Composition Of Gingival Crevicular Fluid Research Articles

Proteomic Analysis of Gingival Crevicular Fluid During Tooth Eruption

Objective: Gingival crevicular fluid (GCF) is a biological fluid that has the unique capacity to reflect changes in periodontium to its protein composition, making it ideal for potential biomarkers. There is limited information about the mechanism of tooth eruption, for which GCF might provide valuable knowledge. This study aimed to provide a proteomic approach to investigate the composition of GCF obtained from two different supraosseous tooth eruption stages of permanent molars changes.
 Methods: GCF samples were taken from a total of 26 healthy children, whose permanent molar just emerged from the gingiva (5-8 years old) and the occlusal equilibrium stage (9-13 years old). Proteins were extracted with Bio-Rad Rehydration Buffer followed by ZebaTM Spin Desalting Column. GCF samples were separated with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) followed by mass spectrometry- based protein identification.
 Results: A new optimized protocol with enriched protein extraction from GCF samples was developed. Several proteins related to tooth eruption were detected. It was determined that keratin type II cytoskeletal 4 and keratin type I cytoskeletal 9 decreased and albumin increased in GCF protein content in erupting teeth compared to those in the occlusal equilibrium phase.
 Conclusion: This methodology, which we have applied for 2D-PAGE of GCF, can also be a source for other studies. There is huge diagnostic potential in mass spectrometry technologies, this study can be carried forward by using other approaches.

MALDI-TOF/MS Profiling of Whole Saliva and Gingival Crevicular Fluid in Patients with the Invisalign System and Fixed Orthodontic Appliances.

The movement of teeth by orthodontic treatment with the Invisalign (IN) system and fixed orthodontic appliances (FOA) is characterized by the reconstruction of periodontal ligaments, alveolar bone, and gingiva. A reflection of these phenomena can be found in the composition of gingival crevicular fluid (GCF). A total of 90 samples from 45 participants (45 whole saliva and 45 GCF), including 15 patients with FOA, 15 patients with IN, and 15 patients with oral health, were subjected to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF/MS) analysis. Mass fingerprints were generated for each sample. Three models were tested: a quick classifier (QC), a genetic algorithm (GA), and a supervised neural network (SNN). For both groups of samples (saliva and GCF), the GA model showed the highest recognition abilities of 88.89% (saliva) and 95.56% (GCF). Differences between the treated (FOA and IN) groups and the control group in saliva and GCF samples were determined using cluster analysis. In addition, we monitored the effect of long-term orthodontic treatment (after 6 months) in the lag phase of orthodontic tooth movement. The results show increased levels of inflammatory markers (α-defensins), which may indicate an ongoing inflammatory process even after 21 days from force application.

Comparing Ionic Profile of Gingival Crevicular Fluid and Saliva as Distinctive Signature of Severe Periodontitis.

Although increasing evidence is emerging on the contribution of chemical elements in periodontal health, no studies have concomitantly evaluated the ionic profile in gingival crevicular fluid (GCF) and saliva in relation to the underlying periodontal status. Our hypothesis is that these biofluids have distinctive ionic content. Therefore, the aim of this cross-sectional study was to analyze the elemental composition of GCF and saliva in order to explore which biological matrix and which combination of elements could discriminate between periodontitis and periodontal health. Twelve ions were analyzed in GCF and unstimulated saliva from 54 subjects (18 periodontally healthy, 18 untreated severe periodontitis and 18 treated severe periodontitis) using inductively coupled plasma–mass spectrometry (ICP-MS) and inductively coupled plasma–optical emission spectroscopy (ICP-OES). These analytical techniques were able to determine levels of sodium (Na), potassium (K), calcium (Ca) and magnesium (Mg), while the other elements were below the detection threshold. Na and K ions were detected at elevated concentration in untreated periodontitis compared with treated periodontitis and healthy periodontium. Ca was increased in untreated periodontitis, but the difference was not significant. In saliva, only Na was significantly associated with periodontitis. The combination of Na and K in GCF enabled the correct assignment of a subject to the periodontitis or healthy group. Based on these preliminary results, GCF demonstrated higher clustering potential than saliva.

