Activity In Gingival Crevicular Fluid Research Articles

Lactate Dehydrogenase Activity in Gingival Crevicular Fluid as a Marker in Orthodontic Tooth Movement

Objectives:This study aims at analyzing the changes in gingival crevicular fluid (GCF) lactate dehydrogenase (LDH) activity during orthodontic movement.Methods:Twenty patients all requiring first premolar extractions were selected and treated with conventional straight wire mechanotherapy. Canine retraction was done using 125 g Nitinol closed coil springs. The maxillary canine on one side served as the experimental site while the contralateral canine served as the control. GCF was collected from the canines before initiation of retraction, then 1 hour after initiating canine retraction, followed by 1 day, 7 days, 14 days and 21 days. GCF LDH levels were estimated and compared with the control site.ResultsThe results revealed significantly higher LDH levels on the 7th, 14th and 21st day at the sites where orthodontic force had been applied. The levels also showed a significant increase from 0 hour to the 21st day. Peak levels were seen on 14th and 21st day following initiation of retraction.Conclusions:The study showed that LDH could be successfully estimated in the GCF and its increased levels could indicate active tooth movement, which could aid the clinician in monitoring active orthodontic tooth movement.

Analysis of matrix metalloproteinase (MMP-8 and MMP-2) activity in gingival crevicular fluid from children with Down’s syndrome

High levels of colonization by periodontopathic bacteria and a high prevalence of chronic inflammatory periodontal disease have been reported in children with Down's syndrome. Matrix metalloproteinases (MMPs) are mediators of extracellular matrix degradation and remodelling, and are deeply involved in the course of periodontal disease. To clarify the relationship between Down's syndrome and periodontitis, we investigated levels of MMP-2 and MMP-8 in gingival crevicular fluid (GCF) and detection of periodontopathic bacteria from subgingival plaque. Samples of GCF and plaque were isolated from central incisors. Levels of MMPs were evaluated by enzyme-linked immunosorbent assay, and periodontopathic bacteria were detected by polymerase chain reaction. Levels of MMP-2 and MMP-8 in Down's syndrome patients were higher than those in healthy control subjects. In the Down's syndrome group, increases in these MMPs were observed in GCF from patients with an oral hygiene index score of < 2 and in GCF from sites that were negative for bleeding on probing. The detection rate of periodontopathic bacteria in Down's syndrome patients was higher than that in the control subjects. Matrix metalloproteinase-2 levels in sites harbouring Porphyromonas gingivalis or Aggregatibacter (Actinobacillus) actinomycetemcomitans were lower than in those without these microorganisms. These results suggest an increase in MMP-2 and MMP-8 in Down's syndrome patients, regardless of whether inflammation of periodontal tissue is present or not.

Gelatinolytic activity in gingival crevicular fluid from teeth with periapical lesions

Many matrix metalloproteinases (MMPs) have been associated with periodontal tissue destruction during chronic and apical periodontitis. The aim of this study was to determine gelatinolytic activity in gingival crevicular fluid (GCF) from teeth with apical lesions and healthy controls. Case-control study. After clinical examination, 30 GCF samples were obtained from 20 affected and 20 control teeth. Subjects with periodontal diseases were excluded. Gelatinolytic activity was determined by gelatin zymography and densitometric analysis. ProMMP-9 levels were significantly increased when compared with controls and were positively correlated with MMP-9 activity in diseased GGF. Similarly, MMP-2 activity, when detected it was exclusively in diseased samples. Additional bands of estimated molecular weights of 60 and 48 kDa were observed. MMP-9 and -2 are highly increased in GCF from teeth with periapical lesions. These gelatinases could represent useful markers in monitoring chronic apical periodontitis in GCF.

