immunohistochemistry. The presence of interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-5, IL-10, IL- 13, and transforming growth factor (TGF)-beta was measured by reverse transcription polymerase chain reaction (RT-PCR) of mRNA extracted from complete gingival biopsies.. Significant differences were found in CD3+ and CD4+ cell counts between both groups. The parameters were lower in the gingival biopsies from AgP patients while CD8+ counts were similar in both groups. The cytokine mRNA analysis showed constant expression of IL-2 and IFN-gamma in all cases. The mRNA of IL-5 and IL-10 was present in the majority of HI/G1 (N = 10, N = 9, respectively) but was not in the AgP group (N = 2, N = 1). IL-13 and TGF-beta were only detected in HI/G1 (N = 2, N = 3) and IL-4 was not detected in any of the individuals.. These results indicate that the role of the CD8+ subpopulation in aggressive periodontitis lesions is limited. On the other hand, cytokines IL-2 and IFN-gamma may not be relevant in the progression of aggressive periodontitis.

Topics: Alveolar Bone Loss; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cytokines; Disease Progression; Gingiva; Gingivitis; Humans; Interferon-gamma; Interleukin-10; Interleukin-13; Interleukin-2; Interleukin-4; Interleukin-5; Lymphocyte Count; Periodontal Attachment Loss; Periodontitis; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

TGFbeta1 is a multifunctional growth factor with both pro- and anti-inflammatory properties. This study aimed to determine levels of TGFbeta1 in gingival crevicular fluid (GCF), serum and plasma in the early stages of gingival inflammation.. A 21-day experimental model of gingivitis employing a split mouth design with a soft vinyl splint used to cover test teeth during brushing.. Ten healthy volunteers (mean age 21 years; five males and five females).. GCF and blood (with and without EDTA) was collected on days 0, 7, 14 and 21. GCF volumes were measured on a precalibrated Periotron 8000TM. Clinical indices of gingival inflammation and plaque levels were obtained after GCF sampling. Normal brushing resumed after GCF collection on day 21 and final samples were collected on day 35. TGFbeta1 and alkaline phosphatase (ALP) levels were determined using enhanced chemiluminescent methods.. Clinical indices and GCF volumes increased at test sites during the 21-day test period. Concentrations of TGFbeta1 and ALP in GCF (test and control), serum and plasma did not change throughout the study (P > 0.3). However, total amounts of TGFbeta1 (pg sample-1) and ALP (mu IU sample-1) in GCF increased at test sites and were significantly higher than baseline values at days 7, 14 and 21 (P < 0.04). Control sites showed no variation in TGFbeta1 or ALP levels throughout the study period (P > 0.35). All parameters at test sites returned to control levels at day 35 (P > 0.3).. The data indicate that GCF TGFbeta1 levels increase early in plaque-induced inflammation. Whether the biological consequence of this site-specific increase is pro- or anti-inflammatory in nature remains to be elucidated.

Topics: Adult; Alkaline Phosphatase; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Male; Statistics, Nonparametric; Transforming Growth Factor beta; Transforming Growth Factor beta1

Evidence of the role of cytokines produced by resident and inflammatory cells during inflammation is well established. The aim of this study was to quantify in healthy and diseased human gingiva the area fraction (AA%) occupied by collagen fibers and the amount of cytokines such as interleukin (IL)-1beta, IL-4, IL-6, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, and epidermal growth factor (EGF) to investigate a possible correlation between such cytokines, collagen degradation, and the gingival index.. Gingival tissue specimens from 6 healthy controls (group 1), 6 patients with mild gingival inflammation (group 2), 6 patients with moderate gingival inflammation (group 3), and 6 patients with severe gingival inflammation (group 4) were cultured for 72 hours, and the cytokines present in the culture media were quantified using an enzyme-linked immunosorbent assay (ELISA). Paraffin gingival sections from the 24 subjects were stained with sirius red F3Ba for visualization of collagen fibers, then the area fraction (AA%) occupied by the gingival fibers was determined by automated image analysis.. The present study revealed significant differences (P < 0.05) between means of AA% in group 1 (53%), group 2 (41%), group 3 (39.5%), and group 4 (35%) for collagen fibers. Compared to controls, there were significant increases of IL-1beta (groups 3 and 4), IL-6, and TNF-alpha (group 3); a significant decrease of IL-4 (groups 2, 3, and 4) and TGF-beta (groups-2 and, 3); and no change of EGF. The collagen AA% was significantly correlated with the amounts of IL-4 and TGF-beta, and significantly inversely correlated with the amounts of IL-1beta for all 3 inflamed groups and IL-6 and TNF-alpha for groups 2 and 3.. The present study showed that EGF was not changed in inflamed gingival tissue and that IL-1beta and IL-4 were particularly and intensively correlated with collagen loss. These 2 cytokines could be markers of clinical severity during active periodontitis.

