Gingivitis Subjects Research Articles

Clonal diversity of Actinobacillus actinomycetemcomitans isolates from young adults with minimal periodontal disease.

Actinobacillus actinomycetemcomitans is a major periodontal pathogen which is associated with both early-onset periodontitis and adult cases refractory to conventional periodontal therapy, although the organism has also been shown to be widely distributed among dentate healthy individuals. The observed disease status may be associated with a variation in virulence of different strains or clones. The aim of the present study was to analyse genotype distribution as assessed by an arbitrarily primed polymerase chain reaction (AP-PCR) among 51 isolates of A. actinomycetemcomitans recovered from more than 200 young adult recruits with no or minor periodontal disease. In addition, isolates from 25 periodontitis patients as well as reference strains were genotyped. Primers amplifying (i) a specific sequence in the ltxA region, (ii) a specific 16S rRNA sequence and (iii) sequences in the leukotoxin promoter region were used to verify species identity of the strains. Three random oligonucleotide primers were employed to analyse genomic polymorphisms of the organism by means of PCR. A total of 19 genotypes could be distinguished, which were grouped by cluster analysis into 5 major clusters based on genetic similarity and a complete linkage sort. Whereas 3 clusters assembled A. actinomycetemcomitans genotypes isolated from both healthy subjects and periodontitis patients, one cluster containing 4 different genotypes exclusively comprised isolates from healthy or gingivitis subjects. Another cluster with 2 genotypes consisted of strains originating from periodontitis patients (p < 0.05). One strain characterized by a specific 530 bp deletion in the promoter region of the leukotoxin region was identified in a Ghanese patient with localized juvenile periodontitis. It was concluded that there is considerable clonal diversity of A. actinomycetemcomitans strains isolated from healthy or periodontally diseased subjects, and that genetically closely related groups might be associated with health or disease.

Antigen-specific T-cell receptor V beta expression in Porphyromonas gingivalis-specific T-cell lines.

FACS analysis was used to determine the expression of 15 T-cell receptor V beta families on CD4 and CD8 cells in Porphyromonas gingivalis specific T-cell lines established from eight P. gingivalis-positive adult periodontitis and seven P. gingivalis-positive healthy or gingivitis subjects. All 15 T-cell receptor V beta families were expressed by the T-cell lines, although a significantly higher proportion of the CD4 cells expressed the 5.2-3 V beta region compared with the other 14 families, including the 5.3 region, suggesting that it is the 5.2 family which is overexpressed. This was also true for the CD8 cells, with the exception of the 3.1 region in adult periodontitis T-cell lines and the 3.1, 13.1/13.3 and 21.3 regions in healthy or gingivitis lines. Between the two clinical groups, a significantly lower percentage of 13.1/13.3-positive CD8 cells was noted in the adult periodontitis lines compared with the healthy or gingivitis lines. There was a significant reduction in DNA synthesis by the lines in the presence of P. gingivalis outer membrane antigens and fixed irradiated lymphoblastoid cell lines compared with cultures containing untreated irradiated lymphoblastoid cell lines and in cultures containing anti-class II major histocompatibility complex antibody in comparison with all other cultures. The results of this study have shown that P. gingivalis preferentially induces the T-cell receptor V beta 5.2 family on CD4 and CD8 cells in P. gingivalis-specific T-cell lines and that activation of T cells by P. gingivalis outer membrane antigens may be by antigen-specific rather than superantigen activity.

Clinical and microbiological findings in elderly subjects with gingivitis or periodontitis.