Collagenases in Gingival Crevicular Fluid in Type 1 Diabetes Mellitus

Studies have demonstrated that high levels of collagenase activity in gingival crevicular fluid (GCF) are associated with degradation of periodontal tissues in progressive periodontitis compared to periodontally healthy tissues. Because the activation of collagenases is an important issue in periodontitis, we have studied the activation of collagenase in gingival crevicular fluid samples of diabetic patients. Collagenase activity was studied in human gingival crevicular fluids. Twenty-two poorly controlled diabetic patients (e.g., blood glucose: 11.0+/-0.7 mmol/l; hemoglobin A1c [HbA1c]: 9.6%+/-0.3%) and five well-controlled diabetic patients were compared to six chronic periodontitis subjects and five healthy controls. Collagenase activity against type I collagen was measured using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis quantitated by laser densitometry. The poorly controlled diabetic patients had more alveolar bone loss than the well-controlled diabetic subjects and controls (P<0.001; t test). The activity of collagenases in GCF in poorly controlled diabetic patients was similar to that seen in chronic periodontitis subjects (P>0.05) but higher than in healthy controls (P<0.01; t test), whereas there was no difference between the well-controlled diabetic subjects and systemically healthy controls (P>0.05; t test). Poorly controlled diabetes is strongly related to periodontal tissue destruction, and collagenases in GCF may mediate and reflect this effect.

Alkaline phosphatase activity in gingival crevicular fluid during canine retraction

The aim of the study was to investigate alkaline phosphatase activity in the gingival crevicular fluid (GCF) during orthodontic tooth movement in humans. Postgraduate orthodontic clinic. Ten female patients requiring all first premolar extractions were selected and treated with standard edgewise mechanotherapy. Canine retraction was done using 100 g sentalloy springs. Maxillary canine on one side acted as experimental site while the contralateral canine acted as control. Gingival crevicular fluid was collected from mesial and distal of canines before initiation of canine retraction (baseline), immediately after initiation of retraction, and on 1st, 7th, 14th and 21st day and the alkaline phosphatase activity was estimated. The results show significant (p < 0.05) changes in alkaline phosphatase activity on the 7th, 14th and 21st day on both mesial and distal aspects of the compared experimental and control sides. The peak in enzyme activity occurred on the 14th day of initiation of retraction followed by a significant fall in activity especially on the mesial aspect. The study showed that alkaline phosphatase activity could be successfully estimated in the GCF using calorimetric estimation assay kits. The enzyme activity showed variation according to the amount of tooth movement.

Alkaline phosphatase activity in gingival crevicular fluid in response to different magnitudes of continuous orthodontic forces

Alkaline phosphatase activity in gingival crevicular fluid in response to different magnitudes of continuous orthodontic forces

Lactate Dehydrogenase Activity in Human Gingival Crevicular Fluid During Orthodontic Treatment: A Controlled, Short‐Term Longitudinal Study

During orthodontic tooth movement, the early response of periodontal tissues to mechanical stress is an acute inflammatory one. This study uses a longitudinal design to examine lactate dehydrogenase (LDH) activity in gingival crevicular fluid (GCF) to determine if GCF LDH can be used as a diagnostic aid in monitoring tooth movement and tissue response during orthodontic treatment. Seventeen patients (mean age: 16.1 years) participated in the study. Each patient was undergoing treatment for distal movement, and an upper first molar served as the test tooth (TT), while the contralateral (CT) and antagonist (AT) teeth were used as controls. The CT was included in the orthodontic appliance, but was not subjected to the distal movement; the AT was free from any orthodontic appliance. The GCF around the experimental teeth was harvested from both mesial and distal tooth sites immediately before appliance activation, and on days 7, 14, and 21. Clinical gingival conditions were also recorded. Gingival crevicular fluid LDH activity was significantly elevated in all sites of the TT and CT, as compared to the AT, where LDH activity remained at the baseline level throughout the study. Enzyme activity levels were also greater in the TT than in the CT, and in the compression sites. Our results suggest that GCF LDH levels reflect the biological activity that takes place in the periodontium during orthodontic movement, and therefore they can be used as a diagnostic tool for monitoring for correct orthodontic tooth movement in clinical practice.

Lactate dehydrogenase activity in gingival crevicular fluid during orthodontic treatment

