Gingival Fluid Flow Research Articles

The immediate efficacy of adjunctive doxycycline in treatment of localized juvenile periodontitis

A randomized, double-blind, placebo-controlled study on the immediate clinical and microbiological efficacy of doxycycline (100 mg for 14 days) was carried out to determine the benefit of adjunctive medication in 16 patients with localized juvenile periodontitis. Measurements of gingival fluid flow, probing depths, bleeding on probing and suppuration were determined at 2 periodontal sites with and 2 without radiographic attachment loss, at weeks 0, 1, 3 and 8. Subgingival bacterial samples were taken with curettes from the same sites. Spirochaetes were searched for by dark-field microscopy. Actinobacillus actinomycetemcomitans, pigmented and non-pigmented Bacteroides spp., Capnocytophaga, Fusobacterium and Actinomyces spp. were cultured on various selective and non-selective media. Bacterial species found at least in 50% of the patients and comprising on average 5% or more of the cultivable flora were included in the analysis. Neither short-term clinical nor microbiological efficacy beyond that of a course of mechanical debridement alone was found by using systemic medication with doxycycline in patients with localized juvenile periodontitis.

Gingival resistance to probing forces. I. Determination of optimal probe diameter.

This paper presents the results of a study designed to determine (1) if advancement of a periodontal probe under a given force against healthy gingiva differed from its advancement under a similar force, against inflamed gingiva, and (2) to select the diameter which best distinguishes healthy gingiva from that which is inflamed to varying degrees. The teeth of eight beagle dogs were cleaned three times per week for two months until a Gingival Index (GI) of 0 and gingival fluid flow (GFF) of 9 or less, as measured by the Periotron, were obtained. Probes of diameters 0.4, 0.6, 0.7, and 0.8 mm were advanced perpendicularly against the attached gingiva in the incisor, premolar, and molar regions by an electromechanical device which simultaneously plotted force versus displacement following initial contact of the probe tip with the gingiva. These forces ranged from 0.1 to 0.9N. This process was repeated in the same animals following cessation of plaque control measures when the GI reached a value of 1 and GFF reached levels of 10 to 19. It was repeated again following placement of an irritating ligature when GI reached 2 and GFF greater than 20. An analysis of covariance showed significant differences between all three levels of health and disease for all probe diameters. F values for the 0.6 mm probe were highest at all probing forces indicating that this diameter most consistently discriminated variation of gingival health status.

Kinin generation in the gingival inflammatory response to topically applied bacterial lipopolysaccharides.

A water-soluble lipopolysaccharide from Salmonella enteritidis and a phenol-soluble lipopolysaccharide from Leptotrichia buccalis were applied topically to the healthy marginal gingivae of beagle dogs. Saline was applied to contralateral areas as an internal control. Increases in vascular permeability were monitored by measurement of gingival fluid, and the collected gingival fluid samples were assayed for kininogenase and kinin activities. Both lipopolysaccharides induced an inflammatory response, as indicated by increased gingival fluid flow. Kininogenase-kinin activities paralleled the increases in gingival fluid flow, with the highest values being associated with peak increases in gingival fluid. The results indicate that both lipopolysaccharides, although different in lipid solubility, penetrate healthy sulcular epithelium and initiate an inflammatory response which is mediated in part by the kallikrein-kinin system. Interrelationships between this system and other inflammatory mediators suggest that kinin generation not only plays a role in the early phases of acute gingival inflammation, but may also contribute to the activation of other mediators appearing later in the response and in chronic inflammatory lesions.

Gas-liquid chromatography of the gingival fluid as an aid in periodontal diagnosis.

