Analysis Of Gingival Crevicular Fluid Research Articles

Association between calcium-channel blockers and gingival enlargement: A case-control study

ObjectivesAs reported by the existing literature, calcium-channel blockers (CCB) can lead to gingival enlargement. The aims of this study were to investigate the factors associated with gingival enlargement in patients on CCB and to assess the saliva and gingival crevicular fluid (GCF) profile of patients on CCB with gingival enlargement. MethodsA total of 131 participants were included. Data were collected from 91 patients taking CCB for treatment of systemic hypertension. The presence of drug-induced gingival enlargement (DIGE) was assessed clinically and associated with patient factors. Patients with DIGE were group-matched for gender and ethnicity with an equal number of consecutive CCB non-DIGE patients (control 1), no-CCB no-DIGE (control 2) and periodontally healthy with no DIGE (control 3) for the saliva and GCF analysis. A bead-based multiplex immunoassay was used to assess a panel of biomarkers. ResultsTwenty-two percent of patients on CCB were diagnosed with DIGE. Lack of daily interdental cleaning and self-reported diagnosis of type II diabetes were associated with the diagnosis of DIGE. When analysing patients only on CCB, those with DIGE had higher GCF levels of vascular endolthelial growth factor (VEGF) (p = 0.032), epidermal growth factor (EGF) (p = 0.030) and matrix metalloproteinase-8 (MMP-8) (p = 0.008). Among the salivary markers, only MMP-8 showed a statistically significant difference across groups (p < 0.001). ConclusionsThis is the first study investigating saliva and GCF biomarkers in patients with DIGE and different control groups, suggesting that causes of the overgrowth might involve inflammatory processes, tissue damage pathways, and potentially an impact on growth factors like VEGF. Future research should verify these results in independent populations and explore the underlying pathogenic mechanisms in-depth. Clinical significanceCalcium-channel blockers (CCB) can lead to gingival enlargement. This study confirms lack of interdental cleaning and type II diabetes as risk factors. Elevated levels of VEGF, EGF, and MMP-8 in gingival crevicular fluid and MMP-8 in saliva suggest inflammatory processes and growth factors might play roles in this condition.

A Comparative Analysis of Alkaline Phosphatase Levels in Gingival Crevicular Fluid of Patients Undergoing Growth Modulation Therapy With Twin Block, Forsus Fatigue Resistant, and Clear Block Appliances Compared to Normal Individuals: An In Vivo Study.

Background In the contemporary era, where science and technology know no boundaries, this in vivo study explores the impact of growth modulation therapy using Twin Block, Forsus Fatigue Resistant, and Clear Block appliances on alkaline phosphatase (ALP) levels in gingival crevicular fluid (GCF). Bone physiology involves modeling and remodeling, with orthodontics applying forces to teeth, influencing tissue reactivity and bone modeling. ALP, a marker of osteoblast function, plays a crucial role in bone growth. GCF reflects immunological and inflammatory responses during orthodontic force application, making it a valuable medium for studying ongoing metabolic processes related to bone turnover. Aim The study aims to comparatively analyze ALP levels in GCF during growth modulation therapy, assessing the efficacy of Twin Block, Forsus Fatigue Resistant, and Clear Block appliances. The research involves 30 experimental samples divided into three study groups and a control group. The samples are collected at various time intervals, and ALP levels are analyzed using a spectrophotometer. Statistical analysis includes paired and unpaired t-tests, one-way analysis of variance (ANOVA), and multiple comparisons. Results Results demonstrate a significant increase in ALP levels during the growth modulation therapy, indicating a positive correlation with bone remodeling. Twin Block appears to be the most effective appliance, exhibiting higher ALP activity compared to Clear Block and Forsus groups. Conclusion In conclusion, this study provides valuable insights into the biochemical responses during growth modulation therapy, emphasizing the potential of GCF analysis in understanding orthodontic treatment effects.

