transforming-growth-factor-beta and Periodontitis

The tooth is stabilized by fiber-rich tissue called the periodontal ligament (PDL). The narrow space of the PDL does not calcify in the physiological state even thought it exists between two calcified tissues, namely, the cementum of the root and alveolar bone. Two situations that require PDL regeneration are periodontitis and dental trauma. Periodontitis induces the loss of PDL and alveolar bone due to inflammation related to infection. Conversely, in PDLs damaged by dental trauma, accelerating bone formation as an overreaction of the healing process is induced, thereby inducing dentoalveolar ankylosis at the tooth root surface. PDL regeneration following dental trauma must therefore be considered separately from periodontitis. Therefore, PDL regeneration in dental trauma must be considered separately from periodontitis. This review focuses on the components involved in avoiding dentoalveolar ankylosis, including oxytalan fibers, aggregated microfibrils, epithelial cell rests of Malassez (ERM), and TGF-β signaling. During root development, oxytalan fibers produced by PDL cells work in collaboration with the epithelial components in the PDL (e.g., Hertwig's root sheath [HERS] and ERM). We herein describe the functions of oxytalan fibers, ERM, and TGF-β signals which are involved in the avoidance of bone formation.

Topics: Ankylosis; Fibrillins; Humans; Periodontal Ligament; Periodontitis; Tooth Ankylosis; Transforming Growth Factor beta

Periodontitis and chronic obstructive pulmonary disease (COPD) are chronic inflammatory diseases with common risk factors, such as long-term smoking, age, and social deprivation. Many observational studies have shown that periodontitis and COPD are correlated. Moreover, they share a common pathophysiological process involving local accumulation of inflammatory cells and cytokines and damage of soft tissues. The T helper 17 (Th17) cells and the related cytokines, interleukin (IL)-17, IL-22, IL-1β, IL-6, IL-23, and transforming growth factor (TGF)-β, play a crucial regulatory role during the pathophysiological process. This paper reviewed the essential roles of Th17 lineage in the occurrence of periodontitis and COPD. The gaps in the study of their common pathological mechanism were also evaluated to explore future research directions. Therefore, this review can provide study direction for the association between periodontitis and COPD and new ideas for the clinical diagnosis and treatment of the two diseases.

Topics: Cytokines; Humans; Interleukin-23; Periodontitis; Pulmonary Disease, Chronic Obstructive; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Transforming Growth Factor beta

Marfan syndrome (MFS) is a systemic connective tissue disorder that is caused by mutations in the extracellular matrix protein fibrillin-1. While MFS patients are considered to be at high risk of dental disorders and cardiovascular complications, little causal relationship has been provided to date. It is well known that an elevated level of active TGF-β in the plasma is a major manifestation of MFS. TGF-β is known to play a critical role in the development of cardiovascular diseases and its levels were also elevated in the serum and saliva of periodontitis patients. These findings may suggest an association between periodontitis and the cardiovascular complications of MFS. In this article, we review the influence of periodontitis in MFS patients with cardiovascular complications in order to identify critical therapeutic targets of TGF-β.

Topics: Humans; Marfan Syndrome; Periodontitis; Transforming Growth Factor beta

Inflammatory periodontal disease is an almost ubiquitous disorder in the adult population. Cases or sites with moderate to advanced disease often continue to show signs of inflammation after non-surgical approach. Our current understanding of periodontal healing is based on a hypothesis by Melcher who proposed that the cell type that repopulates the exposed root surface at the periodontal repair site will define the nature of the attachment/repair that take place. If mesenchymal cells from periodontal ligament/perivascular region of the bone proliferate and colonize the root surface, regeneration occurs. Growth factors are natural cell products that are released or activated when cell division is needed. This action typically occurs during such events as wound healing or tissue regeneration. Activated platelets at the wound margins release several growth factors such as platelet-derived growth factor (PDGF), transforming growth factor (TGF)-alpha, epidermal growth factor etc. Cells adjacent to the injured site also are induced to release growth factors such as insulin-like growth factor-I, PDGF, TGF-alpha and TGF-alpha within a few hours after injury. In periodontal regeneration, the coronal re-establishment of the periodontal ligament (PDL) is required together with corresponding cementum and supporting alveolar bone. Thus, agents which promote periodontal ligament fibroblast (PLF) proliferation and migration as well as collagen biosynthesis would appear to be mediators for enhancing new PDL formation. When combinations or cocktails of different factors are used, greater repair is achieved than when individual factors are applied.

Topics: Adult; Fibroblast Growth Factors; Guided Tissue Regeneration, Periodontal; Humans; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Periodontal Ligament; Periodontitis; Platelet-Derived Growth Factor; Regeneration; Transforming Growth Factor beta

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

The goal of periodontal therapy is to protect and maintain the patient's natural dentition for his or her lifetime. More specifically, after periodontal regenerative surgery, the aim is to achieve complete wound healing and regeneration of the periodontal unit. A recent innovation in dentistry is the preparation and use of platelet-rich plasma (PRP), a concentrated suspension of the growth factors found in platelets. These growth factors are involved in wound healing and are postulated as promoters of tissue regeneration. This clinical update outlines the specific effects of these growth factors, both in vitro and in vivo, on periodontal wound healing. The review focuses on current animal and human trials using PRP to promote tissue regeneration and alveolar bone repair. The article goes on to describe the clinical benefits of PRP and the step-by-step preparation of PRP in the dental office.

Topics: Alveolar Bone Loss; Animals; Blood Platelets; Bone Regeneration; Humans; Insulin-Like Growth Factor I; Periodontitis; Plasmapheresis; Platelet-Derived Growth Factor; Transforming Growth Factor beta; Wound Healing

Soluble proteins that serve as mediators of cell function and are produced by various cell types, such as structural and inflammatory cells, are collectively called cytokines. Several lines of evidence have revealed that cytokines play important roles not only in tissue homeostasis but also in the pathogenesis of many infectious diseases. Recent research on biological activities in normal periodontium and the pathogenesis of periodontal diseases has clarified the involvement of various cytokines in the biological activities observed in the sites. Cytokines play crucial roles in the maintenance of tissue homeostasis, a process which requires a delicate balance between anabolic and catabolic activities. In particular, growth factors--such as fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), transforming growth factor-beta (TGF-beta)--are thought to play important roles in modulating the proliferation and/or migration of structural cells in the periodontium and the production of various extracellular matrices by these cells. On the other hand, there is little doubt that excessive and/or continuous production of cytokines in inflamed periodontal tissues is responsible for the progress of periodontitis and periodontal tissue destruction. Particularly, inflammatory cytokines--such as IL-1 alpha, IL-1 beta, IL-6, and IL-8--are present in the diseased periodontal tissues, and their unrestricted production seems to play a role in chronic leukocyte recruitment and tissue destruction. It is possible that monitoring cytokine production or its profile may allow us to diagnose an individual's periodontal disease status and/or susceptibility to the disease. In addition, although the hypothesis is still controversial, it has been suggested that discrete T-cell subsets (Th1 and Th2) with different cytokine profiles play specific roles in the immunopathogenesis of periodontal diseases.

Topics: Cell Division; Cell Movement; Cytokines; Disease Susceptibility; Extracellular Matrix; Fibroblast Growth Factors; Gene Expression Regulation; Homeostasis; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Leukocytes; Periodontal Diseases; Periodontitis; Periodontium; Platelet-Derived Growth Factor; Somatomedins; T-Lymphocyte Subsets; Transforming Growth Factor beta

In this paper, we review recent work on collagen degradation, 2 main routes of breakdown are described and their relevance during healthy and inflammatory conditions of the periodontium is discussed. Special attention is paid to the possible role of cytokines, in particular interleukin 1 (IL-1) and transforming growth factor beta (TGF-beta), on the modulation of collagen phagocytosis and metalloproteinase production. IL-1 has been shown to have a dual function in collagen digestion. It inhibits the intracellular phagocytic pathway, but at the same time, it strongly promotes extracellular digestion by inducing the release of collagenolytic enzymes like collagenase. TGF-beta has an opposite effect on both pathways and antagonizes IL-1. Collagenase is released in an inactive form, and a considerable fraction of the proenzyme may become incorporated in the extracellular matrix. This reservoir of latent enzyme can be activated (for instance by plasmin), leading to a sudden and extensive breakdown of the collagenous fibre meshwork. It is suggested that this phenomenon may also take place during progressive periodontitis and could explain an episodic nature of collagenolysis, clinically resulting in bursts of attachment loss (burst hypothesis).

Topics: Animals; Collagen; Collagenases; Cytokines; Disease Progression; Enzyme Activation; Humans; Interleukin-1; Metalloendopeptidases; Neutrophil Activation; Periodontitis; Phagocytosis; Transforming Growth Factor beta

Transforming growth factor beta (TGF-beta), a cytokine identified in acute and chronic inflammatory sites, mediates leukocyte recruitment and activation essential to the development of such lesions. Released by platelets upon aggregation and by leukocytes stimulated with bacterial products or inflammatory mediators, TGF-beta has potent chemotactic activity for blood neutrophils, monocytes, and lymphocytes. By augmenting integrin expression, TGF-beta facilitates leukocyte adhesion to the vessel wall and extracellular matrix at the site of inflammation. Once within the inflammatory site, mononuclear cells are stimulated by TGF-beta to release cytokines important in the network of molecules regulating the host response to microorganisms and immunologic challenge. Thus, bacteria and their products, in addition to directly recruiting and activating leukocytes at sites of infection, indirectly influence these events through the induction of cytokines such as TGF-beta. By antagonizing the activity of TGF-beta with neutralizing antibodies, a causal relationship between this cytokine, inflammation, and pathogenesis has been demonstrated. Administration of anti-TGF-beta to sites of chronic destructive inflammation not only blocked leukocyte recruitment and activation, but also inhibited the subsequent destruction of bone and cartilage characteristics of such lesions.

