Gingival crevicular fluid and saliva.

Understanding the structure and function of the mouth, its tissues and secretions is of great interest to physiologists, cell biologists, immunologists and microbiologists but is also of fundamental interest to the dental professional interested in comprehending the aberrant processes associated with oral disease and in the application of effective clinical interventions. The field of periodontology, which has a truly multidisciplinary perspective cutting across leading edge molecular and cellular biology, clinical dentistry, epidemiology and behavioural science, exemplifies this. A paradigm shift in recent years has led to the consideration of the oral cavity (and, thus, oral disease) not in isolation but as a component integrated with systemic physiology, important in maintaining systemic health and reflective of systemic disease; this has served to promote periodontology, in particular, into the forefront of medicine in general. This volume of Periodontology 2000 considers the role of gingival crevicular fluid and saliva in physiological function, maintenance of oral tissue integrity, defense against pathogens and oral disease as well as the many, emerging applications of analysis of these fluids in support of periodontal disease diagnosis, prognosis and epidemiology. However, whilst the emphasis is on periodontal disease, the wider contexts of oral and systemic health are also key considerations.

Metal-ceramic crowns cemented with two luting agents: short-term results of a prospective clinical study.

A prospective, randomized, controlled, split-mouth trial was performed to evaluate the cementation modes for metal-ceramic crowns. A total of 40 fully veneered metal-ceramic crowns were delivered in the posterior jaw segments of 20 patients using either a self-adhesive resin cement (RelyX Unicem Aplicap, 3M ESPE; n = 20) or a zinc oxide phosphate cement (Hoffmann's Cement, Hoffmann; n = 20). Thirteen parameters related to the abutment teeth and their periodontal status were evaluated. A visual analog scale was used to assess the sensitivity of the abutment teeth by patient-based outcomes. Data were statistically analyzed by a single-classification ANOVA (α = 0.05) and logistic regression analysis. The results presented were obtained after a mean observation period of 1.8 years. The dropout rate was 0%. None of the abutment teeth exhibited secondary caries at the restoration margins. No significant differences were demonstrated between the luting agents based on visual analog scale (p > 0.05), hypersensitivity (OR = 1.31), abutment mobility (p > 0.05), or probing depths (p > 0.05). Based on the sulcus fluid flow rates, a significantly greater mean difference was obtained with zinc oxide phosphate cement than with self-adhesive resin cement (9.2 units; p = 0.0006). Significant differences between the baseline examination and the follow-up examinations for sulcus bleeding index (p = 0.0013) and plaque index (p < 0.0001) were observed regardless of the luting agent used. The two cement types showed scarcely any differences between the parameters investigated. The outcomes of cementing fully veneered metal-ceramic crowns were equally good with self-adhesive resin cement as with the clinically proven zinc oxide phosphate cement.

Long-term effects of a single application of a water-cooled pulsed Nd:YAG laser in supplement to scaling and root planing in patients with periodontal inflammation.

The aim of this work was to investigate the long-term effects of a single application of a water-cooled pulsed neodymium yttrium aluminium garnet (Nd:YAG) laser, in combination with scaling and root planing (SRP) for the treatment of periodontal inflammation. Twenty-two patients were included in this split-mouth single blind randomized controlled clinical trial. The parameters of the air and water-cooled Nd:YAG laser were: 4 W, 80 mJ/pulse, 50 Hz and a pulse width of 350 µs. The "test side" was treated with a single application of Nd:YAG laser and SRP; while the "control side " was treated with SRP alone. At baseline, and after a median follow-up time of 20 months (range 12-39), periodontal inflammatory parameters (plaque index [PI], gingival index [GI], probing pocket depth [PPD]), and marginal bone loss (on digital bite-wing radiographs) were measured. Gingival crevicular fluid (GCF) was collected from the teeth 35, 36, 45, and 46 at baseline and at follow-up. Pl (p < 0.01), GI (p < 0.01), and PPD (p < 0.001) were significantly lower on the test side compared to the control side at follow-up. Radiological results showed significantly less bone loss on the test side compared to the control side (p < 0.05). GCF volume was lower on the test side compared to the control side (p < 0.01). In conclusion, a single application of Nd:YAG laser in combination with SRP had a positive long-term effect on periodontal health compared to treatment by SRP alone.