A simplified protocol for deep quantitative proteomic analysis of gingival crevicular fluid for skeletal maturity indicators

Assessment of craniofacial skeletal maturity is of great importance in orthodontic diagnosis and treatment planning. Traditional radiographic methods suffer from clinician subjectivity and low reproducibility. Recent biochemical methods, such as the use of gingival crevicular fluid (GCF) protein biomarkers involved in bone metabolism, have provided new opportunities to assess skeletal maturity. However, mass spectrometry (MS)-based GCF proteomic analysis still faces significant challenges, including the interference of high abundance proteins, laborious sample prefractionation and relatively limited coverage of GCF proteome. To improve GCF sample processing and further discover novel biomarkers, we herein developed a single-pot, solid-phase-enhanced sample-preparation (SP3)-based high-field asymmetric waveform ion mobility spectrometry (FAIMS)-MS protocol for deep quantitative analysis of the GCF proteome for skeletal maturity indicators. SP3 combined with FAIMS could minimize sample loss and eliminate tedious and time-consuming offline fractionation, thereby simplifying GCF sample preparation and improving analytical coverage and reproducibility of the GCF proteome. A total of 5407 proteins were identified in GCF samples from prepubertal and circumpubertal groups, representing the largest dataset of human GCF proteome to date. Compared to the prepubertal group, 61 proteins were differentially expressed (31 up-regulated, 30 down-regulated) in the circumpubertal group. The six-protein marker panel, including ATP5D, CLTA, CLTB, DNM2, HSPA8 and NCK1, showed great potential to predict the circumpubertal stage (ROC-AUC 0.937), which provided new insights into skeletal maturity assessment.

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.&#x0D; 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.&#x0D; 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.&#x0D; 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.

Evaluation of the clinical parameters and volumetric analysis of gingival crevicular fluid for chlohexidine vying silver nanoparticle mouthwash the avant garde for chronic periodontitis

BACKGROUND: Oral cavity ecosystem represents a dynamic pattern. Scaling and root planing (SRP) is the gold standard approach for treatment of chronic periodontitis but used alone it may not be effective in removing periodontal pathogens from sites where access is poor. Although chlorhexidine is the most effective mouthwash against oral microorganisms, the side effects of long- term use suggest the need for an alternative. Hence, this study has been undertaken to know effectiveness of silver nanoparticle mouthwash with chlorhexidine mouthwash which is effective on common microorganisms of oral cavity. OBJECTIVE: To evaluate and compare the clinical and biochemical outcome of chlorhexidine (CHX) and silver nanoparticle as an adjunct to scaling and root planing (SRP) in patients with chronic periodontitis. METHODS: Ninety- two patients with generalized moderate to severe chronic periodontitis from the Dept. of Periodontology who fulfilled the inclusion and exclusion criteria were recruited for the study. In the experimental clinical trial, the clinical parameters likewise; Plaque index (PI), Gingival index (GI), probing depth (PD), clinical attachment loss (CAL) were assessed. The volumetric analysis of gingival crevicular fluid (GCF) levels were assessed at baseline and follow up visits. The IBM SPSS Statistics for Windows software, v. 20.0 (IBM Corp., Armonk, USA), was used. The data was numerically coded and entered into the program. Both the descriptive statistics and the inferential statistics were analyzed to compare the responses between the groups. RESULTS: The statistical analysis was carried out by descriptive as well as inferential statistics and both groups demonstrated significant intragroup reduction in PI, GI, PPD, CAL, GCF from baseline to 3 months follow-up. There were statistically significant intragroup differences between both the groups for all of the parameters. CONCLUSION: Silver nanoparticle mouthwash can be an alternative adjunct for non-surgical periodontal therapy in patients with chronic periodontitis. Results of this study indicated that both the mouth rinse demonstrated an equal range of effectiveness. These conclusions were supported by the clinical parameters and biochemical outcomes.

μ-FTIR, μ-Raman, and SERS Analysis of Amide I Spectral Region in Oral Biofluid Samples during Orthodontic Treatment.

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 periodontal and bone remodeling in response to diseases or mechanical stimuli such as orthodontic tooth movement (OTM). In recent years, the use of vibrational spectroscopies such as Fourier Transform Infrared and Raman microspectroscopy and Surface-Enhanced Raman spectroscopy contributed to characterizing changes in GCF during fixed orthodontic treatment. Amide I band plays a relevant role in the analysis of these changes. The aim of this study was to investigate the spectroscopy response of Amide I depending on the OTM process duration. A model based on Gaussian–Lorentzian curves was used to analyze the infrared spectra, while only Lorentzian functions were used for Raman and SERS spectra. Changes induced by the OTM process in subcomponents of the Amide I band were determined and ascribed to secondary structure modification occurring in proteins. The vibrational spectroscopies allow us to efficiently monitor the effects of the orthodontic force application, thus gaining increasing attention as tools for individual patient personalization in clinical practice.