The objective of the present study was to study the supra- and subgingival microflora by culture and cDNA probe methods in 20 elderly subjects who were between 62 and 93 years of age. 10 of them had gingivitis only, and 10 had periodontitis. B. forsythus (BF), P. gingivalis (PG), P. intermedia (PI), P. nigrescens (PN), A. actinomycetemcomitans (AA), T. denticola (TD), and pathogen-related oral spirochetes (PROS) were studied. Oral hygiene was similar and poor in both groups. The mean probing depth at sample sites was 6.7 mm (S.D+/-1.3) in the periodontitis group and 2.2 mm (S.D.+/-1.5) in the gingivitis group (F=17.75, p<0.001). Mean clinical attachment levels (CAL) were 4.3 mm (S.D.+/-2.0) and 1.7 mm (S.D.+/-0.9) respectively (p<0.001). Total viable counts >1.0x10(5) in supra-gingival plaque samples were found in all periodontitis and in eight gingivitis subjects. 70x more black-pigmented organisms were found in supra-gingival and 185 times more in sub-gingival plaque from the periodontitis group (p<0.01). Culture data showed P. nigresecens in 10% periodontitis and 50% gingivitis subjects (p<0.03). In supra-gingival samples by the Affirm DP test, BF was present in 50% periodontitis and 60% gingivitis while culture data were negative for all subjects. PG was found in 30% periodontitis and 50% gingivitis subjects with TD in 70% periodontitis and in 30% gingivitis subjects. In the sub-gingival plaque samples 80% periodontitis and 70% gingivitis subjects had >1x10(5) anaerobes. The total count of black-pigmented organisms was significantly greater in the periodontitis elders (p<0.001). cDNA probes by the Affirm DP test identified subgingival presence of BF (80%) PG (80%), PI (80%), AA (0%), TD (50%) in periodontitis subjects with BF (70%), PG (40%), PI (30%) and TD (20%) in gingivitis subjects. PROS were found in (80%) samples from periodontitis and in (60%) of gingivitis elderly. Only the quantities of PI (r=0.48, p<0.01) and TD (r=0.37, p<0.01) were associated with the disease definition. The smoking habit in the periodontitis group was significantly higher (p<0.01). A history of smoking may contribute significantly to periodontitis in the presence of pathogens.

Occurrence of Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola in periodontally healthy and diseased subjects as determined by an ELISA technique.

The aim of this study was to assess by means of an ELISA technique, the occurrence of 3 putative periodontopathogens, Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola, in 3 clinically-defined adult periodontal conditions. Thirty systemically-healthy subjects were selected and grouped into 3 categories according to their periodontal health: 1) 10 periodontitis subjects (PS), having moderate adult chronic periodontitis; 2) 10 untreated gingivitis subjects (UGS), exhibiting no signs of periodontal destruction but presenting with clinical signs of mild gingivitis; and, 3) 10 treated gingivitis subjects (TGS), having the same clinical status as UGS, but who received a thorough prophylaxis treatment within the past 7 to 14 days prior to the baseline examination. A total of 60 samples were collected subgingivally from the six Ramfjord teeth per subject in each group and ELISA analysis was carried out to give a semiquantitative estimate of P. gingivalis. B. forsythus, and T. denticola. The immunologic detection method suggested the presence of antigens of P. gingivalis, B. forsythus, and T. denticola in subjects from each of the 3 groups. When a global analysis for the 3 disease groups was performed at one time, statistically significant differences were found among the ELISA scores of the 3 bacterial species. For example, comparisons of the ELISA scores showed that the concentrations of P. gingivalis differed significantly when comparing TGS to UGS and PS, but not when examining UGS/PS. The ELISA scores for B. forsythus were significantly different between TGS and PS. Mean concentrations of T. denticola were significantly different when comparing PS to TGS or UGS, whereas no difference was found between the latter categories. Within the limited scope of this study, the concentration of antigens detectable from putative periodontopathogens like P. gingivalis, B. forsythus, and T. denticola differed among the 3 diseased groups, with periodontitis subjects often showing the greatest level of antigens. Thus, it is reasonable to expect that, when using sensitive immunological detection methods, antigens of suspected periodontal pathogens can be found irrespective of the individual's clinical status. However, while detectable in the periodontal sites, the concentrations of these microorganisms are most likely to be above the threshold necessary to induce clinically-significant disease. Studies with larger sample size and standardized antigens are necessary to determine if the groups we found not to differ, were, in fact, different.

Clinical, microbiological and immunological profile of healthy, gingivitis and putative active periodontal subjects.