Topics: Bacterial Physiological Phenomena; Humans; Leukocytes; Periodontitis; Transforming Growth Factor beta

At present there is limited data concerning the efficacy of non-surgical periodontal therapy supplemented with subantimicrobial dose doxycycline (SDD) in the treatment of severe, generalized periodontitis. The purpose of the present study was to evaluate the effect of adjunctive SDD therapy on clinical periodontal parameters and gingival crevicular fluid (GCF) transforming growth factor-beta1 (TGF-beta1) levels in patients with severe, generalized chronic periodontitis over a 6-month period.. Thirty-five patients with severe, generalized periodontitis and 11 periodontally healthy subjects were included in the present study. Patients received full-mouth supragingival debridment at baseline and randomized to take either SDD b.i.d. or placebo b.i.d. for 3 months. Patients received root planing and oral hygiene instruction once a week for four consecutive weeks. Clinical measurements including probing depth (PD), clinical attachment level, papilla bleeding index and plaque index and GCF sampling were performed at baseline, 3 and 6 months. The GCF TGF-beta1 levels were analysed by enzyme-linked immunosorbent assay.. Thirteen patients in both study groups completed the 6-month trial. Following scaling and root planing (SRP) plus SDD and SRP plus placebo therapy significant improvements in clinical periodontal parameters of both groups were observed (p<0.025). In the SDD group a significantly higher percentage (%73.4) of deep pockets resolved (PD reduction > or =3 mm from baseline) when compared with placebo group (%49.7) at 6 months (p<0.05). At baseline there were no significant differences in GCF TGF-beta1 levels between three groups. Both total amount and concentration of GCF TGF-beta1 in SDD and placebo groups increased when compared with baseline at 3 months. However, only GCF TGF-beta1 levels of SDD group was significantly higher than baseline (p<0.025) and placebo group (p<0.017) at 3 months. At 6 months GCF TGF-beta1 levels of both groups were similar to baseline levels (p<0.025).. These data indicate that combination of SDD with non-surgical therapy improves clinical parameters of periodontal disease and increases GCF TGF-beta1 levels together with a decrease in prevalence of residual pockets in patients with severe, generalized chronic periodontitis. Increased GCF TGF-beta1 levels following SDD therapy might suggest a novell pleiotrophic mechanism for tetracyclines to inhibit connective tissue breakdown.

Topics: Adult; Anti-Bacterial Agents; Chronic Disease; Combined Modality Therapy; Dental Plaque Index; Double-Blind Method; Doxycycline; Female; Follow-Up Studies; Gingival Crevicular Fluid; Humans; Male; Middle Aged; Oral Hygiene; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Placebos; Root Planing; Subgingival Curettage; Transforming Growth Factor beta; Transforming Growth Factor beta1

The aim of this study was to evaluate the effects of Bdellovibrio bacteriovorus HD100 on experimental periodontitis (EP) in rats.. The EP group showed greater alveolar bone loss than EP-HD100 (p < .05). The EP-HD100 group showed higher levels of MCP-1, RANTES, IL-10, and TGF-β, lower levels of TNF-α than the EP group (p < .05). No differences were observed in IL-1β, IL-6, and M-CSF levels between EP and EP-HD100 groups. The C-HD100 group had higher IL-6, TNF-α, RANTES, and MCP-1 levels than the control group (p < .05). Regarding BD, the EP-HD100 group showed a larger immunolabeling pattern for BD-1, BD-2, and BD-3 than the EP group (p < .05). No significant differences in the immunolabeling pattern were observed for TLR-2, TLR-4, CD-4, CD-8, and CD-57 between EP and EP-HD100 groups.. The topical use of B. bacteriovorus HD100 reduces alveolar bone loss, increases expression of BD, and modulates the cytokines levels on periodontal tissues in rats with EP.

Topics: Alveolar Bone Loss; Animals; Bdellovibrio bacteriovorus; Interleukin-10; Interleukin-6; Macrophage Colony-Stimulating Factor; Periodontitis; Rats; Rats, Wistar; Toll-Like Receptor 4; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

To determine the changes of Porphyromonas gingivalis (P. gingivalis) growth and metabolism and identify whether the vascular epithelium change could be induced in diabetic periodontitis.. Maintaining favourable vascular function is a precondition for periodontal regeneration. In diabetic periodontitis, high glucose levels could enhance the metabolism of pathogens, and a complex condition involving inflammation and high glucose levels would disrupt homeostasis of the epithelium and promote fibrosis by endothelial-to-mesenchymal transition (EndMT).. Porphyromonas gingivalis was cultured with glucose to judge its metabolic activity. Human umbilical vein endothelial cells (HUVECs) were treated with P. gingivalis-lipopolysaccharide (LPS) (10 μg/ml) and/or high glucose concentrations (25 mM), and transforming growth factor (TGF)-β inhibitor was used to block EndMT. Inflammation level was assessed by flow cytometry. Multiple biological functions including EndMT, angiopoiesis, and cell migration were analysed. Additionally, gene expressions and protein levels were determined with qPCR and western blot, respectively. Finally, blood vessels were cultured ex vivo, and EndMT and fibrosis markers were detected by immunohistochemistry.. Glucose could promote P. gingivalis growth and biofilm formation as well as the expression of virulence factor genes including FimA, RgpA, RgpB, and Kgp. P. gingivalis-LPS and glucose could increase intracellular reactive oxygen species (ROS) and promote fibrosis via EndMT in HUVECs, along with attenuating angiopoiesis and cell migration, which could be resumed by blocking EndMT with TGF-β inhibitor. Vascular fibrosis was observed after the addition of glucose via EndMT regulation.. Glucose augmented the growth and metabolism of P. gingivalis and promoted fibrosis by the activation of EndMT, as well as the inhibition of angiopoiesis and cell migration.

Topics: Cells, Cultured; Fibrosis; Glucose; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Lipopolysaccharides; Periodontitis; Signal Transduction; Transforming Growth Factor beta

Cytokine-producing B cells play a well-established role in modifying immune responses in chronic inflammatory diseases. We characterized B-cell cytokine responses against periodontitis-associated bacteria in patients with periodontitis.. Blood and saliva samples were collected from patients with periodontitis grade B (N = 31) or grade C (N = 25), and 25 healthy controls (HCs). Mononuclear cells were stimulated with Porphyromonas gingivalis, Fusobacterium nucleatum, Staphylococcus epidermidis, or Cutibacterium acnes, and B-cell production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, interferon (IFN)-γ, IL-10 and transforming growth factor (TGF)-β by B cells was assessed by flow cytometry.. HCs had higher baseline frequencies of B cells producing IFN-γ or TNF-α than grade B patients, but only B cells from grade B patients showed significant differentiation into IFN-γ-, TNF-α-, TGF-β-, or IL-10-producing cells after challenge with P. gingivalis and into IFN-γ-, TGF-β-, or IL-10-producing cells after challenge F. nucleatum. Notably, the baseline frequency of IL-10-producing B cells from grade C patients correlated inversely with clinical attachment loss (AL). The major proportion of the IFN-γ- and TGF-β-producing B cells were CD27. B cells from grade B patients, particularly those harboring P. gingivalis, showed proinflammatory B-cell responses to P. gingivalis. Moreover, the baseline frequency of IL-10-producing B cells in the grade C group correlated inversely with AL, suggesting a diminished immunoregulatory capacity of IL-10-producing B cells in these patients.

Topics: Cytokines; Humans; Interleukin-10; Interleukin-6; Periodontitis; Porphyromonas gingivalis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Antibacterial photodynamic treatment (aPDT) has indispensable significance as a means of treating periodontal disorders because of its extraordinary potential for killing pathogenic bacteria by generating an overpowering amount of reactive oxygen species (ROS). The elevated ROS that may result from the antibacterial treatment procedure, however, could exert oxidative pressure inside periodontal pockets, causing irreparable damage to surrounding tissue, an issue that has severely restricted its medicinal applications. Accordingly, herein, we report the use of black phosphorus nanosheets (BPNSs) that can eliminate the side effects of ROS-based aPDT as well as scavenge ROS to produce an antibacterial effect.. The antibacterial effect of ICG/aPDT was observed by direct microscopic colony counting. A microplate reader and confocal microscope enabled measurements of cell viability and the quantification of ROS fluorescence. BPNS administration regulated the oxidative environment. IL-1β, IL-6, TNF-α, IL-10, TGF-β, and Arg-1 mRNA expression levels were used to assess the inflammatory response after BPNS treatment. In vivo, the efficacy of the combination of BPNSs and ICG/aPDT was evaluated in rats with periodontal disease by histomorphometric and immunohistochemical analyses.. The CFU assay results verified the antibacterial effect of ICG/aPDT treatment, and ROS fluorescence quantification by CLSM indicated the antioxidative ability of the BPNSs. IL-1β, IL-6, TNF-α, IL-10, TGF-β, and Arg-1 mRNA expression levels were significantly decreased after BPNS treatment, confirming the in vitro anti-inflammatory effect of this nanomaterial. The histomorphometric and immunohistochemical analyses showed that the levels of proinflammatory factors decreased, suggesting that the BPNSs had anti-inflammatory effects in vivo.. Treatment with antioxidative BPNSs gives new insights into future anti-inflammatory therapies for periodontal disease and other infection-related inflammatory illnesses and provides an approach to combat the flaws of aPDT.

Topics: Animals; Anti-Bacterial Agents; Interleukin-10; Interleukin-6; Periodontal Diseases; Periodontitis; Photochemotherapy; Photosensitizing Agents; Rats; Reactive Oxygen Species; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Periodontitis is a chronic inflammatory disease characterized by the destruction of tooth-supporting tissues. The gingival epithelium is the first barrier of periodontal tissue against oral pathogens and harmful substances. The structure and function of epithelial lining are essential for maintaining the integrity of the epithelial barrier. Abnormal apoptosis can lead to the decrease of functional keratinocytes and break homeostasis in gingival epithelium. Interleukin-22 is a cytokine that plays an important role in epithelial homeostasis in intestinal epithelium, inducing proliferation and inhibiting apoptosis, but its role in gingival epithelium is poorly understood. In this study, we investigated the effect of interleukin-22 on apoptosis of gingival epithelial cells during periodontitis. Interleukin-22 topical injection and Il22 gene knockout were performed in experimental periodontitis mice. Human gingival epithelial cells were co-cultured with Porphyromonas gingivalis with interleukin-22 treatment. We found that interleukin-22 inhibited apoptosis of gingival epithelial cells during periodontitis in vivo and in vitro, decreasing Bax expression and increasing Bcl-xL expression. As for the underlying mechanisms, we found that interleukin-22 reduced the expression of TGF-β receptor type II and inhibited the phosphorylation of Smad2 in gingival epithelial cells during periodontitis. Blockage of TGF-β receptors attenuated apoptosis induced by Porphyromonas gingivalis and increased Bcl-xL expression stimulated by interleukin-22. These results confirmed the inhibitory effect of interleukin-22 on apoptosis of gingival epithelial cells and revealed the involvement of TGF-β signaling pathway in gingival epithelial cell apoptosis during periodontitis.

