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PubMed2002

Quantitative assessment of apoptotic and proliferative gingival keratinocytes in oral and sulcular epithelium in patients with gingivitis and periodontitis.

BACKGROUND: Periodontal disease is caused by a chronic infection inducing an inflammatory reaction that leads to a breakdown of tooth-supporting tissue. The maintenance of an equilibrium between the host defence and microorganisms in the sulcus is essential to preserve health. All multicellular organisms have mechanisms for killing their own cells, and use physiological cell death for defence, development, homeostasis and ageing. Apoptosis and proliferation are very important phenomena in regulating this and a disturbance is often associated with disease e.g. cancer, AIDS, Alzheimer's disease, rheumatoid arthritis. OBJECTIVE: The aim of this study was to determine whether the number of apoptotic and proliferative gingival keratinocytes differed between patients with gingivitis and those with periodontitis. MATERIAL AND METHODS: The distribution of neutrophil elastase, PCNA/cyclin, DNA fragmentation (apoptosis) and p53 was determined with immunocytochemical techniques. We used paraffin-embedded sections from gingival biopsies and did quantitative analyses. RESULTS AND CONCLUSION: These showed that 5-12% of the keratinocytes in the basal layers of the epithelium proliferated in the two groups. Fewer apoptotic cells were seen in the oral epithelium than in the sulcus in all subjects in both groups. Only in the most apical part of the sulcus, close to the junctional epithelium, did the number of apoptotic keratinocytes exceed the proliferative ones in patients with periodontitis.

Adult↗
PubMed1997

IgG and IgA subclass mRNA-bearing plasma cells in periodontitis gingival tissue and immunoglobulin levels in the gingival crevicular fluid.

The humoral immune response, especially the production of IgG and IgA, is considered to have a protective role in the pathogenesis of periodontal disease, but the precise mechanisms are still unknown. In order to determine local IgG and IgA production, we investigated the presence of human IgG and IgA subclass mRNA-bearing plasma cells within periodontal tissue by in situ hybridization using digoxigenin-labelled oligonucleotide probes in 24 gingival biopsy samples (pocket depth > or = 5 mm) which were obtained from eight patients with adult periodontitis. Furthermore, we examined IgG and IgA subclass proteins and digested IgA1 Fab portions in the gingival crevicular fluid (GCF), corresponding to the sites from which the tissues were taken, by ELISA. IgG and IgA subclass mRNA-expressing cells were detected in all serial formalin-fixed/paraffin-embedded gingival tissue sections sampled. Plasma cells showed strong cytoplasmic staining with a high contrast and a good retention of morphology with these probes. IgG1 mRNA-expressing cells were predominant (mean 63%) and IgG2 mRNA-expressing cells were present at around 23% of total IgG plasma cells, while IgG3 and IgG4 mRNA-expressing cells were present to a much lesser extent (3% and 10%, respectively). Similar proportions of IgG subclass proteins in GCF were detected, which were also consistent with 'normal' serum levels. In terms of IgA subclass, IgA1 mRNA-positive cells were predominant (mean 65.1%, P < 0.001). In contrast, IgA2 protein in the GCF samples were detected at higher concentrations than IgA1 (P < 0.001). The ratio of total IgG to IgA mRNA-positive plasma cells was approximately 7.5:1. There was a good correlation between the amounts of IgG subclass proteins in GCF and the number of IgG subclass mRNA-positive cells in the same sites, but not between IgA subclass proteins and the number of IgA subclass mRNA-positive cells. These data suggest that IgG and IgA subclass proteins can be locally produced in the periodontitis gingiva. In addition, as we detected IgA1 Fab fragments in GCF, this is further confirmation that secreted IgA1 protein in GCF may be digested by periodontal bacteria.

Adult↗
PubMed2004

Proliferative response to phenytoin and nifedipine in gingival fibroblasts cultured from humans with gingival fibromatosis.

The response of gingival fibroblasts cultured from humans with gingival fibromatosis to phenytoin (PHT) and nifedipine (NIF) was investigated. PHT and NIF induced proliferation, and increased the expression of immunoreactive endothelin-1 (ET-1). ET-1 (0.1 nm-1 microm) itself also induced proliferation in a concentration-dependent manner. The proliferation was inhibited by BQ-123 (ETA receptor antagonist; 1 microm) and TAK044 (ETA/ETB receptor antagonist; 1 microm), but not by BQ-788 (ETB receptor antagonist; 1 microm). The proliferation induced by PHT (0.25 microm) and NIF (0.25 microm) was inhibited by BQ-123 (1 microm). In addition, the results of Western blot analysis indicated the presence of ETA and ETB receptors in/on the fibroblasts. These findings suggest that PHT- and NIF-induced gingival proliferation may be mediated by endogenously generated ET-1, possibly via ETA receptors.