During orthodontic treatment, the early response of periodontal tissues to mechanical stress involves several metabolic changes that allow tooth movement. Many studies have evaluated such modifications by analysis of various host metabolites released into the gingival crevicular fluid (GCF). This study used a cross-sectional design to examine the lactate dehydrogenase (LDH) activity in GCF to assess whether GCF LDH can be proposed as a sensitive marker for periodontal tissue modifications during orthodontic tooth movement. Thirty-seven subjects, 16 males and 21 females (mean age 18.7 years, range 14.0 to 26.7 years), participated in this study. Each subject underwent a session of professional oral hygiene and received oral hygiene instructions; 2 weeks later, a fixed orthodontic appliance was placed on the maxillary arch. A randomly selected maxillary canine was considered as the test tooth, and its antagonist, which had no appliance, was used as the control tooth. From 2 to 12 weeks after orthodontic appliance placement, GCF was harvested from both experimental teeth at the mesiobuccal angle, for GCF volume and LDH activity determinations. Clinical monitoring consisted of recording supragingival plaque presence, bleeding on probing, and probing depth at the same collection sites. The results showed that no differences in clinical conditions and GCF volume occurred between the experimental teeth. On the contrary, GCF LDH activity in the test teeth was significantly greater than that of the control teeth ( P < .01). Moreover, no differences were found in the enzymatic activity between the sexes by experimental tooth, and no significant correlation was present between GCF LDH activity and patients’ ages within experimental teeth. Our enzymatic results initially indicated a possible role of GCF LDH during the early phases of orthodontic treatment and therefore warrant further study as a possible diagnostic tool for tissue response during orthodontic treatment.s

Aspartate aminotransferase activity in gingival crevicular fluid during orthodontic treatment. A controlled short-term longitudinal study.

During orthodontic tooth movement, the early response of periodontal tissues to mechanical stress involves an acute inflammatory response, with a sequence characterized by periods of activation, resorption, reversal, and formation in both tension and compression sites. This study used a longitudinal design to examine aspartate aminotransferase (AST) activity in gingival crevicular fluid (GCF) in order to assess whether AST in GCF has potential as a possible diagnostic aid to monitor tooth movement and tissue response during orthodontic treatment. Eighteen patients (mean age, 16.1 years) participated in the study. An upper first molar from each patient undergoing treatment for distal movement served as the test tooth (TT), with its contralateral (CC) and antagonist (AC) first molars used as controls. The CC was included in the orthodontic appliance, but was not subjected to the orthodontic force; the AC was free from any orthodontic appliance. The GCF around the experimental teeth was collected from both mesial and distal tooth sites immediately before appliance activation, 1 hour after, and weekly over the following 4 weeks. Clinical gingival condition was evaluated at baseline and at the end of the experimental period. AST activity was determined spectrophotometrically at 30 degrees C, and the results were expressed as total AST activity (mU/sample). Throughout the experiment, AST levels were significantly elevated in all sites from the TT and CC groups compared to the AC group where, conversely, AST activity remained at the baseline level. However, enzyme levels in the TT group were significantly greater than in the CCs at tension sites on day 14, and in compression sites on days 7 and 14. Moreover, AST activity from the TT group was significantly greater in compression sites than in tension sites on day 7; this was not observed for the CCs. Our results suggest that AST levels in GCF reflect the biological activity which occurs in the periodontium during controlled occlusal trauma and, therefore, should be further evaluated as a diagnostic tool for monitoring correct orthodontic tooth movement in clinical practice.

Relationship between fibrinolytic activity and gingival inflammatory reaction in young individuals.

The fibrinolytic system (the plasminogen activating system) is involved in several physiological and pathological processes. Through the transformation of plasminogen to the aggressive broad spectrum protease plasmin, potent enzymatic activity is released. Plasmin acts directly on connective tissue components, and indirectly by activating proforms of the metalloproteinases. The destructive potential of the fibrinolytic system may thus be of importance for the initiation and progression of periodontal diseases. Earlier studies have shown high concentrations of the plasminogen activator t-PA and its inhibitor PAI-2 in gingival crevicular fluid (GCF) as well as enhanced concentrations in areas of gingival inflammation. The aim of this study was to investigate a possible relationship between the gingival inflammatory reactivity and the fibrinolytic activity in gingival crevicular fluid. Thirty-one young individuals took part in the study. Gingival Index scores and Plaque Index scores were assessed and used to formulate a score expressing an individuals' inflammatory response to microbial plaque levels (Relative G/P score). The fibrinolytic activity of GCF was assessed with a fibrin gel lysis assay, and the levels of t-PA and PAI-2 were assayed with ELISAs. All samples showed fibrinolytic activity. A positive correlation between the fibrinolytic activity and Relative G/P score was found. Thus, in individuals with an enhanced reactivity to dental plaque, a higher plasminogen activating activity in GCF was seen. This indicates a higher potential for tissue proteolysis in these individuals, possibly facilitating spread and deeper involvement of the lesions.