Extracellular neutrophil traps in periodontitis.

BACKGROUND AND OBJECTIVE: Chronic periodontitis, the chronic inflammatory disease of the periodontium, is caused by bacteria and is characterized by an influx of neutrophils into the gingival crevice. Recently, a 'new' extracellular neutrophil defense mechanism - neutrophil extracellular traps - has been described. However, their role in periodontitis has not yet been investigated. MATERIAL AND METHODS: Clinical examinations, transmission and scanning electron microscopy, as well as cytology and confocal laser-scanning microscopy, were employed to analyze gingiva biopsies and crevicular exudate from patients with chronic periodontitis. RESULTS: An abundance of neutrophil extracellular traps and some phagocytic neutrophils was found on the gingival pocket surface and in the purulent crevicular exudate. Finding neutrophil extracellular traps in the spontaneously effused purulent crevicular exudate clearly indicated that they are flushed from the pocket by the crevicular exudate. In cases of dispersal of subgingival plaque bacteria, their trapping by neutrophil extracellular traps in purulent crevicular exudate and on the gingival surface was demonstrated. CONCLUSION: Trapping the crevicular bacteria prevents their adhesion to and invasion of the gingiva. The combination of neutrophil extracellular traps and crevicular exudate outflow appears to be a 'novel' defense mechanism for the clearance of crevicular bacteria in chronic periodontitis.

Fluid dynamics of gingiva in diabetic and systemically healthy periodontitis patients.

OBJECTIVES: The influence of diabetes mellitus (DM) on the fluid dynamics of periodontium has not been reported in periodontal disease. The objectives of this study were (i) to investigate the alterations in the fluid dynamics of periodontium in diabetic periodontitis patients, and present the association of this phenomenon with the metabolic control of DM; (ii) to reveal any correlation between the fluid dynamics of periodontium and clinical signs of periodontal disease in DM and periodontitis. DESIGN: Fifteen well-controlled diabetic chronic periodontitis patients (Group 1), 14 systemically healthy chronic periodontitis patients (Group 2), and 14 systemically and periodontally healthy individuals were included in the study. Gingival crevicular fluid volume (GCF-V) and gingival tissue osmotic pressure (GOP) were used as the parameters of periodontal fluid dynamics. GCF-V was measured by a Periotron device, while GOP was measured by a digital osmometer. Silness-Löe plaque index (PI), Löe-Silness gingival index (GI) and clinical attachment loss (AL) levels were recorded to determine the periodontal health status. RESULTS: PI, GI and AL were higher in Groups 1 and 2 than in Group 3 (P<0.05), but similar between Groups 1 and 2 (P>0.05). Increased GCF-V and GOP were observed in Groups 1 and 2 compared with Group 3 (P<0.01), and the increase in Group 1 was greater than that in Group 2 (P<0.01). There were strong positive correlations between GCF-V and GOP in all three groups: between GI and GCF-V and GI and GOP in Groups 1 and 2; and between AL and GCF-V and AL and GOP in Groups 2 and 3. CONCLUSION: The results suggest that (i) DM may have an additive influence on the fluid dynamics of periodontium in the presence of periodontal disease; (ii) this phenomenon may not be prevented by the metabolic control of DM; (iii) the clinical signs of periodontal disease may be affected by the fluid dynamics of periodontium in both DM and periodontitis.

Biology of teeth and implants: The external environment, biology of structures, and clinical aspects.

