The neutrophil subset defined by CD177 expression is preferentially recruited to gingival crevicular fluid in periodontitis.

In recent years, the concept of distinct subpopulations of human neutrophils has attracted much attention. One bona fide subset marker, exclusively expressed by a proportion of circulating neutrophils in a given individual, and therefore dividing neutrophils in two distinct subpopulations, is the glycoprotein CD177. CD177 is expressed on the plasma and granule membranes of 0-100% of circulating neutrophils depending on the donor. Several in vitro studies have linked CD177 to neutrophil transmigration, yet very few have looked at the role of CD177 for tissue recruitment in vivo. We investigate whether the CD177+ and CD177- neutrophil subsets differ in their propensity to migrate to both aseptic- and microbe-triggered inflamed human tissues. Microbe-triggered neutrophil migration was evaluated in samples of gingival crevicular fluid (GCF) from patients with periodontitis, whereas neutrophil migration to aseptic inflammation was evaluated in synovial fluid from patients with inflammatory arthritis, as well as in exudate from experimental skin chambers applied on healthy donors. We found that the proportion of CD177+ neutrophils was significantly higher in GCF from patients with periodontitis, as compared to blood from the same individuals. Such accumulation of CD177+ neutrophils was not seen in the two models of aseptic inflammation. Moreover, the proportion of CD177+ neutrophils in circulation was significantly higher in the periodontitis patient group, as compared to healthy donors. Our data indicate that the CD177+ neutrophil subset is preferentially recruited to the gingival crevice of periodontitis patients, and may imply that this subtype is of particular importance for situations of microbe-driven inflammation.

Refractory neutrophil activation in type 2 diabetics with chronic periodontitis.

BACKGROUND: Inflammation increases diabetes mellitus type 2 (T2DM) progression and severity. T2DM patients are at high risk of the rapid development of chronic periodontitis (CP). Topical presence, high numbers, and bactericidal effects of immune cells are challenged by augmented antigen-induced inflammation, which promotes both diseases. OBJECTIVES: To investigate gingival cellular inflammatory responses in individuals with previously undiagnosed T2DM with CP or CP alone and in systemically and periodontally healthy controls (H) in vivo and to establish an ex vivo technique permitting quantitative and qualitative assessments of gingival crevicular immune cells. MATERIALS AND METHODS: T2DM + CP, CP, and H individuals (n = 10, each) received a 2-week oral hygiene regimen (OHR). Afterwards, a noninvasive sampling technique was performed to evaluate gingival inflammation induced under standardized conditions in vivo, that is, in the absence of severe periodontal destruction and inflammation at clinically healthy sites. Stimuli (casein/test or phosphate-buffered saline w/o. Ca2+ or Mg2+ , PBS(-/-) /control) were randomly applied contralaterally in the gingival sulci of participants' upper dentes canini. One day after completion of the OHR, gingival crevicular fluid (GCF) was kinetically assayed between the time of the baseline (BL) measurement and 55 minutes. Polymorphonuclear leukocyte (PMN) content (PMNGCF ) was quantitated at an optimum time of 35 minutes. PMNGCF counts reflect local inflammation. Ex vivo samples were fluorimetrically labeled, gated according to the donor's peripheral blood polymorphonuclear neutrophils (PMNPB ), and then counted, employing flow cytometry. RESULTS: PMNGCF counts in unstimulated gingival crevices (at BL) in the T2DM + CP group were higher than those in the CP and H groups. PMNGCF counts were elevated in casein vs PBS(-/-) -stimulated gingival crevices in all groups. Patients with T2DM + CP showed increased PMNGCF counts compared to those with CP (P = .035) according to scatter plots. CD45+ counts in the stimulated sites in T2DM + CP patients were higher than those in CP and H patients (P = .041). Under stimulation conditions, the CD45+ counts differed from those under placebo conditions (P = .019), indicating augmented, inducible inflammatory leukocyte infiltrate in T2DM + CP patients. CONCLUSIONS: This noninvasive technique permits quantitative assessment of (experimental) gingival inflammation in vivo, revealing an influence of T2DM + CP on the number of primary immune cells in the gingival crevice. Patients who are challenged with (local) leukocytosis are likely at risk of collateral damage to the gingival crevice neighboring tissues, favoring the severity and progression of CP and consequently T2DM (www.clinicaltrials.gov NCT01848379).