Alkaline phosphatase activity in gingival crevicular fluid during human orthodontic tooth movement

Bone remodeling that occurs during orthodontic tooth movement is a biologic process involving an acute inflammatory response in periodontal tissues. A sequence characterized by periods of activation, resorption, reversal, and formation has been recently described as occurring in both tension and compression tooth sites during orthodontic tooth movement. We used a longitudinal design to investigate alkaline phosphatase (ALP) activity in gingival crevicular fluid (GCF) to assess whether it can serve as a diagnostic aid in orthodontics. Sixteen patients (mean age, 15.5 years) participated in the study. The maxillary first molars under treatment served as the test teeth (TT) in each patient; in particular, 1 first molar was to be retracted and hence was considered the distalized molar (DM), whereas the contralateral molar (CM) was included in the fixed orthodontic appliance but was not subjected to the distal forces. The DM antagonist first molar (AM), free from any orthodontic appliance, was used as the baseline control. The GCF around the experimental teeth was harvested from mesial and distal tooth sites immediately before appliance activation, 1 hour after, and weekly over the following 4 weeks. The clinical gingival condition was evaluated at the baseline and at the end of the experimental term. ALP activity was determined spectrophotometrically at 30° C, and the results were expressed as total ALP activity (mUnits/sample). GCF ALP activity was significantly elevated in the DMs and the CMs as compared with the AMs at 1, 2, 3, and 4 weeks; conversely, in the AMs, GCF ALP activity remained at baseline levels throughout the experiment. Moreover, the enzyme activity in the DMs was significantly greater than in the CMs. In the DMs, a significantly greater ALP activity was observed in sites of tension compared with sites of compression. This difference was not seen with the CMs, in which the enzyme activity increased to the same extent in tension and compression sites. These results suggest that ALP activity in GCF reflects the biologic activity in the periodontium during orthodontic movement and therefore should be further investigated as a diagnostic tool for monitoring orthodontic tooth movement in clinical practice. (Am J Orthod Dentofacial Orthop 2002;122:548-56)

Relationship of changes in interleukin-8 levels and granulocyte elastase activity in gingival crevicular fluid to subgingival periodontopathogens following non-surgical periodontal therapy in subjects with chronic periodontitis.

To determine the effect of scaling and root planing (SRP) on the interrelations of subgingival periodontopathogens and both interleukin-8 (IL-8) and granulocyte elastase activity in gingival crevicular fluid (GCF), and to assess their relations to the short-term treatment response in management of chronic periodontitis. GCF and subgingival plaque were collected from 16 subjects with untreated chronic periodontitis at baseline and 4 weeks after SRP. IL-8 levels were determined by ELISA. Granulocyte elastase activity was analyzed with a specific substrate, pGluProVal-pNA, and the maximal rate of elastase activity (MR-EA) was calculated. 5 DNA-probes were used to detect the presence of A. actinomycetemcomitans (A. a.), B. forsythus (B.f.), P. gingivalis (P.g.), P. intermedia (P.i.), and T. denticola (T.d.), with a sensitivity = 103 cells/paper point. IL-8 and MR-EA levels in GCF decreased significantly after SRP (p < 0.001) with a corresponding reduction of total count of the species. Of the sites with probing depth (PD) >/= 5.0 mm and co-infection by B.f., P.g., P.i. & T.d. at baseline, the sites without persistent co-infection of these species after SRP exhibited a significant reduction of IL-8 levels (p < 0.02), MR-EA levels (p < 0.02) and PD (p < 0.01). No such change was found in the sites where such a co-infection persisted. Moreover, reduction of IL-8 levels in those pocket sites was accompanied by a concomitant reduction of MR-EA (p < 0.02) and PD (p < 0.01), while no significant change in MR-EA levels and PD was noted in those pocket sites that exhibited an increase of IL-8 levels after SRP. At baseline, the former group of sites showed significantly higher IL-8 levels than the latter group of sites (p < 0.02). IL-8-related granulocyte elastase activity was related to the change in infection patterns of the target periodontopathogens following scaling and root planing. Varying initial IL-8 levels in GCF and a corresponding shifting change of granulocyte elastase activity in GCF may characterize the different short-term treatment responses.

“Acid Phosphatase activity in Gingival Crevicular Fluid - A non-invasive adjunct in assessing orthodontic tooth movement???”

“Acid Phosphatase activity in Gingival Crevicular Fluid - A non-invasive adjunct in assessing orthodontic tooth movement???”