Clinical and laboratory rationale for the effectiveness of transcranial electrostimulation in the treatment of chronic generalized periodontitis in elderly patients

Relevance. Immunomodulation is becoming increasingly relevant in geriatric dentistry. The administration of this therapy cannot be empirical and requires the study of the corresponding immunological parameters. The study of the gingival crevicular fluid composition allows evaluating not only the immune system ageing processes and their influence on the periodontitis course but the effectiveness of the treatment. The study aimed to evaluate the effectiveness of transcranial electrostimulation (TES) for the treatment of periodontitis in elderly patients and to prove the value of gingival crevicular fluid laboratory testing during immunomodulatory therapy. Materials and methods. We conducted a clinical, prospective, controlled, randomized, unblinded, comparative study. We examined two groups of periodontal patients (68 subjects), aged 60-74, and equally treated them. But the patients in the second group additionally received transcranial electrostimulation (TES). The research included the follow-up assessment of IL-1β, TNF-α, IL-10, sIgA and LDH in the crevicular fluid and periodontal index reduction: SBI and PMA (%), before the treatment and on the 7th, 14th, 21st and 30th day.Results. The study established that the periodontal treatment scheme, which includes TES (immunomodulation), is more clinically effective than conventional treatment. There were statistically significant differences in the clinical and laboratory parameters between the 1st and the 2nd patient groups after the 14th day (p < 0,05). On the 30th day of observation, the described trend persisted: the parameters of crevicular fluid in both groups continued to decrease. Moreover, their mean values were significantly lower in the 2nd group than in the 1st. The level of IL-1β (pg/ml) was 16.9 ± 0.33 and 18.8 ± 0.38;TNF-α (pg/ml): 11.9 ± 0.37 and 14.4 ± 0.4; IL-10 (pg/ml): 11.0 ± 0.35 and 12.9 ± 0.36, respectively (p < 0.05).Conclusion. In elderly patients, TES (as a part of a comprehensive treatment) is more effective clinically than the conventional periodontal treatment alone. In geriatric dentistry, it is of primary importance to test gingival crevicular fluid composition and properties in a laboratory and to select the most significant immunological and biochemical components: IL-1β, TNF-α, IL-10, sIgA, LDH. It allows a periodontist to confirm the changes in the manifestations of periodontitis in the elderly patients and to verify the clinical effectiveness of immunomodulation.

Cytomorphometric Analysis of Inflammation Dynamics in the Periodontium Following the Use of Fixed Dental Prostheses.

Cytomorphometry is used in the sampling of biological materials and diagnostic procedures. The use of cytological studies in periodontal diseases is not well described in the literature. Our study aimed to quantitatively assess the inflammation dynamics using cytomorphometric analysis of the periodontium before and after the use of fixed dental prostheses. Following ethics approval, a total of 105 subjects were divided in 3 groups as gingivitis (n = 23), periodontitis (n = 58), and healthy periodontium (control) (n = 24). The fixed dental prostheses (crowns and fixed partial dentures) were fabricated from cobalt-chrome metal-ceramic prostheses using the conventional method (C/M-CoCr), cobalt-chrome metal-ceramic prostheses by the computer-aided design and computer-aided manufacturing (CAD/CAM) technique (C/C-CoCr), and zirconia-based ceramic prostheses by the CAD/CAM technique (C/C-Zr) among subjects with gingivitis and periodontitis. The gingival crevicular fluid (GCF) was obtained from subjects before and after the use of the prostheses. The total count of epithelial cells and the connective tissue cells or polymorphonuclear neutrophils (PMNs) in GCF were studied using cytomorphometric analysis. The Statistical Package Tor the Social Sciences (SPSS), Version 20 (IBM Company, Chicago, IL, USA) was used to analyze the results and the significance level was set at p = 0.05. The data for before and after the use of the prostheses were compared using independent t-Tests. Similarly, the results after the use of prostheses in gingivitis, periodontitis, and control in each type of prostheses were compared using One-way ANOVA with post hoc using Scheffe. The total epithelial cells and the PMNs were determined along with the epithelium/leukocyte index. Regardless of the prostheses type used, no significant change in the parameters was identified among patients with a healthy periodontium, before and after prosthetic treatment. In all study groups, a statistically increase (p value < 0.05) was observed in the oral epithelial cell counts and a statistically decrease (p < 0.05) in the PMNs count following the use of the fixed prostheses. Data on cytomorphometric analysis could enable the selection of the most appropriate prostheses for use in patients with periodontal pathologies. When choosing prostheses, changes in the composition of GCF could be considered as a useful criterion for their use.