Oral extracellular vesicles in early pregnancy can identify patients at risk of developing gestational diabetes mellitus.

AIM: To isolate and characterize oral extracellular vesicles from gingival crevicular fluid at 11-14 weeks and evaluate their capacity to identify patients at risk of developing gestational diabetes mellitus. METHODS: A case-control study was conducted, including patients who developed gestational diabetes mellitus (n = 11) and healthy pregnant controls (n = 23). Obstetric and periodontal histories were recorded at 11-14 weeks of gestation, and samples of gingival crevicular fluid obtained. Extracellular vesicles were isolated from gingival crevicular fluid by ExoQuick. Nanoparticle tracking analysis, ELISA and transmission electron microscopy were used to characterize extracellular vesicles. RESULTS: Total extracellular vesicles isolated from gingival crevicular fluid were significantly higher in patients who developed gestational diabetes mellitus later in pregnancy compared to normoglycemic pregnant women (6.3x109 vs 1.7 x1010, p value = 0.0026), and the concentration of the extracellular vesicles delivered an area under the ROC curve of 0.81. The distribution size of extracellular vesicles obtained using ExoQuick was around 148 ± 57 nm. There were no significant differences in the periodontal status between cases and controls. The exosome transmembrane protein CD63 was also detected in the extracellular vesicles of gingival crevicular fluid. CONCLUSION: We were able to isolate extracellular vesicles from gingival crevicular fluid using a method that is suitable to be applied in a clinical setting. Our results provide an insight into the potential capacity of first trimester oral extracellular vesicles as early biomarkers for the prediction of gestational diabetes mellitus in pre-symptomatic women.

Influence of Dental Restorations on Oxidative Stress in Gingival Crevicular Fluid.

Biocompatibility of dental materials (DM) can be evaluated by gingival crevicular fluid (GCF) oxidative stress (OS) status. The goal of the study was to ascertain influence of dental caries degree, teeth position, and type and amount of applied DM on GCF OS profile. For this purpose, we tested six DMs that were sealed in one session: amalgam (Amg), composites: Tetric EvoCeram and Beautifil (BF), phosphate cement-zinc phosphate and polycarboxylate cements-zinc polycarboxylate cements, and glass ionomer cement (GIC). The study included 88 dental outpatients. Follow-up was scheduled at 7th and 30th day. Oxidative stress parameters (malondialdehyde (MDA) and glutathione (GSH) levels and total superoxide dismutase (tSOD) activity) were measured before (0th day) and after the treatment (7th and 30th day) in GCF. Control teeth were mirror-positioned healthy teeth. The DM accomplished the following effects (listed in descending order): increase of GSH in GCF was realized by ZPoC > BF > GIC > Amg; tSOD activity increase by ZPoC > BF > Amg; and MDA decrease by ZPoC > ZPhC > Amg > TEC. Dental caries provokes insignificant rise of OS in GCF. ZPoC and ZPhC showed the highest antioxidant effect, contrary to GIC. Restorations with antioxidant properties may reduce gum diseases initiated by caries lesion, what is of great clinical relevance in dentistry.

Identification of Gingival Crevicular Fluid Sampling, Analytical Methods, and Oral Biomarkers for the Diagnosis and Monitoring of Periodontal Diseases: A Systematic Review.