Anticonvulsants↗
PubMed2005

Immunosuppression and gingival overgrowth: gene and protein expression profiles of collagen turnover in FK506-treated human gingival fibroblasts.

AIM: FK506 is an immunosuppressive agent that, unlike cyclosporin A (CsA), does not induce gingival overgrowth (GO). CsA-induced GO is caused by quantitative modifications of the extracellular matrix components, particularly collagen (COL). Up to now, clinical trials have only investigated FK506 in relation with GO, so we aimed at analysing the effect of FK506 on COL turnover using a molecular approach, to evaluate the expression of genes and proteins related to this process. MATERIALS AND METHODS: Human gingival fibroblasts were incubated with FK506 or its vehicle (VH) for 24, 48 and 72 h. COL type I (COL-I), matrix metalloproteinases (MMP)-1 and 2, tissue inhibitor of MMP (TIMP)-1 and transforming growth factor (TGF)-beta1 mRNA were assayed by Reverse transcriptase polymerase chain reaction; COL-I protein levels were determined by dot blot, MMP-1 and MMP-2 activity by zymography. RESULTS: Fibroblast proliferation decreased 48 and 72 h after treatment. COL-I gene and protein expression, TGF-beta1 and TIMP-1 mRNA levels were not significantly affected, whereas MMP-1 gene and protein expression and MMP-2 mRNA levels rose significantly in treated fibroblasts compared with VH. CONCLUSIONS: These findings suggest that increased MMP-1 gene and protein expression may be important for regulating COL-I homeostasis in the gingival connective compartment of FK506-immunosuppressed subjects.

Adult↗
PubMed1995

ICAM-1-expressing pocket epithelium, LFA-1-expressing T cells in gingival tissue and gingival crevicular fluid as features characterizing inflammatory cell invasion and exudation in adult periodontitis.

Activated T lymphocytes constitute a major component of inflammatory cells in the early periodontal lesion, and also appear in the gingival crevicular fluid. In an attempt to clarify the relationship between the ICAM-1 (CD54) expression of pocket epithelium in gingiva and the infiltrating lymphocyte population, we carried out an analysis of CD11a+(LFA-1 alpha), CD25+(IL-2R alpha) and CD4+(Th) cells subjacent to ICAM-1-expressing pocket epithelia and CD11a+CD25+CD4+ cells in gingival crevicular fluid (GCF). GCF was collected by crevicular washing from 16 patients with periodontitis (P group) and 3 subjects with healthy gingiva (H group). Peripheral blood (PB) was collected at the same time. Mononuclear cells were isolated by Ficoll-paque gradient centrifugation from GCF and PB. Monoclonal antibodies (mAb) to CD11a, CD25, and CD4 were used for three-color flow cytometry. Gingival biopsies were obtained from 7 patients in P group and 3 subjects in H group. Serial cryostat sections (6 microns in thickness) were prepared from each biopsy, on which a double staining was performed. The number of CD11a+CD25+CD4+ cells and the fluorescence intensity of FITC conjugated anti-CD11a were significantly higher in GCF than in PB (p < 0.001 to p < 0.01). CD11a+CD25+CD4+ cells were not detected in GCF in H group. The pocket epithelia expressed CD54 in P group, but not in H group. The number of CD11a+, CD25+ and CD4+ cells infiltrating the connective tissue subjacent to the upper, middle and lower parts of the CD54 positive pocket epithelium (n = 16) was 141 +/- 26, 38 +/- 13, 144 +/- 29 (cells/0.04 mm2), respectively, whereas in the CD54 negative pocket epithelium, it was (n = 5) 9 +/- 2, 3 +/- 1, 8 +/- 3. In P group, the CD11a+CD25+CD4+ cell number in GCF correlated with CD25+, CD11a+ cells in the connective tissue subjacent to the CD54+ pocket epithelium. These results indicate that expression of ICAM-1 in pocket epithelium is relevant to the migration of CD11a, CD25, CD4 positive cells in connective tissue subjacent to the pocket epithelium into the periodontal pocket. Assessing the relationship of our findings and other adhesion molecules would offer important clues to the understanding of T cell migration in affected gingiva.