Topics: Adolescent; Adult; Analysis of Variance; Case-Control Studies; Child; Culture Techniques; Cytokines; Disease Progression; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Fibrillar Collagens; Gingivitis; Humans; Interleukins; Periodontal Index; Periodontitis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Transforming growth factor-beta (TGF-beta) is composed of a family of multifunctional polypeptide growth factors involved in embryogenesis, inflammation, regulation of immune response, angiogenesis, wound healing, and extracellular matrix formation. TGF-beta1 is the most common isoform found in human tissues. A role of TGF-beta in the pathogenesis of periodontal disease has been suggested. The aim of the present study was a comparative immunohistochemical evaluation of TGF-beta1 in normal keratinized gingiva and in the peri-implant soft tissues surrounding failing non-submerged implants.. Twenty patients participated in this study. Ten biopsies from healthy keratinized mucosa and 10 biopsies from peri-implant soft tissues surrounding failing implants were obtained (one biopsy per patient). The biopsies were obtained from different patients.. In 5 cases of healthy mucosa, the stromal cells were positive between 1 to 5. In 7 cases, the epithelial layers were positive, between 1 and 18 cells. The superficial epithelial layer was negative in all cases. In 9 cases, there was a positivity of the vascular component, between 2 and 16 vessels. In failing implants, the stromal cells were positive in 6 cases, between 1 and 4. In all cases, cells of the epithelial layers were positive, between 15 and 40. The vascular component was positive in all cases, between 12 and 30 vessels. The differences between TGF-beta1 expression in the epithelium around healthy and failing implants were statistically significant (P < 0.0001). The differences between TGF-beta1 expression in the blood vessels in the soft tissues around healthy and failing implants were also statistically significant (P < 0.0001). No statistically significant difference was observed between the 2 groups in the TGF-beta1 expression in the stromal cells (P = 0.88).. TGF-beta1 may be one of the most important factors in the regulation of the infiltrate, and in the production of tissue repair with a stimulation of fibroblasts and endothelial cells.

Topics: Adult; Dental Implants; Dental Restoration Failure; Endothelium; Female; Gingiva; Gingivitis; Humans; Immunohistochemistry; Male; Middle Aged; Mouth Mucosa; Statistics, Nonparametric; Transforming Growth Factor beta; Transforming Growth Factor beta1

To investigate the distribution of transforming growth factor-beta isoforms in chronically inflamed periodontal tissues.. The present study determined, by immunohistochemistry, the expression patterns of TGF-betas and their receptors in the lining epithelia of inflamed gingiva. Frozen sections were obtained from 22 human gingival biopsies.. TGF-beta 1 was not detected in gingival epithelial cells in examined sections. Detection of TGF-beta 2 indicated a progressive reduction of staining from the external oral epithelium through to gingival sulcus and the gingival attachment or pocket epithelium. TGF-beta 3 showed intense staining in all domains of both minimally inflamed gingiva and advanced periodontitis tissues. TGF-beta RI was visualised as focal staining of the spinous layer in the external oral epithelium of both periodontitis lesions and minimally inflamed tissues. TGF-beta RII was present throughout the strata, but with progressive reduction in intensity from the oral epithelium to gingival attachment or pocket epithelium respectively while, conversely, TGF-beta RIII showed an increase in diffuse staining intensity from external oral epithelium to pocket epithelium.. A distinct expression profile was observed within different individuals for TGF-betas and the corresponding receptors. These findings provide a basis for evaluation of the role of these growth factors in the pathogenesis of periodontitis.