Background. Several studies in the last decades have focused on finding a precise method for the diagnosis of periodontal disease in its early stages. Aim. To evaluate from current scientific literature the most common and precise method for gingival crevicular fluid (GCF) sample collection, biomarker analytical methods, and the variability of biomarker quantification, even when using the same analytical technique. Methodology. An electronic search was conducted on in vivo studies that presented clinical data on techniques used for GCF collection and biomarker analysis. Results. The results showed that 71.1%, 24.7%, and 4.1% of the studies used absorption, microcapillary, and washing techniques, respectively, in their gingival crevicular fluid collection. 73.1% of the researchers analyzed their samples by using enzyme-linked immunosorbent assay (ELISA). 22.6%, 19.5%, and 18.5% of the researchers included interleukin-1 beta (IL-1ß), matrix metalloproteinase-8 (MMP-8), and tumor necrosis factor-alpha (TNF-α), respectively, in their studies as biomarkers for periodontal disease. Conclusion. IL-1ß can be considered among the most common biomarkers that give precise results and can be used as an indicator of periodontal disease progression. Furthermore, paper strips are the most convenient and accurate method for gingival crevicular fluid collection, while enzyme-linked immunosorbent assay can be considered the most conventional method for the diagnosis of biofluids.

Optimization of buffer solutions to analyze inflammatory cytokines in gingival crevicular fluid by multiplex flow cytometry

OBJECTIVE: the aim of this study was to test two buffer solutions in order to attain a reliable and reproducible analysis of inflammatory cytokines (IL-Beta, IL-6, TNF-alpha, OPG, OPN and OC), in gingival crevicular fluid (GCF)by flow cytometry. MATERIAL AND METHODS: GCF samples from healthy volunteers were collected with perio-paper strips and diluted either in phosphate buffered saline (PBS) or Tris-HCl buffer, with and without protease inhibitors (PI). Cytokine immunoassays were carried out by flow cytometry (Luminex Xmap 200) generating standard curves. RESULTS: standards curves generated with the use of phosphate-buffered saline (PBS) demonstrated best adjustment for cytokines IL-1beta, IL-6 and TNF- α levels, when using Tris-HCl (p < 0.05). CONCLUSIONS: The use of PBS buffer with the addition of PI provided reliable measurements of inflammatory bi-omarkers in GCF samples of healthy volunteers

Cellular populations of periimplant tissues: cytological analysis with sulcular microbrushing.

PURPOSE: Cellular populations of gingival crevicular fluid cytological analysis of integrated implants sites have been investigated by using sulcular cytological brushing, as a means of providing an objective and reproducible technique for monitoring periimplant tissue health. MATERIALS AND METHODS: A total of 60 patients with osteointegrated implants bearing at least for 2 years were divided in 2 groups, A and B. Group A consisted of 30 subjects who presented scarce oral hygiene. In Group B, 30 subjects with a good oral hygiene were included. RESULTS: Comparative analysis of the data obtained by sulcular microbrushing of the 2 groups put into evidence significative differences in the expression of the microbiological and the cytological parameters. CONCLUSION: Clinical monitoring of parodontal and periimplant tissues makes use of several diagnostic tests ranging from clinical and radiological tests to biological assays. However, none of these techniques allows to evaluate periimplant tissue cytological status. This preliminary study suggested sulcular microbrushing might be a useful tool in the early diagnosis and in the micrological monitoring of peri-implantitis.

Periodontal treatment downregulates protease-activated receptor 2 in human gingival crevicular fluid cells.