Topics: Animals; Apoptosis; Epithelial Cells; Gingiva; Humans; Interleukin-22; Mice; Periodontitis; Porphyromonas gingivalis; Signal Transduction; Transforming Growth Factor beta

This study aimed to examine the levels of transforming growth factor-beta (TGF-β) and inhibitory-Smads (I-Smads) in saliva and gingival crevicular fluid (GCF) in patients with Stage 3 Grade B periodontitis before and after non-surgical periodontal treatment.. Recently, it has been stated that Smads play an active role in all conditions where TGF-β is involved, including periodontal inflammation.. Twenty healthy participants (control) and 20 patients with Stage 3, Grade B periodontitis were recruited. GCF and saliva samples and clinical periodontal recordings were investigated at the baseline and 1 month after treatment. TGF-β and I-Smads (Smads 6 and 7) were determined by ELISA.. Salivary Smad6 and Smad7 levels were significantly lower in the periodontitis group than healthy controls (p < .05), while there was no difference in salivary TGF-β levels between groups at baseline (p > .05). The total amounts and concentrations of GCF TGF-β, Smad6, and Smad7 were significantly lower in the periodontitis group than healthy controls at baseline (p < .05), and then decreased in concentration levels with treatment (p < .001). Positive correlations were found between total amounts and concentrations of GCF TGF-β, Smad6, and Smad7 (p < .05).. Our findings revealed that Smad6 and Smad7 in GCF and saliva decreased in periodontitis and then increased after periodontal treatment. Our study suggests that I-Smads act in parallel with TGF-β in periodontal inflammation and may have a role in the development of periodontitis.

Topics: Gingival Crevicular Fluid; Humans; Inflammation; Periodontitis; Saliva; Transforming Growth Factor beta

Periodontitis (PD) is the sixth most prevalent disease around the world and is involved in the development and progression of multiple systemic diseases. Previous studies have reported that PD may aggravate liver injuries. The objective of this study was to investigate whether and how PD affects liver fibrosis.. Ligature-induced PD (LIP) was induced in male C57/B6J mice, and sub-gingival plaques (PL) from patients with PD were applied to mouse teeth. Liver fibrosis was induced by carbon tetrachloride (CCl. Mice in the CCl. PD aggravates CCl

Topics: Actins; Alanine Transaminase; Animals; Azo Compounds; Carbon Tetrachloride; Hydroxyproline; Liver Cirrhosis; Male; Methylene Blue; Mice; Microbiota; Periodontitis; RNA, Ribosomal, 16S; Transforming Growth Factor beta

Host immunity is crucial during periodontal inflammations. B cells are considered to have a function of immunoregulation, and TLRs are considered to be crucial in this process. The present study illustrates the potential roles and rules of CD25+ B cells during periodontitis, especially its effect on regulating host IL-35 level and Th1, Th17, and Treg differentiation.. The proportion of local and systemic CD25+ B cell subpopulations from periodontitis models were identified by flow cytometry. To illustrate further mechanism, B cells were cultured with a different type of TLR activators. Expression of IL-10, IL-35, and TGF-β was detected by ELISA and real-time PCR. We also set adoptive transfer models by using CD25+ B cells. Alveolar bone erosion, proportion of Th1, Th17, and Tregs, and levels of IFN-γ, TNF-α, IL-1β, and IL-17 were identified.. Periodontitis induces more CD25+ B cell subpopulations and promotes their IL-10, IL-35, and TGF-βproduction. TLR activators enhanced Breg proliferation and function. LPS+CpG obviously induced more CD25+ B cell differentiation and production of IL-10, IL-35, and TGF-β. Adoptive transfer of CD25+ B cells reduces alveolar bone destruction and local Tregs, proportion, especially the local level of IFN-γ and IL-17. In addition, adoptive transfer of CD25+ B cells remedies the pathological change in the proportion of IL-1β and Th1/Th17 in local lesions. We did not find any significant difference in peripheral blood, regardless of group and detected items.. Results of the present study clarify that CD25+ B cells enlarged and produced more IL-10, IL-35 and TGF-β during periodontitis, activation of TLR4 and TLR9 played crucial roles in this process. Also, CD25+ B cells alleviated periodontal inflammation and alveolar bone resorption. Our findings further expanded the potential of B cells during periodontitis.

Topics: Alveolar Bone Loss; Humans; Inflammation; Interleukin-10; Interleukin-17; Lipopolysaccharides; Periodontitis; T-Lymphocytes, Regulatory; Toll-Like Receptor 4; Toll-Like Receptor 9; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Inflammatory imbalance of bone formation/resorption leads to alveolar bone destruction. Astragalus polysaccharide has been confirmed to have anti-inflammatory effects. We sought to disclose the protective effect and its potential mechanisms of astragalus polysaccharide in the periodontitis model. Experimental periodontitis was induced by cotton ligatures for this study. We measured the alveolar bone damage rate, periodontal osteoclasts, proportion of CD4+Foxp3+, CD4+IL-10+, CD4+TGF-β+ subsets in the gingiva, and RANKL, OPG, TGF-β+, and IL-10+ level in the gingiva. We also cultured osteoclast precursor cells in the presence of RANKL and astragalus polysaccharide. Osteoclasto-like cells were identified by TRAP staining, mRNA of RANK, TRAP, and TRAF6 were evaluated by real time PCR. We found that astragalus polysaccharide caused significant protection of the alveolar bone via reducing local osteoclasts. It also decreased the proportion of CD4+Foxp3+ cells and upregulated the level of CD4+IL-10+ cells, reduced RANKL, and remedied IL-10 levels. In cell culture experiments, astragalus polysaccharide prohibited the RANKL mediated osteoclast differentiation. The findings of this study disclose the functions and possible mechanisms of astragalus polysaccharide engaged in local osteoclastogenesis, and reveal the considerable effect of astragalus polysaccharide in alveolar bone homeostasis and its likely contribution to host immuno-regulation in periodontitis.

Topics: Animals; Astragalus Plant; Bone Resorption; Forkhead Transcription Factors; Interleukin-10; Osteogenesis; Periodontitis; Polysaccharides; Rats; Transforming Growth Factor beta

The balance between pro- and anti-inflammatory signals maintains tissue homeostasis and defines the outcome of chronic inflammatory diseases such as periodontitis, a condition that afflicts the tooth-supporting tissues and exerts an impact on systemic health. The induction of tissue inflammation relies heavily on Toll-like receptor (TLR) signaling, which drives a proinflammatory pathway through recruiting myeloid differentiation primary response gene 88 (MyD88) and activating nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB). TLR-induced production of proinflammatory cytokines and chemokines is reined in by anti-inflammatory cytokines, including the transforming growth factor β (TGFβ) family of cytokines. Although Smad6 is a key mediator of TGFβ-induced anti-inflammatory signaling, the exact mechanism by which TGFβ regulates TLR proinflammatory signaling in the periodontal tissue has not been addressed to date. In this study, we demonstrate for the first time that the ability of TGFβ to inhibit TLR-NFκB signaling is mediated by protein arginine methyltransferase 1 (PRMT1)-induced Smad6 methylation. Upon methylation, Smad6 recruited MyD88 and promoted MyD88 degradation, thereby inhibiting NFκB activation. Most important, Smad6 is expressed and methylated in the gingival epithelium, and PRMT1-Smad6 signaling promotes tissue homeostasis by limiting inflammation. Consistent with this, disturbance of Smad6 methylation exacerbates inflammation and bone loss in experimental periodontitis. The dissected mechanism is therapeutically important, as it highlights the manipulation of PRMT1-Smad6 signaling as a novel promising strategy to modulate the host immune response in periodontitis.

Topics: Arginine; Cells, Cultured; Gingiva; Humans; Inflammation; Methylation; Myeloid Differentiation Factor 88; NF-kappa B; Periodontitis; Protein Interaction Domains and Motifs; Protein-Arginine N-Methyltransferases; Repressor Proteins; Signal Transduction; Smad6 Protein; Transforming Growth Factor beta; Ubiquitin-Protein Ligases

Topics: Alveolar Bone Loss; Animals; Gingiva; Interleukin-1beta; Interleukin-6; Myeloid Cells; Periodontitis; Porphyromonas gingivalis; Rats; Rats, Sprague-Dawley; RNA, Messenger; Transforming Growth Factor beta; Triggering Receptor Expressed on Myeloid Cells-1; Tumor Necrosis Factor-alpha

Periodontitis is an inflammatory disease of the supporting tissues of the teeth. Interleukin (IL)-13 is a multifunctional T-helper type2 (Th2) cytokine that can diminish inflammatory responses. I investigated using ELISA the effects of IL-13 on transforming growth factor-beta (TGF-β) and matrix metalloproteinase-1 (MMP-1). MMP-1 was detected using immunohistochemistry. Gingival fibroblasts were stimulated with IL-13 or together with tumor necrosis factor-α (TNF-α). I found that macrophage-like cells, fibroblast-like cells, vascular endothelial cells and gingival epithelial cells were stained more intensely for MMP-1 and were observed more frequently in the periodontitis affected group than in the control group. The cultured gingival fibroblasts with IL-13 produced more TGF-β than unstimulated cells. After stimulation with additional TNF-α, MMP-1 production was diminished. IL-13 may play a role in regulating collagen homeostasis in gingival fibroblasts. IL-13 induces both up-regulation of TGF-β, a cytokine known to stimulate production of collagen, and down-regulation of collagen-destroying MMP-1 production. This effect may be strong during periodontitis when Th2 cells assist T cells.

Topics: Adjuvants, Immunologic; Adult; Cells, Cultured; Fibroblasts; Gene Expression Regulation; Humans; Immunohistochemistry; Interleukin-13; Matrix Metalloproteinase 1; Periodontitis; Staining and Labeling; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Young Adult

The dysbiotic microbiota associated with aggressive periodontitis includes Aggregatibacter actinomycetemcomitans, the only oral species known to produce a cytolethal distending toxin (AaCDT). Give that CDT alters the cytokine profile in monocytic cells, we aimed to test the hypothesis that CDT plays a role in bone homeostasis by affecting the differentiation of precursor cells into osteoclasts. Recombinant AaCDT was added to murine bone marrow monocytes (BMMC) in the presence or absence of RANKL and the cell viability and cytokine profile of osteoclast precursor cells were determined. Multinucleated TRAP(+) cell numbers, and relative transcription of genes related to osteoclastogenesis were also evaluated. The addition of AaCDT did not lead to loss in cell viability but promoted an increase in the average number of TRAP(+) cells with 1-2 nuclei in the absence or presence of RANKL (Tukey, p < 0.05). This increase was also observed for TRAP(+) cells with ≥3nuclei, although this difference was not significant. Levels of TGF-β, TNF-α, and IL-6, in the supernatant fraction of cells, were higher when in AaCDT exposed cells, whereas levels of IL-1β and IL-10 were lower than controls under the same conditions. After interaction with AaCDT, transcription of the rank (encoding the receptor RANK), nfatc1 (transcription factor), and ctpK (encoding cathepsin K) genes was downregulated in pre-osteoclastic cells. The data indicated that despite the presence of RANKL and M-CSF, AaCDT may inhibit osteoclast differentiation by altering cytokine profiles and repressing transcription of genes involved in osteoclastogenesis. Therefore, the CDT may impair host defense mechanisms in periodontitis.

