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PubMed1998

Longitudinal changes in TCRB variable gene expression and markers of gingival inflammation in experimental gingivitis.

The aim of this study was to gain information of the cellular and molecular events which occur during the development of experimental gingivitis and to determine whether such changes occur in the presence or absence of alveolar bone resorption. Clinical, radiographic, biochemical and immunological variables were monitored in a 3-week, single-centre, experimental gingivitis study of 10 healthy volunteers. Following screening and professional prophylaxis to achieve visibly healthy gingival status, subjects abstained from all oral hygiene practises in one maxillary (test) quadrant for a period of 21 days. At days 0 and 21, in test and (contralateral) control quadrants, % bleeding on controlled pressure probing (% BOP) was calculated, and radiographic alveolar bone status was assessed using bilateral standardised vertical bite-wing radiographs and digital subtraction radiography (DSR) analysis. In test quadrants, gingival crevicular fluid (GCF) was sampled from 4 sites per subject with Periopaper strips, and prostaglandin E2 (PGE2) levels measured using an enzyme immunoassay (EIA) kit. At days 0, 7 and 21, one interdental papilla was surgically excised from the test quadrant, and the expression of T cell receptor B variable (TCRBV) genes was investigated using a reverse transcription-polymerase chain reaction (RT-PCR) procedure. At days 0, 7 and 21, peripheral blood lymphocytes (PBL) were isolated and additionally investigated for TCRBV gene expression. Following 21 days of plaque accumulation in test quadrants, a statistically significant increase in % BOP scores confirmed the presence of gingival inflammation (p<0.001). DSR analysis revealed that there were no significant alveolar bone changes in either the test or control quadrants between days 0 and 21 (p>0.05). EIA analysis of GCF samples identified a significant decrease in mean GCF PGE2 concentrations from day 0 to day 21 (p<0.05). RT-PCR analysis indicated that genes from all 3 TCRBV families studied (TCRBV-2, -6, -8) were expressed in the PBL samples at all time points and in healthy gingival tissues at day 0. A restriction in the expression pattern of TCRBV genes similar to those which have previously been reported in chronic periodontitis was noted at gingivitis sites. It is possible that such an event may identify susceptibility to periodontal disease independently of other positive predictive markers such as GCF-PGE2.

Adolescent↗
PubMed1990

Relationship between levels of aspartate aminotransferase in gingival crevicular fluid and gingival inflammation.

Data from several sources demonstrate that disease-active and disease-inactive periodontal pockets exist, but currently available diagnostic procedures do not permit identification of disease-active sites at any given point in time. Using the experimental gingivitis model, we have performed studies aimed at determining whether levels of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid correlate with the presence and extent of periodontal inflammation. Gingival inflammation was assessed using the Gingival Index and the Sulcular Bleeding Index, and enzyme activity was measured using a standard procedure. Our data reveal a statistically significant association between AST values and Gingival Index scores for spontaneously occurring lesions (p less than 0.02-0.04) and experimentally induced lesions (p less than 0.0001), as well as the extent of change in these values during developing experimental gingivitis (p less than 0.0001) and resolving experimental gingivitis (p less than 0.0001). The data demonstrate that AST levels can be used to assess the presence and extent of periodontal inflammation.

Adolescent↗
PubMed2004

[The clinical observations and the measurement of gingival crevicular fluid volume during the experimental gingivitis].

OBJECTIVE: To observe the changes of clinical conditions, and measure the GCF volume during the experimental gingivitis in Chinese for studying the relationship between the GCF volume and the development of gingival inflammation. METHODS: 11 young male subjects with healthy gingiva, who had no systemic diseases, were selected for this study. GCF samples (18 teeth/person) were collected with strips of filter paper and the clinical parameters were recorded at the baseline (0 day), the 7th, 14th, 21st day (without oral hygiene), and 28th day (7 days after reestablishing oral hygiene) during experimental gingivitis. The GCF volume was measured by weighting. RESULTS: During the experimental gingivitis, all of the clinical parameters plaque index (PLI), bleeding index (BI), gingival index (GI) and probing index (PD) increased gradually following the plaque assembling, there were significant differences comparing the data of baseline with the data of afterwards without oral hygiene. On the 28th day, the data reduced to the level of baseline rapidly. The amount of GCF had the same tendency with the clinical parameters during the experimental gingivitis. There was positive correlation between the amount of GCF and clinical parameters. CONCLUSION: The amount of GCF can reflect the development of gingival inflammation.

