Tetracycline inhibition identifies the cellular origin of interstitial collagenases in human periodontal diseases in vivo

Mammalian interstitial collagenases (E.C.3.4.24.7) are considered as key initiators of collagen degradation in periodontal diseases. However, the cellular sources of collagenases present in gingival crevicular fluid have not been completely clarified. Resident fibroblasts and epithelial cells as well as infiltrating neutrophils and monocyte/macrophages are potential sources of the enzymes. We have recently found significant differences in tetracycline inhibition between human neutrophil and fibroblast interstitial collagenases. To address the cellular source of collagenase present in gingival crevicular fluid in 2 distinct periodontal diseases, we studied the tetracycline inhibition of collagenase in gingival crevicular fluid of patients with localized juvenile periodontitis and adult periodontitis. Gingival crevicular fluid samples were collected from deep (greater than 5 mm) periodontal pockets and assayed for collagenase in the presence of 0-1000 microM doxycycline as well as a chemically modified tetracycline devoid of antimicrobial activity (4-de-dimethylaminotetracycline). The drug concentration required to inhibit 50% of collagenase activity (IC50) in localized juvenile periodontitis gingival crevicular fluid was 280 microM for doxycycline and 470 microM for 4-de-dimethylaminotetracycline. Significantly lower values, 10-20 microM, were obtained for collagenase in gingival crevicular fluid of patients with adult periodontitis. We propose that systemic tetracycline levels are efficient inhibitors of collagenase in gingival crevicular fluid in affected sites of patients with adult periodontitis but not of patients with localized juvenile periodontitis and that the fibroblast type interstitial collagenase is the predominant collagenase type in gingival crevicular fluid in affected sites of patients with localized juvenile periodontitis and the neutrophil collagenase in adult periodontitis gingival crevicular fluid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Levels of specific immunoglobulin G to Porphyromonas gingivalis in gingival crevicular fluid are related to site disease status

Titers of immunoglobulin G (IgG) directed against Porphyromonas gingivalis in gingival crevicular fluid of 40 periodontitis patients were measured at three sites in each patient (healthy, gingivitis and periodontitis) by enzyme-linked immunosorbent assay. When paired analyses were performed using Wilcoxon signed-rank tests, periodontitis sites were found to have lower median titers than gingivitis sites. Both systemic and locally-produced antibodies contribute to the overall gingival crevicular fluid antibody profile. Albumin, in contrast, is derived only from serum, and thus this protein serves as an indicator of serum contribution to gingival crevicular fluid. Correction was therefore made for systemic input to the gingival crevicular fluid IgG profile by expressing the results per unit of albumin. When this was done, periodontitis sites were also found to have significantly lower antibody levels than gingivitis sites. These findings suggest that a failure of local antibody production or reduction in quantities, by, for example, degradation by bacterial proteases, may contribute to the change from a gingivitis to a periodontitis lesion.     

Aberrant neutrophil reactions in periodontitis

BACKGROUND: The aim of this study was to compare the activity of neutrophilic granulocytes in patients with severe periodontitis and patients with gingivitis alone. METHODS: The study population comprised 22 patients with gingivitis and 44 with periodontitis. Samples of gingival crevicular fluid (GCF) were collected from untreated patients with gingivitis and from shallow and deep pockets in untreated patients with periodontitis. GCF samples were analyzed for lactoferrin, elastase, matrix metalloproteinase-8 and -9, and collagenolytic activity. RESULTS: The free elastase activity and the neutrophil activity, estimated as the ratio between elastase and lactoferrin, were significantly higher in the samples from the periodontitis patients. These differences were also observed in shallow pockets in periodontitis patients compared to similar pockets in patients with gingivitis. CONCLUSION: This study shows higher levels of free elastase in untreated patients with periodontitis, relative to inflammation-matched controls, which may explain the tissue destruction seen in periodontitis.     

Granulocyte elastase in gingival crevicular fluid

The granulocyte elastase activity and the immuno-reactive (antigenic) granulocyte elastase of gingival crevicular fluid (GCF) were studied in 16 periodontitis patients and in 10 gingivitis patients. The elastase activity was measured with a low molecular weight substrate specific for granulocyte elastase. The antigenic elastase was determined with specific antibodies against granulocyte elastase. Intracrevicular sampling of GCF with paper strips for 30 s seemed to provide representative values of elastase. The elastase activity correlated with probing depth and attachment loss and appeared to be a measure of the degree of tissue destruction. Antigenic elastase represents the number of granulocytes in GCF and should thus be related to the degree of inflammation. The periodontitis patients and the gingivitis patients both had a similar degree of inflammation as measured by antigenic elastase per microliter GCF and gingival index. The elastase activity per microliter GCF, however, was higher in the periodontitis group. Elevated granulocyte elastase activity in GCF seems to be independent of inflammation and could thus be an indicator of patients at risk for periodontitis.     