Protease-activated receptor 2 (PAR2) is implicated in the pathogenesis of chronic inflammatory diseases, including periodontitis; it can be activated by gingipain and produced by Porphyromonas gingivalis and by neutrophil protease 3 (P3). PAR2 activation plays a relevant role in inflammatory processes by inducing the release of important inflammatory mediators associated with periodontal breakdown. The effects of periodontal treatment on PAR2 expression and its association with levels of proinflammatory mediators and activating proteases were investigated in chronic periodontitis patients. Positive staining for PAR2 was observed in gingival crevicular fluid cells and was reflective of tissue destruction. Overexpression of PAR2 was positively associated with inflammatory clinical parameters and with the levels of interleukin-6 (IL-6), IL-8, tumor necrosis factor alpha, matrix metalloprotease 2 (MMP-2), MMP-8, hepatocyte growth factor, and vascular endothelial growth factor. Elevated levels of gingipain and P3 and decreased levels of dentilisin and the protease inhibitors secretory leukocyte protease inhibitor and elafin were also associated with PAR2 overexpression. Healthy periodontal sites from individuals with chronic periodontitis showed diminished expression of PAR2 mRNA and the PAR2 protein (P < 0.05). Furthermore, periodontal treatment resulted in decreased PAR2 expression and correlated with decreased expression of inflammatory mediators and activating proteases. We concluded that periodontal treatment resulted in decreased levels of proteases and that proinflammatory mediators are associated with decreased PAR2 expression, suggesting that PAR2 expression is influenced by the presence of periodontal infection and is not a constitutive characteristic favoring periodontal inflammation.

[Crystallographic picture of the gingival fluid in healthy teeth and by inflammatory periodontal disease].

The research revealed crystallographic structure of the gingival fluid in healthy teeth and by inflammatory periodontal disease. Comparing the results with clinical data both crystallographic patterns in healthy teeth gingival fluid and markers of periodontal tissue pathology were established.

Correlation between probing pocket depth and neutrophil counts in dental plaque, saliva, and gingival crevicular fluid.

OBJECTIVE: Neutrophils play a critical role in the innate immune response. There are no studies that have correlated the neutrophils in plaque, saliva, and gingival crevicular fluid (GCF) to probing pocket depth (PPD) and to each other in periodontally healthy and diseased subjects. The aim of the present investigation was to assess and correlate the neutrophil levels in dental plaque, saliva, and GCF in periodontally healthy and diseased subjects. METHOD AND MATERIALS: Forty-five subjects were recruited. They were divided into three groups based on the Gingival Index (GI) and the Russell Periodontal Index (PI) as clinically healthy (group 1), gingivitis (group 2), and chronic generalized periodontitis (group 3). Saliva and GCF samples were collected using a Durapore filter, and plaque samples were collected using an area-specific subgingival curette. Neutrophils were counted using an improved Neubauer chamber. RESULTS: Neutrophils were present in the plaque, saliva, and GCF of all three samples. There was a statistically significant difference between the neutrophil numbers in all the samples with respect to the severity of periodontal disease. The strength of association was the strongest between PPD and plaque neutrophils. CONCLUSION: The neutrophils in dental plaque samples correlated positively with PPD in periodontally healthy and diseased subjects.

Fibrin mimics neutrophil extracellular traps in SEM.

Neutrophil extracellular traps (NETs) are extracellular web-like structures produced by activated polymorphonuclear neutrophils. NETs kill bacteria extracellularly, but their role in human pathology remains largely unclear. One possible way of studying NETs is through the SEM approach. However, web-like structures observed with SEM in sites of inflammation have been interpreted either as NETs or as fibrin. Thus, the question arises whether a reliable SEM discrimination between NETs and fibrin is at all possible. NET samples were collected as purulent crevicular exudate from periodontal pockets. DNase-digested controls for SEM were employed to demonstrate the DNA backbone and immuno-staining for confocal laser scanning microscopy was used to show the citrullinated histones of NETs. Blood clot samples were treated in the same way as the exudate samples to demonstrate that fibrin and fibrinolysis can mimic NETs and DNA digestion, respectively. No discrimination between fibrin and NETs based on morphological criteria in SEM was possible. Furthermore, only a vague distinction between DNA digestion and fibrinolysis could be made. These findings unambiguously indicate that the discrimination between NETs and fibrin by means of SEM is untrustworthy for samples of inflammatory exudate.

Neutrophil fate in gingival crevicular fluid.