Oral lichen planus identification by mid-infrared spectroscopy of oral biofluids: A case-control study

Background and AimsThis study aims to access the effectiveness of mid-infrared (MIR) spectroscopy on the identification of the reticular form of OLP, following the assessment of gingival crevicular fluid (GCF) and oral mucosa transudate (OMT). Material and MethodsThe trial follows a case-control design. Samples were characterized through MIR spectroscopy and chemometric tools were applied to distinguish between case and control participants, further identifying the spectral regions with the highest contribution to the developed models. ResultsMIR spectroscopy was capable to discriminate between OLP patients and controls with 95.1% and 85.4% of correct predictions, regarding GCF and OMT samples, respectively. Additionally, the spectral regions mostly contributing to the successful prediction were identified, and possibly related with the distinctive presence of amino acids/proteins and oxidative stress mediators in oral biofluids, supporting the role of the immune-inflammatory activation on OLP etiology and disease course. ConclusionMIR spectroscopy analysis of GCF and OMT may be regarded as an innovative, non-invasive, low cost and sensitive technique, contributing to the identification of the reticular from of OLP.

Analysis of gingival crevicular fluid biomarkers in patients with metabolic syndrome

ObjectivesTo assess associations between gingival crevicular fluid (GCF) markers in patients with metabolic syndrome, with or without concomitant periodontitis. MethodsA total of 95 patients with Metabolic Syndrome (MetS) had a periodontal examination and gingival crevicular fluid samples taken. Proteomic analysis of gingival crevicular fluid (GCF) was carried out by Human XL Cytokine protein arrays in 12 selected patients, followed by multiplex ELISA of 11 analytes in 95 participants. ResultsIncreased levels of Aggrecan, IL-6 and IL-8 were found in patients with periodontal health compared with moderate and severe periodontitis. The inverse stepwise association between severity of periodontitis and reduced Aggrecan levels was also observed at adjusted linear regression analysis. Diagnosis of diabetes was associated with higher GCF levels of IL-8 and MMP-8. ConclusionDiabetes may affect GCF levels of cytokines, irrespective of periodontal status. Periodontal status may be associated with Aggrecan levels in the GCF of patients affected by metabolic syndrome. Clinical significanceInvestigation of GCF biomarkers may potentially help have diagnostic potential in patients with MetS.

Liquid chromatography method with tandem mass spectrometry and fluorescence detection for determination of inflammatory biomarkers in gingival crevicular fluid as a tool for diagnosis of periodontal disease

The new ultra-high performance liquid chromatography method with tandem mass spectrometry and fluorescence detection allowing fast, selective, and high-throughput analysis of neopterin, kynurenine, tryptophan, and creatinine in gingival crevicular fluid (GCF) has been optimized. Defining the pathophysiology of periodontal disease and identification of potential diagnostic test for active periodontitis remains a significant challenge in the field of oral disease diagnosis. Analysis of GCF provides a non-invasive means of evaluating the role of the host response in periodontal disease. In addition, the analysis of GCF provides an information about current inflammation level of sampled site/tooth. Determination of GCF inflammatory biomarkers such as neopterin, kynurenine, and tryptophan can contribute to diagnosis, evaluation of treatment, and progression of periodontal diseases such as gingivitis and periodontitis. The separation of target analytes was carried out using a column Kinetex™ Polar C18 100 Å, (100 × 3.0 mm) packed with 2.6 µm core-shell particles applying an elution with a gradient formed from 0.2% aqueous formic acid and 90% aqueous acetonitrile. Kynurenine, tryptophan, and creatinine were detected using mass spectrometry with electrospray ionization to improve the sensitivity while neopterin was detected using fluorescence detection. The separation of these four substances was achieved after using a very simple sample preparation technique convenient for small amount of biological sample. Only less than 20 µL sample was needed and the separation was completed in 4 min. MS/MS analysis was performed using multiple reaction monitoring (MRM) under a positive ionization mode. Deuterium labeled internal standard was used for the more precise quantification. The lower limits of quantification (LLOQ) for target analytes were 0.50 × 10–3 µmol/L for neopterin, 0.10 µmol/L for kynurenine, and 0.20 µmol/L for tryptophan and creatinine. The within-run and between-run accuracy were in a range of 96.67–114.77% for all quality controls and LLOQ of all analytes. Matrix effect, extraction recovery, and stability testing have all been investigated. The method was tested with real-life samples using GCF collected from patients suffering from periodontitis and from healthy controls. Neopterin levels in patients were significantly higher (P = 0.020) than in healthy subjects and indicate good potential of this method for using in evaluation of periodontal pathogenesis and healing outcomes following a treatment.