The relationship between elastase and lactoferrin in healthy, gingivitis and periodontitis sites

OBJECTIVES: To compare the relative amounts of elastase (primary polymorphonuclear leucocyte granule constituent) and lactoferrin (secondary PMN granule constituent) in the gingival crevicular fluid (GCF) of healthy, gingivitis and periodontitis sites.
DESIGN: This cross-sectional study looked at the two GCF constituents in three categories of disease status within the same subject.
MATERIALS AND METHODS: Patients with chronic adult periodontitis were screened and those exhibiting all three types of sites ie periodontally healthy, gingivitis and periodontitis sites were recruited (n = 10) and had GCF collected from the three sites. Lactoferrin and elastase were measured in eluates of GCF by enzyme-linked immunosorbent assay.
RESULTS: The absolute amount of lactoferrin measured in ng per 30 s samples was significantly lower in healthy and gingivitis sites as compared to periodontitis sites however this difference failed to reach significance when the concentration of lactoferrin in GCF was used as the analytical unit. No significant differences were found for elastase levels at any sites when expressed as either absolute amounts or concentrations. Secondary granule release, as evidenced by lactoferrin levels, occurs during cell migration and the process is independent of primary granule release, which is thought to correlate with PMN activation. The relationship between granule constituents in the samples showed significant differences, the highest lactoferrinlelastase ratio being at periodontitis sites (P < 0.001).
CONCLUSIONS These findings imply a change in the relative amounts of elastase and lactoferrin released at different disease level sites, with an almost 10-fold increase in the proportion of lactoferrin to elastase in periodontitis sites over healthy and gingivitis sites. This variation in the release by PMNs of primary and secondary granule constituents may indicate alterations in PMN function in different disease environments.     

Two-dimensional gel electrophoresis of dog crevicular fluid proteins

The ligature-induced periodontitis model was used in beagle dogs to compare and contrast profiles of crevicular fluid (CF) proteins collected from gingivitis and periodontitis sites. The protein profiles of CF and serum were determined by 2-dimensional gel electrophoresis (2-D PAGE) using a silver stain. 2-D PAGE showed that CF contained proteins with molecular weight 16 K or less and many proteins with molecular weights between 64 K and 16 K in the isoelectric pH between approximately 5.8 and 6.8. The number of such proteins was greater in samples collected from the ligated (periodontitis) side compared to the non-ligated (gingivitis) side. Thus, analysis by 2-D PAGE revealed differences between CF samples from gingivitis and ligature-induced periodontitis sites. This study suggests that analysis of human CF by 2-D PAGE may be useful in diagnosis and investigation of the pathogenesis of periodontitis.     

Increased interleukin-18 in gingival crevicular fluid from periodontitis patients

INTRODUCTION: This study aimed to measure the levels of interleukin-18 (IL-18) in inflamed shallow sites and inflamed deep sites in patients with periodontitis and to compare the data with results from inflamed shallow sites in patients with gingivitis. A secondary aim was to examine the composition of the subgingival microbiota in the sampled sites. METHODS: Gingival crevicular fluid was collected from five gingivitis sites and five periodontitis sites from 18 patients with chronic periodontitis, and from five gingivitis sites from 15 patients with gingivitis. Samples from each site category were pooled and IL-18 levels were measured using an enzyme-linked immunosorbent assay. The subgingival microbiota was analyzed by checkerboard DNA-DNA hybridization. RESULTS: All clinical parameters and gingival crevicular fluid volumes were higher in periodontitis sites compared with gingivitis sites from patients with periodontitis and gingivitis. The total amount of IL-18 was higher in periodontitis sites than gingivitis sites in both periodontitis (P = 0.018) and gingivitis (P = 0.002) patients and was higher in gingivitis sites from periodontitis patients than in those from gingivitis patients (P = 0.015). There were higher levels of Tannerella forsythia, Porphyromonas gingivalis, and Treponema denticola (red complex species) in periodontitis sites compared with gingivitis sites in both the periodontitis and gingivitis patients (P < 0.001). CONCLUSION: Levels of IL-18 were higher in patients with chronic periodontitis compared with patients with gingivitis, even at sites with similar pocket depths. The presence of similar levels of red complex species in gingivitis sites from periodontitis patients and from gingivitis patients suggested that the higher levels of IL-18 were not associated with a different microbial challenge.     