Adult↗
PubMed2004

Effects of scaling and root planing on the amounts of interleukin-1 and interleukin-1 receptor antagonist and the mRNA expression of interleukin-1beta in gingival crevicular fluid and gingival tissues.

OBJECTIVE: The purpose of this study was to evaluate the relationship between the clinical changes after non-surgical periodontal therapy and interleukin 1 (IL-1) in gingival crevicular fluid (GCF) and gingival tissues from patients with chronic periodontitis. BACKGROUND: The inflammatory responses mediated by IL-1 play an important role in periodontal tissue destruction. Although numerous studies have attempted to elucidate the dynamic movement involved in chronic periodontitis, the results have often conflicted. Such discrepancies may have been due to the inability to determine clinical disease activity. METHODS: Seven patients with chronic periodontitis were examined. The severity of periodontal inflammation was expressed using clinical parameters before and after a scaling and root planing (SRP) procedure. The amounts and concentrations of IL-1alpha, IL-1beta and IL-1 receptor antagonist in GCF were measured by enzyme-linked immunosorbent assay (ELISA) and IL-1 activity index was calculated. A needle biopsy in matching gingival tissues was also performed before and after the SRP procedure. The localization and mRNA expression of IL-1beta were determined using histological methods. RESULTS: Clinical parameters improved slightly after the SRP procedure. Only the probing pocket depth (PPD) was reduced significantly (p < 0.05). However, the amount of IL-1beta in GCF was slightly increased. The localization and mRNA expression of IL-1beta could still be observed after the SRP procedure. Therefore, none of the clinical parameters showed a high sensitivity or specificity for evaluating subgingival inflammation. CONCLUSION: These observations suggest that IL-1 is effective for evaluating in detail the state of subgingival inflammation.

Adult↗
PubMed1991

Single-dose concentrations of tinidazole in gingival crevicular fluid, serum, and gingival tissue in adults with periodontitis.

Previous studies have shown that metronidazole is effective in the treatment of subgingival microflora associated with destructive periodontitis. The aim of this study was to determine whether tinidazole, a close analogue of metronidazole, would reach sufficient concentrations in serum, gingival crevicular fluid, and gingival tissue, to inhibit putative periodontopathic bacteria. Ten adult patients with moderate to advanced periodontitis took a single 2-g dose of tinidazole orally. Samples were assayed by high-performance liquid chromatography. The concentrations of tinidazole in serum and GCF were in a similar range (3.2-46.5 micrograms/mL). Tinidazole was not detected in the GCF in three of the patients. The drug was found in gingival tissue obtained at two h (0.17 +/- 0.14 micrograms/mg) and six h (0.15 +/- 0.18 micrograms/mg) after oral administration. The mean concentration of tinidazole in serum at 24 h (13 +/- 3.0 micrograms/mL) is greater than the minimum inhibitory concentration for anaerobic bacteria as reported by others. The present data suggest that a single 2-g oral dose of tinidazole may lead to the presence of potentially bactericidal levels of tinidazole for up to 24 h in the periodontal pockets of some patients with periodontitis.

Administration, Oral↗
PubMed1982

The effect of waxed and unwaxed dental floss on gingival health. Part I. Plaque removal and gingival response.

The purpose of this study was to determine the effectiveness of waxed and unwaxed floss in plaque removal and on gingival health when used in a home oral hygiene program. Eighty patients, having previously received periodontal therapy, were divided into four similar groups, according to the S-OHI. Each group represented four different types of dental floss being tested: Butler waxed, Butler unwaxed, Johnson and Johnson waxed, and Johnson and Johnson unwaxed. After receiving a thorough prophylaxis, each patient received oral hygiene instruction with a video tape, and was given a toothbrush and a quantity of test floss. At 0, 28 and 56 day intervals, The patients were scored for plaque and gingivitis. The data were then analyzed statistically using analysis of variance. It was found that there was no statistical difference among the four different types of tested floss as far as their plaque removal ability or prevention of gingivitis is concerned.

Adult↗
PubMed1992

Treatment of gingival lichen planus with a free gingival graft: a case report.