Topics: Acid Phosphatase; Aggregatibacter actinomycetemcomitans; Animals; Bone Marrow Cells; Bone Resorption; Cathepsin K; Cell Differentiation; Cell Survival; Cells, Cultured; Interleukin-10; Interleukin-1beta; Interleukin-6; Isoenzymes; Macrophages; Male; Mice; Mice, Inbred C57BL; Monocytes; NFATC Transcription Factors; Osteoclasts; Pasteurellaceae Infections; Periodontitis; RANK Ligand; Tartrate-Resistant Acid Phosphatase; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Marfan syndrome (MFS) is a systemic connective tissue disorder that is caused by mutations of fibrillin-1. While MFS patients are at a high risk of periodontitis and aortic diseases, little causal information has been provided to date. To clarify the relationship, their oral condition and sinus of Valsalva (SoV) were evaluated.The subjects were patients with MFS (n = 33) who attended the University of Tokyo Hospital. We divided them into two groups; MFS patients with highly dilated (the diameters were equal to or more than 39 mm) SoV (high group, n = 18) and MFS patients with mildly dilated (less than 39 mm) SoV (mild group, n = 15). Blood examinations, echocardiograms, and full-mouth clinical measurements, including number of teeth, probing pocket depth (PPD), bleeding on probing (BOP), and community periodontal index (CPI) were performed.We found that the high group patients had greater rates of BOP compared to that of the mild group. Furthermore, the high group tended to have higher serum levels of C-reactive protein, matrix metalloproteinase-9, and transforming growth factor-β compared to the mild group.Periodontitis may deteriorate SoV dilatation in MFS patients.

Topics: Adult; Biomarkers; C-Reactive Protein; Dilatation, Pathologic; Female; Hospitals, University; Humans; Japan; Male; Marfan Syndrome; Matrix Metalloproteinase 9; Periodontal Index; Periodontitis; Predictive Value of Tests; Sensitivity and Specificity; Sinus of Valsalva; Transforming Growth Factor beta

The interaction between human gingival fibroblasts (HGFs) and Porphyromonas gingivalis plays an important role in the development and progression of periodontitis. Porphyromonas gingivalis possesses several virulence factors, including cysteine proteases, the arginine-specific (Rgp) and lysine-specific (Kgp) gingipains. Studying the mechanisms that P. gingivalis, and its derived virulence, use to propagate and interact with host cells will increase the understanding of the development and progression of periodontitis. In this study, we aimed to elucidate how P. gingivalis influences the inflammatory events in HGFs regarding transforming growth factor-β1 (TGF-β1 ), CXCL8, secretory leucocyte protease inhibitor (SLPI), c-Jun and indoleamine 2,3-dioxygenase (IDO). HGFs were inoculated for 6 and 24 h with the wild-type strains ATCC 33277 and W50, two gingipain-mutants of W50 and heat-killed ATCC 33277. The P. gingivalis regulated CXCL8 and TGF-β1 in HGFs, and the kgp mutant gave significantly higher immune response with increased CXCL8 (P < 0.001) and low levels of TGF-β1 . We show that HGFs express and secrete SLPI, which was significantly suppressed by P. gingivalis (P < 0.05). This suggests that by antagonizing SLPI, P. gingivalis contributes to the tissue destruction associated with periodontitis. Furthermore, we found that P. gingivalis inhibits the expression of the antimicrobial IDO, as well as upregulating c-Jun (P < 0.05). In conclusion, P. gingivalis both triggers and suppresses the immune response in HGFs. Consequently, we suggest that the pathogenic effects of P. gingivalis, and especially the activity of the gingipains on the inflammatory and immune response of HGFs, are crucial in periodontitis.

Topics: Adhesins, Bacterial; Cells, Cultured; Cysteine Endopeptidases; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gingipain Cysteine Endopeptidases; Gingiva; Humans; Indoleamine-Pyrrole 2,3,-Dioxygenase; Inflammation; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Proto-Oncogene Proteins c-jun; Real-Time Polymerase Chain Reaction; Secretory Leukocyte Peptidase Inhibitor; Transforming Growth Factor beta

Periodontitis is the most prevalent infectious disease caused by periodontopathic bacteria and is also a chronic inflammatory disease. Gingival crevicular fluid (GCF) is an inflammatory exudate that seeps into the gingival crevices or periodontal pockets around teeth with inflamed gingiva, and contains various materials including leukocytes and cytokines. Since gingival epithelial cells, which form a barrier against bacterial challenges, are affected by GCF, cytokines or other materials contained within GCF are engaged in the maintenance and disruption of the epithelial barrier. Accordingly, its compositional pattern has been employed as a reliable objective index of local inflammation. Transforming growth factor β1 (TGF-β1) levels in GCF were previously shown to be markedly higher in patients with periodontitis than in healthy subject. However, it currently remains unclear how TGF-β1 affects gingival epithelial cell growth or apoptosis; therefore, elucidating the mechanism responsible may lead to a deeper understanding of pathogenic periodontitis. In the present study, the human gingival epithelial cell line, OBA9 cells were stimulated with recombinant TGF-β1. Apoptosis-related protein levels were determined by Western blotting. Caspase-3/7 activity was measured with a Caspase-Glo assay kit. Surviving and apoptotic cells were detected using an MTS assay and TUNEL staining, respectively. TGF-βRI siRNA and smad2 siRNA were transfected into cells using the lipofectamine RNAiMAX reagent. TGF-β1 elevated caspase-3 activity and the number of TUNEL-positive apoptotic cells in OBA9 cells. Furthermore, while the levels of the pro-apoptotic proteins Bax, Bak, Bim, and Bad were increased in OBA9 cells stimulated with TGF-β1, the TGF-β1 treatment also decreased the levels of anti-apoptotic proteins such as Bcl-2 and Bcl-xL in a time-dependent manner. Additionally, TGF-β1 up-regulated the protein levels of cleaved caspase-9. These results indicated that TGF-β1-induced apoptosis was involved in a mitochondria-related intrinsic pathway. TGF-β1 phosphorylated smad2 in OBA9 cells and this phosphorylation was clearly reduced by SB431542 (a TGF-β type I receptor inhibitor). Consistent with this result, SB431542 or smad2 siRNA-induced reductions in smad2 protein expression levels attenuated TGF-β1-induced apoptosis. On the other hand, the ligation of TGF-β1 on its receptor also stimulated the phosphorylation of Erk and Akt, which are smad2-independent pathways. However, the

Topics: Acetylcysteine; Apoptosis; Benzamides; Caspase 3; Caspase 7; Cell Line; Cell Line, Tumor; Cell Proliferation; Cell Survival; Dioxoles; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gingiva; Humans; Inflammation; MAP Kinase Signaling System; Periodontitis; Phosphorylation; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Signal Transduction; Smad2 Protein; Transforming Growth Factor beta; Transforming Growth Factor beta1

Multiple studies have shown that diabetes mellitus is an established risk factor for periodontitis. Recently mesenchymal stem cells derived from periodontal ligament (PDLSCs) have been utilized to reconstruct tissues destroyed by chronic inflammation. However, impact of periodontitis with diabetes mellitus on PDLSCs and mechanisms mediating effects of complex microenvironments remain poorly understood. In this study, we found multiple differentiation potential of PDLSCs from chronic periodontitis with diabetes mellitus donors (D-PDLSCs) was damaged significantly. Inhibition of NF-κB signaling could rescue osteogenic potential of PDLSCs from simple chronic periodontitis patients (P-PDLSCs), whereas did not promote D-PDLSCs osteogenesis. In addition, we found expression of DKK1 in D-PDLSCs did not respond to osteogenic signal and decreased osteogenic potential of D-PDLSCs treated with DKK1 could be reversed. To further elucidate different character between P-PDLSCs and D-PDLSCs, we treated PDLSCs with TNF-α and advanced glycation end products (AGEs), and find out AGEs which enhance effect of TNF-α in PDLSCs might mediate special personality of D-PDLSCs. The adverse effect of AGEs in PDLSCs could be reversed when PDLSCs were treated with DKK1. These results suggested DKK1 mediating WNT signaling might be a therapy target to rescue potential of PDLSCs in periodontitis with diabetes mellitus.

Topics: Adult; Aged; Animals; beta Catenin; Cell Differentiation; Cells, Cultured; Core Binding Factor Alpha 1 Subunit; Diabetes Mellitus, Type 2; Glycation End Products, Advanced; Humans; Intercellular Signaling Peptides and Proteins; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred NOD; Mice, SCID; Middle Aged; NF-kappa B; Osteogenesis; Periodontal Ligament; Periodontitis; Transforming Growth Factor beta

Variations in the expression of cytokines during the progression of periodontitis remain ill-defined. We evaluated the expression of 19 cytokine genes related to T-cell phenotype/function during initiation, progression and resolution of periodontitis, and related these to the expression of soft and bone tissue destruction genes (TDGs).. A ligature-induced periodontitis model was used in rhesus monkeys (M. mulatta) (n = 18). Gingival tissues were taken at baseline pre-ligation, 2 weeks and 1 month (Initiation) and 3 months (progression) post ligation. Ligatures were removed and samples taken 2 months later (resolution). Total RNA was isolated and the Rhesus Gene 1.0 ST (Affymetrix) used for gene expression analysis. Significant expression changes were validated by qRT-PCR.. Disease initiation/progression was characterized by overexpression of Th17/Treg cytokine genes (IL-1β, IL-6, TGFβ and IL-21) and down-regulation of Th1/Th2 cytokine genes (IL-18 and IL-25). Increased IL-2 and decreased IL-10 levels were seen during disease resolution. Several Th17/Treg cytokine genes positively correlated with TDGs, whereas most Th1/Th2 genes exhibited a negative correlation.. Initiation, progression and resolution of periodontitis involve over- and underexpression of cytokine genes related to various T-helper subsets. In addition, variations in individual T-helper response subset/genes during disease progression correlated with protective/destructive outcomes.