Elastase and lactoferrin in gingival crevicular fluid: possible indicators of a granulocyte-associated specific host response

Periodontitis affects a limited number of susceptible humans. The aim of this study was to determine whether there is a difference in the inflammatory reaction between patients with gingivitis and those with periodontitis. For this purpose the levels of elastase and lactoferrin were measured in gingival crevicular fluid (GCF) from three types of sites: i) inflamed sites in patients with gingivitis alone, inflamed sites both ii) with and iii) without tissue destruction in patients with periodontitis. Elastase activity, measured with a chromogenic substrate was significantly higher in the two types of sites in periodontitis patients. Lactoferrin levels, measured with ELISA were the same in the three types of sites. in vitro activation of granulocytes from healthy volunteers with Fc-receptor stimulation showed that the entire release of lactoferrin occurred immediately. In contrast, elastase release was time-dependent and continued throughout the experiment. Thus, the degranulation of the specific (lactoferrin) and azurophil granule (elastase) are under separate control and the two parameters can be combined in a ratio in order to characterize the granulocytes of a given patient. Assuming an immediate release of lactoferrin from the activated granulocytes in vivo , similar amounts of lactoferrin in the three types of sites can be regarded as reflecting similar numbers of granulocytes in the three types of sites. Consequently, a higher elastase activity in GCF from patients with periodontitis indicates a higher rate of release from the cells per se and a granulocyte-associated specific host response.     

Lactoferrin and other markers from gingival crevicular fluid and saliva before and after periodontal treatment

OBJECTIVES: The aim of the study was to verify (i) if crevicular fluid defence variables reflect the changes after surgical periodontal treatment and (ii) if they are in correspondence with changes of these variables in the unstimulated and stimulated whole saliva. MATERIAL AND METHODS: For 12 male and 13 female volunteers with chronic periodontitis lactoferrin concentration as well as the lysozyme and peroxidase activities were determined in crevicular fluid as well as in unstimulated and stimulated saliva before and 14 days after surgical periodontal treatment by a minimal invasive flap technique. RESULTS: The lactoferrin concentrations decreased significantly in the crevicular fluid eluting solution from 1.63 to 1.23 mg/l reflecting a decrease in the total amount collected, in unstimulated saliva from 10.54 to 8.96 mg/l, and in stimulated saliva from 9.00 to 7.11 mg/l after treatment. No significant change could be found for lysozyme. Peroxidase activity was significantly reduced from 269.06 to 186.15 U/l only in the crevicular fluid. CONCLUSION: The results of this study suggest that (i) the defence factor lactoferrin is suitable for monitoring of periodontal treatment results and (ii) changes of the lactoferrin concentration in crevicular fluid are related with significant changes in unstimulated and stimulated saliva.     

Increased interleukin‐18 in gingival crevicular fluid from periodontitis patients

Introduction: This study aimed to measure the levels of interleukin‐18 (IL‐18) in inflamed shallow sites and inflamed deep sites in patients with periodontitis and to compare the data with results from inflamed shallow sites in patients with gingivitis. A secondary aim was to examine the composition of the subgingival microbiota in the sampled sites. Methods: Gingival crevicular fluid was collected from five gingivitis sites and five periodontitis sites from 18 patients with chronic periodontitis, and from five gingivitis sites from 15 patients with gingivitis. Samples from each site category were pooled and IL‐18 levels were measured using an enzyme‐linked immunosorbent assay. The subgingival microbiota was analyzed by checkerboard DNA–DNA hybridization. Results: All clinical parameters and gingival crevicular fluid volumes were higher in periodontitis sites compared with gingivitis sites from patients with periodontitis and gingivitis. The total amount of IL‐18 was higher in periodontitis sites than gingivitis sites in both periodontitis (P = 0.018) and gingivitis (P = 0.002) patients and was higher in gingivitis sites from periodontitis patients than in those from gingivitis patients (P = 0.015). There were higher levels of Tannerella forsythia, Porphyromonas gingivalis, and Treponema denticola (red complex species) in periodontitis sites compared with gingivitis sites in both the periodontitis and gingivitis patients (P < 0.001). Conclusion: Levels of IL‐18 were higher in patients with chronic periodontitis compared with patients with gingivitis, even at sites with similar pocket depths. The presence of similar levels of red complex species in gingivitis sites from periodontitis patients and from gingivitis patients suggested that the higher levels of IL‐18 were not associated with a different microbial challenge.     