Thirteen periodontally healthy subjects were monitored clinically for 6-12 months. Clinical measurements at 6-weekly intervals included duplicate PD measurements, presence of plaque, redness, and bleeding on probing. Baseline measurements consisted of 2 visits 1 wk apart. Microbial samples were taken from 11 of the subjects who had completed at least 8 months of monitoring. Levels of serum antibodies to 12 periodontal species were determined from 10 subjects. Standard deviations of replicate PD measurements, computed for each subject, ranged from 0.2-0.3 mm over the monitoring period. Plaque and redness increased during monitoring, and showed a weak association with PD change. Baseline and follow-up distributions of PD changes indicated that changes of > 1.5 mm could reasonably be considered to represent active sites. Five subjects demonstrated at least 1 site deepening by 1.5 mm over the period monitored, and these were considered putative active subjects. Sites from 2 subjects showed PD increases in the 6 wk just before sampling, and these were considered to represent active sites. Species associated with putative active subjects included Actinomyces naeslundii, Veillonella parvula, Selenomonas noxia and Prevotella nigrescens. Streptococcus sanguis, S. gordonii and Peptostreptococcus micros were associated with inactive subjects. S. gordonii and S. oralis were associated with health, whereas P. nigrescens was associated with gingivitis. Elevated serum antibodies were detected to A. actinomycetemcomitans in 4 subjects. The predominant microbiota of putative active subjects included some species previously associated with gingivitis, and some species previously associated with progressing periodontitis.

Protein composition of whole and parotid saliva in healthy and periodontitis subjects

Cystatins are physiological inhibitors of cysteine proteinases and they are widely distributed in human tissues and body fluids including saliva. We previously reported an increased cystatin activity in whole saliva of gingivitis and periodontitis subjects. Based on this result we decided to investigate the type and origin of cystatins involved in this increased cystatin activity by collecting both whole and parotid saliva of 25 healthy and 30 periodontitis subjects. Saliva samples were quantified for cystatins S and C by enzyme-linked immunosorbent assay and cystatin activities were measured toward papain. Besides, three other salivary proteins were determined: the plasma protein albumin, the typical parotid derived amylase and the salivary immunoglobulin IgA. The present investigation shows that levels of total protein and cystatin activity as well as the levels of glandular derived proteins amylase and cystatin C were significantly higher in whole and parotid saliva of subjects with periodontitis than in healthy controls. Cystatin S, the major salivary cystatin, however was higher in the whole saliva of the healthy group. Whole saliva concentrations of albumin and IgA, originating from sources other than the glandular cells, were not different between healthy and periodontitis subjects and were also not correlated with the typical salivary gland proteins. In conclusion, this study provides additional evidence that the human salivary glands may respond to an inflammatory disease of the oral cavity, periodontitis, by enhanced synthesis of some acinar proteins.

Antibody responses of Porphyromonas gingivalis infected gingivitis and periodontitis subjects

Porphyromonas gingivalis demonstrates a strong association with adult periodontitis although some individuals with the infection do not experience attachment loss. Therefore differences in the immune response to this organism may be of importance to the outcome of the disease. The aim of this study was to determine whether P. gingivalis positive subjects with and without periodontal breakdown, reacted differently to P. gingivalis antigens as assessed by the pattern of serum antibody reactivity. Two highly defined groups of subjects were chosen for this study. Both demonstrated P. gingivalis in their plaque and both had responded to P. gingivalis as shown by the presence of serum antibodies. The two groups differed only in their apparent clinical susceptibility to periodontal breakdown. Western blots of P. gingivalis membrane antigens were probed with sera from the two groups to determine their reactivity to specific antigens. Analysis of the immunoblots showed that there were no differences in either the total numbers of bands, or bands recognized by the majority of subjects in the gingivitis and adult periodontitis groups. There were however, four bands recognized by the majority of the gingivitis group and not by the majority of the adult periodontitis group, there being a significant difference (P = 0.03) in the recognition of the 91.4-kDa antigen band. A further five antigens of lower molecular weight were seen by the majority of the adult periodontitis group and not by the majority of the gingivitis group. When sera were tested against purified P. gingivalis LPS, the results indicated that the five antigens seen by the majority of the adult periodontitis group had molecular weights which were in the range exhibited by the LPS antigens. These results suggest that gingivitis and adult periodontitis subjects with P. gingivalis infection, may recognize different P. gingivalis antigens.

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.