There is increasing evidence that rod‐shaped Gram negative anaerobic bacteria, as those belonging to black‐pigmented Bacteroides, may play a role in the pathogenesis of periodontal diseases. Anaerobic bacteria are known to produce, as end‐products of their metabolism, several volatile fatty acids, whose detection in clinical specimens by direct gas liquid chromatography (GLC) has been usefully utilized to demonstrate their presence in the infected site. The results obtained analyzing by GLC the gingival fluid collected from patients suffering from periodontal diseases are reported here. A good statistical correlation of the GLC profile has been found with the gingival index and the colonization by gram‐negative bacteria at each selected site.The data obtained strongly suggest that butyric acid may be considered a reliable marker of the severity of the disease, and its quantitative changes may be useful in following the clinical course of the process. The data correlate well with the biological activities of butyric acid, which have been reported to be abundant in plaque and to exert a cytotoxic activity on a variety of cells including gingival fibroblasts.No correlation has been found between gingival fluid flow rate and fatty acids profile in GLC. It can be concluded that GLC measures the overall metabolic activity of the bacterial population colonizing the subgingival area, while the rate of gingival fluid flow reflects the degree of the inflammatory response of the host. The combined use of the two laboratory tests should provide the clinician with more quantitative and reproducible parameters for monitoring the disease activity.

Inhibition of plaque formation in humans by octenidine mouthrinse.

Octenidine, a new bispyridine antimicrobial compound, has significant antiplaque activity in animals. The present study was designed to assess the effectiveness of this agent in inhibiting plaque formation in humans. Sixty males (age 22–36 years) were randomly assigned 4 formulations which included 1) 0.1% octenidine in vehicle, 2) 0.1% octenidine in aqueous solution, 3) 0.05% octenidine in vehicle, and 4) vehicle only. After establishment of minimal levels of plaque and gingivitis, each subject rinsed twice daily under supervision with his assigned test agent. Over the next seven days, the subjects were not permitted any other means of tooth cleaning. Plaque accumulation was measured daily and gingival fluid and stain levels determined on days 0 and 7.Analysis of the data showed that both formulations of 0.1% octenidine almost completely inhibited plaque formation over the 7 day period. The vehicle had no effect on the agent's activity. The 0.05% concentration allowed some plaque accumulation, but all octenidine groups had significantly less plaque than the control group. Gingival fluid flow was significantly lower in all octenidine groups than in the control group. Tooth stain increased in all groups using the agent but remained unchanged in the group using the vehicle alone.Results of this short term clinical trial suggest that octenidine may be a useful topical antiplaque agent.

The gingival bleeding time index.

Recent studies have supported an emphasis on gingival bleeding indices for detection of inflammation. The purposes of the present study were: (1) to evaluate the relationships between the time necessary for stimulated gingival bleeding to occur and both gingival fluid flow and Löe's gingival index, and (2) to develop a clinical index based on gingival bleeding time. In the first part of the study, the elapsed time for gingival bleeding to first occur after sulcular stimulation was correlated with the gingival index and fluid flow on 150 gingival units. A high correlation was found between bleeding time with both the gingival index and exudate flow. A bleeding time index based on a scattergram of the data obtained in part I of the study was then formulated and evaluated in a similar fashion on an additional 172 gingival units. Results indicate a high correlation between the bleeding time index and the other indices. It is concluded that the bleeding time index may be used by itself as an accurate and objective clinical tool to evaluate gingival inflammation.

Sulcular polymorphonuclear leucocytes and gingival exudate during experimental gingivitis in man.

The number of polymorphonuclear leucocytes (PMNs) in the products of gingival washing and the rate of gingival fluid flow were measured in eight volunteers during a period of experimental gingivitis on the upper dental arch. Both parameters increased significantly during the 21 days of no brushing. However, the concentration of PMNs increased, between day 0 and day 21, by a factor of 2.1, whereas the flow of gingival fluid collected on teeth no 13 and 21 showed a 5.4 fold increase during the same period. When the amount of gingival fluid was very low (at days 0 and 28), the number of migrating PMNs was still rather high. For each volunteer, the concentration of PMNs in the washings through the period of experimental gingivitis was not always correlated to the rate of gingival fluid flow. These data confirm previous findings on dogs that sulcular PMNs emigration and flow of gingival fluid could be two independent phenomena.

Sonicated and passively dispersed hydrogen peroxide in periodontitis.