For the past several thousand years, until development of the titanium dental implant, only a few missing teeth were replaced successfully in a very small number of individuals. Nowadays, placement of dental implants has become sufficiently commonplace that there is a need to interchange information between what we know about periodontal health and disease and what we know about health and disease involving dental implants. This review discusses the similarities and differences between teeth and dental implants with regards to anatomy, biology, physiology, and pathologic processes. The concept of biologic width is discussed in the context of interaction of periodontal and peri-implant tissues with microbial products produced by periodontal biofilms. The periodontal microbiome is discussed as networks of organisms interacting not only with periodontal and peri-implant tissues, but also with each other as networks of competing organisms. Overall, the transfer of biologic knowledge from what we know about peri-implantitis and what we know about periodontitis should help to develop new directions for biologic understanding about both health and disease of teeth and dental implants.

Fluid dynamics of gingiva and gingival health in children with end stage renal failure.

OBJECTIVES: Impaired homeostasis and fluid balance are important physiopathological alterations in patients with chronic renal failure which may adversely affect the fluid dynamics and health status of tissues and organs. There are insufficient data about this phenomenon in periodontal tissues. The aim of this study was to evaluate the fluid dynamics of gingiva in children with end stage renal failure (ESRF), correlating this entity with gingival health in the same patient group. DESIGN: Fifteen paediatric ESRF patients undergoing peritoneal dialysis (test group) and 15 systemically healthy children (control group) who were without periodontitis participated in the study. Fluid dynamics of gingiva were assessed via the gingival crevicular fluid (GCF) volume and tissue osmotic pressure (GOP) levels in the groups. GCF volume was measured using a Periotron 8000, whereas GOP was measured using a digital osmometer. Silness and Löe Plaque index (PI) and, Löe and Silness gingival index (GI) scores were utilized to determine the gingival health status in the study population. RESULTS: There were increases in the GCF volume and GOP of the test group compared to those of the control group (p<0.01). The PI and GI scores were higher in the test group than in the control group (p<0.01). Strong and positive correlations were found between GI and GCF volume, GI and GOP and, GCF volume and GOP in both groups (p<0.01). CONCLUSIONS: Our findings suggest that the fluid dynamics of gingiva may alter in children with ESRF, and this phenomenon may consequently affect the gingival health of these patients.

Patients with dental calculus have increased saliva and gingival crevicular fluid fetuin-A levels but no association with fetuin-A polymorphisms.

Fetuin-A is a potent inhibitor of calcium-phosphate precipitation and of the calcification process, therefore it can also be related with dental calculus. Thus, we aimed to investigate a possible relationship between fetuin-A gene polymorphism and the presence of dental calculus. A possible relationship between serum, saliva and gingival crevicular fluid (GCF) levels of fetuin-A was also investigated. Fetuin-A c.742C > T and c.766C > G polymorphisms were investigated in 103 patients with or without dental calculus. Additionally, serum, saliva and GCF fetuin-A levels of patients were compared according to dental calculus presence. A significant difference was not observed in the distribution of the fetuin-A c.742C > T and c.766C > G polymorphisms between patients with or without dental calculus. Saliva and GCF fetuin-A concentrations of patients with dental calculus were statistically higher than those without dental calculus (P=0.001, P=0.036 respectively). According to our results, fetuin-A c.742C > T and c.766C > G polymorphisms were not associated with presence of dental calculus. However, higher GCF and saliva fetuin-A levels were detected in patients with dental calculus than in patients without dental calculus, which may result from an adaptive mechanism to inhibit mineral precipitation and eventually calculus formation.

Oxidative Stress and Periodontal Disease in Obesity.