Identification of IL-18 and Soluble Cell Adhesion Molecules in the Gingival Crevicular Fluid as Novel Biomarkers of Psoriasis.

Psoriasis is a chronic immunoinflammatory skin disease. Although its diagnosis is clinical, differences in the appearance and severity of lesions pose a challenge for clinicians worldwide. The use of accessible biomarkers for psoriasis could aid in the early diagnosis and treatment of the disease. To date, evidence on the analysis of gingival crevicular fluid (GCF) molecules as novel, accessible, and reliable biomarkers for psoriasis is limited. This cross-sectional study compared the GCF levels of IL-18, soluble (s)ICAM-1, and sE-selectin in psoriatic patients (n = 42) and healthy controls (n = 39). Individuals with psoriasis not undergoing treatment and healthy individuals were included independent of periodontal status. GCF samples were collected, and a multiplex bead immunoassay was performed to quantify the levels of the target molecules. Psoriatic patients presented higher concentrations of IL-18 and lower concentrations of sE-selectin compared to controls (p < 0.05). No differences were found in the levels of sICAM-1 between the two groups (p > 0.05). Psoriasis was associated with IL-18 and E-selectin levels regardless of periodontal status, age, and smoking habit (p < 0.05). The areas under the receiver operating characteristic curve (ROC) for IL-18 and sE-selectin were 0.77 and 0.68, respectively. In conclusion, IL-18 and sE-selectin levels in the GCF could be promising biomarker for psoriasis.

Application of Vibrational Spectroscopies in the Qualitative Analysis of Gingival Crevicular Fluid and Periodontal Ligament during Orthodontic Tooth Movement

Optical vibrational techniques show a high potentiality in many biomedical fields for their characteristics of high sensitivity in revealing detailed information on composition, structure, and molecular interaction with reduced analysis time. In the last years, we have used these techniques for investigating gingival crevicular fluid (GCF) and periodontal ligament (PDL) during orthodontic tooth treatment. The analysis with Raman and infrared signals of GCF and PDL samples highlighted that different days of orthodontic force application causes modifications in the molecular secondary structure at specific wavenumbers related to the Amide I, Amide III, CH deformation, and CH3/CH2. In the present review, we report the most relevant results and a brief description of the experimental techniques and data analysis procedure in order to evidence that the vibrational spectroscopies could be a potential useful tool for an immediate monitoring of the individual patient’s response to the orthodontic tooth movement, aiming to more personalized treatment reducing any side effects.

Label-free analysis of gingival crevicular fluid (GCF) by surface enhanced Raman scattering (SERS).

Gingival crevicular fluid (GCF) is an interesting biofluid reflecting the physiological and pathological states of a single dental element. Due to this unique feature, in recent years, metabolomic analysis of GCF has gained attention as a biometric tool for the diagnosis and therapy of periodontal disease. Traditional methods are, however, too slow, cumbersome and expensive for a health-care routine. Surface enhanced Raman scattering (SERS) can offer rapid and label-free detailed molecular fingerprints that can be used for biofluid analysis. Here we report the first SERS characterization of GCF using an easy and quick sample preparation. The dominant features in the SERS spectrum of GCF are ascribed to very few metabolites, in particular to uric acid, hypoxanthine, glutathione and ergothioneine. Additionally, we succeeded in differentiating between the SERS signal of GCF collected from healthy volunteers and the one collected from patients with periodontal disease.