The purpose of this study was to determine the effect of hydrogen peroxide delivered into periodontal pockets on gingival fluid flow and myeloperoxidase activity in the gingival fluid. Twenty systemically healthy human subjects presenting with moderate to advanced periodontitis were utilized. One side of the maxilla served as the control and the contralateral segment served as the experimental side. Measurements were made 0.5, 24, and 168 hours postoperatively. The experimental side was treated with 0.03 % hydrogen peroxide and the control side received distilled water. Five milliliters of fluid was delivered into the periodontal pockets either by sonication or passive dispersion. The 30 minute results indicated a mean increase in gingival fluid myeloperoxidase of 8.08 units per milliliter (U/ml) for the sonicated hydrogen peroxide, 2.05 U/ml for the passively dispersed hydrogen peroxide, 1.31 U/ml for the sonicated distilled water, and 0.60 U/ml for the passively dispersed distilled water. The values for hydrogen peroxide both sonicated and passively dispersed were significantly greater than baseline readings (p < 0.01), and the sonicated value was significantly larger than the passively dispersed value.

Studies of host responses during experimental gingivitis in humans

The aim of the study was to find out whether autologous supragingival plaques collected at various times during the development of experimental gingivitis in men differ in their potential to induce PMN responses in vitro. Ten subjects refrained from all oral hygiene for a period of 21 days. At day 0, Day 3 and Day 21 after cressation of oral hygiene clinical indices (plaque index, gingival index, gingival crevicular fluid flow) were assessed plaque was collected at day 3 and Day 21 after cessation of Oral hygiene clinical indices (plaque index, gingival index, gingival crevicular fluid flow) were assessed plaque was collected at Day 3 and Day 21. PMNs, isolated from peripheral blood 1–2 months after the non‐brushing period were incubated with the various autologous plaques in the presence or the absence of autologous serum. Supernatants of cell cultures were monitored for extracellular release of lysosomal constituents, viz. lysozyme, myeloperoxidase and lactoferrin. There was no difference in the amounts of lysozyme, myeloperox dase and lactoferrin. There was no difference in the amounts of lysozyme or myeloperoxidase released by PMNs. The inclusion of fresh serum enhanced PMN release reactions but there was no significant differences in the relative capacity of Day 0, Day3 and Day 21 sera to modulate PMN responses. A statistically significant positive correlation was established betwen the magnitude of lactoferrin release from PMNs exposed to Day 3 plaque and the gingival fluid flow recorded on Day3. Lactoferrin is segregated in the specific granules of the PMNs which also contain other cationic proteins, collagenases and neutral proteinases. These data suggest that a possible causal relationship may exit between the release of PMN constituents and the development of inflammation during the early phases of gingivitis.

Effect of experimental neutropenia on initial gingivitis in dogs

The role of neutrophilic granulocytes in the loss of gingival collagen has been studied by inducing experimental neutropenia during initial gingivitis in beagle dogs. Neutropenia was induced for 4 d in three animals with normal gingiva by repeated injections of rabbit anti-neutrophil serum. During neutropenia microbial plaque was allowed to form on the teeth. Samples of junctional (crevicular) leukocytes and gingival fluid were taken on days 0 and 4. Block biopsies of buccal gingiva were obtained on day 4. Stained semi- and ultrathin sections were used for histometric and stereologic tissue analysis. Gingival fluid flow increased from day 0 to day 4 in all dogs while junctional leukocytes increased in one dog only. Subgingival plaque had formed in most biopsies, and in the junctional epithelium very few neutrophilic granulocytes were present. In the coronal connective tissue subjacent to the junctional epithelium lymphoid cells, structurally abnormal neutrophilic granulocytes and monocytes/macrophages were diffusely scattered. The gingival collagen appeared mainly displaced by the inflammatory cells rather than dissolved. The data suggest that neutrophilic granulocytes may contribute to the loss of gingival collagen during initial gingivitis in dogs. The neutrophils also seem to be of importance for the limitation of subgingival plaque growth along the tooth surface.

The effect of levamisole on experimental gingivitis in juvenile periodontitis. Sequential clinical, immunological, and haematological changes.