Topics: Blotting, Western; Chronic Disease; Epithelial Cells; Fluorescent Antibody Technique, Indirect; Gingiva; Gingivitis; Humans; Periodontitis; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta

In inflammatory gingival diseases, cytokines have been demonstrated to play critical roles by coordinating the stimulation of immunological and connective tissue cells. The activities of these cells, degrading and remodeling extracellular matrices, constitute the major pathological and repair processes. Thus, elucidating cellular and molecular events occurring in inflamed connective tissues is crucial for the understanding and treatment of inflammation. In order to test a hypothesis that proinflammatory cytokines affect metabolism of major extracellular matrix molecules, we studied metabolism of proteoglycans (PGs) by human gingival fibroblasts (HGF) under the influence of interleukin-4 (IL-4) as a model of gingivitis. HGF in cell culture were metabolically radiolabeled using [3H]glucosamine and [35S]sulfate in the presence or absence of IL-4, and the labeled PGs were analyzed by chromatographic techniques. The incorporation of 35S into PGs increased with IL-4 both in media and cell layer. At 100 ng/ml of IL-4, the increment of 35S incorporation over control culture was 16-39% (p<0.001) in media and 12-35% (p=0.01) in cell layer. The 35S-labeled macromolecules were PGs containing heparan sulfate (HS) and chondroitin sulfate (CS) chains. From the molecular weight and glycosaminoglycan composition analyses, versican and perlecan-type and biglycan and decorin-type were very likely to be the major PG constituents both in media and cell layer. IL-4 stimulated synthesis of versican and perlecan-type more potently than biglycan and decorin-type. With IL-4 treatment, the ratio of CSPG/HSPG decreased in media and increased in cell layer. This ratio suggested that syndecan family HSPGs were also present in HGF. In conclusion, IL-4 stimulated accumulation of CS/HSPGs in human gingival fibroblasts.

Topics: Adult; Biglycan; Cell Culture Techniques; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Chromatography, Agarose; Chromatography, Ion Exchange; Decorin; Extracellular Matrix Proteins; Female; Fibroblasts; Gingiva; Gingivitis; Glucosamine; Glycosaminoglycans; Heparan Sulfate Proteoglycans; Humans; Interleukin-4; Lectins; Lectins, C-Type; Membrane Glycoproteins; Molecular Weight; Proteoglycans; Radiopharmaceuticals; Sulfur Radioisotopes; Syndecans; Transforming Growth Factor beta; Tritium; Versicans

The aim of the present study was to investigate the level of transforming growth factor-beta 1 (TGF-beta 1) in gingival crevicular fluid (GCF) samples of cyclosporin A (CsA)-treated patients and to compare the results with control groups.. Fourteen renal transplant patients exhibiting severe CsA-induced gingival overgrowth, 10 patients with chronic gingivitis, and 10 subjects with clinically healthy periodontium were included in the study. In CsA-treated patients, GCF samples were harvested from sites exhibiting gingival overgrowth (CsA GO+) and sites not exhibiting gingival overgrowth (CsA GO-). The TGF-beta 1 levels in a total of 96 GCF samples from the 34 participants were analyzed by enzyme-linked immunosorbent assay. The results were expressed in terms of total amount (pg/2 sites) and concentration (ng/ml).. TGF-beta 1 total amounts in CsA GO+ and CsA GO- sites were similar and significantly higher than that of healthy sites (P < 0.02 and P < 0.01, respectively). The total amount of TGF-beta 1 was also higher in gingivitis sites compared to the healthy sites, but the difference was not statistically significant (P > 0.05). CsA GO+ and CsA GO- sites exhibited higher total amount and concentration of TGF-beta 1 than that of gingivitis sites, but the differences were insignificant (P > 0.05).. The results of the present study support the theory that CsA increases the synthesis of TGF-beta 1 in GCF. However, since the difference between CsA GO+ and CsA GO- sites was not statistically significant, it seems unlikely that GCF TGF-beta 1 level is the sole factor responsible for the CsA-induced gingival overgrowth. Complex interactions between various mediators of inflammation and tissue modeling are possibly involved in the pathogenic mechanisms of this side effect.

Topics: Adolescent; Adult; Confidence Intervals; Cyclosporine; Enzyme-Linked Immunosorbent Assay; Female; Gingiva; Gingival Crevicular Fluid; Gingival Overgrowth; Gingivitis; Humans; Immunosuppressive Agents; Inflammation Mediators; Kidney Transplantation; Male; Statistics, Nonparametric; Transforming Growth Factor beta; Transforming Growth Factor beta1