Estimation of Calcium and Iron Levels in Gingival Crevicular Fluid and Serum in Periodontal Health and Disease

Introduction: Gingival Crevicular Fluid (GCF) has been referred to as a promising medium for detection of markers for periodontal disease activity. Analysis of GCF shows minute changes in biomarker levels well before the onset of clinical signs and symptoms; which helps to even predict a person’s predisposition towards periodontal disease occurrence. The elemental analysis of human blood serum is noteworthy in routine clinical practice as well as in medical research. Aim: This study was done to determine the changes in calcium and iron levels in GCF and serum in human subjects with normal periodontal health and those with disease. Materials and Methods: This was a cross-sectional study conducted from March 2019 to December 2019. Eight study subjects (4 healthy subjects and 4 periodontitis cases) were selected from the patients reporting to the Department of Periodontics at Tagore Dental College and Hospital, Chennai. The subjects were chosen based on inclusion and exclusion criteria and all patients were subjected to a clinical examination wherein the Probing Depth (PD) and Clinical Attachment Level (CAL) were recorded by a single examiner using William’s Periodontal probe. The GCF samples were collected by Capillary Tubing method. Blood was collected by venipuncture and centrifuged to provide serum samples. Dual viewing Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) was used to estimate Calcium and Iron in GCF and serum. SPSS version 21.0 was used for statistical analysis. Mann Whitney U test was used for comparing the groups. p-value less than 0.05 was considered statistically significant. Results: Serum iron levels were significantly less in periodontitis patients than healthy subjects (p-value 0.043). GCF iron level (p-value 0.386), GCF calcium level (p-value 0.149) and serum calcium level (p-value 0.564) did not show any major variation among subjects with normal periodontal health and those with disease. Conclusion: The findings of this study showed that iron and calcium are present in GCF and serum samples of healthy persons and patients with chronic periodontitis which can be detected using ICP-OES. A significant difference in serum iron levels between health and disease could indicate a patient’s predisposition towards developing periodontitis. Calcium levels in GCF and serum do not point towards periodontal disease activity.

Identification of galectin‑10 as a biomarker for periodontitis based on proteomic analysis of gingival crevicular fluid.

Periodontitis is a chronic inflammatory disease caused by the gradual breakdown of tissues surrounding the teeth due to various factors. The disease has been frequently noted in dental outpatients for a number of years. Improvements are required to current diagnostic methods, which have limitations in assessing the condition and progression of periodontitis. The development of diagnostic biomarkers for periodontitis to increase the sensitivity and accuracy of diagnosis is important for the management of periodontitis. In the present study, whole gingival crevicular fluid(GCF) from patients with periodontitis and healthy individuals was characterized via liquid chromatography with tandem mass spectrometry. Label‑free quantification was used to identify the differentially abundant protein biomarkers. A total of 1,295proteins were identified from the whole GCF of patients with periodontitis and healthy individuals via proteomic analysis. When analyzing biological processes, 'metabolic process' and 'cell organization and biogenes' were identified to play important roles in GCF under periodontitis conditions according to GeneOntology. When analyzing molecular functions, 'catalytic activity' and 'protein binding' were the terms most enriched with differentially abundant proteins under periodontitis conditions. Galectin‑10(Gal‑10) was one of the most upregulated proteins in the GCF of patients with periodontitis. The levels of prostaglandin E2 were increased in oral keratinocytes and gingival fibroblasts treated with recombinant (r)Gal‑10. The levels of interleukin‑8, matrix metalloproteinase 9 and C‑reactive protein were increased in the conditioned media (CM) of rGal‑10‑treated gingival fibroblasts. In addition, the CM of rGal‑10‑treated gingival fibroblasts induced osteoclast differentiation. These results suggested that Gal‑10 expression was increased in the GCF of patients with periodontitis and contributed to the process of osteoclastogenesis. Therefore, Gal‑10 may be a candidate biomarker for periodontitis.

Label-free quantitative proteomic analysis of gingival crevicular fluid to identify potential early markers for root resorption