In four patients with juvenile periodontitis (group A), five adults with advanced periodontitis (group B), and in four controls (group C) the concentration of serum immunoglobulins, serum complement (C3C4, and C5) T and B lymphocyte ratio, leukocyte migration inhibition and lymphocyte transformation responses to dental plaque bacteria (A. viscosus, B. melaninogenicus, F. nucleatum, and V. parvula) and PPD or PHA, and lymphocyte ATP‐ase activity were assessed before and during development of experimental gingivitis.All the patients in group A and B Had completed extensive periodontal treatment, and at the time of the experiment the periodontal conditions were excellent. Plaque accumulation and gingival inflammation were assessed and blood samples were drawn: at day 0; after plaque accumulation for 14 d (no oral hygiene) (period I); after 14 d with optimal oral hygiene (period II); after plaque accumulation for 14 d (no oral hygiene) and concomitant immunostimulatory treatment with levamisole (period III); after 14 d with optimal oral hygiene. After period I, gingival inflammation and increased gingival fluid flow developed in all groups; however, in group C some of the clinical and immunological responses were slightly enhanced as compared wiht group A and group B. On the other hand, after period III (plaque accumulation + levamisole) more pronounced clinical and immunological changes were recorded in group A and B than in group C, including a significant rise in gingival fluid flow, LIF production, and lymphocyte ATP‐ase activity. The present study has supported the asumption that a state of immunodeficiency can be an underlying, predisposing factor for juvenile periodontitis. The results, however, give only vague information on the nature of this deficiency

Relative distribution of bacteria at clinically healthy and periodontally diseased sites in humans*

Abstract Samples of subgingival bacteria were collected with a clean curette from two relatively normal and two periodontally diseased sites in each of 12 patients with advanced periodontal disease. The samples were suspended in physiologic saline containing 1 % gelatin and examined within 1 hour by darkfield microscopy. From 100‐200 bacteria were classified on a percentage basis into one of the following categories: (1) coccoid cells, (2) straight rods, (3) filaments, (4) fusiforms, (5) curved rods, (6) small spirochetes, (7) medium‐sized spirochetes, (8) large spirochetes, and (9) motile rods. For each area sampled the following clinical criteria were also recorded: (1) Gingival Index, (2) Plaque Index, (3) probing depth and (4) gingival fluid flow. For each patient separate mean values were calculated for the normal and the diseased sites. The results indicated that significant differences existed in the microbial flora of clinically normal and diseased sites using a paired t‐test comparison (2α= 0.001), with coccoid cells more predominant at normal sites (74.3 % vs. 22.3 %), while at diseased sites motile rods were more frequent (12.7 % vs. 0.3 %), as well as curved rods (1.7 % vs. 0 %), small spirochetes (12.6 % vs. 1.1 %), medium‐sized spirochetes (18.5 % vs. 0.5 %) and large spirochetes (6.7 % vs. 0.2 %). The ratio of motile to non‐motile cells in the normal was 1:49, whereas at diseased sites the ratio was in the vicinity of 1:1. These results clearly show that a different flora is associated with healthy and periodontally diseased sites in the same patient population and that these differences can be detected by means of a technique which is simple and readily adaptable to a clinical setting. The data obtained in this fashion may be useful in monitoring the effect of various treatment modalities on the periodontal flora and possibly in determining the presence or absence of active disease.

A comparison of the scalloped and linear incisions as used in the reverse bevel technique

Using a split-mouth technique in 20 patients, a comparison was made of the linear and scalloped incisions as used in the reverse bevel technique. Forty full thickness periodontal flap procedures were evaluated. The PlI and GI were recorded and gingival fluid collected at operation and at weeks 1, 2, 3, 4 and 12 after operation. Pocket depths and attachment levels were noted and gingival contour assessed, using the Contour Index, pre-operatively and 12 weeks after operation. Pain experience during the first week after operation was evaluated. No significant changes in PlI occurred throughout the study. The GI and gingival fluid flow showed comparable trends. More inflammation was present after the linear incision during the first 2 weeks after operation. Clinically, satisfactory results were achieved for both scalloped and linear incision procedures as shown by the improvement in gingival contour, reduction in pocket depth, slight gain in attachment, lack of gingivitis and low plaque scores.

