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The aim of this study was to evaluate the efficacy of topical subgingival application of doxycycline hyclate (DH) gel adjunctive to non-surgical periodontal therapy on gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 levels in chronic and aggressive periodontitis patients. Forty teeth of 10 chronic periodontitis patients and 32 teeth of eight aggressive periodontitis patients were screened for 6 months. Scaling and root planing (SRP) was applied to the control sites and DH gel adjunctive to SRP was applied to the test sites of each patient simultaneously. GCF MMP-8 levels were analysed at baseline, 7 days; and at 1, 3 and 6 months by Sandwich Elisa Method. At 1, 3 and 6 months, probing depth ( P < 0.0051) and plaque scores and bleeding on probing values ( P = 0.000) significantly decreased in each group when compared with the baseline, but there was no statistically significant difference between the test and control sites. GCF MMP-8 levels reduced presenting statistically significant differences on 7 days, 1, 3 and 6 months in four of the groups ( P < 0.05); however, intergroup differences were not statistically significant. Developing functional and immunological-based chair-side MMP tests might serve as useful adjunctive diagnostic tools when monitoring the effects of DH gel application. [ABSTRACT FROM AUTHOR]

1. Gingival fluid from severely inflamed periodontal tissue represents a 15 to 30% dilution of serum with respect to the proteins studied, with the exception of C3 and in some subjects, C4. 2. A marked decrease in C3 levels is found in most, if not all, gingival fluids, and a marked decrease of C4 levels is found in some gingival fluids. This suggests that complement might be activated during periodontal inflammation. 3. The concentrations of proteins in the gingival fluid are not related to their molecular weight.

Crevicular fluid from gingivitis patients contains significant levels of a cysteine protease which was characterized as the lysosomal protease cathepsin B, as judged by substrate specificity, thiol dependence, pH optimum, kinetic parameters, pH stability, and inhibitor sensitivities. A highly-sensitive fluorometric assay procedure was used to establish the mean level of cathepsin B activity for 25 gingivitis patients.

The concentration of tetracycline in gingival crevice fluid and blood was determined using a sensitive bioassay after oral administration of repeated doses of tetracycline. Crevicular fluid was sampled by an intracrevicular technique from four gingival sites in each individual and blood was obtained by finger puncture. Four volunteers received doses of 250 mg of tetracycline-HCl either every 6 hours or every 12 hours and were sampled at hours 0 to 15, 21 to 36, 48 to 60 and 96 to 102. Volunteers given 250 mg every 6 hours had average crevicular fluid concentrations between 4 to 8 micrograms/ml and blood concentrations between 2 to 2.5 micrograms/ml after 48 hours. The levels in crevicular fluid and blood of volunteers who received 250 mg every 12 hours were 2 to 4 micrograms/ml and 0.3 to 1.4 micrograms/ml respectively after 48 hours. The results demonstrated that after repeated doses of tetracycline the crevicular fluid levels were typically 2 to 4 times the blood levels.

An agar diffusion assay in capillaries, with Fusobacterium nucleatum M2 as the indicator strain, has been developed to measure metronidazole concentrations in gingival crevice fluid. The assay allows the measurement of 1.9 micrograms of metronidazole per ml in sample amounts of 0.6 microliters.

In order to develop a rapid and simple method based on chemical alterations in gingival pockets during inflammation, the HEMASTIX strips were used to record the degree of gingivitis. This dehydrated test strip records the presence of both hemoglobin derived from erythrocytes and myeloperoxidase from PMN-leukocytes. The intensity of the blue color that appeared and the amount of adsorbed fluid were recorded. The correlation of the Strip Index with Gingival Index was highly significant (P is less than 0.001) and with Gingival Bleeding Index significant (P is less than 0.01). When determined both from the left side of the upper jaw and from the Ramfjord teeth, the correlation between these two areas was highly significant (P is less than 0.001) in the case of Strip Index but only slightly significant (P is less than 0.05) with Gingival Index. According to the present results the Strip Index can be used for the assessment of biochemical changes in gingival pockets resulting from the development of gingivitis.

Protein concentrations in gingival fluid exudate were obtained in both the clinically minimal and severe inflammatory states. A comparison of the results indicated no significant differences in the two groups studied. Since total protein concentrations do not appear to reflect accurately the clinical inflammatory status of the gingival tissues, specific components of the protein molecule should be studied.

This investigation was undertaken to compare the Gindex saliva test scores with the Gingival Index scores and crevicular fluid flow scores, as proposed by Löe and Holm-Pedersen. Eighty-one patients were tested using the Gindex chemical analysis kit. Crevicular fluid was collected from the six teeth advocated by Ramfjord, followed by a clinical assessment of these teeth using the G.I. Of the original 81 subjects tested, 11 were given a prophylaxis consisting of scaling, root planing, polishing, and oral hygiene instruction; and were retested 14 days later. Results of the two-sample t test show that there were no significant differences between males and females in any of the three tested parameters. The sample was pooled and tested for multiple correlations between the G.I., crevicular fluid, and Gindex scores. Nonparametric rank correlation coefficient tests indicated a statistically significant correlation between the three parameters. A pair wise "t" test for the pre- and post-treatment sample demonstrated a significant reduction in the three parameters. A pair wise rank analysis also showed a significant reduction in the G.I., crevicular fluid, and Gindex scores.