Novel and known periodontal pathogens residing in gingival crevicular fluid are associated with rheumatoid arthritis.

BackgroundPeriodontitis is a suspected environmental risk factor for the development of rheumatoid arthritis (RA). However, correlation mechanisms between the two pathologies remain elusive. This study examined potential correlations between detached subgingival bacteria collected in gingival crevicular fluid (GCF) and RA parameters.MethodsRA patients (n = 52, F:M = 40:12), patients with Behcet's disease (BD, n = 40, F:M = 29:11) as another systemic inflammatory disease were studied along with a systemically healthy control group (HC, n = 57, F:M = 40:17). All participants were non‐smokers. Full mouth periodontal parameters were recorded. RA activity was assessed using the 28‐joint Disease Activity Score (DAS‐28). Rheumatoid factors (RFs)‐IgM and ‐IgA were measured by ELISA. GCF samples were investigated by means of fluorescent in situ hybridization for 10 different bacterial taxa.ResultsThe taxa TM7, Synergistetes cluster B, Leptotrichia, Megasphaera, Anaeroglobus geminatus, and Tannerella forsythia displayed significantly differential abundances between the groups. Whereas abundances of Megasphaera and A. geminatus were significantly increased in the RA group, only Porphyromonas gingivalis displayed significant correlations with plaque scores, bleeding on probing, and RF‐IgA. RA patients displaying RF‐IgA levels >75 IU/mL exhibited five‐fold more abundant P. gingivalis levels than patients below the threshold. This association with RF‐IgA levels appeared even more pronounced, by six‐fold more P. gingivalis (P = 0.025), in patients with a DAS‐28 score >3.2, indicative of moderate/very active RA.ConclusionsUnattached GCF bacteria may mediate the association between periodontitis and RA, and monitoring the bacterial composition of GCF might inform on RA activity. The role of newly identified bacterial taxa in RA warrants further investigations.

Biomarker for Asymptomatic Apical Periodontitis in Gingival Crevicular Fluid: aMMP-8.

Objectives Asymptomatic apical periodontitis (AAP) is one of the most widespread chronic inflammatory diseases in the field of dental medicine. Active matrix metalloproteinase (aMMP-8) previously demonstrated diagnostic potential as a biomarker for AAP in gingival crevicular fluid (GCF). The aim of this study was to determine the levels and diagnostic accuracy of aMMP-8 in GCF from teeth with AAP. Materials and Methods In this cross-sectional study, GCF samples were obtained from teeth with AAP (sample group, n = 31) and their contralateral teeth (control group, n = 31). Clinical and cone beam computed tomography (CBCT) parameters were used for the diagnosis and assessment of AAP and the determination of clinically healthy marginal periodontal tissues. One pool GCF sample per tooth was obtained from the whole crevice’s perimeter. aMMP-8 levels were determined by enzyme-linked immunosorbent assay (ELISA). Statistical Analysis Wilcoxon signed ranks test and Spearman rank correlation coefficient (rs) were used as statistical tools. The significance level was set at p < .05. Results The two groups demonstrated biomarker levels corresponding to a healthy marginal periodontal tissue. aMMP-8 levels were statistically and significantly higher in the samples collected from teeth with AAP. Lesions with greater volume showed correspondingly larger diameters. No statistically significant correlation between aMMP-8 levels and lesions’ volume or diameter was discovered. Conclusion GCF composition is modified by AAP only to a minimal extent. Further research is needed to substantiate the utilization of aMMP-8 as a potential biomarker for the diagnosis of the disease as well as to explore its relationship with other biomarkers.

Age effect on orthodontic tooth movement rate and the composition of gingival crevicular fluid : Aliterature review.

To evaluate and form acomprehensive understanding of the effect of patient age on bone remodeling and consequently on the rate of orthodontic tooth movement (OTM). Asystematic search in PubMed and Embase from 1990 to December 2017 was performed and completed by ahand search. Prospective clinical trials which investigated the rate of OTM and/or studies assessing age-related changes in the composition of gingival crevicular fluid (GCF) in older compared to younger study groups were included. Study selection, data extraction and risk of bias were assessed by two authors. Eight studies fulfilled the inclusion criteria. Among them, four evaluated the rate of OTM and six investigated mediators in the GCF (prostaglandin E2, interleukin [IL]-1β, IL‑6, IL‑1 receptor antagonist, receptor activator of nuclear factor kappa‑Β ligand, osteoprotegerin, granulocyte-macrophage colony-stimulating factor, pentraxin3). Patient age ranged between 16 and 43years for older and <16years for younger groups. In most of the studies, the younger patients showed faster OTM in the first phase of treatment and more pronounced cytokine levels. Older patients had adelayed reaction to orthodontic forces. The small number of included studies and large heterogeneity in study design give limited clinical evidence that the older patients are less responsive to orthodontic force in comparison to younger patients. The initial cellular response to orthodontic force is expected to be delayed in older patients. Control intervals during orthodontic treatment should be adjusted to the individual's treatment response.