The role of matrix metalloproteinases (MMPs) in cell migration was studied by measuring cell growth, migration, and production of MMP-2 and -9 in oral mucosal and skin keratinocytes cultured in the presence of synthetic MMP inhibitors. MMP-2 was the major gelatinolytic MMP produced by these cells while MMP-9 was produced at a low basal level. Inhibitor effects on MMP-9 production were therefore studied in keratinocytes stimulated by tumor necrosis factor alpha (TNFalpha). Tetracycline analogues at concentrations that inhibited the production of MMP-2 but not MMP-9 were able to drastically inhibit migration of both mucosal and skin keratinocytes. Tetracycline analogues also inhibited keratinocyte growth, an effect not found for the other inhibitors tested. Heterocyclic carbonate-derived compounds (LWs) that inhibited MMP-9 but not MMP-2 production had no effect on cell migration. Batimastat, a potent MMP inhibitor, did not have any effect on MMP production or cell growth but did inhibit keratinocyte migration. Tumor growth factor beta (TGFbeta) increased keratinocyte migration as well as both cell-associated and secreted MMP-2 production in wounded cell cultures. The secreted enzyme was partially converted into an active form. In this model batimastat totally blocked TGFbeta-promoted keratinocyte migration. Immunostaining of keratinocytes advancing into the wound revealed that MMP-2 was localized in extracellular matrix contactlike structures against the endogenously produced laminin-5-rich matrix. MMP-9 was localized diffusely along the cell membranes. Using in situ hybridization we observed that in chronically inflamed human gingiva MMP-2 is expressed in epithelium extending into subepithelial connective tissue. These results suggest that MMP-2 plays a specific role in epithelial migration, possibly by detaching the advancing cells from the pericellular matrix or by activating other MMPs.

Topics: Cell Adhesion Molecules; Cell Division; Cell Line; Cell Movement; Cell Size; Cells, Cultured; Collagenases; Dose-Response Relationship, Drug; Gelatinases; Gene Expression Regulation, Enzymologic; Gingivitis; Humans; In Situ Hybridization; Kalinin; Keratinocytes; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Metalloendopeptidases; Mouth Mucosa; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing

TGF-beta 1 is a multifunctional molecule which has unique and potent effects on many target cells and tissues. TGF-beta 1 may promote inflammatory reaction by certain intercellular interaction. TGF-beta 1 at extremely low concentrations shows strong chemotatic activity for mononuclear phagocytes and stimulates bone resorption by enhancing production of PGE2. On the other hand, TGF-beta 1 plays a very important role in the regulation of extracellular matrix turnover presumably by modulating the action of other growth factors on matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) expression. TGF-beta 1 was identified intra- and extracellularly in the inflamed gingival tissues and the distribution was associated with areas of inflammation. Sixteen miniature swines were used in this experimental gingivitis/periodontitis study. The ligatures were placed in situ for periods of 3, 5, 8 and 13 weeks and peroral innoculations of Porphyromonas gingivalis/Actinomyces viscosus into the ligatures were carried out only in the experimental group. ELISA was used to measure the levels of TGF-beta 1 in gingival tissues from the experimental and control groups. Recording of the clinical periodontal parameters was performed and the proportion of black-pigmented Bacteroides in the ligature (plaques) removed immediately prior to the biopsies was recorded. The results revealed that the concentration of TGF-beta 1 of the experimental group was higher and significantly different in statistics on the period of third week than that of the control group. The concentration of TGF-beta 1 was significantly different between the third week and the thirteenth week in the experimental group, and was negatively related to the time-length of ligatures. Furthermore, the concentration of TGF-beta 1 was negatively related to the changes of the calculus index and gingival index. These data indicated that the concentration of TGF-beta 1 of gingival tissue exhibited dynamic changes associated with the progression of experimental periodontal inflammation. The levels of TGF-beta 1 in gingival tissue may be valuable in detecting the inflammatory reaction of periodontal tissues.

Topics: Animals; Disease Progression; Gingivitis; Periodontitis; Swine; Swine, Miniature; Transforming Growth Factor beta

Transforming growth factor-beta (TGF-beta) represents a family of polypeptide growth factors, involved in embryogenesis, inflammation, regulation of immune responses and wound healing. To determine whether TGF-beta contributes to the evolution of periodontal disease, we assayed TGF-beta levels in gingiva and crevicular fluid of patients with gingivitis and periodontitis. In parallel, TGF-beta was quantified in gingival fluid and serum of beagles with experimentally-induced periodontitis. Disease was monitored by several clinical parameters including Plaque Index, Gingival Index, probing depth, and epithelial attachment loss. Gingival tissues were obtained from 9 patients at the time of periodontal surgery, and gingival fluid samples were collected from an additional population of 10 periodontal patients. In 14 beagles, experimental periodontitis was induced and gingival fluids collected 6 months later. Fluid was collected by paper strips and volume measured by Periotron. Additionally, sera was collected before and 9 months after the ligature-induced periodontitis in 7 beagles. The levels of TGF-beta 1 were measured by ELISA. In the patients, a significantly higher concentration of TGF-beta 1 was observed both in the gingival tissues and fluid samples obtained from the sites with deeper periodontal pockets than in the less involved sites. In beagles, TGF-beta 1 levels measured in gingival fluid were elevated in moderate disease, declining in fluid samples obtained from the pockets during more advanced experimental periodontitis. Furthermore, with the progression of experimental periodontitis, a decrease in TGF-beta 1 occurred in the sera of the beagle dogs. These data suggest that TGF-beta 1 may play a rôle in the pathogenesis and diagnosis of periodontal disease, and that its actions can be further explored in an animal model.