A review of current information was undertaken examining suggested roles for bacterial endotoxin in periodontal disease. Evidence for the presence of endotoxin in plaque, gingival crevicular fluid and periodontally involved cementum is discussed. The biological effects of endotoxin are numerous, and its abilities to induce tissue inflammation, resorb rat foetal bone, activate the complement pathway, stimulate macrophages and activate immune responses are examined in relation to the suggestion that endotoxin is an important factor in the pathogenesis of periodontal disease.

The clinical effects of a periodontal dressing after modified Widman flap surgery were studied. A total of thirty surgical sites in 13 patients were included in the study. Half of the sites were not covered by a periodontal dressing while the other half were. To effect a blind evaluation, the dressings were placed and removed by someone other than the examiner. Gingival crevicular fluid flow and gingival inflammation were measured prior to surgery, 2 weeks, 1 month and 2 months after surgery. Clinical attachment level and pocket depth were measured prior to surgery, 1 month and 2 months after surgery. All patients were instructed in oral hygiene procedures and received rubber cup prophylaxes and flossing at 1 week, 2 weeks, 1 month and 2 months postoperatively. At the conclusion of the study, a questionnaire was administered to each patient. No significant differences were found between dressed and undressed segments regarding changes in clinical attachment levels, pocket depth, or gingival inflammation. The omission of a dressing did not result in a greater incidence of discomfort. More patients stated preference for no dressing.

A number of factors which might affect the readings of the HAR-600 GCFM were studied: 1. The readings were higher when the filter paper strips were placed between the fron halves of the upper and lower counterparts than when they were placed between the rear halves. No differences in readings were observed when the strips were placed in the middle, the left, or the right areas of the counterparts. 2. The higher the fluid viscosity the lower the reading. Salt water (0.9% NaCl) gave a lower reading than deionized water. 3. Increased room temperature or relative humidity led to higher readings, especially when the fluid volume exceeded 0.2 microliter. 4. More reading time was required for high viscosity fluid, low temperature, and high relative humidity. The HAR-600 GCFM readings and the lengths of ninhydrin-stained areas were compared. Both methods were able to reflect the amounts of serum absorbed.

Thirty patients took part in a 4 week 'double-blind' clinical trial. Following initial oral hygiene instruction and scaling, half the patients were instructed to brush with a 1% Chlorhexidine gel in the evenings, the other half to use placebo. A standard dentifrice was used in the morning. In both groups there was a marked improvement in all parameters recorded, and although final plaque scores showed a statistically significant difference in favor of the test group, there was virtually no difference in rate or degree of resolution of gingivitis. The use of standard dentifrice was only partially successful in preventing staining.

On the whole, the studies on GCF have demonstrated that the flow of this fluid is sufficiently indicative of the inflammatory state that it can be used under a variety of clinical conditions to monitor and control gingival inflammation. Since gingivitis is extremely common, and since some cases of gingivitis presumably do not progress to periodontitis, the question could be posed whether or not a concerted effort to control inflammation (i.e. trying to achieve a GCF flow as near to zero as possible) would be clinically significant. Until there is evidence to the contrary, the answer must be "yes", since few cases are known where periodontitis occurs without being preceded by gingivitis. In other words, the control of all gingivitis, if feasible, should prevent most cases of periodontitis. Although control of all gingivitis would mean the treatment of many cases that would not progress to periodontal breakdown, such efforts would be worth-while if most periodontal destruction were prevented. Even the early destructive lesion exhibiting little or no inflammation may soon be identified, mainly because the minute volume of fluid collected from the gingival crevice can now be measured accurately. Accordingly, the concentration of various constituents in the GCF can be determined, which should lead to the development of tests to differentiate between pockets undergoing active destruction with minimal inflammation from the majority of active lesions that are intimately involved with frank inflammation. Thus, a clinician would measure sub-clinical gingival inflammation by measuring GCF flow, then differentiate destructive from quiescent lesions by analyzing the GCF sample for some constituent(s), chemical or microbial (Listgarten et al. 1975) indicative of the periodontal destructive process. Monitoring the flow of GCF might be of value in other clinical situations. For example, one could monitor the response of gingival tissues to various restorative and prosthetic procedures (Strauss et al. 1975) to ensure that these procedures do not aggravate the periodontal tissues and induce gingivitis or periodontitis. The education of the patient should be easier since patients can read their own numbers on the GCF meter at each examination and self-evaluate their personal periodontal condition and the effectiveness of their home care. Even the education of the dental student should be easier since he or she would have the means of self-evaluating the effectiveness of treatment, and not be as dependent upon the subjective assessment of his efforts by an instructor. Finally, monitoring GCF for various components could provide the dentist with a valuable means of easily screening patients for systemic disease. Obviously, this area of investigation is in its infancy, but does promise an exciting future for the oral diagnostician.