Studies on periodontal pathogenic proteinases from Porphyromonas gingivalis and host cells

Progressive periodontal disease is characterized by acute progressive lesions of gingival connective tissues, excessive leukocyte infiltration, and occurrence of a characteristic microflora. A variety of proteolytic enzymes derived from oral bacteria and host cells are found in gingival crevices and thought to play an important role in the onset and development of progressive periodontal disease. The anaerobic bacterium Porphyromonas gingivalis has been implicated in the etiology of the disease. Recently, we have purified a novel arginine-specific cysteine proteinase, termed "argingipain", from the culture supernatant of the organism. The enzyme was shown to have two important abilities related to the virulence of the organism. One is direct association with periodontal tissue breakdown through its abilities to degrade physiologically important proteins such as human collagens (type I and IV) and to evade inactivation by internal protease inhibitors. The other is associated with disruption of the normal host defense mechanisms through its abilities to degrade immunoglobulins and to inhibit the bactericidal activity of polymorphonuclear leukocytes. The virulence of argingipain was further substantiated by disruption of argingipain-encoding genes on the chromosome by use of suicide plasmid systems. On the other hand, we have studied roles of host cell-derived proteinases in the periodontal tissue breakdown. Levels of lysosomal proteinases such as cathepsins B, H, L, G and medullasin were determined in gingival crevicular fluid from periodontitis patients and experimental gingivitis subjects by activity measurement and sensitive immunoassay. The results suggested that all of these enzymes would be involved in the development of both gingivitis and periodontitis.

Adhesion molecule expression in chronic inflammatory periodontal disease tissue.

Differences in lymphocyte populations have been demonstrated in gingivitis and periodontitis lesions. A differential expression of adhesion molecules may play a role in lymphocyte trafficking in these tissues. An indirect avidin biotin immunoperoxidase technique was used to stain a range of adhesion molecules in tissue sections of 21 gingival biopsies from both gingivitis and periodontitis subjects. These specimens were placed into three groups according to the size of the infiltrate. ICAM-1, PECAM-1 and LECAM-1 expression on mononuclear cells in the inflammatory infiltrates increased significantly with increasing size of infiltrate. Approximately 50% of these mononuclear cells were LFA-1+ and CD29+. When specimens were grouped according to their putative disease status there were no significant differences between mononuclear cell adhesion molecule expression in small infiltrates from either gingivitis or adult periodontitis subjects. This was also the case with larger lesions from both clinical groups. Therefore there does not appear to be a differential expression of adhesion molecules on lymphocytes in gingivitis and periodontitis tissue. Endothelial cells were positive for ICAM-1, PECAM-1, CD29, GMP-140 but negative for ELAM-1. Keratinocyte expression of ICAM-1 increased with increasing size of infiltrate although in heavy infiltrates, cells in the region of the junctional epithelium which were positive in small lesions, became ICAM-1 negative. The upper layers of the oral epithelium were positive for LECAM-1 in small infiltrates and with increasing size of infiltrate, the lower layers and many of the sulcular and junctional epithelium keratinocytes were positive.(ABSTRACT TRUNCATED AT 250 WORDS)

Host responses in patients with generalized refractory periodontitis.

Although patients with refractory periodontitis have been widely reported, no clear biologic profile of these patients has been noted. The purpose of the present study was to evaluate host responsiveness of a well-defined group of refractory periodontitis patients by determining the effect of a lipopolysaccharide (LPS) challenge on monocyte surface receptor density and on the release of inflammatory mediators. Venous blood was obtained from 7 refractory periodontitis, 8 stable periodontal maintenance, and 8 gingivitis patients with no evidence of periodontitis. Mononuclear cells were cultured in either control media or media treated with Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), or Salmonella typhimurium (S. typh) LPS. At 0 and 24 hours supernatants were assayed for prostaglandin-E2 (PGE2) and interleukin-1 beta (Il-1 beta) release by ELISA. Using flow cytometry the density of specific monocyte surface receptors were assayed with Mo3e and LeuM3 monoclonal antibodies (mAb); T-cell CD4/CD8 ratios were assayed with OKT-3, OKT-4, and OKT-8 mAb. After 24 hours incubation with Pg or S. typh LPS, the upregulation of the Mo3e receptor was significantly decreased for refractory periodontitis patients (P < 0.05) when compared to gingivitis and to stable maintenance patients. In refractory periodontitis patients the T-cell CD4/CD8 ratio was decreased. Upon stimulation with Pg or S. typh LPS, monocytes from stable maintenance and refractory periodontitis patients released more Il-1 beta (P < 0.05) and PGE2 (P = 0.13 and 0.15) than monocytes from gingivitis subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Reproducibility of an electronic probe in relative attachment level measurements.