Topics: Animals; Cathepsin K; Cytokines; Disease Models, Animal; Disease Progression; Female; Gene Expression Profiling; Interleukin-10; Interleukin-17; Interleukin-18; Interleukin-1beta; Interleukin-2; Interleukin-6; Interleukins; Macaca mulatta; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Periodontitis; RANK Ligand; T-Lymphocytes; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells; Transforming Growth Factor beta

Marfan syndrome is an autosomal dominant disease characterized by aneurysm and dilatation of the aortic root, tall stature, and ectopia lentis. These manifestations reflect excessive signaling of transforming growth factor beta (TGF-β). Moreover, cases are frequently associated with severe periodontitis, which is a chronic inflammation of the gingiva, periodontal ligament, and alveolar bone. Recently, angiotensin II receptor blockers (ARBs) were discovered to be an effective drug class that can prevent aortic aneurysm and dilation in Marfan syndrome by inhibiting TGF-β signaling. To investigate the effect of ARB on the progression of periodontitis, the application of a potent ARB, telmisartan, was examined in a mouse model of Marfan syndrome (MgΔ). Six-week-old male heterozygous MgΔ and wild-type mice were challenged with Porphyromonas gingivalis, which causes chronic periodontitis, with and without telmisartan application. After infection, alveolar bone resorption was measured by micro-computed tomography (μCT), and inflammatory cytokine levels were examined. Infection of Porphyromonas gingivalis induced alveolar bone resorption in both MgΔ and wild-type mice. The amount of resorption was significantly larger in the former than the latter. Immunoarray and enzyme-linked immunosorbent assay (ELISA) analyses demonstrated that interleukin-17 (IL-17) and tumor necrosis factor alpha (TNF-α) levels were significantly higher in infected MgΔ mice than infected wild-type mice. Telmisartan treatment significantly suppressed the alveolar bone resorption of infected MgΔ mice. Telmisartan also significantly decreased levels of TGF-β, IL-17, and TNF-α in infected MgΔ mice to levels seen in infected wild-type mice. This study suggests that ARB can prevent the severe periodontitis frequently seen in Marfan syndrome.

Topics: Alveolar Bone Loss; Angiotensin Receptor Antagonists; Animals; Bacteroidaceae Infections; Benzimidazoles; Benzoates; Bone Resorption; Disease Models, Animal; Inflammation; Interleukin-17; Male; Marfan Syndrome; Mice; Osteoclasts; Periodontitis; Porphyromonas gingivalis; Telmisartan; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Periodontitis is a polymicrobial oral infection characterized by the destruction of tooth-supporting structures that can be linked to systemic diseases such as cardiovascular disease, diabetes or rheumatoid arthritis. Porphyromonas gingivalis, a bacterium implicated in the etiology of periodontitis, has shown variation in inducing T-cell responses among different strains. Therefore, in this study we investigated the strain-specific immune response using a murine experimental model of periodontitis. Periodontitis was induced by P. gingivalis strains A7A1-28, W83 and W50, and later confirmed by the presence of P. gingivalis in the oral microflora and by alveolar bone resorption. Splenocytes were evaluated for gene expression, cellular proteins and cytokine expression. Dendritic cells were stimulated in vitro for T helper cell-cytokine profiling. Results showed that P. gingivalis had the ability to alter the systemic immune response after bacterial exposure. Strains W50 and W83 were shown to induce alveolar bone loss, whereas the A7A1-28 strain did not significantly promote bone resorption in mice. Splenocytes derived from mice infected with strains W50 and W83 induced expression of high levels of interleukin-4 (IL-4) but A7A1-28 stimulated increased IL-10. Stimulation of dendritic cells in vitro showed a similar pattern of cytokine expression of IL-12p40, IL-6 and transforming growth factor-β among strains. A distinct systemic response in vivo was observed among different strains of P. gingivalis, with IL-10 associated with the least amount of alveolar bone loss. Evaluation of pathogen-driven systemic immune responses associated with periodontal disease pathogenesis may assist in defining how periodontitis may impact other diseases.

Topics: Alveolar Bone Loss; Animals; Bacteroidaceae Infections; Cytokines; Dendritic Cells; Disease Models, Animal; Flow Cytometry; Gene Expression Regulation, Bacterial; Interleukin-10; Interleukin-12 Subunit p40; Interleukin-4; Interleukin-6; Male; Mice; Mice, Inbred DBA; Mice, Inbred Strains; Periodontitis; Porphyromonas gingivalis; Spleen; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta; X-Ray Microtomography

The underlying mechanism and the therapeutic regimen for the transition of reversible gingivitis to irreversible periodontitis are unclear. Since transforming growth factor (TGF)-β has been implicated in differentially regulated gene expression in gingival fibroblasts, we hypothesized that TGF-β signaling is activated in periodontitis-affected gingiva, along with enhanced collagen degradation, that is reversed by TGF-β inhibition. A novel three-dimensional (3D) gel-culture system consisting of primary human gingival fibroblasts (GF) and gingival epithelial (GE) cells in collagen gels was applied. GF populations from patients with severe periodontitis degraded collagen gels, which was reduced by TGF-β-receptor kinase inhibition. Up-regulation of TGF-β-responsive genes was evident in GF/GE co-cultures. Furthermore, the TGF-β downstream transducer Smad3C was highly phosphorylated in periodontitis-affected gingiva and 3D cultures. These results imply that TGF-β signaling is involved in fibroblast-epithelial cell interaction in periodontitis, and suggest that the 3D culture system is a useful in vitro model for therapeutic drug screening for periodontitis.

Topics: Adult; Aprotinin; Cell Culture Techniques; Coculture Techniques; Collagen; Culture Media; Epithelial Cells; Fibroblasts; Gels; Gene Expression Regulation; Gingiva; Humans; Hydroxamic Acids; Matrix Metalloproteinase 3; Matrix Metalloproteinase Inhibitors; Periodontitis; Phosphorylation; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Serine Proteinase Inhibitors; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta; Up-Regulation

Previous studies reported that bone morphogenetic protein (BMP)-2 induced periodontal regeneration in animals. However, the effects of local host factors on bone formation when using recombinant human (rh)BMP-2 are unknown. The purpose of this study was to evaluate local conditions in recipient sites that affected periodontal regeneration following BMP implantation in experimentally induced horizontal defects in dogs.. Experimental periodontitis was induced in the maxillary and mandibular premolars of six male beagles. The recipient sites were divided into four quadrants (maxillary buccal, maxillary palatal, mandibular buccal, and mandibular lingual sites). A polymer-coated gelatin sponge (PGS; 3 x 3 x 2 mm) was impregnated with 7.2 mul rhBMP-2 solution. The recipient sites in each quadrant were treated with physiologic saline/PGS and rhBMP-2/PGS (rhBMP-2 at 7.2 mug/7.2 mul). The dogs were sacrificed 12 weeks post-surgery, and healing was evaluated histologically.. Bone formation and connective tissue attachment were observed following rhBMP-2/PGS implantation. In the four recipient sites of the BMP-treated group, significant positive correlations were found between the width of residual bone and the height and area of regenerated bone (r = 0.791; r = 0.828; P <0.0001). The height of regenerated bone was also significantly correlated to the area (r = 0.891; P <0.0001).. The width of residual bone was one of the clinical host factors that affected bone regeneration following BMP implantation. However, it did not affect connective tissue attachment, cementum regeneration, and downgrowth of junctional epithelium.

Topics: Alveolar Bone Loss; Alveolar Process; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Connective Tissue; Dental Cementum; Dogs; Drug Carriers; Epithelial Attachment; Gelatin Sponge, Absorbable; Gingiva; Guided Tissue Regeneration, Periodontal; Humans; Male; Mandible; Maxilla; Osteogenesis; Periodontal Attachment Loss; Periodontitis; Recombinant Proteins; Root Planing; Transforming Growth Factor beta; Wound Healing

Periodontitis was shown to have an impact on glucose levels in prediabetic and diabetic rats. The Zucker fatty rat (ZFR) is a well-characterized model of prediabetes presenting with impaired glucose tolerance, hyperinsulinemia, dyslipidemia, and moderate hypertension. The aim of the present study was to investigate whether periodontitis influences kidney changes in ZFRs.. Male adult ZFRs (N = 19) and their lean littermates (N = 18) were studied. Periodontitis was induced with ligatures in half of the ZFRs and lean rats, whereas the other half served as controls. After 4 weeks, the rats were sacrificed, and the kidneys, liver, and heart were removed and weighed. Kidneys were evaluated histologically for glomerular volume and renal mRNA levels of vascular endothelial growth factor (VEGF), VEGF receptor 2, transforming growth factor-beta, connective tissue growth factor, collagen IValpha1, fibronectin, and nephrin. Urinary albumin excretion and creatinine clearance were also evaluated.. In prediabetic ZFRs, periodontitis was associated with kidney hypertrophy (P = 0.03) and a tendency for increased glomerular volume (P = 0.06). In lean littermates, elevated fibronectin mRNA levels (P = 0.03) were noted in the presence of periodontitis.. Our findings suggest the participation of periodontitis in the development of early renal changes in ZFRs.

Topics: Albuminuria; Animals; Collagen Type IV; Connective Tissue Growth Factor; Creatinine; Fibronectins; Hypertrophy; Immediate-Early Proteins; Insulin-Like Growth Factor Binding Proteins; Intercellular Signaling Peptides and Proteins; Kidney; Kidney Diseases; Kidney Glomerulus; Male; Membrane Proteins; Organ Size; Periodontitis; Prediabetic State; Random Allocation; Rats; Rats, Zucker; RNA, Messenger; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2

The responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis has been found to play a significant role for susceptibility and resistance to periodontal disease. In the present study we have investigated the effects of two different treatment strategies, which have been found to down-regulate the HPA axis, on ligature-induced periodontitis.. In experiment 1, newborn rats were treated with the synthetic glucocorticoid hormone dexamethasone-21-phosphate, which permanently down-regulates HPA axis responsiveness. In experiment 2, adult rats were treated with the novel antidepressant drug tianeptine, which opposes the action of stress. Periodontitis was inflicted upon all rats. Just before decapitation the animals received gram-negative bacterial lipopolysaccharide (LPS) to induce a robust immune and HPA axis response.. Compared to the saline-treated control rats, dexamethasone-treated rats had significantly less periodontal bone loss (p < 0.01), reduced expression of glucocorticoid receptors in the hippocampus (p < 0.001), lower corticosterone (p=0.01) and higher plasma levels of the cytokine tumor necrosis factor (TNF)-alpha (p < 0.05) after LPS challenge. Also the tianeptine-treated rats showed significantly reduced periodontal bone loss (p=0.01), enhanced plasma levels of TNF-alpha (p < 0.05), and transforming growth factor-1beta (p < 0.01), whereas no significant difference was found in corticosterone levels.. An individual's responsiveness to danger signals, whether they are of immunological, chemical, or psychological origin, may be an important factor for explaining variability in susceptibility to periodontal disease. The results may provide new insight into the mechanisms of periodontal disease development, and open new vistas for disease prevention.