The fate of the neutrophils within the inflammatory exudate in the periodontal crevice and their possible participation in the formation of neutrophil extracellular traps (NETs) are of clinical interest. However, the cytological analysis of clinical samples of inflammatory exudate is restricted by the obtainable quantities, which do not enable employing the routine approaches. Clinical examinations, ACLAR strip sampling, scanning electron microscopy, and confocal laser scanning microscopy were employed to analyze purulent crevicular exudate and gingival crevicular fluid in periodontitis. Bacteria, neutrophil activation, NETosis stages, and NETs were identified by molecular probe, expression of citrullinated histone H3, enzymatic digestion, and ultrastructurally. Crevicular neutrophils, all in diverse NETosis stages marked by the histone citrullination, and an abundance of NETs were found in both purulent crevicular exudate and gingival crevicular fluid. Largely varying quantities of dispersed crevicular bacteria were entrapped by NETs, but no phagocytized bacteria were evident in gingival crevicular fluid. The offered method enables for the first time the demonstration NETs in gingival crevicular fluid. The histone citrullination of all the floating crevicular neutrophils indicates that they all undergo NETosis.

Influence of amine fluoride/stannous fluoride mouthwashes with and without chlorhexidine on secretion of proinflammatory molecules by peri-implant crevicular fluid cells.

AIM: Patients with dental implants need optimal plaque control. Peri-implantitis is an inflammation of soft and hard tissues around implants characterized by bone loss mediated by proinflammatory molecules such as IL-1beta, PGE(2), vascular endothelial growth factor (VEGF). The aim of this study was to evaluate the influence of amine fluoride/stannous fluoride (AmF-SnF(2)) vs chlorhexidine 0.12% (CHX) combined with Am-SnF(2) on IL-1beta, PGE(2) and EGF secretion by cells of crevicular peri-implant fluid. METHODS: Thirty patients with dental implants were included in this study. The test group used AmF-SnF(2) rinsing for 14 days, the control group used CHX rinsing during the first 7 days and AmF-SnF(2) during the following 7 days. Crevicular samples were collected using filter paper strips and assayed for level of IL-1beta, PGE(2) and VEGF with ELISA test. Data were analyzed with paired and unpaired t test. RESULTS: IL-1beta, VEGF and PGE(2) levels were significantly lower in test compared to control group. Comparing first with second week of treatment, a greater decrease of IL-1beta and VEGF was evident in sample group during the second week. There was a lower decrease of IL-1beta and VEGF during the entire treatment in control group. Differences of PGE(2) levels after 7 days in both the groups were not significant while there was a significant difference during the second week. CONCLUSION: The following data suggest that the use of AmF-SnF(2) could decrease the production of IL-1beta, PGE(2) and VEGF by inflammatory cells.AmF-SnF(2) could be an alternative to CHX mouth rinses in plaque control of patients with implants.

Effect of smoking on crevicular polymorphonuclear neutrophil function in periodontally healthy subjects.