Periodontal disease is a chronic inflammatory disease of the jaws and is more prevalent in obesity. Local and systemic oxidative stress may be an early link between periodontal disease and obesity. The primary aim of this study was to detect whether increased periodontal disease susceptibility in obese individuals is associated with local and systemic oxidative stress. Accordingly; we analyzed periodontal status and systemic (serum) and local (gingival crevicular fluid [GCF]) oxidative status markers in young obese women in comparison with age-matched lean women.Twenty obese and 20 lean women participated. Periodontal condition was determined by clinical periodontal indices including probing depth, clinical attachment level, gingival index, gingival bleeding index, and plaque index. Anthropometric, hormonal, and metabolic measurements were also performed. Blood and GCF sampling was performed at the same time after an overnight fasting. Serum and GCF total antioxidant capacity (TAOC), and total oxidant status (TOS) levels were determined, and oxidative stress index (OSI) was calculated.Clinical periodontal analyses showed higher gingival index and gingival bleeding index in the obese group (P = 0.001 for both) with no significant difference in probing depth, clinical attachment level, and plaque index between the obese and the lean women. Oxidant status analyses revealed lower GCF and serum TAOC, and higher GCF and serum OSI values in the obese women (P < 0.05 for all). GCF TOS was higher in the obese women (P < 0.05), whereas there was a nonsignificant trend for higher serum TOS in obese women (P = 0.074). GCF TAOC values showed a negative correlation with body mass index, whereas GCF OSI was positively correlated with fasting insulin and low-density lipoprotein-cholesterol levels (P < 0.05 for all). Clinical periodontal indices showed significant correlations with body mass index, insulin, and lipid levels, and also oxidant status markers.Our results suggest that young obese, otherwise healthy, women show findings of early periodontal disease (gingival inflammation) compared with age-matched healthy lean women, and that local/periodontal oxidative stress generated by obesity seems to be associated with periodontal disease.

Determination of the pH of peri-implant crevicular fluid in successful and failing dental implant sites: a pilot study.

The aims of this pilot study were to assess if a standard technique which is used to determine the pH of dental plaque around natural teeth (the 'touch electrode' technique) could be modified for use to determine the pH of crevicular fluid around dental implants, and to evaluate any possible changes in the peri-implant crevicular fluid pH in successful and failing implants. pH measurements of a sample of subjects' (n=17) peri-implant crevicular fluid of both successful and failing dental implants present in the same oral cavities were performed using iridium/iridium oxide (Beetrode) electrodes with 100 microm sensing tips, connected to an Orion 720 A pH meter. The technique appeared to be satisfactory for the purpose intended. The mean pH of the successful implants was 6.80 [STD+/-0.4; 95% confidence intervals (CI), 6.50-7.0], and that for the failing dental implants was 7.20 (STD+/-0.6; 95% CI, 6.90-7.50). The results showed that the technique described could be satisfactorily used to determine the pH of peri-implant crevicular fluid at dental implant sites, and that there was a significant difference between the pH of successful and failing dental implants (P<0.05). However, the results obtained should be interpreted with caution in view of the small sample size used in this pilot study.

Serendipity or luck: stumbling on gingival crevicular fluid.

The word "serendipity" comes from the tale "The Three Princes from Serendip" by Horace Walpole. It is defined as an aptitude for making fortunate discoveries accidentally." The demonstration of crevicular fluid was truly an accidental discovery. Niels Brill, a successful private practitioner in Copenhagen with prosthodontics as a specialty, and Bo Krasse, associate professor of cariology, were examining the microflora of gingival pockets in a dog. Before the samples were taken, gingiva and teeth were disinfected with an iodine solution. The solution disappeared from different pockets at different speeds. After intravenous injection of a fluorescein solution, the occurrence of fluorescein could be recorded on filter paper strips. Thus, after stumbling on the disappearance of iodine from the cervical areas of the teeth, two non-periodontologists managed to demonstrate why it disappeared. This article describes the details of the discovery and subsequent studies, and offers some reflections by one of the original authors 37 years later.

Phagocytosis and binding via complement receptors by salivary polymorphonuclear leukocytes. Modulation by saliva and gingival exudate.

The ability of oral polymorphonuclear leukocytes (PMNs) to phagocytose Candida albicans cells and bind Salmonella typhi via complement receptors was investigated. A significantly higher percent of oral PMNs could phagocytose and bind via complement receptors as compared to peripheral blood PMNs. While treatment of peripheral blood PMNs with the donor's saliva caused an increase in the number of complement-receptor bearing cells, as well as a partial increase in phagocytosis, PMNs treated with gingival crevicular fluid (CF) showed a decrease both in phagocytosis and binding. The complexity of environmental conditions and factors, and its role in PMN functions in inflammatory sites is discussed.

Bacteriolytic activity of human gingival exudate.