Topics: Age Factors; Alveolar Bone Loss; Animals; Animals, Newborn; Anti-Inflammatory Agents; Antidepressive Agents, Tricyclic; Dexamethasone; Disease Susceptibility; Down-Regulation; Glucocorticoids; Gram-Negative Bacteria; Hypothalamo-Hypophyseal System; Lipopolysaccharides; Periodontitis; Pituitary-Adrenal System; Rats; Rats, Wistar; Thiazepines; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

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

Bone morphogenetic protein (BMP) is a potent differentiating agent for cells of the osteoblastic lineage. It has been used in the oral cavity under a variety of indications and with different carriers. However, the optimal carrier for each indication is not known. This study examined a synthetic bioabsorbable carrier for BMP used in osseous defects around dental implants in the canine mandible.. Twelve canines had their mandibular four premolars and first molar teeth extracted bilaterally. After 5 months, four implants were placed with standardized circumferential defects around the coronal 4 mm of each implant. One-half of the defects received a polylactide/glycolide (PLGA) polymer carrier with or without recombinant human BMP-2 (rhBMP-2), and the other half received a collagen carrier with or without rhBMP-2. Additionally, one-half of the implants were covered with a non-resorbable (expanded polytetrafluoroethylene [ePTFE]) membrane to exclude soft tissues. Animals were sacrificed either 4 or 12 weeks later. Histomorphometric analysis included the percentage of new bone contact with the implant, the area of new bone, and the percentage of defect fill. This article describes results with the PLGA carrier.. All implants demonstrated clinical and radiographic success with the amount of new bone formed dependent on the time and presence/absence of rhBMP-2 and presence/absence of a membrane. The percentage of bone-to-implant contact was greater with rhBMP-2, and after 12 weeks of healing, there was approximately one-third of the implant contacting bone in the defect site. After 4 weeks, the presence of a membrane appeared to slow new bone area formation. The percentage of fill in membrane-treated sites with rhBMP-2 rose from 24% fill to 42% after 4 and 12 weeks, respectively. Without rhBMP-2, the percentage of fill was 14% rising to 36% fill, respectively.. After 4 weeks, the rhBMP-2-treated sites had a significantly higher percentage of contact, more new bone area, and higher percentage of defect fill than the sites without rhBMP-2. After 12 weeks, there was no significant difference in sites with or without rhBMP-2 regarding percentage of contact, new bone area, or percentage of defect fill. In regard to these three outcomes, comparing the results with this carrier to the results reported earlier with a collagen carrier in this study, only the area of new bone was significantly different with the collagen carrier resulting in greater bone than the PLGA carrier. Thus, the PLGA carrier for rhBMP-2 significantly stimulated bone formation around dental implants in this model after 1 month but not after 3 months of healing. The use of this growth factor and carrier combination appears to stimulate early bone healing events around the implants but not quite to the same degree as a collagen carrier.

Topics: Absorbable Implants; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Collagen; Dental Implantation, Endosseous; Dental Implants; Dogs; Drug Carriers; Guided Tissue Regeneration, Periodontal; Humans; Implants, Experimental; Lactic Acid; Male; Membranes, Artificial; Models, Animal; Osseointegration; Periodontitis; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Polytetrafluoroethylene; Recombinant Proteins; Statistics, Nonparametric; Transforming Growth Factor beta

Genetic polymorphisms in the TGF-beta1 gene were shown to interfere with the transcriptional activity of the TGF-beta1 gene, and this influences the production, secretion or activity of the TGF-beta1 growth factor. Transforming growth factor-beta1 (TGF-beta1) gene polymorphism is associated with risk of inflammatory diseases.. The aim of this study was to evaluate TGF-beta1 gene polymorphisms in a Turkish population with different periodontal diseases and to investigate the association between TGF-beta1 genotype and clinical periodontal parameters.. A total of 134 subjects were included in this study. Genomic DNA was isolated from 51 patients with chronic periodontitis (CP), 43 with generalized aggressive periodontitis (G-AgP) and 40 healthy controls. Three TGF-beta1 gene polymorphisms were identified by PCR-RFLP and MS-PCR at positions +915G/C, Thr263Ile and 713/8delC. Probing pocket depth (PPD), clinical attachment loss (CAL), plaque accumulation (plaque %) and bleeding of probing were obtained. Chi-square, Mann-Whitney U test and logistic regression analysis were used.. There was a slightly significant difference in +915C positive genotype distributions between CP and control groups (OR: 2.46, CI: 1.010-6.005, p = 0.047). No significant differences were present between G-AgP and controls in +915C positive genotype. Thr263Ile and -713/8delC genotype distributions were not different between study groups. There were significant differences in PPD and CAL scores between +915C positive and negative CP patients.. These findings suggest that the TGF-beta1 (+915C) polymorphic allele might be associated with chronic periodontitis in the Turkish population.

Topics: Adult; Alleles; Chronic Disease; Female; Gene Frequency; Humans; Male; Middle Aged; Periodontal Diseases; Periodontitis; Polymorphism, Genetic; Regression Analysis; Transforming Growth Factor beta; Turkey

We previously reported evidence that patients with periodontitis have serum antibodies to oral Gram positive bacteria that are cross-reactive with epithelial antigens, including CD24. High level expression of CD24 was confined to the reactive periodontal epithelium and inflamed gingival attachment. As a model for the reactive epithelium of chronic periodontitis, H413 epithelial cells derived from a human oral squamous cell carcinoma were cloned and lines expressing high levels of CD24 were selected. RNA interference protocols were designed to determine if CD24 could modulate intercellular interactions and regulate the biology of these epithelial cells. Knock-down of CD24 protein was demonstrated by Western blot and flow cytometry. The level of mRNA for CD24 was reduced 90% by RNAi treatment as assayed by real-time, reverse transcriptase (RT)-PCR. Gene products known to be important in epithelial biology, including E-cadherin and TGF-beta3 that were demonstrated to undergo altered expression patterns in the periodontal lesion, were investigated. Down-regulation of CD24 mRNA was associated with reduced e-cadherin expression and up-regulated expression of snail, twist, and tgf-beta3. The cells were treated with monoclonal antibodies to CD24 to mimic the action of auto-reactive antibodies to CD24 detected in affected patients. Relative to isotype control antibody, stimulation by anti-CD24 antibodies induced up-regulated expression of e-cadherin and down-regulation of tgf-beta3 as assessed by real-time RT-PCR. No consistent changes for expression of beta-catenin, connexins, integrins, icam-1, tgf-beta1 or tgf-beta2 were observed. CD24 could play an important role in modulating expression of genes that regulate epithelial differentiation in periodontal disease.

Topics: Cadherins; CD24 Antigen; Cells, Cultured; Epithelial Cells; Gene Expression Regulation; Gingiva; Humans; Models, Genetic; Periodontitis; Plasmids; RNA Interference; RNA, Small Interfering; Time Factors; Transforming Growth Factor beta; Transforming Growth Factor beta3

The reconstruction of bone on exposed implant surfaces requires an almost complete elimination of attached bacteria. Only then is new bone growth possible. This case report describes a new way to kill biofilms with photodynamic therapy and to augment bone by using recombinant human bone morphogenetic protein-2. Though the new technique of bone tissue engineering causes some problems, new bone growth on uncovered implant surfaces is successful to a large extent.

Topics: Adult; Alveolar Bone Loss; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Dental Implants; Drug Carriers; Female; Follow-Up Studies; Gelatin Sponge, Absorbable; Humans; Laser Therapy; Osteogenesis; Periodontitis; Photochemotherapy; Transforming Growth Factor beta

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

The function of T cells infiltrating periodontitis lesions is complex and has not been fully elucidated. Here, we established T-cell clones from the gingival tissues of periodontitis patients and examined their gene expression. A total of 57 and 101 T-cell clones were established by means of immobilized anti-CD3 antibody and IL-2 from gingival tissues and peripheral blood, respectively. The gingival T-cell clones were derived from three patients, and the peripheral blood T-cell clones from two of these patients and a further patient whose gingival T-cell clones were not established. Gingival tissues were also obtained from a further 19 periodontitis patients. The expression of cytokines and molecules related to both regulatory function and tissue destruction were examined by means of reverse-transcription polymerase chain reaction. All the gingival T-cell clones expressed mRNA for TGF-beta1, CTLA-4, and CD25, and all the T-cell clones from peripheral blood expressed IFN-gamma and TGF-beta1 mRNAs. Most but not all the T-cell clones from gingival tissues and peripheral blood expressed mRNA for IFN-gamma and, CD25 and CTLA-4, respectively. The frequency of T-cell clones and gingival tissues expressing FOXP3, a possible master gene for mouse CD4(+)CD25(+) regulatory T cells, was very high (97%, 93%, and 100% for gingival T-cell clones, peripheral blood T-cell clones, and gingival tissues, respectively). Whereas the frequency of IL-4-expressing T-cell clones was lower for gingival T-cell clones (70% vs. 87%), the frequency of the gingival T-cell clones expressing IL-10 and IL-17 was higher than peripheral blood T-cell clones (75% vs. 62% for IL-10, 51% vs. 11% for IL-17). A similar expression profile was observed for gingival T-cell clones compared with gingival tissue samples with the exception of IL-4 expression, where the frequency of positive samples was lower in the gingival tissues (70% vs. 11%). These results suggest that the individual T cells infiltrating gingival lesions can express mRNA for both Th1 and Th2 cytokines as well as regulatory cytokines simultaneously.

Topics: Antigens, CD; Antigens, Differentiation; CD4 Antigens; CD4-Positive T-Lymphocytes; Clone Cells; CTLA-4 Antigen; Forkhead Transcription Factors; Gene Expression; Gene Expression Profiling; Gingiva; Humans; Immunoglobulin Fc Fragments; Interferon-gamma; Interleukin-10; Interleukin-17; Interleukin-4; Periodontitis; Receptors, Interleukin-2; Th1 Cells; Th2 Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1

Previous studies have shown a limited potential for bone augmentation following guided bone regeneration (GBR) in horizontal alveolar defects. Surgical implantation of recombinant human bone morphogenetic protein-2 (rhBMP-2) in an absorbable collagen sponge carrier (ACS) significantly enhances bone regeneration in such defects; however, sufficient quantities of bone for implant dentistry are not routinely obtained. The objective of this study was to evaluate the potential of rhBMP-2/ACS to enhance GBR using a space-providing, macro-porous expanded polytetrafluoroethylene (ePTFE) device.. Bilateral, critical size, supra-alveolar, peri-implant defects were surgically created in four Hound Labrador mongrel dogs. Two turned and one surface-etched 10-mm titanium dental implant were placed 5 mm into the surgically reduced alveolar ridge creating 5-mm supra-alveolar defects. rhBMP-2/ACS (rhBMP-2 at 0.2 mg/ml) or buffer/ACS was randomly assigned to left and right jaw quadrants in subsequent animals. The space-providing, macro-porous ePTFE device was placed to cover rhBMP-2/ACS and control constructs and dental implants. Gingival flaps were advanced for primary wound closure. The animals were euthanized at 8 weeks postsurgery for histologic and histometric analysis.. Bone formation was significantly enhanced in defects receiving rhBMP-2/ACS compared to control. Vertical bone gain averaged (+/- SD) 4.7 +/- 0.3 and 4.8 +/- 0.1 mm, and new bone area 10.3 +/- 2.0 and 8.0 +/- 2.5 mm2 at turned and surface-etched dental implants, respectively. Corresponding values for the control were 1.8 +/- 2.0 and 1.3 +/- 1.3 mm, and 1.8 +/- 1.3 and 1.2 +/- 0.6 mm2. Bone-implant contact in rhBMP-2-induced bone averaged 6.4 +/- 1.4% and 9.6 +/- 7.5% for turned and surface-etched dental implants, respectively (P=0.399). Corresponding values for the control were 14.6 +/- 19.4% and 23.7 +/- 9.7% (P=0.473). Bone-implant contact in resident bone ranged between 43% and 58% without significant differences between dental implant surfaces.. rhBMP-2/ACS significantly enhances GBR at turned and surface-etched dental implants. The dental implant surface technology does not appear to substantially influence bone formation.