Adult↗
PubMed2004

Effect of cyclosporin A on human gingival fibroblast collagen turnover in relation to the development of gingival overgrowth: an in vitro study.

In a significant number of cases (25-81%) immunosuppressant treatment with cyclosporin A (CsA) is associated with gingival overgrowth, seriously interfering with the functions of mastication and speech. In CsA-induced gingival enlargement, quantitative modifications of the extracellular matrix components occur, and collagen (COL) metabolism and matrix metalloproteinases (MMPs) have been suggested as being the main targets. Since the mechanisms at the basis of CsA-induced gingival overgrowth are not yet completely understood, our aim was to analyze the effect of CsA on COL turnover in cultured human gingival fibroblasts. Cultured human gingival fibroblasts from four healthy volunteers were incubated with CsA (800 ng/ml) or with its vehicle (VH) for variable intervals of time (24, 48, 72 h). Fibroblast morphology was studied by light and electron microscope. Collagen type I (COL-I), MMP-1, MMP-2, TIMP-1 and TGF-beta1 mRNA were determined by RT-PCR; COL-I and MMP-1 by dot blot, and MMP-2 by zymography. Our results evidenced an up-regulation of COL-I and TGF-beta1 gene expression 72 h after CsA treatment. MMP-1, MMP-2 and TIMP-1 mRNA levels are affected but not significantly. Protein analysis revealed COL-I increase at all the considered times and, 72 h after CsA treatment, reduced collagenolytic levels. Our data suggest that COL accumulation during CsA-induced gingival overgrowth may be mainly sustained by an altered COL-I degradation due to decreased MMP-1 activity. However, interindividual differences of collagenase levels after CsA treatment suggest that a genetic predisposition to develop gingival overgrowth may be relevant.

Cells, Cultured↗
PubMed2002

Hepatocyte growth factor in gingival crevicular fluid and the distribution of hepatocyte growth factor-activator in gingival tissue from adult periodontitis.

Hepatocyte growth factor (HGF), also known as scatter factor, is a broad-spectrum and multifunctional cytokine required for the development, growth and regeneration of various organs and tissues. The expression of HGF in human gingival fibroblasts is induced by inflammatory cytokines such as interleukin 1. Thus, although it is possible that content of HGF in gingival crevicular fluid (GCF) in periodontitis is increased, this has not so far been reported because the volume of GCF is too small to determine HGF by the available enzyme-linked immunosorbent assay (ELISA). A recently developed, highly sensitive ELISA for HGF, with a detection limit of 1 pg/ml sample, has now enabled HGF to be measured in GCF.The mean HGF content in GCF from sites with clinically healthy gingiva, defined by the absence of overt signs of gingival inflammation and a probing depth (PD) <3 mm, was 1.7 ng/ml, and that of periodontitis, defined by obvious alveolar bone loss detected by radiographic examination and a PD> or =3 mm, was 3.23 ng/ml. Although treating the periodontitis did not significantly decrease the HGF concentration despite significantly improved clinical scores such as PD and Gingival Index, the total amount of HGF in GCF did decrease significantly after treatment. HGF was expressed by gingival fibroblasts and inflammatory cells as determined by in situ hybridization. HGF-activator (HGFA), which converts inactive pro-HGF to active mature HGF, was detected in gingival epithelial cells by immunostaining. The expression of HGFA was also confirmed in gingival tissue by reverse transcription-polymerase chain reaction (RT-PCR). These findings indicate that HGF is synthesized and activated in gingiva that is clinically healthy or associated with periodontitis.

Adult↗
PubMed1994

Effect of pre-experimental maintenance care duration on the development of gingivitis in a partial mouth experimental gingivitis model.

