Identification of a cathepsin b-like protease in the crevicular fluid of gingivitis patients.

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.

Detection of functional complement components in gingival crevicular fluid from humans with periodontal diseases.

Crevicular fluid was collected from patients with periodontitis by a capillary tube procedure. Complement component activities were determined by functional assay systems, with human complement, and partially purified human first complement component (C1) as controls. The complement-fixing properties of the dental plaque of each patient were also examined C1 activity in the crevicular fluid of all patients was approximately 1/8 of whole serum C1 and diminished rapidly with time after collection. There was no significant relationship between C1 concentration and crevicular fluid flow rate. Hemolytic activity of whole complement was also invariable detected when sufficient amounts (8 micronl) of crevicular fluid could be obtained. Dental plaque was found to fix C1. A role for crevicular complement in inflammatory periodontal disease is suggested.

Effect of two antibacterial mouth sprays and dentifrices on dental plaque and gingivitis in beagle dogs.

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.

Sulcular exudate protein levels as an indicator of the clinical inflammatory response.

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.

Clinical significance of gingival fluid measurement by "Periotron".

The accuracy of gingival fluid measurement with an electric device was compared with such clinical parameters as Gingival Index scores and pocket depths, and the treatment effect was evaluated by changes in the gingival fluid amount. After the initial examination 10 patients received oral hygiene instruction, scaling, root planing and soft tissue curettage. The operated areas were protected by a periodontal dressing for 1 week and all patients used a 0.2% chlorhexidine mouthrinse twice daily during that period and for the next week. Four weeks later the patients were reexamined for the same factors that were evaluated at the initial examination. The amount of gingival fluid correlated with the Gingival Index (r = 0.65), the pocket depth (r = 0.74) and the total amount of acid phosphatase in the gingival fluid (r = 0.92). Following treatment, a significant improvement in periodontal status was demonstrated by all of the parameters utilized.

Evaluation of cleansing devices in the maintenance of interproximal gingival health.

This study consisted of 24 dental students, who 10 days prior to the start of the experimental period, were thoroughly scaled and given instructions in the use of a rubber tip stimulator, unwaxed dental floss, and a modified Bass brushing technique. On day 0, the subjects had reached a high level of interproximal gingival health as measured by intracrevicular exudate flow and löe's Gingival Index. Subjects were then randomly divided into 4 groups which were to brush; brush and floss; brush and rubber tip; and brush, floss and rubber tip. Evaluation was performed on days 0, 9, 15, and 33 using Löe's Gingival Index, Podchadley's Plaque Index, and gingival exudate flow. The results of this study indicate that interproximal gingival health can be maintained in motivated patients with initially healthy gingivae (for at least several weeks) with proper use of the modified Bass brushing technique alone.

The effect of axial crown overcontour on adolescents.

The relationship between axial crown overcontour and the health of the subjacent gingival investing tissues was investigated by placing standardized axial over-contoured facings on the buccal, axiogingival aspect of 42 premolar teeth. The gingival tissues were evaluated on the basis of clinical criteria, evaluation of standardized photographic records and by comparing gingival sulcular fluid production at test and control sites. Of the 42 test sites examined, 27 showed clinical signs of gingival inflammation and alteration of normal soft tissue architecture after a period of 42 to 49 days. There was a significant correlation as measured by die Spearman rank correlation technique between data derived from clinical examination and data obtained from interpretation of standardized photographic material. Evaluation of Test Site to Control Site Gingival sulcular fluid score quotients was suggestive of some degradation of the gingival tissues at the test sites, but no significant correlation was found. It may be concluded that alteration of normal crown form by overcontouring the buccal, axial third of a tooth may be a factor which predisposes the subjacent gingival tissues to inflammatory disease.

Comparison of results following modified Widman flap surgery with and without surgical dressing.

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.

Antibodies to herpes simplex virus in human gingival fluid.

Antibodies to herpes simplex virus were found by immunofluorescent techniques in the gingival fluid of 84.0% of the persons tested. A mixture of IgG and IgA antibodies was demonstrated in 52.3% of the specimens while in 47.7% only IgG antibodies were found. Neutralization of herpes simplex virus infection in Vero cells by the gingival fluid was also demonstrated. No correlation between recurrences of herpes infection and the type of antibodies produced could be established.

Tetracycline: levels achievable in gingival crevice fluid and in vitro effect on subgingival organisms. Part I. Concentrations in crevicular fluid after repeated doses.

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.

Evaluation of an electronic device for gingival fluid quantitation.

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.

The effect of intrasulcular toothbrushing on the amount of sulcular fluid.