Methodological evaluation of gingival crevicular fluid volume and neutrophil elastase levels: sequential sampling, length of sampling time and two different sampling methods

Objectives: The mechanisms underlying the formation and composition of gingival crevicular fluid (GCF) and its flow into and from periodontal pockets are not understood very well. The aim of this study was to evaluate the length of sampling time and sequential sampling of GCF neutrophil elastase (NE) enzyme levels by using intracrevicular and orifice methods.Material and methods: Twenty adults (mean age of 41.8 years, ranged 31–60 years, 18 males and 2 females) with chronic periodontitis were enrolled and all completed the 3-d study. GCF was collected by both intracrevicular and intrasulcular methods, 720 samples of GCF were collected. In first, second and third day, the length of sampling time in seconds (s) and order were ‘5- 10-30-s’; ‘10- 30- 5-s’ and ‘30- 5- 10-s,’ respectively. GCF elastase levels were determined by hydrolysis of neutrophil specific substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-p-nitroanilide.Results: NE activity (µU) and NE activity/volume (µU/µl) were significantly different for order of sampling (p < .05), but not for the length of sampling time (p>.05).Conclusions: Within the limits of this study, the choice of sampling technique in GCF-profile studies seems to be a critical decision as it has the potential to affect the GCF volume and NE activity.

Infrared microspectroscopy characterization of gingival crevicular fluid during orthodontic treatment

The gingival crevicular fluid (GCF) is a site-specific exudate deriving from the epithelium lining of the gingival sulcus. GCF analysis provides a simple and noninvasive diagnostic procedure to follow-up the periodontal ligament and bone remodeling in response to diseases or mechanical stimuli as the orthodontic tooth movement. The aim of this study was to use infrared (IR) microspectroscopy to assess GCF composition changes at different phases of fixed orthodontic treatment. GCF samples were collected using sterile paper cones from young patients before starting the orthodontic treatment (control) and after 2, 7 and 14 days of fixed orthodontic treatment. Infrared spectra were obtained on GCF extracted from cones. In these spectra, the contributions of main functional groups and the changes related to the different phases of orthodontic treatment were present. Different normalizations to significant peaks were performed to better evidence the changes occurring at different times of the orthodontic treatment. Useful information was also obtained by evaluating the ratio between the areas of selected bands related to protein, lipid and carbohydrate contents. The results here reported show that IR microspectroscopy analysis can give an important contribution for monitoring GCF changes during the early phases of the orthodontic tooth movement.

Monitoring early phases of orthodontic treatment by means of Raman spectroscopies.

Gingival crevicular fluid (GCF) is a site-specific exudate in the gingival sulcus. GCF composition changes in response to diseases or mechanical stimuli, such as those occurring during orthodontic treatments. Raman microspectroscopy (μ-RS) and surface-enhanced Raman spectroscopy (SERS) were adopted for a GCF analysis during different initial phases of orthodontic force application. GCF samples were pooled from informed patients using paper cones. SERS spectra were obtained from GCF extracted from these cones, whereas μ-RS spectra were directly acquired on paper cones without any manipulation. The spectral characteristics of the main functional groups and the changes in cytochrome, amide III, and amide I contributions were highlighted in the different phases of orthodontic treatment with both SERS and μ-RS analysis. μ-RS directly performed on the paper cones together with proper statistical methods can offer an effective approach for the development of a tool for monitoring the processes occurring during orthodontic treatments, which may help the clinician in the choice of type of treatment individually for each patient and accelerate and improve the orthodontic therapy.

Diagnostic accuracy for apical and chronic periodontitis biomarkers in gingival crevicular fluid: an exploratory study.