Influence of “time of day”, pocket depth and scaling on gingival fluid flow

Gingival fluid was sampled from the orifices of the gingival crevices in five male subjects with clinically healthy gingiva. The sampling was performed at 0915 h, 1215 h and 1515 h of the same day. Variation in "time of day" did not seem to influence the rate of gingival fluid flow. A total of 144 pockets (2-11 mm) in 19 patients were sampled for gingival fluid at the orifices. The adjacent gingiva was evaluated accordingly to the Gingival Index (GI). Gingival fluid flow was much more influenced by the degree of gingival inflammation than by pocket depth. Scaling with curettes was performed in five patients. Gingival fluid was sampled at the orifices immediately before scaling and on days 1, 14 and 28 after scaling. The flow rate decreased to a minimum on day 14 after scaling for all pocket depths and remained at almost the same level until day 28. Pockets less than 5 mm deep were more frequently without measurable amounts of fluid than pockets deeper than 5 mm on days 14 and 28.

Initial gingivitis in dogs.

The gingival characteristics after four days of gingivitis development have been studied in three Beagle dogs. Gingival normality was induced by regular tooth cleaning and gingivitis was provoked by abolishing oral hygiene procedures and giving the animals a soft diet. Measurements of the gingival fluid flow and the amount of crevicular leukocytes served to clinically assess the gingival condition. Semi‐ and ultra‐thin sections from contralateral biopsies of buccal gingival tissues from premolars obtained on day 0 and day 4 respectively were subjected to stereologic analysis based on morphometric point counting procedures. On day 0 of the experiment two dogs revealed minute amounts of gingival fluid and crevicular leukocytes and the gingival tissues of these dogs were normal. The third dog had somewhat higher amounts of the gingival parameters and the connective tissue adjacent to the junctional epithelium harboured a small lymphoid infiltrate occupying 8.3 ± 2.8% of the gingival tissues. On day 4 the gingival fluid flow and the amounts of crevicular leukocytes were increased in all dogs compared to their values on day 0.The coronal con nective tissue in the two dogs with initially normal gingiva revealed on day 4 an infiltrated portion which occupied 6.1 ± 2.4% of the gingival tissues. The infiltrate was predominated by neutrophilic granulocytes, monocytes, macrophages, immunoblasts and small lymphocytes. A 60% loss of the collagen density in the infiltrated fraction was noted between biopsies from day 0 and day 4 in these two dogs. The tissues of the dog which entered the experiment with a preestablished lymphoid infiltrate reacted with similar but more intense inflammatory processes on day 4 than those of the dogs with normal gingiva on day 0.The results indicate that initial gingivitis in dogs is characterized by acute exudative inflammation and immunopathological processes as well as by a marked loss of collagen fibers in the connective tissue located beneath the coronal part of the junctional epithelium. Possible mechanisms responsible for the observed alterations have been discussed.

Clinical and histologic characteristics of normal gingiva in dogs.

The aims of the present study were to establish normal gingiva in dogs, to characterize the clinical conditions prevailing and to stereologically describe the structural composition of the normal gingival tissues. Three beagle dogs were subjected to regular oral hygiene procedures during 15 weeks. Measurements of gingival fluid flow and the amounts of crevicular leukocytes served to clinically assess the gingival conditions. Semi‐and ultrathin sections from biopsies of normal buccal gingival tissues from premolars were subjected to stereologic analysis based on morphometric point counting procedures. Gingival normality was achieved in two of the dogs. The normal gingival tissue in these dogs was characterized clinically by abscense of gingival fluid flow and by a minute amount of crevicular leukocytes. A gingival sulcus was most often absent. The junctional epithelium was without rete pegs, and the entire gingival connective tissue was densely filled by homogeneous collagen fiber bundles. A few isolated inflammatory cells were present in the junctional epithelium and the adjacent connective tissue. No clusters of inflammatory cells forming an infiltrate could be observed.Stereologically, the gingival tissue comprised 48% epithelium and 52% connective tissue. The junctional epithelium occupied 10% of the gingival tissue and included a fraction of 2.8% occupied by leukocytes. The latter by volume comprised 50% neutrophilic granulocytes and mononuclear cells each. The connective tissue was composed of 67% collagen fibers, 14% free cells and 19% residual tissue. The composition of the connective tissue adjacent to the junctional epithelium differed somewhat from that of more central connective tissue fractions.

Dressing after reverse bevel flap procedures.