Nineteen young adult Caucasian males had an average Gingival Score of 0.47. Each was asked to demonstrate his present method of toothbrushing. Thirteen used a scrub method, and six combined the scrub and roll methods. Sulcular fluid measurements were taken from the facial aspect of the mandibular left second bicuspid using filter paper strips stained with ninhydrin. An average value of 0.94 mm was obtained. Intrasulcular brushing was performed in the mandibular left posterior segment, for 30 seconds daily for 40 days. The brushing was done by the investigator on 28 days and by the subjects on 12 days. The average Gingival Index score following intrasulcular brushing was 0.26. Sulcular fluid measurements were again taken from the mandibular left second bicuspid. A decreased amount of sulcular fluid was found in six subjects, while nine exhibited an increase and four showed no change. An average sulcular fluid measurement of 0.87 mm resulted following the experimental brushing period. From these data, it can be concluded that, within the limitations of this study, intrasulcular brushing had no significant effect on the amount of gingival fluid.

Lysozyme and lactoferrin quantitation in the crevicular fluid.

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.

Gingival fluid fucose to protein ratios as indicators of the severity of periodontal disease.

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.

The effect of levamisole on experimental gingivitis in the beagle dog.

The effect of Levamisole on gingivitis development was studied in three beagle dogs. Plaque accumulation was first allowed on one side of the jaws whereas the other side was subjected to careful tooth cleaning procedures. After 2 weeks' experimental gingivitis the dogs were given Levamisole and plaque accumulation was permitted on the other side of the jaws for another two weeks. The following parameters were investigated in the two gingivitis experiments: Plaque and Gingival Indices, Gingival Exudate measurements and histologic measurements. The biopsies were prepared both for light and electron microscopy. The clinical parameters did not show any differences between the two gingivitis experiments. The biopsies, however, demonstrated a larger area of infiltrated connective tissue and an increased number of leukocytes in the junctional epithelium in Levamisole-treated compared to normal animals.

Hydrogen sulfide production from gingival crevicular fluid.

A METHOD was developed and evaluated in a pilot study to determine a relationship between gingival health, crevicular fluid flow and the production of H2S from the crevicular fluid. A moderate degree of correlation was observed between the crevicular fluid volume and the gingival index. A stronger correlation was obtained between the crevicular fluid volume and H2S production by the crevicular fluid. The method described could prove useful in evaluating gingival health and disease objectively and for monitoring the activity of periodontitis.

A rapid chairside test for the severity of periodontal disease using gingival fluid.

A study was done in an attempt to develop a simple test for the severity of periodontal disease. Gingival fluid collected on filter paper was tested for protein content, and the resulting color was compared to standard color filters. Tissue was excised and prepared for histologic examination. The inflammatory cell infiltrate on each slide was graded on a scale of zero to three. Zero was least and three was the highest number of inflammatory cells. The results indicate that the white cell infiltrate graded 0 or 1 on the histologic inflammatory index has a color index of B 1/8 TO B1 whereas the tissue graded 2 or 3 HII has a range of B2 to B6 on the color scale. This test can give the dental practitioner a general idea of the severity of the inflammation.

The effect of a 1% chlorhexidine gel in the initial therapy of chronic periodontal disease.

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.

Determination of periodontal disease activity.

The determination of disease activity has a direct impact on therapeutic measures in periodontics. Evaluations that may indicate disease activity have been reviewed under the topics of traditional evaluations, gingival crevicular fluid contents, tissue changes, circulating factors, and sulcular microbiota. The traditional methods, such as probing, radiographs, and clinical indices, were static evaluations that had limited validity in determining disease activity. When used in longitudinal tests, these methods may be used retrospectively to detect disease activity. Evaluation of the contents of gingival crevicular fluid has promise as a noninvasive method of determining tissue changes in the periodontium. To date, none of the crevicular fluid constituents has been a reliable indicator of disease activity. Tissue changes in the periodontium are important in understanding the disease processes but have had limited value as indicators of disease activity. The ability to determine disease activity through identification of substances or changes in the systemic circulation would prove quite useful to the clinician. Although some reports found lymphocyte transformation indicative of active disease, others had conflicting results. Finally, examinations of sulcular microbiota provided evidence suggesting that active periodontal disease was associated with specific groups of pathogens and with elevated counts of motile organisms. The evaluation of such pathogens and motile organisms currently shows the most promise for determining periodontal disease activity. However, long-term assessments and further clinical trials are necessary before this type of evaluation can be widely accepted. It is hoped that such studies will enable the clinician to predict the natural history of active and inactive periodontal disease.

Tetracycline: levels of achievable in gingival crevice fluid and in vitro effect on subgingival organisms. Part II. Susceptibilities of periodontal bacteria.

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.