The aim of this study was to assess the levels and diagnostic accuracy of a set of potential biomarkers of periodontal tissue metabolism in gingival crevicular fluid (GCF) from patients with chronic periodontitis (CP) and asymptomatic apical periodontitis ( AAP). Thirty one GCF samples from 11 CP patients, 44 GCF samples from 38 AAP patients and 31 GCF samples from 13 healthy volunteers were obtained (N = 106). Matrix metalloproteinases (MMPs) -2 and -9 were determined by zymography; levels of MMP-8 by ELISA and IFMA and MPO by ELISA. IL-1, IL-6, TNFα, DKK-1, Osteonectin, Periostin, TRAP-5 and OPG were determined by a multiplex quantitative panel. Statistical analysis was performed using linear mixed-effects models. The MMP-9 and MMP-8 were higher in CP, followed by AAP, versus healthy individuals (p < 0.05). ProMMP-2, MPO, IL-1, IL-6, PTN, TRAP-5 and OPG were significantly higher in CP when compared with AAP and healthy patients (p < 0.05). The highest diagnostic accuracies were observed for ProMMP-2, ProMMP-9, MMP-8 and TRAP-5 (AUC > 0.97) in CP, and for the active form of MMP-9 and MMP-8 (AUC > 0.90) in AAP. Gingival crevicular fluid composition is modified by CP and AAP. MMP-9 and MMP-8 show diagnostic potential for CP and AAP, whereas MMP-2 and TRAP-5 are useful only for CP.

Correlation between magnesium and alkaline phosphatase from gingival crevicular fluid periodontal diseases

Background: Magnesium is one of the alkaline phosphatase (ALP) cofactor. The amount of magnesium contained infoods affect ALP activity. Increased ALP activity will indicate the level of inflammation in periodontal disease. Elevated inflammation in periodontal disease will change gingivitis to periodontitis, where there has been damage to the bone ssupporting the teeth, and an increasing number of gingival crevicular fluid (GCF). The content of GCF consists of enzymatic and non-enzymatic. Changes in the composition of GCF occurs when the inflammation gets worse. Purpose: This study was aimed to prove the correlation between magnesium and ALP from GCF on periodontal disease. Method: This research involved 60 Minangkabaunese people with 20 healthy samples, 20 mild gingivitis samples, and 20 mild periodontitis samples. GCF was collected by absorbing method. Then ALP level in GCF was measured by using ELISA technique. Magnesium level in Minangkabaunese food was tested by Food Frequency Questionnaire (FFQ).Univariate analysis was performed to describe each variable. To see a normal distribution, Kolmogorov Smirnof Test was used (p>0.05). Unpaired T-test and Pearson correlation test was used to see correlation between ALP and magnesium level in Minangkabaunese food. Result: There is a significant correlation between the levels of ALP and magnesium level in Minangkabaunese food with periodontal disease (p=0.005). ALP is higest on mild periodontitis (137.74±23.01 ng/dl). Magnesium level normal control group is highest (250.14±32.34 mg) and in mild periodontitis is the lowest (110.83±21.04 mg). Corelation between ALP and magnesium level indicates strong correlation with negative direction (r=- 0.907). Conclusion: There is correlation between the levels of alkaline phosphatase and magnesium level on periodontal disease. Increasing inflamation rate will elevate the ALP level.

Biomarkers in orthodontic tooth movement.

Tooth movement by orthodontic treatment is characterized by remodeling changes in the periodontal ligament, alveolar bone, and gingiva. A reflection of these phenomenons can be found in the gingival crevicular fluid (GCF) of moving teeth, with significant elevations in the concentrations of its components like, cytokines, neurotransmitters, growth Factors, and a arachidonic acid metabolites. GCF arises at the gingival margin and can be described as a transudate or an exudate. Several studies have focused on the composition of GCF and the changes that occur during orthodontic tooth movement (OTM). GCF component analysis is a non-invasive method for studying the cellular response of the underlying periodontium. Clinically, GCF can be easily collected using platinum loops, filter paper strips, gingival washings, and micropipettes. A number of GCF biomarkers involve in bone remodeling during OTM. The data suggest that knowledge of all the biomarkers present in the GCF that can be used to mark the changes in tooth that is undergoing orthodontic treatment may be of clinical usefulness leading to proper choice of mechanical stress to improve and to shorten treatment time and avoid side effects.

Gingival crevicular fluid proteomes in health, gingivitis and chronic periodontitis.