BackgroundOrthodontically-induced root resorption is an iatrogenic effect and it cannot be examined regularly due to the harmful effects of sequential doses of radiation with more frequent radiography. This study aims to compare protein abundance (PA) of pre-treatment and during orthodontic treatment for root resorption and to determine potential early markers for root resorption.MethodsTen subjects (n = 10) who had upper and lower fixed appliances (MBT, 3 M Unitek, 0.022″ × 0.028″) were recruited for this study. Human gingival crevicular fluid (GCF) was obtained using periopaper strips at pre-treatment (T0), 1 month (T1), 3 months (T3), and 6 months (T6) of orthodontic treatment. Periapical radiographs of the upper permanent central incisors were taken at T0 and T6 to measure the amount of root resorption. Identification of changes in PA was performed using liquid chromatography-tandem mass spectrometry. Student’s t-test was then performed to determine the significance of the differences in protein abundance before and after orthodontic treatment.ResultsOur findings showed that all ten subjects had mild root resorption, with an average resorption length of 0.56 ± 0.30 mm. A total of 186 proteins were found to be commonly present at T0, T1, T3, and T6. There were significant changes in the abundance of 16 proteins (student’s t-test, p ≤ 0.05). The increased PA of S100A9, immunoglobulin J chain, heat shock protein 1A, immunoglobulin heavy variable 4–34 and vitronectin at T1 suggested a response to stress that involved inflammation during the early phase of orthodontic treatment. On the other hand, the increased PA of thymidine phosphorylase at T3 suggested growth promotion and, angiogenic and chemotactic activities.ConclusionsThe identified proteins can be potential early markers for root resorption based on the increase in their respective PA and predicted roles during the early phase of orthodontic treatment. Non-invasive detection of root resorption using protein markers as early as possible is extremely important as it can aid orthodontists in successful orthodontic treatment.

Top down proteomic analysis of gingival crevicular fluid in deciduous, exfoliating and permanent teeth in children

Gingival Crevicular Fluid (GCF), a plasma-derived exudate present in the gingival crevice was collected from deciduous, exfoliating and permanent teeth from 20 children (60 samples) with the aim to characterize and quantify by a mass spectrometry based top-down proteomic approach, the peptide/proteins in the fluid and verify possible variations occurring during the exfoliating process.The results obtained confirmed the presence in GCF of α-Defensins 1–4, Thymosin β4 and Thymosin β10, as described in previous works and revealed the presence of other interesting peptides never described before in GCF such as specific fragments of α-1-antitrypsin, α-1-antichymotrypsin; fragments of Thymosin β4 and Thymosin β10; Fibrinopeptide A and its fragments and Fibrinopeptide B; S100A8 and S100A9, LVV Hemorphin-7 (hemoglobin chain β fragment), as well as some other peptides deriving from α and β subunits of hemoglobin. Statistical analysis evidenced different levels in 5 proteins/peptides in the three groups. Our study demonstrate that an in-depth analysis of a biological fluid like GCF, present in small amount, can provide useful information for the understanding of different biological processes like teeth eruption. Data are available via ProteomeXchange with identifier PXD016010 and PXD016049. SignificanceGCF due to his site-specific nature has a great potential in containing factors that are specific for action at a given site and might have diagnostic value to detect qualitative and quantitative variations of proteins/peptides composition linked to physiological or pathological conditions.

Microbial and metabolomic analysis of gingival crevicular fluid in general chronic periodontitis patients: lessons for a predictive, preventive, and personalized medical approach

ObjectivesGeneral chronic periodontitis (GCP) is a bacterial inflammatory disease with complex pathology. Despite extensive studies published on the variation in the oral microbiota and metabolic profiles of GCP patients, information is lacking regarding the correlation between host-bacterial interactions and biochemical metabolism. This study aimed to analyze the oral microbiome, the oral metabolome, and the link between them and to identify potential molecules as useful biomarkers for predictive, preventive, and personalized medicine (PPPM) in GCP.MethodsIn this study, gingival crevicular fluid (GCF) samples were collected from patients with GCP (n = 30) and healthy controls (n = 28). The abundance of oral microbiota constituents was obtained by Illumina sequencing, and the relative level of metabolites was measured by gas chromatography-mass spectrometry. Full-mouth probing depth, clinical attachment loss, and bleeding on probing were recorded as indices of periodontal disease.ResultsThe relative abundances of 7 phyla and 82 genera differed significantly between the GCP and healthy groups. Seventeen differential metabolites involved in different metabolism pathways were selected based on variable influence on projection values (VIP > 1) and P values (P < 0.05). Through Spearman’s correlation analysis, microorganisms, metabolites in GCF, and clinical data together showed a clear trend, and clinical data regarding periodontitis can be reflected in the shift of the oral microbial community and the change in metabolites in GCF. A combination of citramalic acid and N-carbamylglutamate yielded satisfactory accuracy (AUC = 0.876) for the predictive diagnosis of GCP.ConclusionsDysbiosis in the polymicrobial community structure and changes in metabolism could be mechanisms underlying periodontitis. The differential microorganisms and metabolites in GCF between periodontitis patients and healthy individuals are possibly biomarkers, pointing to a potential strategy for the prediction, diagnosis, prognosis, and management of personalized periodontal therapy.