Topics: Absorbable Implants; Alveolar Bone Loss; Animals; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Collagen; Dental Implantation, Endosseous; Dental Implants; Dogs; Guided Tissue Regeneration, Periodontal; Humans; Male; Membranes, Artificial; Osseointegration; Periodontitis; Polytetrafluoroethylene; Radiography; Recombinant Proteins; Transforming Growth Factor beta

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

Patients who smoke are at increased risk for chronic periodontitis (CP). Also, CP patients who smoke exhibit significantly less reduction of probing depths and gains in clinical attachment compared to non-smokers following periodontal therapy. Several studies suggest that the effects of smoking on the host response may be paramount in regulating the basal systemic inflammatory status and therapeutic outcomes in this cohort. Growth factors, specifically transforming growth factor beta1 (TGF-beta1), are critical in regulating the wound healing response by controlling cell division, differentiation, and motility. The hypothesis to be tested was that gingival crevicular fluid (GCF) TGF-beta1 production was altered in smokers compared to non-smokers with CP.. GCF was collected from smokers and non-smokers with CP, both at baseline and 1 to 2 weeks after initial therapy. GCF volume was determined using an electronic device and TGF-beta1 concentration was measured by enzyme-linked immunosorbent assay (ELISA).. Smokers exhibited a higher mean concentration of GCF TGF-beta1 at baseline compared to non-smokers (P = 0.03). After initial therapy, smokers exhibited significantly less reduction in mean GCF volume compared to non-smokers (P = 0.04).. Augmented constitutive production of GCF TGF-beta1 in smokers may explain the clinical appearance of fibrotic gingival tissue exhibited by this patient cohort. A diminished reduction in GCF volume in smokers following root instrumentation suggests a chronic inflammatory status in conjunction with an ineffective host response. These findings support the concept that smokers with CP display an altered local inflammatory response after initial therapy, perhaps symptomatic of colonization by residual periodontal pathogens.

Topics: Analysis of Variance; Case-Control Studies; Chronic Disease; Dental Scaling; Enzyme-Linked Immunosorbent Assay; Female; Gingival Crevicular Fluid; Humans; Male; Middle Aged; Periodontitis; Smoking; 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

A polymorphism in the promoter region of the transforming growth factor beta-1 (TGF-beta1) gene was described at position -509. This polymorphism represents a C-to-T base exchange, which creates a YY1 consensus sequence in an area involved with down transcription regulation. This polymorphism has been associated with risk for asthma and allergies. In this study we investigated the association between this polymorphism and chronic periodontitis severity.. Genomic DNA from oral mucosa of 87 Caucasian subjects was amplified by PCR, and digested with Eco81I restriction endonuclease. The alleles were separated by polyacrylamide gel electrophoresis. The differences in genotype distribution from those expected by Hardy-Weinberg equilibrium, and the significance of the differences in observed frequencies of the polymorphism in moderate and severe disease and healthy groups were assessed by the chi2 test.. There was a difference in the presence of the different alleles and genotypes among the healthy, moderate and severe periodontitis groups. The allele T was seen at 57.7% in the group with severe periodontitis and 37.8% and 35.4% in the healthy group and moderate periodontitis group, respectively (p=0.0387). The genotype T/T was found at 38.5% in the group with severe periodontitis, and at a frequency of 8% in the healthy group (p=0.0258).. These results demonstrate that the polymorphism at bp -509 in the TGF-beta1 promoter may have a small effect on the modulation of the inflammatory process during periodontitis.

Topics: Adult; Alleles; Case-Control Studies; Chronic Disease; Cytosine; Female; Gene Frequency; Genetic Markers; Humans; Male; Mouth Mucosa; Periodontitis; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Severity of Illness Index; Thymine; Transforming Growth Factor beta; Transforming Growth Factor beta1

The regeneration of periodontal tissues lost due to periodontal disease requires cell migration, differentiation and proliferation. Several procedures have been proposed to promote wound healing events such as the application of growth factors including PDGF-BB, TGF-beta1 and rhBMP-2. The purpose of this study was to evaluate the mitogenic responses of human periodontal ligament cells and gingival fibroblasts to PDGF-BB, TGF-beta1 and rhBMP-2. Human periodontal ligament cells were isolated from the mid root of three maxillary third molars extracted from three adult patients with moderate periodontitis and gingival fibroblasts were obtained from two patients also affected by moderate periodontitis, who underwent periodontal surgery. Cells were grown in 24-well dishes. On day 2 of quiescence, new medium was added with PDGF-BB or TGF-beta1 or rhBMP-2 at the concentration of 10 ng/ml. To determine the effects of the test agents on cell proliferation, DNA synthesis was estimated by measuring [3H] thymidine incorporation. After 48h of incubation the cells were processed and subject to scintillation counting. Counts per minute (cpm/ well) were determined for each sample. The results of this study demonstrated that PDGF-BB acts like a strong mitogenic agent for human periodontal ligament cells and gingival fibroblasts, TGF-beta1 mostly supports the proliferation of these cells and rhBMP-2 had an opposite effect on cell mitosis.

Topics: Adult; Analysis of Variance; Becaplermin; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Cell Culture Techniques; Cell Division; DNA; Fibroblasts; Gingiva; Humans; Mitogens; Mitosis; Periodontal Ligament; Periodontitis; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-sis; Recombinant Proteins; 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

Transforming growth factor-beta (TGF-beta) represents a family of polypeptide growth factors, involved in inflammation and regulation of immune responses. The purpose of this study was to determine whether polymorphisms in the TGF-beta 1 gene may confer susceptibility to adult periodontitis.. We studied 90 patients with adult periodontitis together with 108 unrelated subjects. 3 polymorphisms located in the 5'region at positions -988 (C/A), -800 (G/A) and -509 (C/T) and 2 polymorphisms located at codons 10 (L10P) and 25 (R25P) of exon 1 were investigated by PCR methods.. There was no statistically-significant difference in genotype or allele frequency distributions between patients and reference group for the -800G/A, -509C/T, L10P and R25P polymorphisms (p>0.05 in all cases). The -988 A polymorphism was present neither in our patients nor in unrelated subjects. Upon stratification for smoking status no significant differences were found in the TGF-beta 1 genotype or allele frequencies either between adult periodontitis smokers compared to control smokers, or between periodontitis non-smokers and control non-smokers.. These data indicate that the mentioned polymorphisms of the TGF-beta 1 gene do not influence susceptibility to adult periodontitis. There was no association between any polymorphisms in the TGF-beta 1 gene, severity of periodontitis and the smoking status in our study.

Topics: Adult; Alleles; Case-Control Studies; Chromosomes, Human, Pair 19; Codon; Confidence Intervals; Exons; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Haplotypes; Humans; Male; Middle Aged; Monte Carlo Method; Odds Ratio; Periodontitis; Polymerase Chain Reaction; Polymorphism, Genetic; Smoking; Statistics as Topic; Transforming Growth Factor beta; Transforming Growth Factor beta1

The host immune response in chronic marginal periodontitis (CMP) raised against bacteria colonizing the dentogingival area is modulated by cytokines. This study examines the distribution of the transforming growth factor-beta1 containing (TGF-beta1+) cells in formalin-fixed and paraffin-embedded gingival specimens from 11 patients with chronic marginal periodontitis and 7 persons with healthy gingiva. Inflamed periodontal tissue contained a 100-fold more TGF-beta1+ cells than healthy gingiva. Diverse morphological TGF-beta1+ cell types were discerned. Double immuno-enzymatic and -fluorescence staining revealed that TGF-beta1+ cells comprised 21-29% macrophages 2-3% T-cells, 3-9% B-cells, 34-35% neutrophilic granulocytes and 7-10% mast cells. The densities of all TGF-beta1+ cell types in CMP were strongly increased in the connective tissue adjacent to the pocket epithelium, in the lamina propria and adjacent to the oral epithelium. In lesions with extensive inflammation, expression was also marked in pocket epithelium. TGF-beta1 is an immunosuppressive cytokine that stimulates wound healing. Upregulation of the cytokine in inflamed gingiva may counterbalance for destructive gingival inflammatory responses that are simultaneously taking place in patients with CMP.

Topics: Case-Control Studies; Cell Count; Chronic Disease; Connective Tissue Cells; Epithelial Cells; Fluorescent Antibody Technique; Gingiva; Granulocytes; Humans; Immunoenzyme Techniques; Lymphocytes; Macrophages; Mast Cells; Periodontitis; Statistics, Nonparametric; Transforming Growth Factor beta; Up-Regulation

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

Semisynthetic tetracyclines used in the adjunctive treatment of inflammatory periodontal disease enhance collagen expression in induced periodontal lesions of rats. Polypeptide growth factors regulate key cellular events in tissue repair. The physiologically active androgen 5 alpha-dihydrotestosterone (DHT) stimulates bone and connective tissue turnover. It was relevant to study the effects of transforming growth factor beta (TGF-beta)/platelet-derived growth factor (PDGF) and minocycline alone and in combination on the formation of biologically effective androgens which can influence repair.. Confluent monolayer cultures of human gingival fibroblasts of the fifth through the ninth passage were incubated in Eagle's minimum essential medium, with 14C-testosterone/14C-4-androstenedione in the presence or absence of optimal concentrations of TGF-beta/PDGF/minocycline (M), alone and in combination. At the end of a 24-hour incubation period, the medium was analyzed for steroid metabolites and quantified using a radioisotope scanner.. The androgen substrates 14C-testosterone (14C-T) and 14C-4-androstenedione (14C-4-A) were metabolized to DHT and 4-androstenedione/testosterone respectively. There were significant increases in the formation of DHT from 14C-T in response to M, TGF-beta, and PDGF, alone and in combination (13 to 48%), compared with controls (n = 4; P<0.01). The yields of 4-androstenedione were also greater in response to these agents (31%; 3-fold). When 14C-4-A was used as substrate, there were 21 to 80% increases in the formation of DHT in response to these agents alone and in combination (n = 4; P<0.01).. The biologically effective androgen metabolites formed in response to minocycline, TGF-beta, and PDGF can contribute to reparatory events in the inflamed periodontium. Judicious, adjunctive usage of the chemically-modified tetracyclines in the treatment of periodontal diseases can obviate the risk of microbial resistance, with potential applications of their anti-inflammatory and proanabolic effects in regenerative technology.