The aim of this study was to compare the proteome composition of gingival crevicular fluid obtained from healthy periodontium, gingivitis and chronic periodontitis affected sites. Owing to its site-specific nature, gingival crevicular fluid is ideal for studying biological processes that occur during periodontal health and disease progression. However, few studies have been conducted into the gingival crevicular fluid proteome due to the small volumes obtained. Fifteen males were chosen for each of three different groups, healthy periodontium, gingivitis and chronic periodontitis. They were categorized based on clinical measurements including probing depth, bleeding on probing, plaque index, radiographic bone level, modified gingival index and smoking status. Gingival crevicular fluid was collected from each patient, pooled into healthy, gingivitis and chronic periodontitis groups and their proteome analyzed by gel electrophoresis and liquid chromatography electrospray ionization ion trap tandem mass spectrometry. One hundred and twenty-one proteins in total were identified, and two-thirds of these were identified in all three conditions. Forty-two proteins were considered to have changed in abundance. Of note, cystatin B and cystatin S decreased in abundance from health to gingivitis and further in chronic periodontitis. Complement proteins demonstrated an increase from health to gingivitis followed by a decrease in chronic periodontitis. Immunoglobulins, keratin proteins, fibronectin, lactotransferrin precursor, 14-3-3 protein zeta/delta, neutrophil defensin 3 and alpha-actinin exhibited fluctuations in levels. The gingival crevicular fluid proteome in each clinical condition was different and its analysis may assist us in understanding periodontal pathogenesis.

Mass spectrometry analysis of gingival crevicular fluid in the presence of external root resorption

In this study, we used liquid chromatography-mass spectrometry (LC-MS) to investigate the differences in the composition of gingival crevicular fluid between resorbing deciduous molars and nonresorbing permanent teeth. The main goal was to identify novel biomarkers associated with root resorption. Eleven children (4 boys, 7 girls) in the mixed dentition were selected to participate in this split-mouth design study, in which a deciduous secondmolar with radiographic evidence of root resorption served as the experimental site, and the permanent first molar on the contralateral quadrant was the control site. Gingival crevicular fluid was collected using absorbing strips. A total of 22 samples (11 root resorption, 11 control) were each analyzed with 1-dimensional LC-MS. The remaining samples were then pooled across the 11 patients and analyzed by 2-dimensional LC-MS. The output files were converted to mascot generic format, which can be used to perform protein identification with conventional search engines. The 2-dimensional LC-MS protocol was able to identify 2789 and 2421 proteins in the control and resorption pooled samples, respectively. In this population, we detected significantly upregulated and downregulated proteins in the teeth with root resorption. Interestingly, many of these proteins are characteristically found in exosomes. We identified novel proteins that might prove to be useful biomarkers of root resorption, individually or as part of a panel.

Differences in the gingival crevicular fluid composition between adults and adolescents undergoing orthodontic treatment

To investigate differences in the gingival crevicular fluid (GCF) composition between adolescent and adult patients undergoing orthodontic treatment with fixed appliances. Ten adolescents (14.4 ± 1.43) and 10 adults (28.5 ± 7.83) with Class I malocclusions and minor upper incisor crowding were allocated to two different age groups. Brackets were bonded only in the upper arch over the 20-week period of the experiment. Samples of GCF were collected from the labial sides of the upper incisors (experimental sites) and lower incisors (control sites) of each subject at five time points. Aliquots from diluted GCF were screened for the presence of receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), interleukin-1 (IL-1), interleukin-1 receptor antagonist (IL-1RA), and metalloproteinase-9 (MMP-9) using a microarray technique. The values were statistically analyzed. In adults, the ratio of IL-1 to IL-1RA decreased significantly (P = .033) in experimental sites 3 weeks after appliance placement and first archwire activation. In adolescents, the ratio of RANKL to OPG peaked 6 weeks after the insertion of the first rectangular archwire. This ratio peak found in adolescents was a consequence of a decrease in the mean concentration of OPG. No significant changes over time were observed in the concentration of MMP-9. This study demonstrates age trends in the GCF levels of IL-1, IL-1RA, RANKL, and OPG that may be used to track differences in tissue response between adults and adolescents undergoing orthodontic treatment.