Abstract Effects of withholding a dressing and applying Coe‐Pak after reverse bevel flap procedures in periodontal surgery were compared in 24 patients. Maxillary and mandibular posterior segments were treated using a “split‐month” technique. Slight differences occurred between dressed and undressed segments. Gingival Index scores for the undressed segments were lower than those for the dressed segments at 7 days after operation, but by 21 and 28 days post‐operatively the position was reversed. Changes in gingival fluid flow followed a similar pattern but were not statistically significant. The 0.18 mm difference in crevice depth of the dressed as opposed to the undressed segments noted 28 days post‐operatively was statistically significant. Clinically satisfactory results were achieved in both instances. Post‐operative dressing was associated with a greater incidence of pain and swelling, but a reduced incidence of bleeding, sensitivity and eating difficulty. More patients stated a preference for no dressing should the operation have to be repeated. After reverse bevel flap procedures, no differences between dressed and undressed segments which would be likely to have practical significance were demonstrated, and it is therefore suggested that application of a dressing should be a matter of individual preference.

Gingival reaction to well-fitted subgingival proximal gold inlays.

Abstract Proximal inlays were seated in contralateral teeth in eight subjects with clinically healthy gingivae. After 23 days, the subgingival area and part of the supragingival surface of one of the inlays was roughened with an abrasive instrument up to 1 mm above the gingival margin; the other inlay remained polished. Gingival fluid flow rate was buccally and lingually measured in the papillary area close to the proximal roughened and polished inlay surfaces and also at papillae beside sound proximal surfaces of the same teeth during a period of 85 days. In addition, plaque was scored and also assessed planimetrically.The mean gingival fluid flow rate was significantly higher adjacent to both the rough and polished filling surfaces when compared with proximal surfaces without restoration. Plaque indices were similar in the vicinity of filled and sound proximal areas irrespective of the gold surface texture. Planimetrically more plaque was formed on the sites of roughened inlays.

Alterations in crevicular fluid flow during healing following gingivectomy and flap procedures.

Gingival fluid was collected from the mesial aspects of the anterior teeth of twelve patients before initial preparation and two weeks after completion of this. All pockets from which fluid was collected were measured prior to initial preparation. On twelve occasions pockets were eliminated by conventional gingivectomy and on eight flaps were reflected followed by curettage and osteoplasties. Gingival fluid was collected immediately prior to operation and at two, three and four weeks post‐operatively. A strong correlation (r =+0.93: P < 0.001) was found between pocket depth and the amount of the gingival fluid exudate. Initial preparation reduced the amount of fluid flow, the extent of the reduction being significant (P < 0.01) where pockets were deep. No significant change in the amount of fluid flow had occurred two weeks after either type of operation but there were significant reductions (P < 0.01) between the third and fourth post‐operative weeks when flap procedures had been performed and in the fourth post‐operative week in the case of gingivectomies. The level of gingival fluid flow was the same in both groups four weeks after the operations even though it was twice as great in the flap procedures group as in the gingivectomy group before operation.

Influence of sex hormones on gingival exudation in gingivitis-free female dogs.

This investigation was undertaken to study the influence of an increased level of estrogen and progesterone upon gingival exudation in gingivitis‐free female dogs. Four female dogs constituted the experimental material. For three weeks prior to the hormone study all were supplied with a hard diet consisting of raw bovine tracheas. In addition, the teeth were cleaned every second day with rubber cups and pumice. During the experimental period gingival fluid was collected from the buccal areas.The intra‐crevicular method was applied in three quadrants and the extra‐crevicular in one quadrant. A total of seven strips from six teeth were measured in each quadrant. Gingival fluid was sampled five times during the course of this study; once immediately before and once seven days after each of the two hormone periods and twice during the course of hormone administration. During the first hormone period comprising 52 hours, each dog was twice given 1 mg estrogen and 25 mg progesterone. During the second hormone period, which also lasted for 52 hours, the dose of each hormone was doubled. The hormones were injected intramuscularly in the hind leg. The results show that when the intra‐crevicular technique of sampling was used, the administration of estrogen and progesterone induced an increased flow of gingival fluid. The increased gingival exudation was observed in all four dogs and occurred in both of the hormone periods. At the end of the period of sex hormone administration the gingival exudation returned to normal. The present study also showed that no influence of sex hormones on gingival exudation could be demonstrated by the extra‐crevicular sampling method.