Association of early onset periodontitis microbiota with aspartate aminotransferase activity in gingival crevicular fluid.

The objective of this study was to determine the relationship between the activity of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) using the colorimetric PerioGard (PTM) test and the subgingival microflora in early onset periodontitis lesions. The study population consisted of 25 otherwise healthy individuals exhibiting early onset periodontitis (EOP). In each patient four experimental sites were identified comprising one deep periodontal pocket (PD >5 mm) randomly chosen in each quadrant. Bacterial samples were obtained from the experimental sites, consecutively cultured anaerobically and in 10% CO(2) using selective and nonselective media. Isolates were characterized to species level by conventional biochemical tests and various identification kits. Clinical measurements as well as AST activity, assessed either as positive or negative using the PTM, were recorded at the same sites. Sixty-two sites exhibited AST positive and 38 AST negative activity. Analysis of bacterial counts using the ANOVA (Mann Whitney U-test) showed that Streptococcus intermedius, Peptostreptococcus micros, Campylobacter concisus, Bacteroides forsythus, Camplobacter gracilis, Campylobacter rectus and Selenomonas sputigena were significantly higher in sites with AST-positive activity. The odds ratio of having high prevalence of S. intermedius, P. micros, C. concisus, B. forsythus, C. gracilis, C. rectus and S. sputigena in the presence of a positive AST site was very high (range: 3.5-17.0). Streptococcus sanguis, Actinomyces naeslundii, Gemella morbillorum, Capnocytophaga gingivalis, Veillonella parvula, Fusobacterium varium, Eubacterium lentum and Prevotella oralis were detected in significantly higher proportions in sites with AST negative activity and manifested a negative odds ratio in the presence of AST positive sites. The logistic regression analysis revealed that smoking and bleeding upon probing showed a significant association with AST activity, while plaque and suppuration were not found to be significant predictors of AST activity. The co-infection of Porphyromonas gingivalis, B. forsythus and P. micros, or P. gingivalis, B. forsythus and C. rectus were found to be significantly associated with the AST activity (p<0.001). AST positive sites revealed significantly higher occurrence of co-infections by P. gingivalis, B. forsythus, S. sputigena or by P. gingivalis, B. forsythus, S. intermedius than AST negative sites (p<0.001). P. gingivalis, B. forsythus, A. naeslundii co-infection was found significantly higher in the AST negative sites (p<0.001). The present study found a high level of agreement between the presence of putative periodontal pathogens and positive AST scores at periodontal sites that clinically were considered to be potentially disease active. Prospective studies should be performed to confirm the findings.

Association of early onset periodontitis microbiota with aspartate aminotransferase activity in gingival crevicular fluid

AbstractObjectives: The objective of this study was to determine the relationship between the activity of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) using the colorimetric PerioGardTM (PTM) test and the subgingival microflora in early onset periodontitis lesions.Material and methods: The study population consisted of 25 otherwise healthy individuals exhibiting early onset periodontitis (EOP). In each patient four experimental sites were identified comprising one deep periodontal pocket (PD &gt;5 mm) randomly chosen in each quadrant. Bacterial samples were obtained from the experimental sites, consecutively cultured anaerobically and in 10% CO2 using selective and nonselective media. Isolates were characterized to species level by conventional biochemical tests and various identification kits. Clinical measurements as well as AST activity, assessed either as positive or negative using the PTM, were recorded at the same sites.Results: Sixty‐two sites exhibited AST positive and 38 AST negative activity. Analysis of bacterial counts using the ANOVA (Mann Whitney U‐test) showed that Streptococcus intermedius, Peptostreptococcus micros, Campylobacter concisus, Bacteroides forsythus, Camplobacter gracilis, Campylobacter rectus and Selenomonas sputigena were significantly higher in sites with AST‐positive acitivity. The odds ratio of having high prevalence of S. intermedius, P. micros, C. concisus, B. forsythus, C. gracilis, C. rectus and S. sputigena in the presence of a positive AST site was very high (range: 3.5–17.0). Streptococcus sanguis, Actinomyces naeslundii, Gemella morbillorum, Capnocytophaga gingivalis, Veillonella parvula, Fusobacterium varium, Eubacterium lentum and Prevotella oralis were detected in significantly higher proportions in sites with AST negative activity and manifested a negative odds ratio in the presence of AST positive sites. The logistic regression analysis revealed that smoking and bleeding upon probing showed a significant association with AST activity, while plaque and suppuration were not found to be significant predictors of AST activity. The co‐infection of Porphyromonas gingivalis, B. forsythus and P. micros, or P. gingivalis, B. forsythus and C. rectus were found to be significantly associated with the AST activity (p&lt;0.001). AST positive sites revealed significantly higher occurence of co‐infections by P. gingivalis, B. forsythus, S. sputigena or by P. gingivalis, B. forsythus, S. intermedius than AST negative sites (p&lt;0.001). P. gingivalis, B. forsythus, A. naeslundii co‐infection was found significantly higher in the AST negative sites (p&lt;0.001).Conclusions: The present study found a high level of agreement between the presence of putative periodontal pathogens and positive AST scores at periodontal sites that clinically were considered to be potentially disease active. Prospective studies should be performed to confirm the findings.