Gingival crevicular fluid of patients with gingivitis or periodontal disease: evaluation of elastase-α, proteinase inhibitor complex

Elastase-alpha 1 proteinase inhibitor (E alpha 1PI) concentrations were assessed in gingival crevicular fluids and evaluated in relation to the clinical signs of periodontal disease. 7 gingivitis patients (group G), 38 patients with adult periodontitis and clinically stable lesions (group AP), 21 patients with rapidly progressive periodontitis and clinically stable lesions (group RPP) and 11 patients with either adult periodontitis or rapidly progressive periodontitis and clinically progressive lesions (group Pr) were studied. 6 healthy subjects served as the control group (group H). Significantly differences were observed in the E alpha 1PI concentration between the healthy, gingivitis, clinically stable periodontitis and clinically progressive periodontitis group. In the control group, no E alpha 1PI was detected. Groups G, AP and RPP showed mean E alpha 1PI concentrations of 10.95 +/- 4.96 micrograms/ml, 35.55 +/- 18.64 micrograms/ml and 38.56 +/- 20.89 micrograms/ml, respectively. In these groups, high enzyme levels were correlated with clinical signs of inflammation. The highest E alpha 1PI levels were observed in the clinically progressive lesions. However, they were not necessarily associated with bleeding on probing or clinical evidence of inflammation. These data suggest that a significant increase in crevicular E alpha 1PI levels may be an early manifestation of a progressive or potentially progressive periodontal lesion.     

Biomarkers of periodontitis in oral fluids

Matrix metalloproteinases (MMPs), the key enzymes responsible for matrix degradation, are derived from polymorphonuclear leukocytes during the early stages of periodontitis. The present study determined the levels of GCF matrix metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9) and salivary MMP-8 in patients with gingivitis and periodontitis and in healthy controls. Levels of crevicular MMP-2, MMP-9 and salivary MMP-8 were determined by ELISA in subjects with healthy gingiva (n = 15), gingivitis (n = 18) and periodontitis (n = 20). Significantly higher salivary MMP-8 and crevicular MMP-9 were observed in cases of periodontitis compared to gingivitis and healthy adults. On the other hand, crevicular MMP-2 levels in periodontitis subjects were lower than those in gingivitis and healthy subjects. The three MMP levels were highly correlated to probing depth, and bleeding on probing. Salivary MMP-8, crevicular MMP- 2 and 9 may serve as biomarkers of periodontal disease and aid in early detection of periodontitis or gingivitis.     

A longitudinal study of crevicular fluid in periodontal disease in beagles

A longitudinal study on the composition of crevicular fluid (CF) and serum was carried out in seven beagle dogs during a period from health to 16 weeks of gingivitis and then 8–10 weeks of ligature induced periodontitis (LIPD). The fluids were collected at weekly intervals and the pH and the concentrations of Ca, Mg, Na, and K and the protein profile were determined. Also at weekly intervals, the plaque index, gingival index, calculus index, and pocket depth and attachment loss were recorded for the teeth from which crevicular fluid was collected. Radiographs were taken on the 0 day and after the dogs were sacrificed. Histopathological studies were carried out following sacrifice of the dogs, and the distances between the cemento-enamel junction and the coronal end of the junctional epithelium and the cemento-enamel junction and the crest of the alveolar bone were measured. A two-way ANOVA analysis of the two fluids (CF and serum) and of the two diseases (gingivitis and periodontitis) showed significant differences between the two fluids and between the two diseases. Further statistical analysis showed that in serum none of the constituents studied differed significantly between gingivitis and LIPD, but a significant rise in the concentrations of Ca, Mg, and K was observed in CF during LIPD. The pH of CF was significantly lower during LIPD, but no significant change was observed in the concentration of Na ion. An apparent relationship was observed only between the concentration of K ion in CF and the attachment loss during LIPD. The serum protein profile remained unchanged throughout the period of this study. During gingivitis the CF protein profile was identical to that of serum, but during LIPD a new protein peak appeared. This material had a molecular weight of < 200,000 daltons. Histopathological and radiographic analysis showed bone loss in all but one dog during LIPD. Histopathological study also showed no significant difference in the distance between CEJ and the coronal end of the junctional epithelium between the ligated and the non-ligated sides. This study showed that in beagle dogs the CF differed significantly from that of serum. Also, LIPD caused significant changes in both ionic concentrations and protein profile of crevicular fluid.     