The present study was designed to evaluate if different periods of supervised oral hygiene prior to the experimental gingivitis trial have an effect on the amount of inflammation which develops in the 'toothshield' model. Two groups were compared. One group (Group A) was supervised over a 5-month period before experimental gingivitis. Six months later Group A participated in a second trial, this time receiving only a 1-month pretrial. A second group (Group B) which served as a control was supervised over a pre-trial period of 1-month prior to the start of the experimental gingivitis phase. During the pre-trial period all subjects undertook a regime of vigorous oral hygiene which included polishing of the test quadrant in the upper jaw and supervised subgingival brushing, interdental taping or the use of toothpicks. The presence of plaque and bleeding were assessed. Following the pre-trial period all plaque control measures in the experimental area were prevented during oral hygiene periods by the temporary placement of soft, loosely fitting vinyl toothshield guard. During this period they accumulated plaque rapidly and developed a generalized gingivitis. The results of this study indicate that, irrespective of the length of the pretrial period, subjects by group develop a comparable mean level of plaque and gingival bleeding. However, the extent to which gingivitis develops differs among individuals and was for a number of subjects not consistent. Therefore, in designing an experimental gingivitis trial care should be taken to include a sufficient number of subjects to account for individual variation.

Adult↗
PubMed2004

Treatment of chronic desquamative gingivitis using tissue-engineered human cultured gingival epithelial sheets: a case report.

Human cultured gingival epithelial sheets were used as an autologous grafting material for regenerating gingival tissue in the maxillary left and mandibular right quadrants of a patient with chronic desquamative gingivitis. Six months post-surgery in both treated areas, there were gains in keratinized gingiva and no signs of gingival inflammation compared to presurgery. In the maxillary left quadrant, preoperative histopathologic findings revealed the epithelium was separated from the connective tissue and inflammatory cells were extensive. After grafting with the gingival epithelial sheets, inflammatory cells were decreased and separation between epithelium and connective tissue was not observed. The human cultured gingival epithelial sheets fabricated using tissue engineering technology showed significant promise for gingival augmentation in periodontal therapy.

Cell Culture Techniques↗
PubMed1995

The pharmacokinetics of phenytoin in gingival crevicular fluid and plasma in relation to gingival overgrowth.

The aim of this study was to determine whether phenytoin (PHT) could be detected in gingival crevicular fluid (GCF), and to relate its concentration to both plasma level and degree of gingival overgrowth. 23 patients medicated with phenytoin for at least 6 months were clinically examined for signs of periodontal disease and gingival overgrowth. 12 patients out of these demonstrated clinically significant overgrowth and their plaque scores and gingival inflammation were greater than for the non-overgrowth group (p < 0.001). Phenytoin concentrations were determined by high performance liquid chromatography, and was detected in GCF. There was a significant correlation between the GCF and plasma phenytoin concentrations (p < 0.05), but it was not related to the extent of gingival overgrowth. Inflammation increased the GCF volume, but was not a determinant of GCF phenytoin concentration. It is concluded that effusion of phenytoin into GCF is regulated by the plasma levels of the drug, but its concentration in GCF is not related to the incidence of gingival overgrowth.

Adult↗
PubMed1987

Hyaluronic acid and hyaluronidase activity in gingival exudate from sites of acute ulcerative gingivitis in man.

Gingival exudates from sites of acute ulcerative gingivitis (AUG) and chronic gingivitis (CG) in adults were investigated by cellulose-acetate electrophoresis for the hyaluronic acid (HA) content and assayed for the levels of HA-degrading enzymes. HA was the only glycosaminoglycan (GAG) in gingival exudate from CG sites. HA was not detected at untreated AUG sites but was evident, at increasing levels, after two and seven days of effective antibacterial treatment. In AUG exudates, the total HA-degrading enzyme activity, of bacterial origin, decreased to approx. 30 and 10 per cent of the high initial levels after two and seven days of treatment respectively, to that level found at sites of CG. The specific activity of HA-degrading enzyme of lysosomal origin was low initially and increased with treatment to a level comparable to CG. The notable absence of HA from gingival exudate from sites of untreated AUG thus appears to result from the increased levels of bacterial hyaluronidase. Electrophoresis of gingival exudate may be an indirect method of monitoring the rate of response of AUG to different antibacterial treatments.

Acute Disease↗
PubMed2000

Gingival crevicular fluid inflammatory mediators and bacteriology of gingivitis in nonhuman primates related to susceptibility to periodontitis.