Concentration of interleukin-1beta and neutrophil elastase activity in gingival crevicular fluid during experimental gingivitis.

The aim of the present study was to measure interleukin-1beta concentrations and neutrophil elastase activity in gingival crevicular fluid (GCF) during experimental gingivitis in humans. 12 healthy young men participated. After prophylaxis, they performed optimal hygiene to reach plaque and gingivitis indices of or approaching zero. All oral hygiene measures were then ceased for a period of 18 days. The Quigley-Hein plaque index (PLI) and Saxer & Mühlemann papillary bleeding index (PBI) were assessed. GCF samples were taken from the mesiobuccal site of two contralateral teeth in the upper jaw by means of periopapers at baseline and on days 3, 7, 14 and 18. After measuring the gingival crevicular fluid volume (GCFV) with the Periotron 8000, the samples were analyzed in our laboratory for the detection of IL-1beta concentration by ELISA. PLI and PBI showed a reduction prior to baseline reaching almost zero, both increasing from day 0 to day 18 (PLI=from 0.1 to 2.9, PBI=from 0 to 2.0). IL-1beta concentration increased from 229.25 ng/ml (day 0) to 526.13 ng/ml (day 18). Clinical data and IL-1beta concentrations were correlated with elastase activity (EA). No significant correlation could be demonstrated between the clinical parameters assessed and IL-1beta or EA (Spearman rank correlation coefficient). A correlation between GCFV and PBI from day 0 to day 18 could be demonstrated. Overall, both IL-1beta and EA showed an increase from baseline throughout the whole study.

Adjunctive treatment with subantimicrobial doses of doxycycline: effects on gingival fluid collagenase activity and attachment loss in adult periodontitis

The therapeutic effects of doxycycline and other tetracyclines in the treatment of periodontitis involve, at least in part, mechanisms that are unrelated to their antimicrobial activity. Previous clinical studies have shown that doxycycline administered orally, at doses below those needed for antimicrobial efficacy, to human subjects with adult periodontitis resulted in significantly reduced collagenase activity in gingival crevicular fluid (GCF) and in extracts of inflamed gingival tissues. The purpose of the present study was to identify clinically effective dosing regimens using subantimicrobial dose doxycycline (SDD) as an adjunctive therapy in patients with adult periodontitis. A total of 75 adult men and women qualified for enrollment into the three-part, placebo-controlled, double-blind, parallel-group study. Patients were stratified based on repeatedly exhibiting pathologic levels of periodontal attachment (ALv) and GCF collagenase activity at several appointments prior to baseline. Patients were administered a scaling and prophylaxis, then 1 of 5 treatment schedules for 12 weeks (part I), followed by a 12-week period of no drug therapy (part II), a second scaling and prophylaxis, and 12 additional weeks of treatment (part III). Primary determinants of efficacy included reductions in GCF collagenase activity and changes in relative ALv. 66 patients completed the 1st 12 weeks (part I) of the 3-part, 36-week study; 51 patients completed the entire 36-week study. From baseline to week 12 (part I), treatment with specially formulated SDD capsules (20 mg) 2x daily (1 x every 12 h) for up to 12 weeks was shown to significantly reduce GCF collagenase activity and to improve ALv, effects not seen in patients treated with placebo. Continuous drug therapy over the 12-week treatment period was needed to maintain and maximize the reduction in GCF collagenase and the improvement in ALv. Improvements in periodontal disease parameters occurred without the emergence of doxycycline-resistant micro-organisms. In patients administered an "on-off-on" regimen of SDD over 36 weeks (parts I-III), essentially no attachment loss occurred in patients receiving the highest of these SDD regimens (20 mg 2x daily during part I and 20 mg 1 x daily in part III), whereas patients administered placebo capsules experienced a mean attachment loss of approximately 0.8 mm at the 24- and 36-week time periods. Doxycycline administered at subantimicrobial doses led to improvements in disease parameters, with no apparent side effects, and appears to have significant potential as an oral adjunctive therapy in the long-term management of adult periodontitis.