4 probing designs have been employed to investigate the reproducibility of the Florida Probe. 3 groups (each composed of 10 subjects) were selected for the study: healthy adults, gingivitis subjects, and periodontitis subjects. The 4 probing designs were as follows: (a) the probe tip was left in the sulcus between successive probings; (b) the probe tip was removed from the gingival margin between probings but the next probing followed immediately; (c) successive whole-mouth probings were interrupted by a 5-min interval and a mouthrinse; (d) there was a 4-week interval between each probing. 3 measurements were taken for each design. The main purpose of this study was to identify variance components in the attachment level variation. The maximum probing error standard deviation was found to be around 0.3 mm, which is considerably smaller than that found in most previous studies. The errors associated with the periodontal condition and probing effect were also estimated. The variance components obtained here can be used for determining the sample size in controlled clinical studies.

Granulocyte medullasin levels in gingival crevicular fluid from chronic adult periodontitis patients and experimental gingivitis subjects

The concentration of medullasin, an elastase-like serine proteinase, in gingival crevicular fluid (GCF) from chronic adult periodontitis patients and experimental gingivitis subjects was determined by the highly sensitive immunoassay method. In periodontitis patients, the medullasin content increased with increase of the GCF volume and then attained a maximum value at a relatively mildly inflamed stage. The value was maintained through more serious stages of disease activity. However, the medullasin content was independent of the probing depth. The medullasin content of the patients was markedly decreased after periodontal treatment, indicating that the enzyme participates in the development of the chronic periodontitis. Large amounts of medullasin were also detected in GCF from experimental gingivitis subjects, although it was not detected by the activity measurements. There was a rapid increase in the medullasin content during the 4-day period after abstention from oral hygiene measures, which corresponded to those of severely inflamed periodontitis patients. The peak value decreased up to the 7th-d followed by a gradual increase during the 21-d experimental period. The increased medullasin level rapidly decreased following resumption of oral hygiene measures. The results suggest that medullasin plays important roles both in the defence mechanism against the gingival inflammation and in the development of the acute inflammation.

Accessible sialic acid content of oral epithelial cells from healthy and gingivitis subjects

The purpose of the present study was to determine if there were differences in the quantity of accessible sialic acid on superficial epithelial cells collected from different areas of the mouth, and from healthy subjects with good oral hygiene, as compared to subjects with gingivitis. Superficial epithelial cells were collected by gently scraping the tongue dorsum, hard palate, free gingiva and buccal epithelium. The cells were washed and treated with clostridial neuraminidase to release accessible sialic acid; this was quantitated using a fluorometric assay. Buccal cells released an average of 62.6 ng sialic acid per 10,000 cells, which was nearly 3-fold more than cells from the hard palate (24.1 ng), free gingiva (21.9 ng), or tongue (15.4 ng). Buccal and free gingival cells collected from 5 healthy subjects had significantly higher levels of accessible sialic acid on their surface than cells collected from 5 subjects with gingivitis. These differences were significant at the p less than 0.001 and p less than 0.01 levels, respectively. The data obtained suggest that the oral hygiene status of an individual can influence the quantity of accessible sialic acid residues on oral epithelium; this would be expected to influence the attachment and colonization of bacteria which bind to sialic acid-containing receptors.

Cathepsins B, H and L activities in gingival crevicular fluid from chronic adult periodontitis patients and experimental gingivitis subjects

To clarify roles of lysosomal cysteine proteinases cathepsins B, H and L in pathological destructive process of periodontal tissues, levels of their enzymatic activities were determined in gingival crevicular fluid (GCF) from chronic adult periodontitis patients and from experimental gingivitis subjects. In periodontitis patients, higher levels of cathepsins B, H and L activities were found at sites with more serious signs of the disease activity. The total activity of each enzyme (per unit time) was positively correlated with the GCF volume. However, it had little or no correlation with the probing depth (PD). In contrast, the specific activity of each enzyme in GCF (activity units per mg protein), which reflects the selectivity of enzyme exudation, was negatively correlated with the GCF volume. These results suggest that the cysteine proteinases are selectively released into gingival crevices at a relatively mild stage of periodontitis. In experimental gingivitis subjects, no significant activity of each enzyme was detected in GCF, even when the quantity of GCF was comparable to that from periodontitis patients. These data suggest that no significant amounts of these enzymes are released at experimental gingivitis sites or that a homeostatic mechanism, including regulation by protease inhibitors, may control activities of these enzymes in GCF with acute inflammation.