The hypothesis to be tested was that the microbiota and resulting local host inflammatory response characteristics in oral conditions of high levels of chronic gingival inflammation increases susceptibility to progressing periodontitis. This study used cynomolgus monkeys, Macaca fascicularis (nonhuman primates), with high and low levels of long-standing gingival inflammation to define the profiles of gingival crevicular fluid mediators, cytokines and immunoglobulins; describe the subgingival microbiota; and evaluate their susceptibility to ligature-induced periodontitis. Sixteen nonhuman primates were stratified into two groups (HI, LO) based upon Bleeding Index as a measure of the natural level of inflammation (HI = 1.26 +/- 0.45; LO = 0.22 +/- 0.16). The host mediator levels, subgingival microbiota, and clinical characteristics of the LO and HI groups were compared after 30 days of oral hygiene, during a 30 day experimental gingivitis (7, 14, and 30 days), and during periodontitis (30, 60, and 90 days). The results demonstrated that nonhuman primates with high levels of long-standing gingival inflammation when compared to those nonhuman primates with low inflammation show: 1) different inflammatory mediator profiles in gingival crevicular fluid (particularly for immunoglobulin A (IgA) and IgG levels), 2) a different quantitative and qualitative subgingival microbiota; and 3) a similar progression of periodontitis. Thus, while variations in host inflammatory responses to local factors exist in the nonhuman primates, an extensive subgingival challenge (such as ligation) may negate these individual differences.

Animals↗
PubMed2000

Detection of tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 2(PAI-2) in gingival crevicular fluid from healthy, gingivitis and periodontitis patients.

BACKGROUND: The regulation of plasminogen activation is a key element in controlling proteolytic events in the extracellular matrix. Our previous studies had demonstrated that in inflamed gingival tissues, tissue-type plasminogen activator (t-PA) is significantly increased in the extracellular matrix of the connective tissue and that interleukin 1beta(IL-1beta) can up regulate the level of t-PA and plasminogen activator inhibitor-2 (PAI-2) synthesis by human gingival fibroblasts. METHOD: In the present study, the levels of t-PA and PAI-2 in gingival crevicular fluid (GCF) were measured from healthy, gingivitis and periodontitis sites and compared before and after periodontal treatment. Crevicular fluid from106 periodontal sites in 33 patients were collected. 24 sites from 11 periodontitis patients received periodontal treatment after the first sample collection and post-treatment samples were collected 14 days after treatment. All samples were analyzed by enzyme-linked immunosorbent assay (ELISA) for t-PA and PAI-2. RESULTS: The results showed that significantly high levels of t-PA and PAI-2 in GCF were found in the gingivitis and periodontitis sites. Periodontal treatment led to significant decreases of PAI-2, but not t-PA, after 14 days. A significant positive linear correlation was found between t-PA and PAI-2 in GCF (r=0.80, p<0.01). In the healthy group, different sites from within the same subject showed little variation of t-PA and PAI-2 in GCF. However, the gingivitis and periodontitis sites showed large variation. These results suggest a good correlation between t-PA and PAI-2 with the severity of periodontal conditions. CONCLUSION: This study indicates that t-PA and PAI-2 may play a significant rôle in the periodontal tissue destruction and tissue remodeling and that t-PA and PAI-2 in GCF may be used as clinical markers to evaluate the periodontal diseases and assess treatment.

Adult↗
PubMed1998

Pre-transplant gingival hyperplasia predicts severe cyclosporin-induced gingival overgrowth in renal transplant patients.

The relationship between the pre-transplant periodontal status and the development of post-transplant gingival overgrowth was investigated in a longitudinal study. The periodontal condition of 35 patients was examined on 2 occasions while they were on the transplant waiting list and then at 4-6, 10-12, 16 and 20 weeks post-transplant. At each visit the plaque index, the bleeding index and a pocket index (CPITN) were measured. Dental impressions were taken of the pre- and post-transplant gingival condition and used to make stone models which were used to score the gingival overgrowth index (GOI). The patients divided into 3 distinct groups having severe (n=13), mild (n=16) or no post-transplant gingival overgrowth (n=6). Only 1 of the patients had taken cyclosporin prior to inclusion into the study. All the patients who developed severe overgrowth had evidence of gingival hyperplasia before the transplant. There was no difference in the serum cyclosporin levels between the three groups (chi2<2.28, p>0.319). Furthermore, there was no statistical difference for any of the periodontal indices. This study indicates that the hyperplastic gingival inflammatory response of some individuals appears to be potentiated by cyclosporin resulting in severe post-transplant overgrowth. In other patients the same reaction may allow the fibroblastic activity to occur to an extent where it produces a mild clinically apparent overgrowth.