Characterization of bone resorbing activity in gingival crevicular fluid from patients with periodontitis.

In attempts to elucidate factors stimulating bone resorption in patients with different inflammatory diseases in the vicinity of the skeleton, e.g., peridontal disease and rheumatoid arthritis, we are investigating the presence of bone-resorbing activity in a variety of inflammatory exudates. The aim of the present study was to characterize the bone-resorbing activity present in patients with periodontitis. Bone-resorbing activity was assessed in gingival crevicular fluids (GCFs) collected from patients with periodontitis and from patients with no signs of gingivitis. Bone-resorbing activity was evaluated by analyzing the capacity of GCFs to stimulate the release of minerals and the breakdown of bone matrix proteins in cultured neonatal mouse calvariae. The concentrations of IL-1alpha, IL-1beta and PGE2 were determined with ELISA and RIA techniques, respectively. GCF eluates from 24 different healthy sites caused a 1.23+/-0.05 fold stimulation of 45Ca release, whereas GCF eluates from 45 different diseased (periodontitis) sites caused a 2.46+/-0.10 fold stimulation. The effect on 45Ca release was time- and concentration-dependent, inhibited by 3 different osteoclast inhibitors and associated with enhanced release of 3H from [3H]-proline-labelled bones. The activity in GCF causing enhanced 45Ca release was unaffected, or in some samples partially reduced, by ultrafiltration using a filter with a molecular weight cut-off of 3000 Daltons. The bone-resorbing activity was temperature sensitive (+90degrees C, 10 min). The concentrations of prostaglandin E2 (PGE2) in the diluted GCF eluates, used in the bone resorption bioassay, were too low to be responsible for the release of 45Ca. Antisera specifically neutralizing human IL-1a inhibited the stimulatory effect of GCF pooled from several diseased sites. The specific, recombinant human IL-1 receptor antagonist completely inhibited the effect of pooled GCFs. GCF eluates from diseased sites contained human IL-1alpha and IL-1beta at concentrations of 1838+/-294 pg/ml and 512+/-91 pg/ml, respectively. These data show that GCF contains activity(ies) stimulating osteoclastic bone resorption in vitro. The factor primarily responsible for this activity seems to be IL-1alpha, but IL-1alpha is not the sole activator of bone resorption in GCF.

Long-term monitoring of adult periodontitis patients in supportive periodontal therapy: correlation of gingival crevicular fluid proteases with probing attachment loss.

The aim of this retrospective study was to determine if a chairside assay for neutral protease activity in gingival crevicular fluid (GCF) could provide an early indication of site-specific disease activity, as defined by probing attachment loss. Clinical measures and assay data were collected at 6-month intervals over an average period of 26 months (range from 24 to 36 months). 38 subjects were selected from a pool of previously-treated chronic adult periodontitis patients who were in periodontal maintenance at intervals of 3 to 6 months. GCF samples were collected from a total of 71 tooth sites exhibiting prior attachment loss, and analyzed for neutral protease activity (NPA). Positive BOP and positive NPA scores were classified as true positives if sites subsequently lost at least 1 mm of attachment over the next 12 months, as confirmed by linear regression analysis. As a predictor of breakdown, the NPA assay had an accuracy of 94% and a risk ratio of 37.6 as compared to values of 58% and 1.5 for BOP. When only the subset of sites > or = 7 mm were considered, the NPA assay had a calculated accuracy of 92% versus a value of 50% for BOP. These results indicate that the assay appears to differentiate between bleeding at sites exhibiting only chronic inflammation with no attachment loss and bleeding at sites undergoing active attachment loss.

Measurement of proteinase activity in gingival crevicular fluid following periodontal therapy.

Measurement of proteinase activity in gingival crevicular fluid following periodontal therapy.