We investigated the bacteriolytic activity of gingival crevicular fluid (CF) on 14C-labeled Streptococcus faecalis, Streptococcus mutans, Staphylococcus aureus, and on whole dental plaque. CF was collected from 100 healthy donors pooled and centrifuged at 200 g. CF supernate and a frozen and thawed extract of the pellet were interacted with the different bacterial strains, while Streptococcus faecalis and Staphylococcus aureus released 60% and 75% of the radioactive label, only 38% of it was solubilized from Streptococcus mutans, following their incubation with the CF supernate. The findings agreed with results obtained by interacting bacteria with a frozen and thawed lysate of human peripheral blood leukocytes. On the other hand, extracts from frozen and thawed CF pellet were inactive. Further, lipoteichoic acid and lipopolysaccharide were released by CF from Gram-positive and Gram-negative bacteria, respectively. The role of bacteriolytic factors, present in CF, as a result of the interaction between microorganisms and leukocytes at inflammatory sites is discussed.

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.

The relationship between gingival tissue temperatures and various indicators of gingival inflammation.

IN PERIODONTICS there is a need for objective measurements in monitoring disease processes and in assessing the effectiveness of treatment. The purpose of this study was to compare gingival temperatures with indicators of gingival inflammation in order to examine the application of temperature as a diagnostic aid. In six patients the temperature of interproximal sulcular tissue was measured and compared with bleeding upon probing, crevicular fluid flow, pocket depth, plaque accumulation and calculus formation. The results show significantly higher temperatures with bleeding upon probing and with the presence of dental plaque and calculus. Temperature increase correlated with increase in crevicular fluid flow, but a relationship to pocket depth is not clear. The thermocouple microprobe is a very sensitive instrument. The method used is noninvasive, safe and efficient. It can be concluded from our data that differences in gingival temperature reflect not only a regional tissue variability but also can indicate an inflammatory state.

Gingival crevicular pH in experimental gingivitis and occlusal trauma in man.

The purpose of this study was to investigate the fluctuation of gingival crevicular pH during experimentally evoked gingivitis and occlusal trauma, and to examine the relationship between the pH and periodontal health status using a newly developed pH sensor. Maxillary first premolars with clinically healthy gingiva were selected as test teeth in 10 volunteers. In the first study phase, experimental gingivitis was evoked, and gingival index, plaque index, and crevicular pH were recorded during the study period. In the second study phase, experimental occlusal trauma was created with metal onlays having occlusal interferences in lateral movements, and tooth mobility and pH were recorded during the study period. In the gingivitis phase, GI, PlI, and pH values fluctuated significantly during the study period (P < 0.01) and positive correlations were observed between both GI and pH values (P < 0.05), and PlI and pH values (P < 0.01). In the occlusal trauma phase, significant fluctuation was found among tooth mobilities (P < 0.05) during the study phase, but not in pH values. Statistically significant correlations were not observed between tooth mobilities and pH values. These data suggest that the crevicular pH level may not be influenced by experimental occlusal trauma, but shifts toward alkaline with experimental gingivitis.

Effect of Achromycin ointment on healing following periodontal surgery.

This study evaluated the effect of applying 3% Achromycin on healing following periodontal surgery. Twelve patients participated, and 15 contralateral sides with similar disease were compared. The parameters assessed included amounts of gingival crevice fluid quantitated with the Periotron, gingival bleeding tendencies, mobility scores and attachment levels. No significant differences in bleeding tendency, mobility or attachment levels were found. Significant changes were noted on amounts of gingival crevicular fluid, however. The amounts of gingival crevicular fluid collected were significantly lower on the side not treated with Achromycin throughout the observation period. Applying 3% Achromycin did not have a beneficial effect on the healing of periodontal wounds. Whether the application of an antibiotic on sutured flaps limited the bacterial contamination of the gingival tissues through a "wicking effect" of the silk suture material, or if this bacterial contamination had any consequence on gingival healing or reattachment of the periodontal tissues adjacent to the tooth, cannot be determined from this study.