Proteomic analysis of gingival crevicular fluid for discovery of novel periodontal disease markers

The protein composition of gingival crevicular fluid (GCF) may reflect the pathophysiology of periodontal diseases. A standard GCF proteomic pattern of healthy individuals would serve as a reference to identify biomarkers of periodontal diseases by proteome analyses. However, protein profiles of GCF obtained from apparently healthy individuals have not been well explored. As a step toward detection of proteomic biomarkers for periodontal diseases, we applied both gel-based and gel-free methods to analyze GCF obtained from healthy subjects as compared with supragingival saliva. To ensure optimized protein extraction from GCF, a novel protocol was developed. The proteins in GCF were extracted with high yield by urea buffer combined with ultrafiltration and the intensity of spots with supragingival saliva and GCF was compared using agarose two-dimensional electrophoresis. Eight protein spots were found to be significantly more intense in GCF. They included superoxide dismutase 1 (SOD1), apolipoprotein A-I (ApoA-I), and dermcidin (DCD). Moreover, GCF proteins from healthy subjects were broken down into small peptide fragments and then analyzed directly by LC-MS/MS analysis. A total of 327 proteins including ApoA-I, SOD1, and DCD were identified in GCF. These results may serve as reference for future proteomic studies searching for GCF biomarkers of periodontal diseases.

Proteome data set of human gingival crevicular fluid from healthy periodontium sites by multidimensional protein separation and mass spectrometry

Gingival crevicular fluid has been of major interest for many decades as a valuable body fluid that may serve as a source of biomarkers for both periodontal and systemic diseases. Owing to its very small sample size, submicroliter volumes, identification of its protein composition by classical biochemical methods has been limited. The advent of highly sensitive mass spectrometric technology has permitted large-scale identification of protein components of many biological samples. This technology has been employed to identify the protein composition of gingival crevicular fluid from inflamed and periodontal sites. In this report, we present a proteome data set of gingival crevicular fluid from healthy periodontium sites. A combination of a periopaper collection method with application of multidimensional protein separation and mass spectrometric technology led to a large-scale documentation of the proteome of gingival crevicular fluid from healthy periodontium sites. The approaches used have culminated in identification of 199 proteins in gingival crevicular fluid of periodontally healthy sites. The present gingival crevicular fluid proteome from healthy sites was compared and contrasted with those proteomes of gingival crevicular fluid from inflamed and periodontal sites, as well as serum. The cross-correlation of the gingival crevicular fluid and plasma proteomes permitted dissociation of the 199 identified gingival crevicular fluid proteins into 105 proteins (57%) that can be identified in plasma and 94 proteins (43%) that are distinct and unique to the gingival crevicular fluid microenvironment. Such analysis also revealed distinctions in protein functional categories between serum proteins and those specific to the gingival crevicular fluid microenvironment. Firstly, the data presented herein provide the proteome of gingival crevicular fluid from periodontally healthy sites through establishment of innovative analytical approaches for effective analysis of gingival crevicular fluid from periopapers both at the level of complete elusion and with removal of abundant albumin, which restricts identification of low-abundant proteins. Secondly, it adds significantly to the knowledge of gingival crevicular fluid composition and highlights new groups of proteins specific to the gingival crevicular fluid microenvironment.

Impact of orthodontic retainers on periodontal health status assessed by biomarkers in gingival crevicular fluid

To evaluate whether biomarkers of inflammation and periodontal remodeling are differentially expressed in the gingival crevicular fluid (GCF) of patients wearing different types of orthodontic retainers. Thirty-one adult subjects (17 men and 14 women with an age range of 20 to 35 years) were allocated to three different groups. Group 1 consisted of 10 patients wearing fixed retainers, group 2 included 11 patients using lower removable retainers, and group 3 comprised 10 patients without retainers (control). Periodontal health assessment and GCF collection were carried out at two sites per subject: the lingual side of a central lower incisor and the lingual side of a lower second premolar. Aliquots from diluted GCF were screened for the presence of biomarkers using a microarray technique. Group 1 patients exhibited a higher percentage of sites with visible plaque in the incisor region than the other groups (P = .03); no differences were noted in gingival bleeding and probing depths. The median concentrations (pg/mL) of interferon-gamma and interleukin-10 were significantly higher in the premolar sites of patients in group 2 (P = .01 and P = .04, respectively), whereas the concentration of matrix metalloproteinase-9 was significantly higher at the incisors of patients wearing fixed retainers (P = .02). A significant difference between the two sites was seen only in group 2. The presence of different orthodontic retainers may promote specific alterations in the GCF composition. With retention periods potentially becoming longer, this finding may be of clinical significance.