BACKGROUND: Polymorphonuclear neutrophils (PMNs) represent the first line of cellular defences in the gingival crevice. Smoking, as probably the most important environmental risk factor for periodontitis, has been shown to adversely affect many neutrophil functions. OBJECTIVE: The aim of this study was to investigate the influence of smoking on PMN numbers and function in periodontally healthy smokers and non-smokers. METHODS: Sixty subjects were recruited: 15 non-smokers, 15 light smokers (< 5 cigarettes/day), 15 moderate smokers (5-15 cigarettes/day) and 15 heavy smokers (> 15 cigarettes/day). Full mouth plaque index, sulcus bleeding index and probing depths were measured. Crevicular washings were obtained from all subjects to harvest PMNs. Numbers of PMNs, percentage viability, and percentage phagocytosis of opsonized Candida albicans were recorded. RESULTS: Mean plaque scores and probing depths were (non-significantly) increased in smokers compared to non-smokers. Mean sulcus bleeding index scores were significantly lower in moderate (0.10 +/- 0.10) and heavy (0.07 +/- 0.11) smokers compared to non-smokers (0.14 +/- 0.13) (p < 0.05). Compared to non-smokers (1.73 +/- 1.08 x 10(6)/ml), the numbers of PMNs were higher in light (1.98 +/- 0.96 x 10(6)/ml) and moderate (2.03 +/- 1.43 x 10(6)/ml) smokers and were lower in heavy smokers (1.68 +/- 1.18 x 10(6)/ml), though there were no significant differences in PMN counts between the groups (p > 0.05). Percentage viability of PMNs was significantly lower in light (77.6 +/- 7.8%), moderate (76.5 +/- 8.2%) and heavy (75.0 +/- 6.5%) smokers compared to non-smokers (85.5 +/- 6.0%) (p < 0.05). Furthermore, the ability of PMNs to phagocytose was significantly impaired in light (58.3 +/- 4.1%), moderate (51.9 +/- 2.33%) and heavy (40.9 +/- 3.5%) smokers compared to non-smokers (74.1 +/- 4.1%) (p < 0.05), with evidence of a dose-response effect. CONCLUSION: Cigarette smoking adversely affected PMN viability and function in this periodontally healthy population.

Aberrant neutrophil reactions in periodontitis.

BACKGROUND: The aim of this study was to compare the activity of neutrophilic granulocytes in patients with severe periodontitis and patients with gingivitis alone. METHODS: The study population comprised 22 patients with gingivitis and 44 with periodontitis. Samples of gingival crevicular fluid (GCF) were collected from untreated patients with gingivitis and from shallow and deep pockets in untreated patients with periodontitis. GCF samples were analyzed for lactoferrin, elastase, matrix metalloproteinase-8 and -9, and collagenolytic activity. RESULTS: The free elastase activity and the neutrophil activity, estimated as the ratio between elastase and lactoferrin, were significantly higher in the samples from the periodontitis patients. These differences were also observed in shallow pockets in periodontitis patients compared to similar pockets in patients with gingivitis. CONCLUSION: This study shows higher levels of free elastase in untreated patients with periodontitis, relative to inflammation-matched controls, which may explain the tissue destruction seen in periodontitis.

The association between neutrophil numbers and interleukin-1alpha concentrations in gingival crevicular fluid of smokers and non-smokers with periodontal disease.

OBJECTIVES: To test whether neutrophil numbers are directly correlated with interleukin-1alpha (IL-1alpha) concentrations in gingival crevicular fluid (GCF) of patients with periodontitis, and to investigate the effects of smoking on these parameters. MATERIALS AND METHODS: A total of 99 GCF samples from 33 patients (14 smokers) suffering from severe chronic periodontitis were collected using Durapore filter strips. Polymorphonuclear leucocyte (PMN) numbers were counted using a Coulter cell counter and IL-1alpha levels were determined by ELISA. Total GCF protein was measured by Bio-Rad assay as a surrogate measure of GCF volume. RESULTS: Mean IL-1alpha concentrations were significantly reduced in smokers compared with non-smokers (non-smokers: 3.29+/-2.02 pg/microg protein, smokers 1.59+/-1.13 pg/microg protein). There was no association between PMN numbers and IL-1alpha concentrations found when analysed either by site or by patient. PMN numbers were not significantly different between the two groups (non-smokers: 1.16 x 10(6)+/-1.04 x 10(6); smokers: 7.30 x 10(5)+/-8.07 x 10(5)). Smoking did not affect mean total protein concentration of samples. CONCLUSIONS: Smoking significantly decreased IL-1alpha concentrations in GCF without affecting GCF volume sampled. The lack of association between IL-1alpha concentration and neutrophil numbers suggests that the reduced IL-1alpha concentrations seen in smokers is independent of any possible effect of smoking on neutrophil chemotaxis, and further suggests that smoking may directly inhibit IL-1alpha production.