Chi-Square Distribution↗
PubMed1993

Disposition of nifedipine in plasma and gingival crevicular fluid in relation to drug-induced gingival overgrowth.

The present study investigates the relationship between the pharmacokinetic variables of nifedipine with the incidence and severity of gingival overgrowth in 9 adult male patients medicated with the drug for at least 6 months. Five of the patients had experienced significant gingival changes and were thus designated "responders". The remaining four patients exhibited no gingival overgrowth, and thus acted as a control. A baseline periodontal examination (plaque scores, bleeding index and gingival overgrowth assessment) was carried out on each patient, and confined to the upper and lower anterior teeth. Serial blood and gingival crevicular fluid samples were collected over an eight-hour investigation period. Samples were analyzed for nifedipine by gas chromatography. No significant difference (p > 0.05) was seen between responders and non-responders with regard to drug therapy, periodontal parameters or plasma pharmacokinetics of nifedipine. Nifedipine was detected in the gingival crevicular fluid of seven subjects (all responders, and two non-responders). The peak concentration of nifedipine in crevicular fluid was 15-90 fold greater than levels observed in plasma.

Adult↗
PubMed1995

Identification and possible function of cathepsin G in gingival crevicular fluid from chronic adult periodontitis patients and from experimental gingivitis subjects.

The levels of cathepsin G in gingival crevicular fluid (GCF) from chronic adult periodontitis patients and experimental gingivitis subjects were determined both by activity measurement using N-benzoyl-(DL)-phenylalanine-2-naphthyl ester as a substrate and by enzyme immunoassay using anti-human cathepsin G IgG. The activity level of cathepsin G in GCF of both periodontitis and experimental gingivitis has no significant correlation with all measured clinical parameters. Western immunoblotting using antibodies specific for cathepsin G or alpha 1-proteinase inhibitor revealed that the difficulty in demonstrating the association of its activity with the severity of these diseases was due largely to formation of the enzyme-inhibitor complexes. By contrast, statistically significant positive correlation was found between cathepsin G content in GCF of periodontitis, which was determined by enzyme immunoassay, and such clinical parameters as the GCF volume, the gingival index and probing depth. The increased cathepsin G content with increasing severity of periodontal inflammation was markedly diminished by the initial treatment. Although no significant activity was detectable in GCF of experimental gingivitis, a rapid increase of the immunoreactive cathepsin G was found in GCF at 3-5 d after refraining from oral hygiene measures, which rapidly decreased by 10 d. The progressively increased cathepsin G between 10th and 21st d rapidly decreased by cleaning of the teeth. The results indicate that cathepsin G is involved in the host's defensive mechanism against the invasion of etiologic microbes and/or the development of either periodontitis or gingivitis.

Adult↗
PubMed1982

Increased thymocyte-activating factor in human gingival fluid during gingival inflammation.

In this study, human gingival fluid from normal subjects was shown to contain a low-molecular-weight factor (molecular weight, 10,000 to 25,000) which augmented murine thymocyte proliferation, enhanced the production of interleukin 2 by T lymphocytes, and augmented the proliferation of fibroblasts. These biochemical and biological properties are characteristic of both macrophage-derived interleukin 1 and epidermal cell-derived thymocyte-activating factor. In addition, we have established that epidermal thymocyte-activating factor or interleukin 1 or both are present to a greater extent in gingival fluid obtained from sites manifesting gingival inflammation. In fact, thymocyte-activating activities were found to be greater in gingival fluid from inflamed than from noninflamed gingival sites from the same subjects. These findings suggest that human gingival fluid contains epidermal thymocyte-activating factor or interleukin 1 or both, which may amplify the inflammatory response in periodontal tissues.

Cell Division↗
PubMed1992

The relationship of gingival fluid leukocyte elastase activity to gingival fluid flow rate.