Associations of cytokine levels in gingival crevicular fluid of mobile teeth with clinical improvement after initial periodontal treatment.

Studies suggest that analysis of gingival crevicular fluid (GCF) is useful for evaluating periodontal status. In this study, clinical variables related to tooth mobility, and multiple cytokine levels in proximate GCF, were measured at four time points during initial periodontal treatment: before treatment (baseline), after supragingival scaling, after occlusal adjustment, and after scaling and root planing (SRP); 20 teeth from 13 patients with periodontitis were included. Baseline interleukin (IL)-10 level in GCF was significantly higher around teeth that showed substantial improvement in periodontal epithelial surface area (PESA) after SRP than around teeth without PESA improvement. IL-3 and IL-16 levels in GCF at baseline were significantly higher around teeth with a periodontal inflamed surface area (PISA) of 0 mm2 after SRP than around teeth without PISA improvement. In addition, baseline IL-7, IL-11, and IL-12p40 levels in GCF were significantly lower around teeth with decreased mobility after occlusal adjustment than around teeth without decreased mobility. These results suggest that pre-treatment cytokine levels in GCF are useful in predicting the effects of initial periodontal treatment.

Current Status of Proteomic Technologies for Discovering and Identifying Gingival Crevicular Fluid Biomarkers for Periodontal Disease.

Periodontal disease is caused by bacteria in dental biofilms. To eliminate the bacteria, immune system cells release substances that inflame and damage the gums, periodontal ligament, or alveolar bone, leading to swollen bleeding gums, which is a sign of gingivitis. Damage from periodontal disease can cause teeth to loosen also. Studies have demonstrated the proteomic approach to be a promising tool for the discovery and identification of biochemical markers of periodontal diseases. Recently, many studies have applied expression proteomics to identify proteins whose expression levels are altered by disease. As a fluid lying in close proximity to the periodontal tissue, the gingival crevicular fluid (GCF) is the principal target in the search for periodontal disease biomarkers because its protein composition may reflect the disease pathophysiology. Biochemical marker analysis of GCF is effective for objective diagnosis in the early and advanced stages of periodontal disease. Periodontal diseases are also promising targets for proteomics, and several groups, including ours, have applied proteomics in the search for GCF biomarkers of periodontal diseases. This search is of continuing interest in the field of experimental and clinical periodontal disease research. In this article, we summarize the current situation of proteomic technologies to discover and identify GCF biomarkers for periodontal diseases.

Optimal frequency of tooth brushing in mechanically ventilated adults: Study protocol for a randomized controlled trial.

The optimal frequency of tooth brushing in the critically ill has not been experimentally determined. For mechanically ventilated patients, optimal frequency of tooth brushing is an important unresolved issue as there is little evidence to judge the benefits or associated risks of tooth brushing. Given this knowledge gap, the project's overall goal is to determine the optimal frequency of tooth brushing (once, twice, or three times daily) for mechanically ventilated adults. A prospective, randomized, experimental design is used. Specifically, 345 subjects, with 207 expected to complete the study, are randomly assigned within 36 hr of intubation to one of three groups (once, twice, or three times tooth brushing daily). Tooth brushing with a soft compact-head toothbrush takes approximately 2 min. Dental plaque assessment, gingival crevicular fluid (GCF) determination for IL-1β, and data regarding healthcare-acquired infection (HAI) and clinical status are collected at baseline and daily in-hospital. The clinical impact of daily frequency of tooth brushing in relation to extent of dental plaque and inflammation reflected by GCF analysis will be compared by the three treatment arms. In addition, evaluation of safety for HAIs will be compared based on calculation of the number needed to harm. The trial will empirically determine the optimal frequency of tooth brushing in mechanically ventilated adults, balancing benefits and risks. This contribution is significant because it will have immediate impact on bedside nursing practice, and is a final component necessary for specific evidence-based guidelines for the common nursing intervention of oral care in mechanically ventilated adults.