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.

These preliminary investigations seem to indicate that the periotron may be useful in assessment of the presence of gingival inflammation in the clinical periodontal pockets. Successive recordings of the same volume varied from 5 to 11%. Data from the periotron showed a high correlation with increased known volumes of fluid and with assessment of gingival inflammation using the GI. Assessment with the periotron varied with the dissipation constant of fluids. Since the dissipation constant of gingival fluid varied among individuals, this constant must be evaluated when the instrument is used in clinical and research efforts.

Gingival hyperplasia was studied in 13 boys with epilepsy living in a state hospital. Boys were selected on the basis of having gingival hyperplasia, having all teeth between cuspids (upper and lower), having no occlusal abnormality and being cooperative. After gingivectomy, regrowth of gingiva was compared around lateral incisors on one side of the mouth having operator-assisted oral hygiene with that around lateral incisors on the other side of the mouth without operator-assisted oral hygiene. Regrowth of tissue was documented by precise photogrammetry. Oral hygiene, gingival inflammation and crevicular fluid were monitored. Less inflammation, less crevicular fluid and less regrowth of gingival tissues occurred around teeth subjected to good oral hygiene. Precise periodic photographic documentation of the clinical status of patients during studies such as this is considered very valuable.

Gingival fluid from eight control subjects and ten diabetics was collected and the content of cAMP and protein in the fluid was assayed to determine the nature of the biochemical changes occurring in the gingival fluid due to diabetes. The gingival fluid of the control subjects had a cAMP concentration of 2.4 X 10(-6) M, which was a hundredfold greater than that seen in serum, thus suggesting that the cAMP in the fluid resulted from active synthesis by the gingival cells and was not merely a transudate from the blood. The gingival fluid of the diabetics contained only one-seventh the level of cAMP seen in the control group. It is suggested that the decreased level of cAMP seen in the givgival fluid of diabetics may be a manifestation of a defect in the cAMP forming mechanism of the gingival tissue, which may reflect the systemic etiology of diabetes. It was also found that in the control subjects the content of cAMP in the gingival fluid was in inverse proportion to the volume of exudate in the gingival crevice. No such relationship was seen in the diabetic group. It appears that the level of cAMP present in the gingival fluid of normal individuals without any generalized endocrine deficiencies may be used as an additional indicator of the inflammatory status of the gingival tissues, along with the clinical evaluation based on gingival index.

The sensitivity to tetracycline of 345 bacterial isolates from periodontal lesions was determined. Most species of bacteria, including those thought to be involved in the initiation and progress of destructive periodontal disease, were inhibited in vitro by tetracycline concentrations of 4 to 8 micrograms/ml. This concentration is equivalent to crevicular fluid levels of tetracycline at dosages of 1 gm/day. These data indicate that tetracycline is inhibitory at levels achieved in crevicular fluid for bacteria currently implicated in destructive periodontal disease.

This study was designed to determine if quantitation of lysosomal products in crevicular fluid may be useful as a diagnostic test to evaluate clinical status in periodontal disease. Levels of lysozyme and lactoferrin were quantitated in crevicular fluid from patients with gingivitis, generalized adult periodontitis, localized juvenile periodontitis and normals. Crevicular fluid (CF) was collected from each patient by standardized filter paper strips and evaluated for lysozyme and lactoferrin by rocket immunoelectrophoresis. Levels of lysozyme (micrograms of protein per microliter of CF) were significantly higher in localized juvenile periodontitis patients as compared to gingivitis and adult periodontitis. On the other hand, levels of lactoferrin (micrograms of protein per microliter of CF) did not show significant differences between gingivitis, adult periodontitis and localized juvenile periodontitis. These results indicate that a lysozyme to lactoferrin ratio could be of value as a diagnostic test for localized juvenile periodontitis patients.

The effects of mechanical tooth cleaning and 0.2% chlorhexidine mouthrinses on healing after gingivectomy were compared in 8 patients by means of a "split mouth" technique. The experiment began after periodontal pack removal on day 7. Gingival fluid was sampled immediately before and on days 14, 21, 28, 35 after gingivectomy. The results showed that mechanical and chemical plaque control were equally effective in promoting healing after gingivectomy as evaluated by gingival fluid measurements.

Total fucose and protein was measured in gingival fluid samples collected from 16 human subjects. A significantly higher total protein concentration was found for the control (clinically normal) group when compared to the experimental (severe clinical inflammation) group. This is probably due to the higher gingival flow rate found in the experimental group. The total fucose analyses showed no statistical difference between both groups, but when fucose to protein ratios were studied, a significantly higher ratio was found for the experimental group when compared to the controls. This probably represents the breakdown of plasma and tissue glycoproteins which may occur as a result of inflammation.

Plaque inhibition by dilute mouth sprays and dentrifrices containing benzethonium chloride and chlorhexidine gluconate was compared in beagle dogs. Agents with chlorhexidine gluconate produced less plaque than their benzethonium chloride or placebo counterparts, but the differences were not significant when compared to mean control scores registered during interexperimental recovery periods.