Recent studies have suggested that leukocyte elastase activity (EA) in tissue exudates is an indicator of inflammatory disease. We assayed gingival fluid (GF) EA with a selective peptide substrate and compared it to GF flow rate with regard to its ability to detect differences in the clinical status of existing inflammatory periodontal disease in 56 human subjects. Compared to healthy sites (Gingival Index = 0, 1 to 3 mm) and mild gingivitis sites (GI = 1, 2 to 5 mm), mean GF EA was significantly (P < 0.05) higher at periodontitis sites with deep probing depths (GI = 2, 6 to 9 mm depth), but not at periodontitis sites with intermediate probing depths (GI = 2, 4 to 5 mm). When expressed as specific EA (i.e., normalized to GF protein content), mean EA was also significantly higher at deep periodontitis sites compared to healthy sites and mild gingivitis sites. In addition, specific EA was significantly higher at periodontitis sites with intermediate probing depths than at healthy sites. As predicted by previous studies, these significant increases in specific EA were associated with significant increases in mean GF flow rate. In contrast to specific EA, however, mean GF flow rate was significantly higher at gingivitis sites than at healthy sites. A strong correlation was observed between GF flow rate and specific EA (rs = 0.737, P = 0.0006). Thus, GF flow rate and GF EA appear to be related indicators of inflammation, but GF flow rate may be more sensitive to early inflammatory changes leading to mild gingivitis.

Alveolar Bone Loss↗
PubMed1998

Gingival crevicular pH in experimental gingivitis and occlusal trauma in man.

The purpose of this study was to investigate the fluctuation of gingival crevicular pH during experimentally evoked gingivitis and occlusal trauma, and to examine the relationship between the pH and periodontal health status using a newly developed pH sensor. Maxillary first premolars with clinically healthy gingiva were selected as test teeth in 10 volunteers. In the first study phase, experimental gingivitis was evoked, and gingival index, plaque index, and crevicular pH were recorded during the study period. In the second study phase, experimental occlusal trauma was created with metal onlays having occlusal interferences in lateral movements, and tooth mobility and pH were recorded during the study period. In the gingivitis phase, GI, PlI, and pH values fluctuated significantly during the study period (P < 0.01) and positive correlations were observed between both GI and pH values (P < 0.05), and PlI and pH values (P < 0.01). In the occlusal trauma phase, significant fluctuation was found among tooth mobilities (P < 0.05) during the study phase, but not in pH values. Statistically significant correlations were not observed between tooth mobilities and pH values. These data suggest that the crevicular pH level may not be influenced by experimental occlusal trauma, but shifts toward alkaline with experimental gingivitis.

Adult↗
PubMed1999

A comparison of the efficacy of 2 powered toothbrushes in affecting plaque accumulation, gingivitis, and gingival bleeding.

BACKGROUND: The purpose of this 30-day blinded, parallel-design study was to compare the effect of 2 powered toothbrushes, the Rowenta MH700 and the Braun Plak Control Ultra, on reducing plaque accumulation, gingivitis, and gingival bleeding in a cohort of 60 healthy adults. METHODS: After baseline evaluation of plaque, gingivitis, and gingival bleeding, subjects were randomly assigned to one of the experimental groups, provided written and verbal toothbrushing instructions, and had their teeth polished. At 2 weeks (follow-up 1) and 4 weeks (follow-up 2), all clinical parameters were again evaluated. RESULTS: The Braun group demonstrated a nearly significant reduction in gingival index (GI) and a significant reduction in GI at follow-up 2. The Rowenta group demonstrated significant reductions in GI, plaque index (PI), and bleeding index (GBI) at both follow-up 1 and 2 examinations. At follow-up 1, the Braun group demonstrated a nearly significant reduction in GI, a significant reduction in PI, and a non-significant reduction in GBI. At follow-up 2, the Braun group demonstrated a significant reduction in GBI, but not a significant reduction in PI. The reduction in GI for the Rowenta group was significantly greater (P values of 0.0001 and 0.0001, respectively) than that demonstrated in the Braun group. However, the Rowenta group did not demonstrate a significantly greater reduction in PI (P values of 0.7135 and 0.3184 for follow-up 1 and follow-up 2, respectively) or GBI than the Braun group at either examination (P values of 0.0663 and 0.3397 for follow-up 1 and 2, respectively). CONCLUSIONS: The results of this study support the findings of numerous other studies that powered toothbrushes have great potential to remove plaque and improve gingival health and that the improvement can be demonstrated in a relatively short period of time.

Adolescent↗