The evaluation of the periotron.

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.

Fluid dynamics of gingival tissues in transition from physiological condition to inflammation.

BACKGROUND: An increase in gingival sulcular fluid filtration is a common clinical sign of early gingivitis. The aim of this study was to describe the fluid dynamics of the gingival interstitial tissues at the level of the sulcus in the transition towards inflammation. METHODS: In 13 anesthetized rabbits, a silk ligature was placed around incisors close to the gingival margin, in order to prevent mechanical cleaning of plaque deposition. After 2 to 7 days, animals were anesthetized and interstitial fluid pressure measured using glass micropipettes connected to a servonull pressure system at the level of the free and attached gingiva. RESULTS: Interstitial pressure was 3.8 +/- 2.9 cmH2O, significantly higher than the normal physiological value (about -1 cmH2O). Colloid osmotic pressure of interstitial fluid samples collected using the wick technique was measured using an osmometer whose membrane had a molecular cut-off of 30 kD and averaged 12.8 +/- 2.8 cmH2O (unchanged relative to control). Mean gingival sulcular fluid flow, measured by placing a PE tube (0.5 mm OD, 0.28 mm ID) in the sulcus, was 0.16 +/- 0.12 microl/h; the mean colloid osmotic pressure was 13.6 +/- 6.6 cmH2O, corresponding to a protein concentration of approximately 2.8 g/dl. Proteins of gingival fluid may leak from inflamed gingival interstitium or derive from bacteria of dental plaque. Histological analysis of gingival biopsies showed neutrophilic polymorphonuclear leukocyte infiltrates in both the dermis and epidermis layer. CONCLUSIONS: Based on hydraulic and colloid osmotic data, the Starling pressure gradient favored filtration from gingival interstitium to the sulcular space, a condition opposite to that observed in healthy gingiva where fluid filtration is absent.

The role of gingipains in the pathogenesis of periodontal disease.

Gingipains are trypsin-like cysteine proteinases produced by Porphyromonas gingivalis, a major causative bacterium of adult periodontitis. HRgpA (95 kDa) and RgpB (50 kDa), products of 2 distinct but related genes, rgpA and rgpB, respectively, are specific for Arg-Xaa peptide bonds. Kgp, a product of the kgp gene, is specific for Lys-Xaa bonds. HRgpA and Kgp are non-covalent complexes containing separate catalytic and adhesion/ hemagglutinin domains, while RgpB has only a catalytic domain with a primary structure essentially identical to that of the catalytic subunit of HRgp. HRgpA and RgpB induce vascular permeability enhancement through activation of the kallikrein/kinin pathway and activate the blood coagulation system, which, respectively, are potentially associated with gingival crevicular fluid production and progression of inflammation leading to alveolar bone loss in the periodontitis site. Kgp is the most potent fibrinogen/fibrin degrading enzyme of the 3 gingipains in human plasma and is involved in the bleeding tendency at the diseased gingiva. HRgpA activates coagulation factors and degrades fibrinogen/fibrin more efficiently than RgpB due to the adhesion/hemagglutinin domains, which have affinity for phospholipids and fibrinogen. Gingipains degrade macrophage CD14, thus inhibiting activation of the leukocytes through the lipopolysaccharide (LPS) receptor, and thereby facilitating sustained colonization of P. gingivalis. Gingipains play a role in bacterial housekeeping and infection, including amino acid uptake from host proteins and fimbriae maturation. Based on the important activities of gingipains in the bacterial infection and the pathogenesis of periodontitis, the bacterial proteinases can be targets for periodontal disease therapy. Immunization with RgpB, HRgpA, or a portion of HRgpA catalytic domain attenuated P. gingivalis induced disorders in mice. In addition, a trypsin-like proteinase inhibitor retarded P. gingivalis growth specifically. Gingipains are potent virulence factors of P. gingivalis, and are likely to be associated with the development of periodontitis. It is, therefore, suggested that gingipain inhibition by vaccination and gingipain-specific inhibitors is a useful therapy for adult periodontitis caused by P. gingivalis infection.