Lichen planus is an inflammatory dermatosis affecting the skin and mucous membrane. In oral lichen planus, the gingival tissue is often involved. Treatment is symptomatic and depends mostly on the administration of corticosteroids and tranquilizers. This report presents the case of a patient with gingival lichen planus who was treated with free gingival graft. Followup after 3 1/2 years showed that the lesions had completely disappeared.

Adult↗
PubMed2005

[Ultrastructural characteristics of gingival mucous cellular composition in experimental gingivitis].

The aim of the work was to estimate morphological changes in cells of gingival mucous membrane in the experimental gingivitis model. Fifty rats with the body weight of 120-160 gr. were used in the experiment. Gingivitis was modeled by front teeth ligation after a preliminary exfoliation of mucous membrane in the dental neck area. The specimens were examined by histological method and electron microscopy on the 3rd, 5th, 14th and 28th days of the experiment. It has been shown that cellular population of plasmocytes, lymphocytes, fibroblasts and mast cells take part in the inflammatory process. In all the areas of microcirculation occurs transformation of endotheliocytes like papillas; the process of roughening of collagenous fibrils indicates their significant role in the inflammatory damage of gingiva.

Animals↗
PubMed1996

Treatment of severe physiologic gingival pigmentation with free gingival autograft.

Melanin is normally found in the skin of all people, and normal pigmentation of the intraoral tissues is a relatively frequent finding. The highest rate of gingival pigmentation has been observed at the incisors. Until recently, however, very little attention has been paid to the esthetic treatment of severe gingival pigmentation. To answer the cosmetic demand for a pleasing smile, pigmented gingiva was replaced with an unpigmented free gingival autograft in 10 patients. At least two areas in each patient were grafted. In all 10 areas in which the recipient site received full-thickness bed preparation, no evidence of repigmentation was found after 4.5 years. Of the 10 areas that received partial-thickness bed preparation, only one exhibited repigmentation (after 1 year).

Adolescent↗
PubMed1982

The effect of waxed and unwaxed dental floss on gingival health. Part II. Crevicular fluid flow and gingival bleeding.

Eighty patients, having received periodontal therapy previously, were divided into four groups, corresponding to one of four different types of dental floss being used. Seven to ten days after receiving a thorough prophylaxis, each patient had oral hygiene instruction by video tape, was given a toothbrush and a quantity of the test floss. At 0, 28 and 56 day intervals, the patients were scored for crevicular fluid flow and gingival bleeding. Data were analyzed statistically. Johnson and Johnson unwaxed dental floss was found to be slightly less effective in reducing gingival bleeding at the 56th day. It was also found that crevicular fluid flow was least with waxed floss use at the 56 day session. However, the range of variations was so minimal that no clinical significance could be ascribed to either finding. There were patients in each of the four floss groups with no fluid flow or bleeding during all three sessions.

Adult↗
PubMedDate not supplied

Some characteristics of collagenase activity in gingival crevicular fluid and its relationship to gingival diseases in humans.

Collagenase activity in human gingival fluid was measured using a radioactive collagen fibril assay. The activity was positively correlated with the severity of gingival disease. The fluid collagenase seemed to be controlled by alpha 2-macroglobulin, based on its activation by NaSCN, and to be present solely in the extracellular fraction. Examination of the collagen breakdown products by acrylamide gel electrophoresis indicated that the fluid collagenase was of tissue rather than bacterial origin.

Adult↗
PubMed2006

Cell interactions between human gingival fibroblasts and monocytes stimulate the production of matrix metalloproteinase-1 in gingival fibroblasts.

BACKGROUND: Matrix metalloproteinase-1 (MMP-1) plays an important role in inflammatory diseases including periodontitis, which is characterized by tissue destruction and dense infiltration of mononuclear cells. OBJECTIVES: The aim of this study was to investigate the effect of cell interactions between human gingival fibroblasts and human monocytes on the production of MMP-1 in a coculture model. METHODS: The fibroblasts were cultured in either cell-to-cell contact with monocytes or in separated cocultures using a microporous membrane to prevent cell-to-cell contact. The mRNA expression of MMP-1 was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) and the protein levels of MMP-1 in the cell medium were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: Coculturing gingival fibroblasts with monocytes in cell-to-cell contact increased the mRNA expression of MMP-1 in both fibroblasts and monocytes. The protein levels of MMP-1 increased in the culture media of the cocultures and correlated to the number of fibroblasts as well as to the number of monocytes. When fibroblasts were cultured with monocytes in separated cocultures, the mRNA expression and protein level of MMP-1 increased in the fibroblasts. In addition, treatment of fibroblasts with conditioned medium from monocytes also stimulated the production of MMP-1 in the fibroblasts. Moreover, the levels of the MMP-1 inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), increased in cocultures with cell-to-cell contact, but not in fibroblasts of separated cocultures. The glucocorticoid dexamethasone and the tetracycline doxycycline reduced the enhanced level of MMP-1 in the cocultures with cell-to-cell contact. CONCLUSION: The current study demonstrates that monocytes stimulate the production of MMP-1 in gingival fibroblasts by cell interactions, which may contribute to the maintenance of MMP-mediated tissue destruction in periodontitis.