Biochemical markers of the periodontal ligament.

For many years the diagnosis of Periodontal Disease has been based on clinical and radiographic methods. Other more recent methods have the objective of studying the inflammatory response of the host. That way, immunologic and biological methods determine the free mediators in the periodontal infection. The components of the gingivo-crevicular liquid or fluid are used to identify or to diagnose the active disease, to anticipate the risk of acquiring the disease and to determine its progress. For it to be clinically useful important changes should be registered the way a specific site turns active or that a previously disease affected site improves its conditions as a result of periodontal therapy. The response of the neutrophillic granulocytes play an important role in the detection of Periodontal Disease. The unspecific defense system in the gingivo-crevicular fluid can be determined through cytokines and/or interleukines that serve to identify sites at risk on the patient. In Periodontal Disease, the cytokines are not only defense mediators of the gingival sulcus fluid, but are also an indicator of tissue destruction. The liberation of high levels of lysosomal enzymes by neutrophils, proteolytic enzymes as the collagenases, or intercytoplasmatic enzymes as dehydrogenase lactate and aspartate amino transferase can equally help monitor the progress of the Periodontal Disease.

Amplified crevicular leukocyte activity in aggressive periodontal disease.

Exaggerated neutrophil responses are a critical component in the pathogenesis of periodontal disease. We investigated whether leukocyte activity in aggressive periodontitis (AP) is increased compared with that in chronic periodontitis (CP) by gingival crevicular fluid (GCF) analysis of myeloperoxidase (MPO), beta-N-acetyl-hexosaminidase (beta-NAH), cathepsin D (CD), and elastase-alpha-1-proteinase inhibitor complex (alpha-1-EPI) before and 6 months after therapy. Initial AP neutrophil responses were significantly amplified compared with those in CP (MPO, 3.2-fold; beta-NAH, 37.5-fold; CD, 2.2-fold; alpha-1-EPI, 1.4-fold; p < 0.05). Surgical therapy resulted in a significant reduction of GCF markers compared with non-surgical treatment. However, the changes in clinical parameters were not different between AP and CP (P > 0.05). Analysis of the results suggests that the local inflammatory response in AP is characterized by increased release of inflammatory mediators of neutrophil origin into the GCF. Analysis of the data further suggests that surgical therapy is a more predictable method for removal of the pro-inflammatory etiology.

PMN responses following use of 2 biodegradable GTR membranes.

OBJECTIVES: In the present prospective trial, the PMN response following resorbable GTR barrier placement was evaluated in mandibular class II furcation lesions. MATERIALS AND METHODS: In 10 patients with treated chronic periodontitis, we randomly selected the 1st molars in the mandible with buccal degree II furcation involvement for either polylactic-citric-acid-ester (PLA) or glycolide-lactic-copolymer (PGL) GTR membrane therapy. We examined contralateral healthy molar sites as untreated controls. We then evaluated the PMN-derived inflammatory tissue response at baseline, weekly up to 6 weeks post-therapy and at 12 and 24 weeks using GCF myeloperoxidase (MPO), beta-glucuronidase (betaG) and beta-N-acetyl-hexosaminidase (betaNAH). RESULTS: The enzyme levels increased from baseline to the 6-week examination. After the 6-week reappointment, enzyme levels dropped reaching the baseline scores at both the 12- and 24-week visit. At PGL sites, the enzyme levels decreased earlier. Compared with healthy control sites, the MPO, betaNAH and betaG tests revealed different maximum levels at week 2 and 3 (PGL) and week 4, 5 and 6 (PLA). For both of the barriers the clinical parameters revealed a sustained improvement following therapy. CONCLUSION: The release of PMN enzymes following placement of bioabsorbable membranes reflects the early soft tissue healing process. Our results suggest that the PMN response is barrier-dependent with the maximum response occuring at different times. However, the host response did not measureably affect the course of clinical healing.