Collagenolytic activity in crevicular fluid from patients with chronic adult periodontitis, localized juvenile periodonitis and gingivitis, and from healthy control subjects

Collagenolytic activity in gingival crevicular fluid (GCF) sampled from 25 healthy control subjects, 25 gingivitis, 25 chronic adult periodontitis (CAP) and 8 LJP patients was correlated with clinical disease parameters using both the site and the patient as sampling units. Among patients collagenase activity increased with the severity of the disease in the order: healthy < gingivitis < periodontitis. Among sites , significant correlation was found between GCF collagenase activity and pocket depth in CAP and LJP, but not in gingivitis patients. Enzyme activity was also correlated with GI score in LJP, but not in CAP and gingivitis patients. In a subset of 10 patients in each of the healthy, gingivitis and CAP groups the association of enzyme activity and crevicular fluid volume (flow) was examined. Significant correlation was found between fluid volume and pocket depth in CAP patients, and between fluid volume and GI score in gingivitis patients, but no association was observed between collagenase activity and fluid volume. The Collagenolytic enzyme was shown to be a genuine vertebrate collagenase derived from unidentified host cells. The concentration of the enzyme in crevicuar fluid from CAP patients was in the order of 10 μg/ml.     

The gingival crevicular fluid ciprofloxacin level in subjects with gingivitis and periodontitis, and its effects on clinical parameters

OBJECTIVES: The purpose of this study, conducted on patients with gingivitis and periodontitis, was twofold: to find out the serum and gingival crevicular fluid concentration of ciprofloxacin, which is a common drug used effectively against Actinobacillus actinomycetemcomitans and to determine the effects of ciprofloxacin administration on clinical parameters. METHOD: A total of 32 adult patients, consisting of 16 subjects with gingivitis and 16 subjects with untreated chronic periodontitis, were included in the study. The subjects were divided into four groups: group I included eight subjects with chronic gingivitis who had not previously received any ciprofloxacin; group II included eight subjects with chronic gingivitis to whom three doses of ciprofloxacin were administered (Siprosan 500 mg) to establish adequate gingival crevicular fluid and serum concentrations of the agent; group III consisted of eight subjects with chronic periodontitis who had not received any ciprofloxacin; group IV included eight subjects with chronic periodontitis to whom three doses of ciprofloxacin were administered to establish adequate gingival crevicular fluid and serum concentrations of the agent. All patients were systemically healthy, free of pain and reported no current medication usage. Each patient was treated with scaling and/or root planing using specific hand instruments under local anesthesia. Gingival index, plaque index and clinical attachment levels of the teeth were used to determine the clinical condition of the subjects and findings were recorded at the beginning, seventh day, 21st day and third month of the study. Serum ciprofloxacin level was measured in venous blood. Approximately 5 ml of venous blood was drawn from subjects in groups II and IV using a standard venipuncture technique. Gingival crevicular fluid samples were sampled from six interproximal sites with six paper strips in the posterior region of upper jaw (excluding third molar) and all gingival crevicular fluid and serum samples were evaluated by high-performance liquid chromatography. RESULTS: The serum concentrations of ciprofloxacin at the first and 72nd hour were not significantly different in subjects with periodontitis compared to subjects with gingivitis. But the gingival crevicular fluid concentrations of ciprofloxacin at the same hours were significantly high in subjects with periodontitis compared to subjects with gingivitis. Both subjects with gingivitis and periodontitis had significantly higher ciprofloxacin levels in the gingival crevicular fluid than in serum. The application of ciprofloxacin did not have any positive or statistically significant effect upon the clinical parameters of the subjects with gingivitis. On the other hand, a significant decrease in the clinical attachment level scores of the subjects with periodontitis (group IV) was observed compared to group III in the 21st day and third month. CONCLUSION: According to these results, the use of ciprofloxacin as an alternative drug in subjects with periodontitis but not gingivitis can be recommended. However, long-term studies are also needed to assess the effects of ciprofloxacin on clinical parameters.     

Microbiological study of localized juvenile periodontitis in Panama

The occurrence of subgingival Actinobacillus actinomycetemcomitans and Capnocytophaga in 12 localized juvenile periodontitis and 10 gingivitis patients from Panama was determined using selective culture techniques. A actinomycetemcomitans was present in all localized juvenile periodontitis lesions studied and was, on average, recovered in hundred-fold-higher numbers from localized juvenile periodontitis lesions than from gingivitis lesions. Capnocytophaga was only recovered in approximately threefold-higher numbers from localized juvenile periodontitis than from gingivitis. The study confirms and extends previous data indicating a close relationship between A actinomycetemcomitans and localized juvenile periodontitis. It is proposed that identification of A actinomycetemcomitans may be a valuable adjunct in the diagnosis of localized juvenile periodontitis.     

Gingival crevicular fluid lactoferrin levels in adult per iodontitis patients

The present study was designed to determine in a cross-sectional study whether there was any relationship between the levels of lactoferrin in gingival crevicular fluid and clinical periodontal parameters. Crevicular fluid was collected from individual sites using standardized filter paper strips (clinically healthy sites, N=23; periodontitis sites, n=66) and evaluated for lactoferrin by enzyme-linked immunosorbent assay. The data showed that: (1) the total amounts of lactoferrin were 0.003-0.021 ng (30 second sample) (average 0.009±0.005 ng) in a clinically healthy periodontium group and 0.016-3.847 ng (30 second sample) (average 0.575±0.069 ng) in adult periodontitis patients (statistically significantly higher in adult periodontitis patients); and (2) the total amounts of lactoferrin were significantly correlated with clinical parameters,     

Collagenase activity in gingival crevicular fluid of patients with juvenile periodontitis

The nature and origin of collagenases in gingival crevicular fluid of juvenile periodontitis patients was investigated. Gingival crevicular fluid collected from deep untreated periodontal pockets of juvenile periodontitis patients was found to contain only vertebrate collagenase (EC 3.4.24.7) activity that cleaved soluble type-I and -III collagens into 3/4 and 1/4 length fragments, as analyzed by SDS-polyacrylamide gel electrophoresis. Type II collagen was degraded at a markedly slower rate. This substrate specificity is indicative of collagenases produced by fibroblasts, epithelial cells and macrophages. We have previously found that collagenase in gingival crevicular fluid of adult periodontics patients appears to be mainly derived from polymorphonuclear leukocytes (PMN). The reasons for the apparent difference in collagenase source between the groups were investigated. We examined whether the pathogen characteristic for juvenile periodontitis, Actinobacillus actinomycetemcomitans, can release collagenase from normal human PMNs. All 10 A. actinomycetemcomitans strains tested, freshly isolated from the subgingival plaque of juvenile periodontitis patients, caused release of collagenase from PMNs in vitro. These results suggest that the lack of normally functioning PMNs in the periodontium of juvenile periodontitis patients may result in a colonization of bacteria that activate the resident periodontal cells to produce increased amounts of collagenase.     

Gingival crevice neutrophil function in periodontal lesions

Polymorphonuclear neutrophils (PMN) may play an important role in protection of the host from pathogenic microorganisms associated with periodontal tissue destruction. The purpose of the present study was to test the hypothesis that unusually severe periodontitis may be associated with defective PMN function at the local disease site. The patients studied included young patients with rapidly progressive periodontitis (including juvenile periodontitis), age-matched patients with gingivitis, and older patients with chronic periodontitis. Gingival crevice (GC) PMN were collected from 10 lesions of each patient by a crevicular washing technique. The number and viability of GC PMN recovered were determined. Their phagocytic capacity was assayed in vitro as a percentage of cells capable of engulfing latex particles. No differences were observed between the periodontitis groups in the number or viability of GC PMN recovered. A statistically significant reduction in mean phagocytic capacity was observed in PMN recovered from lesions of rapidly progressive periodontitis when compared with lesions of chronic periodontitis (12.9 ± 2.1 % vs. 83.7 ± 4.8 %). GC PMNs recovered from non-diseased sites of patients with localized juvenile periodontitis did not show this decreased function. These results suggest that locally diminished PMN function in rapidly progressive and juvenile periodontitis is associated with the environment of these lesions.     

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.