Adolescent↗
PubMed1988

The total protein concentration of gingival crevicular fluid. Variation with sampling time and gingival inflammation.

The total protein concentration of gingival crevicular fluid (GCF), sampled repeatedly over a 10-min period with the minimum of physical irritation to the sulcus, was evaluated in a group of 32 healthy adolescents. The mean concentration of the 1st sample was comparable to that of normal tissue fluids and lymph, irrespective of the state of inflammation of the sample site. However, during repeated sampling, the values rose to resemble serum protein levels, except at those sites with no clinically detectable inflammation. Gel electrophoretic analysis confirmed the increasing proportion of serum-derived molecules in the more proteinaceous GCF samples. The results demonstrate the extreme sensitivity of the gingival vasculature to GCF sampling and consequently the need for accurate standardisation of GCF collection protocols. This will apply particularly when compositional data is to be normalised with respect to the total protein content or when the levels of a serum constituent are being examined.

Adolescent↗
PubMed1989

Gingival health and gingivitis development during puberty. A 4-year longitudinal study.

The purpose of this investigation was to follow the development of the gingival conditions during puberty and to correlate oral clinical parameters with chronological age as well as with parameters used for the determination of the pubertal development. In 22 boys and 20 girls pubertal and skeletal development, as well as plaque index (PlI) and gingival index (GI) were monitored at 1-year intervals between the ages of 11 and 15 years. During this time, the papillary bleeding index (PBI) was assessed 10 times in all interdental spaces of the dentition. The bleeding tendency, represented by whole mouth mean PBI values, as well as the % of bleeding interdental sites, was found to increase significantly with the start of the pubertal phase. It reached a peak value after 1-5 years in 35% of the children. A significant trend of decrease was noted after the age of 14 years in boys and girls. In boys, mean PBI and the % of interdental sites with bleeding were correlated with testes growth, in girls with the Tanner index for secondary sex characteristics (breast development). PlI and GI, which were only recorded annually, did not show a significant trend of increase or decrease.

Adolescent↗
PubMed1997

Gingival crevicular neutrophils: membrane molecules do not distinguish between periodontitis and gingivitis.

We have earlier reported hyperreactive peripheral neutrophils in adult periodontitis, measured as respiratory burst after Fc gamma receptor-mediated activation in vitro, but we have not been able to relate this increased activity to aberrations in the expression of relevant membrane molecules. Various types of inflammatory conditions involving the gingiva should affect membranes differently. We therefore collected crevicular neutrophils from three types of inflammatory sites: (i) with and (ii) without tissue destruction in the same periodontitis patients and (iii) inflamed sites in controls with gingivitis alone and compared the expression of membrane molecules by flow cytometry. The % of positively stained cells and their mean intensities of fluorescence (IFL) were similar in the three types of sites for CD15, CD11a, CD11b and CD16. Peripheral neutrophils studied with the same markers were not activated. This was verified by similar plasma concentrations of lactoferrin and L-selectins in the periodontal and control groups. Compared to peripheral cells, the crevicular neutrophils showed a significantly lower percentage of stained cells, while the stained cells increased their IFL. In conclusion, hyperreactive peripheral neutrophils in periodontitis show the same expression of membrane molecules after migration through different types of inflammatory lesions as do normal neutrophils in gingivitis.

Adult↗
PubMed1978

Variability of gingival bleeding in experimental gingivitis trials.

Gingival bleeding is a difficult parameter to assess in experimental trials. The Confirmed Bleeding Day was recently developed as a measurement for this purpose. It was observed that considerable variations existed between the measurements made in a pilot study and an experimental trial, and these variations were subjected to a statistical analysis. The reasons for the variations are discussed, and recommendations made for the design of experimental trials involving gingival bleeding.

Chlorhexidine↗