Topics: Adult; Aged; Animals; Dental Plaque Index; Disease Models, Animal; Disease Progression; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Gingiva; Gingival Crevicular Fluid; Gingivitis; Humans; Male; Middle Aged; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Transforming Growth Factor beta

The effects of TGF-beta and PDGF on the metabolic conversion of 14C-testosterone by human gingival tissue (HGT) from 5 subjects was investigated. The metabolic conversions in response to TGF-beta and PDGF were also studied in 4-6 cell-lines of cultured gingival fibroblasts, using 14C-testosterone and 14C-4-androstenedione as substrates. Duplicate incubations of HGT were performed in Eagle's MEM + 10% FCS and optimal stimulatory concentrations of TGF-beta/PDGF for 24 h. Similar incubations were performed in duplicate with cell-lines of cultured gingival fibroblasts, TGF-beta/PDGF, 14C-testosterone/14C-4-androstenedione in Eagle's MEM + 10% FCS. The radioactive metabolites were extracted, separated and quantified. With HGT, TGF-beta and PDGF caused 2.5/2-fold increases in DHT synthesis (p < 0.1; Wilcoxon signed rank test) and 3.4/2-fold increases in 4-androstenedione formation (p < 0.1) from 14C-testosterone. PDGF increased DHT and testosterone synthesis from 14C-4-androstenedione by 3-fold in gingivae (p < 0.1). With cell-lines, average values of duplicate incubations showed 2.8/2-fold increases in DHT synthesis from 14C-testosterone in response to TGF-beta/PDGF (p < 0.1; p < 0.2) and 2.4/2-fold increases in 4-androstenedione synthesis (p < 0.1; p < 0.2). With 14C-4-androstenedione as substrate, TGF-beta/PDGF caused 1.6/1.9-fold increases in DHT synthesis compared with controls (p < 0.05; p < 0.1) and 1.7/1.5-fold increases in testosterone formation from this substrate (p < 0.05; p < 0.1). Due to the strong implications of TGF-beta/PDGF and anabolic androgens on matrix repair, significant increases in DHT synthesis from 2 androgenic substrates in response to TGF-beta and PDGF are of particular relevance to inflammatory repair processes.

Topics: Adult; Androgens; Androstenedione; Cells, Cultured; Dihydrotestosterone; Extracellular Matrix Proteins; Female; Fibroblasts; Gas Chromatography-Mass Spectrometry; Gingiva; Gingivitis; Humans; Male; Platelet-Derived Growth Factor; Regeneration; Statistics, Nonparametric; Testosterone; Transforming Growth Factor beta; Up-Regulation

Cellular and biochemical observations were made of fibroblasts harvested from ligature-induced periodontitis and treated gingivitis areas in four adult female cynomolgus monkeys (Macaca fascicularis) to define the changes that occur in the early periodontitis lesion. Compared with fibroblasts from the treated sites, fibroblasts from the diseased areas had a significantly higher rate of proliferation, produced about two-thirds the amount of total protein and collagen, and failed to respond to TGF-beta, which normally stimulates extracellular matrix formation in mesenchymal cells. The diseased cells were also deficient in the activity of poly(ADP-ribose) synthetase, an enzyme involved in the repair of DNA breaks such as occur from the insults of superoxide and other active radicals present in inflamed areas. Although the precise nature of these biochemical defects is not fully elucidated, they may have an important bearing on chronic periodontitis cases with a "downhill" course.

Topics: Animals; Cell Division; Cells, Cultured; Collagen; Disease Models, Animal; Disease Progression; DNA Repair; Female; Fibroblasts; Gingivitis; Ligation; Macaca fascicularis; Periodontitis; Poly Adenosine Diphosphate Ribose; Protein Biosynthesis; Proteins; Transforming Growth Factor beta

Topics: Animals; Cytokines; Fibroblasts; Gingiva; Gingivitis; Interleukin-1; Phospholipases A; Phospholipases A2; Rats; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha