Analysis Of Gingival Crevicular Fluid Research Articles

Enzyme activity in crevicular fluid for detection and prediction of clinical attachment loss in patients with chronic adult periodontitis. Six month results.

Previous reports have described a method by which multiple constituents can be analyzed from a sample of gingival crevicular fluid (GCF) collected with a precut filter paper strip. In this study the relationship of changes in GCF levels of the vertebrate (lysosomal) enzymes beta-glucuronidase (BG) and arylsulfatase (AS) and the cytoplasmic enzyme lactate dehydrogenase (LDH) was evaluated longitudinally in reference to loss of clinical attachment in patients with existing chronic adult periodontitis. Thirty-six patients were followed for six months. Clinical attachment loss was recorded as the change between the baseline and three month examinations, and the three- and six-month examinations. GCF analysis was performed at baseline and three months. Three groups of patients were identified based on disease progression. Group I patients (N = 5) displayed a generalized form of disease activity. In these patients we observed clinical attachment loss of at least 2.0 mm at a minimum of three unrelated sites. Group II patients (N = 4) displayed a localized form of disease activity. In these patients clinical attachment loss of at least 2.5 mm occurred at one site, or two anatomically related sites. Group III patients (N = 27) did not display clinical attachment loss as defined here. Enzyme analysis was evaluated as a whole mouth score (the per cent of samples from a patient in which enzyme activity was at least twice the population mean) and at individual samples. Group I patients could be identified by elevated whole mouth scores for BG, while Group II patients could not be identified by whole mouth scores for any of the enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparison of 4 methods of data presentation for lysosomal enzyme activity in gingival crevicular fluid

In previous studies, we have emphasized the importance of considering the methods used for analysis of gingival crevicular fluid (GCF). This study evaluated 4 different approaches for data presentation of lysosomal enzyme activity in GCF. GCF was collected from patients displaying at least 2 mm of clinical attachment loss at a minimum of 3 sites in the mouth (DA), and patients who did not display clinical attachment loss of 2 mm or more at any site in the mouth (DI), during a 3-month interval following entry into a longitudinal trial. GCF was collected by the timed intrasulcular placement of precut filter paper strips. 16 to 28 individual GCF samples were collected from each patient. The lysosomal enzymes studied were B-glucuronidase (BG) and arylsulfatase. The mean values for the DA and DI groups at baseline and 3 months are reported. The results indicate that when the data is expressed as total enzyme activity (unit activity) per 30-s collection (UA) or UA x GCF volume (microliter) per mm of probing depth, the DA group demonstrated significantly greater mean values than the DI group at baseline and 3 months. In contrast, when the data was expressed as concentration (UA/microliter), or UA per mm of probing depth, differences between the DA and DI groups were observed only at the 3-month evaluation. The difficulty in using concentration when reporting GCF lysosomal enzyme activity is emphsized by comparison of the data from the DA group and the high and low enzyme activity subsets of the DI group.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of treatment on IgG, IgA, IgM and α-2-macroglobulin in gingival crevicular fluid from patients with chronic adult periodontitis

With a technique for sampling, processing and analysis of gingival crevicular fluid (GCF) that allows multiple constituents to be analysed from a sample collected on a filter paper strip, we have examined IgG, IgA, IgM and α-2-macroglobulin (α2M) in GCF from patients with chronic adult periodontitis. Clinical data and GCF were collected before and 3 months after root planing and scaling, and analysed to determine trends for the population. A statistically-significant decrease in the percentage of sites with bleeding on probing, erythema and supragingival plaque was observed 3 months after therapy. The mean amount of each glycoprotein in GCF decreased dramatically at 3 months. In contrast, the mean volume of GCF was virtually identical at the two evaluations. The IgG/IgA and IgG/IgM ratios in GCF were elevated when compared with human serum suggesting the preferential occurrence of IgG in GCF. Correlation of the four glycoproteins with GCF volume and with enzyme markers of the acute inflammatory response in GCF revealed a relationship between arylsulphatase (a lysosomal enzyme), fluid influx, and the passage of larger molecular-weight glycoproteins (α2M, IgM) into the gingival crevice.

Lysosomal and cytoplasmic enzyme activity, crevicular fluid volume, and clinical parameters characterizing gingival sites with shallow to intermediate probing depths.

The biochemical analysis of gingival crevicular fluid (GCF) may offer a sensitive means of determining periodontal disease activity, including the transition of gingivitis to periodontitis. To continue our evaluation of the relationship between clinical and GCF parameters, 552 sites with shallow to intermediate (2.0-5.0 mm) probing depths (PD) were examined. The data were collected at baseline from 33 periodontitis patients participating in a longitudinal trial examining the relationship of changes in GCF biochemistry to attachment loss. Mesiobuccal sites were scored for dichotomous measures of bleeding on probing, gingival redness, suppuration, and plaque accumulation. In addition, GCF was collected using filter paper strips inserted into the sulcus for 30 seconds, eluted in buffer and assayed for activity of the enzymes beta-glucuronidase (BG), arylsulfatase (AS), and lactate dehydrogenase (LDH), markers for ground substance-degradation and cellular necrosis, respectively. Clinical and GCF parameters were evaluated by increasing PD. Plaque accumulation and bleeding on probing increased with increasing PD, although there was considerable overlap across groups. Suppuration was present in only a very small number of sites and the proportion of sites displaying gingival redness was not related to PD. GCF volume was grouped in 0.25-microliter increments, revealing a progressive shift with increasing PD toward a normal distribution around the median range of 0.51 to 0.75 microliter at 5.0 mm. Mean enzyme activities of BG, and to a lesser extent AS and LDH increased sharply from 2.0 to 3.0 mm, were relatively stable from 3.5 to 4.5 mm, and were significantly higher in 5.0 mm than 4.5 mm sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Enzyme activity in human gingival crevicular fluid: considerations in data reporting based on analysis of individual crevicular sites.

Using a reproducible approach to collection, processing and analysis of gingival crevicular fluid (GCF), this study examined 284 fluid samples from individual crevicular sites for the presence of the enzymes lactate dehydrogenase (LDH), B-glucuronidase (BG) and arylsulfatase (AS). 88 of the sites were from periodontally healthy individuals (probing depth 1-3 mm), while 98 sites from patients with periodontitis were examined before and 2 weeks after scaling and root planing (probing depths 1-3 mm, 4-6 mm and 7-10 mm). This study demonstrated the sensitivity of the enzyme assays. When GCF was collected with a 30-s insertion of the filter strip, 90% of the sites from the control subjects demonstrated LDH activity, 85% demonstrated BG activity and 73% demonstrated AS activity. For the 1-3 mm sites from the patients with periodontitis, 100% of sites from which fluid was collected demonstrated LDH and BG activity, and 90% of sites had AS activity before therapy. After therapy, 100% of sites demonstrated LDH activity, 90% had BG activity and 83% had AS activity. All sites in the 4-6 mm and 7-10 mm categories demonstrated activity of all 3 enzymes. The data were analyzed in terms of enzyme activity/30-s sample and as concentration of enzyme in a standard volume of GCF. Enzyme activity/30-s sample was a different and possibly more sensitive indicator of periodontal pathology than standard clinical parameters. There was a disassociation between clinical parameters and the data for enzyme analysis when it was reported as concentration.

Development of a biochemical profile for gingival crevicular fluid. Methodological considerations and evaluation of collagen-degrading and ground substance-degrading enzyme activity during experimental gingivitis.

The potential application of gingival crevicular fluid (GCF) analysis to periodontal diagnosis has been examined for more than 25 years. Unfortunately, the information available has not provided the clinician with a more sensitive means of diagnosing periodontal disease or an effective means of monitoring periodontal therapy. A careful review of the literature on GCF, however, suggests that discrepancies occur in the method of GCF collection, the use of GCF for analysis from pooled or isolated crevicular locations, the method of analyzing the samples and the way in which the data is reported. Studies in our laboratory have suggested a technique for GCF analysis that collects GCF from individual crevices with a filter paper strip inserted for a standard time, determines the volume of GCF collected with a calibrated electronic meter and elutes the material into a larger volume of diluent. This approach allows for detection of site-to-site and patient-to-patient differences in GCF volume while providing sufficient samples to analyze GCF for multiple constituents. We have used this approach to evaluate GCF for vertebrate forms of the enzymes collagenase (latent and active forms), beta-glucuronidase and arylsulfatase during the development of experimental gingivitis in man. Interproximal and midradicular areas were studied. Our results indicate that during the 4 weeks of the gingivitis, the absolute amount of active collagenase in GCF increased 550% at the interproximal sites and 190% in the midradicular sites, and the per cent of active collagenase increased from 15 to 71% at the interproximal sites, and from 16 to 36% at the midradicular sites.(ABSTRACT TRUNCATED AT 250 WORDS)