Topics: Adult; Androstenedione; Anti-Bacterial Agents; Carbon Radioisotopes; Cells, Cultured; Dihydrotestosterone; Female; Fibroblasts; Gingiva; Humans; Male; Minocycline; Periodontitis; Platelet-Derived Growth Factor; Statistics, Nonparametric; Testosterone; Transforming Growth Factor beta

Periodontitis is a chronic inflammatory disease characterized by a progression that is very much dependent on host response. The gingiva can be considered to be in a constant state of wounding (pathologic wounding by bacterial plaque) and a constant state of maintenance/repair. In this context, any metabolic disturbance in the host which compromises tissue repair/wound healing will exacerbate the progression of periodontitis. Diabetes presents an interesting example because two major complications of diabetes are delayed wound healing and periodontitis. Our previous studies indicate that delayed wound healing and periodontitis may be manifestations of a general systemic deficit in diabetes involving alteration of macrophage cytokine gene expression. The present study was designed to determine whether: 1) diabetes-induced metabolic alterations affect gingival cytokine levels; and 2) diabetes-induced metabolic alterations modify the gingival cytokine profile in periodontitis. Sprague-Dawley rats (N=12/group) were injected with streptozotocin (65 mg/kg) into the tail vein to induce diabetes (defined by blood glucose levels > 250 mg/dl) or received the injection vehicle or no treatment as controls. Periodontitis was induced in additional groups of diabetic and control rats by gavage with Porphyromonas gingivalis A7436. After 90 days, serum glucose was analyzed to document diabetes; alveolar bone level was measured to document severity of periodontitis; gingiva was harvested circumferentially from the first and second molars; and cytokines in gingival homogenates were assayed by ELISA using commercial kits. Cytokine levels were expressed as mean+/-SEM pg/microg protein. Diabetes alone did not alter the gingival cytokine profile for platelet-derived growth factor B (PDGF-B), interleukin 1-beta (IL-1beta), transforming growth factor-beta (TGF-beta), and tumor necrosis factor-alpha (TNF-alpha). Periodontitis alone demonstrated a significant increase (P < 0.05) in levels of PDGF-B and IL-1beta. Diabetes superimposed on periodontitis prevented these increases. Thus, diabetes-induced metabolic alterations do not affect gingival cytokine levels per se; however, they do alter the normal host response to periodontitis through blockage of periodontitis-induced increases in PDGF-B and IL-1beta.

Topics: Alveolar Bone Loss; Animals; Bacteroidaceae Infections; Blood Glucose; Dental Plaque; Diabetes Mellitus, Experimental; Disease Models, Animal; Disease Progression; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Gingiva; Interleukin-1; Macrophages; Male; Periodontitis; Pharmaceutical Vehicles; Platelet-Derived Growth Factor; Porphyromonas gingivalis; Protein-Tyrosine Kinases; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-sis; Random Allocation; Rats; Rats, Sprague-Dawley; Streptozocin; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing

The purpose of this study was to examine the regeneration of periodontal tissue after the application of recombinant human bone morphogenetic protein-2 (rhBMP-2) to horizontal circumferential defects created by experimental periodontitis. Twelve mandibular second premolars in 6 adult beagle dogs were subjected to experimental periodontal breakdown by placing silk ligatures around the teeth until the bone loss exceeded half of the root length. Flap surgery was then performed and the exposed cementum removed. The distance between the bone crest and cemento-enamel junction (CEJ) was about 5 mm. RhBMP-2 (40 micrograms/100 microliters) with a sponge-type carrier material made of gelatin and polylactic acid polyglycolic acid copolymer was placed in the furcation area (5 mm x 5 mm x 5 mm) and around the roots (10 mm x 5 mm x 2.5 mm x 2 pieces). In the control group, the same carrier material without rhBMP-2 was placed in the same manner. The flaps were replaced and sutured to cover these materials completely. Twelve weeks after surgery, the animals were sacrificed and serial sections were prepared in a bucco-lingual plane. Considerable new bone formation was observed in the rhBMP-2-treated sites. New cementum with Sharpey's fibers was observed on the instrumented root surface. On histometric analysis, the amount of new bone, new cementum, and connective tissue attachment was significantly greater in the rhBMP-2-treated group (paired t test; P < 0.01). These results indicate that suitable application of rhBMP-2 can produce considerable periodontal tissue regeneration, even in cases of horizontal circumferential defects.

Topics: Alveolar Bone Loss; Animals; Biocompatible Materials; Bone Morphogenetic Protein 2; Bone Morphogenetic Proteins; Bone Regeneration; Dental Cementum; Dogs; Drug Carriers; Guided Tissue Regeneration, Periodontal; Humans; Lactic Acid; Periodontal Ligament; Periodontitis; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Recombinant Proteins; Regeneration; 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

Fibronectin is a major component of the extracellular matrix and is considered to have an important role in chronic inflammatory periodontal disease. The fibronectin gene product has been shown to be subject to alternative splicing in 3 regions, each generating different mRNA transcripts associated specifically with normal adult tissue, embryogenesis, tissue regeneration, and wound healing. In the present study, using the reverse-transcribed polymerase chain reaction to examine splicing profiles of the primary transcript, we found that healthy periodontal tissue expressed all alternatively spliced embryonic isoforms, indicative of the extensive and ongoing rebuilding processes which occur in these tissues. In marked contrast, only the exon-skipped transcripts were generated in tissue from chronic inflammatory periodontal disease patients. The loss of the high molecular weight isoforms in lesional tissues may be due to the excess production of inflammatory mediators in this disease, since we observed that high concentrations of the cytokine IL-1beta caused down-regulation of these transcripts in normal periodontal cells in tissue culture. Moreover, we also demonstrated that growth factors likely to be involved in periodontal regeneration and repair, such as PDGF, IGF-1 and TGF-beta, elicited pronounced upregulation of the embryonic isoforms of fibronectin in these cells. Although the functional activities of the antigens corresponding to the alternatively spliced variants of fibronectin are not yet known, our finding that they are selectively expressed suggests that they have highly specific roles in both periodontal breakdown and repair.

Topics: Adult; Alternative Splicing; Antigens; Cells, Cultured; Chronic Disease; Culture Techniques; Down-Regulation; Embryonic and Fetal Development; Exons; Extracellular Matrix; Fibronectins; Gene Expression Regulation; Gene Expression Regulation, Developmental; Humans; Inflammation Mediators; Insulin-Like Growth Factor I; Interleukin-1; Molecular Weight; Periodontitis; Periodontium; Platelet-Derived Growth Factor; Polymerase Chain Reaction; Regeneration; RNA, Messenger; Transcription, Genetic; Transforming Growth Factor beta; Up-Regulation; Wound Healing

Monocytes have recently been recognized as a precursor of Langerhans cells. This study examined the regulatory influence of the epithelial environment on the putative first step of the transition towards a Langerhans cell phenotype--the induction of CD1a antigen. The keratinocyte-derived cytokines granulocyte-macrophage-colony-stimulating factor, tumour necrosis factor-alpha, interleukin-6, and interleukin-1 beta induced CD1a expression, as did supernatants of keratinocytes extracted from inflammatory sites (periodontitis). Induction was abrogated by transforming growth factor-beta and a keratinocyte-derived interleukin-1 inhibitor. The optimal temperature for induction was 34 degrees C, not 37 degrees C. These results demonstrate that the components of the epithelial environment (cytokines and lower temperature) exert important influences, which may be part of local regulation of Langerhans cell development.

Topics: Adult; Antigens, CD; Antigens, CD1; Antigens, Surface; Epithelial Cells; Gene Expression Regulation; Gingiva; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-1; Interleukin-6; Keratinocytes; Langerhans Cells; Middle Aged; Monocytes; Periodontitis; Phenotype; Stem Cells; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

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

Gingival mononuclear cell production of interleukin 1 (IL-1), interleukin 6 (IL-6) and transforming growth factor-beta (TGF-beta) after stimulation with the putative periodontopathic bacteria, Porphyromonas gingivalis and Fusobacterium nucleatum was investigated. Using an ELISA method, gingival mononuclear cells extracted from 18 adult periodontitis subjects were found to be producing IL-1. However, IL-1 activity could only be detected in 5 out of these 18 cases when tested using a thymocyte proliferation bio-assay, suggesting the presence of IL-1 inhibitors. Depletion of monocytes from peripheral blood cultures resulted in a significant decrease in IL-1 activity following P. gingivalis stimulation while there was no effect in the level of IL-1 activity following stimulation with F. nucleatum. This suggests that P. gingivalis and F. nucleatum stimulate different cell types to produce IL-1. Like IL-1, IL-6 production by gingival mononuclear cells was significantly greater than that produced by the control peripheral blood mononuclear cells. Following P. gingivalis and F. nucleatum stimulation, higher levels of IL-6 could be detected; however, both organisms stimulated similar levels. Intracytoplasmic immunofluorescence staining demonstrated a lower percent TGF-beta+ cells in bacterial stimulated peripheral blood mononuclear cell cultures compared with cells in medium alone. In the gingival mononuclear cell cultures, the percentage TGF-beta+ cells peaked at day 1 in F. nucleatum-stimulated, whereas in P. gingivalis-stimulated cultures the peak TGF-beta+ cells occurred at day 3, again suggesting stimulation of different cell subsets. These results illustrate that different periodontopathic bacteria may stimulate different cell types to produce cytokines which may have synergistic or antagonistic effects.

Topics: B-Lymphocytes; Cytokines; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique; Fusobacterium nucleatum; Gingiva; Humans; Interleukin-1; Interleukin-6; Leukocytes, Mononuclear; Periodontitis; Porphyromonas gingivalis; Transforming Growth Factor beta

Topics: Alveolar Bone Loss; Bone Regeneration; Collagen; Concanavalin A; Connective Tissue; Enzyme Activation; Extracellular Matrix Proteins; Fibroblasts; Fibronectins; Gingiva; Glycoproteins; Humans; Metalloendopeptidases; Osteonectin; Periodontitis; Phenotype; Tissue Inhibitor of Metalloproteinases; Transforming Growth Factor beta