Arylsulphatase activity in human gingival crevicular fluid

Arylsulphatase activity in fluid collected from non-inflamed ( n = 5), gingivitis ( n = 5) and periodontitis ( n = 5) subjects was assayed. The mean volume activity for each group was: non-in-flamed = 768 ± 165 nm/ml per h; gingivitis = 2431 ± 1118 nm/ml per h; periodontitis = 2860 ± 1839 nm/ml per h. The mean total unit activity for each group was: non-inflamed = 0.326 ± 0.076 nm; gingivitis = 1.394 ± 0.411 nm; and periodontitis = 3.571 ± 1.700 nm. Analysis of fluid from isolated sites in periodontitis suggests that reporting total unit activity (absolute amount of enzyme activity) is more meaningful than reporting volume activity (enzyme concentration) for arylsulphatase activity.

Association between ABO blood groups, HL-A antigens and periodontal diseases in young adults: a follow-up study.

Two hundred and thirty-eight Caucasians were studied for ABO blood typing and for HL-A antigens. The subjects were divided into normal, necrotizing ulcerative gingivitis, chronic gingivitis, periodontosis and periodontitis groups. Results showed the chronic gingivitis group was significantly different in ABO grouping than the control group with the gingivitis subjects having a larger percentage of AB types and a smaller percentage of O types. The periodontosis group showed a trend toward more A and B blood groups and a smaller percentage of O groups than the controls. Compared to the normal group there was a significant reduction in HL-A2 antigen frequency in the periodontitis groups and a trend in reduced frequency in the periodontosis group.

Bradykininase activity in gingival fluid in the periodontal disease (author's transl)

The subjects in various periodontal conditions were examined based on the clinical symptoms and roentgenograms, and divided into five groups: clinically normal, gingivitis, slight, moderate and advanced periodontitis. The gingival fluid was collected from gingival sulcus by filter paper strips. The amount of gingival fluid and its bradykininase activity were assayed according to Takeya's method. The bradykininase activity was expressed as the amount of gingival fluid (mg) to produce 50% inactivation of added bradykinin (1μg/ml).The following results were obtained;1) The quantity of gingival fluid in clinically normal subjects averaged 1.74±0.38mg (mean±S. D.). Subjects with gingivitis, slight periodontitis, moderate periodontitis and advanced periodontitis showed the values of 4.68±3.13mg, 7.56±2.81mg, 10.68±7.67mg, and 19.18±12.89mg, respectively. The quantity of gingival fluid was increased with the progression of periodontal disease.2) The bradykininase activity in gingival fluid in clinically normal subjects expressed as the amount of gingival fluid to produce 50% inactivation of added bradykinin (1μg/ml) averaged 0.31±0.14mg (mean±S. D.). Subjects with gingivitis, slight periodontitis, moderate periodontitis and advanced periodontitis showed the values of 1.05±0.90mg, 1.47±0.59mg, 2.61±2.59mg and 3.61±2.87mg, respectively. The bradykininase activity was decrease with the progression of periodontal disease.3) The bradykininase activities in subjects with advanced periodontitis were significantly lower than those of clinically normal (P<0.05) and gingivitis subjects (P<0.05). No statistically significant difference was found between the normal and disease groups.4) Statistically significant correlations were obtained between the bradykininase activity expressed as the amount of gingival fluid to produce 50% inactivation of added bradykinin (1μg/ml) in diseases, and oral examination; gingival score (P<0.01), plaque score (P<0.05), calculus score (P<0.05), sulcus depth (P<0.001), and quantity of gingival fluid (P<0.001), except bone loss score.5) THe degrees of gingival score, oral hygiene score, sulcus depth, bone loss score, and quantity of gingival fluid were increased according to the disease process.It is concluded from these results that the measurement of the kininase activity which is thought to be an important factor for occurence and progression of periodontal disease may be a valuable mean.