Project description:We profiled miRNAs in gingival crevicular fluid (GCF) by a PCR-based method that yielded quantitative measures of more than 600 miRNAs. We found that miRNA profiles in GCF of periodontitis patients are distinct from those of healthy controls.
Project description:The human gingival crevicular fluid proteome and metaproteome of periodontitis are investigated, to shed light on the factors that mediate the host-microbiota interactions in the pathogenesis of periodontitis.
Project description:Inflammatory periodontal disease (periodontitis) is widespread in dogs. This study aimed to evaluate site-specific changes in the canine gingival crevicular fluid (GCF) proteome during the longitudinal progression from very mild gingivitis to mild periodontitis. Periodontitis diagnosis in dogs requires anaesthesia, our ultimate aim was to develop a periodontitis diagnostic that could be applied to samples taken from conscious dogs. The objective of this work was to identify potential biomarkers of periodontal disease progression in the GCF of dogs.
Project description:Complement inhibition has been successfully used to treat periodontitis in animal models. However, proteome studies that investigated the role of complement C3 were missing. The objective of this study was to characterize local tissue exudate (“gingival crevicular fluid”; GCF) changes after administration of a potent C3 inhibitor (Cp40, also known as AMY-101) to cynomolgus monkeys (Macaca fascicularis) with established periodontitis. Cp40 was administered locally in the maxillary gingival tissue either once per week (n=5 animals) or three times per week (n=10 animals), for six weeks followed by another six weeks of observation in the absence of treatment. The GCF were collected for liquid chromatography–mass spectrometry analysis (LC-MS/MS).
Project description:Gingival Crevicular Fluid, 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 Gingival Crevicular Fluid 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 Gingival Crevicular Fluid, such as specific fragments of α-1-antitrypsin, α-1-antichymotrypsin, Thymosin β4 and Thymosin β10 fragments, Fibrinopeptide A, Fibrinopeptide B, S100A, 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 in particular with higher level in exfoliating teeth. Our study demonstrate that an in-depth analysis of a biological fluid like Gingival Crevicular Fluid, present in small amount, can provide useful information for the understanding of different biological processes like teeth eruption.
Project description:Thirteen healthy men diagnosed with periodontitis were enrolled. Progression of periodontitis was monitored by clinical attachment level (CAL) changes at the site level over twelve weeks via weekly clinical evaluations and gingival crevicular fluid (GCF) samples were collected using cellulose strips for subsequent mass spectrometry analysis. Progression group (PG) included 18 GCF samples matching sites with disease progression (CAL ≥ 2 mm), while the non-Progression group (NP) included 18 GCF samples from stable sites (No CAL changes) matched in the same patients. Clinical metrics, HPRLC-MS/MS, in silico methods, and bioinformatics tools, were employed to analyze the proteome of GCF, revealing the significant occurrence of ferroptosis during clinical progression of periodontitis.
Project description:Analysis of gingival crevicular fluid (GCF) samples may give information of the identity of unattached (planktonic) subgingival bacteria, the 35 forefront candidates for systemic dispersal via ulcerated periodontal pocket epithelium. Our study represents the first one targeting the identity of bacteria in gingival crevicular fluid. Methodology/Principal findings: We determined bacterial species diversity in GCF samples of a group of periodontitis patients and delineated contributing bacterial and host-associated factors. Subgingival paper point (PP) samples from the same sites were taken for comparison. After DNA extraction, 16S rRNA genes were PCR amplified and DNA-DNA hybridization was performed using a microarray for over 300 bacterial species or groups. Altogether 133 species from 41 genera and 8 phyla 45 were detected with 9 to 62 and 18 to 64 species in GCF and PP samples, respectively, 46 per patient. Projection to latent structures by means of partial least squares (PLS) was applied to the multivariate data analysis. PLS regression analysis showed that species of genera including Campylobacter, Selenomonas, Porphyromonas, Catonella, Tannerella, Dialister, Peptostreptococcus, Streptococcus and Eubacterium had significant positive correlations and the number of teeth with low-grade attachment loss a significant negative correlation to species diversity in GCF samples. OPLS/O2PLS discriminant analysis revealed significant positive correlations to GCF sample group membership for species of genera Campylobacter, Leptotrichia, Prevotella, Dialister, Tannerella, Haemophilus, Fusobacterium, Eubacterium, and Actinomyces. Conclusions/Significance: Among a variety of detected species those traditionally classified as Gram-negative anaerobes growing in mature subgingival biofilms were the main predictors for species diversity in GCF samples as well as responsible for distinguishing GCF samples from PP samples. GCF bacteria may provide new prospects for studying dynamic properties of subgingival biofilms. The microbial profiles of GCF and subgingival plaque were analyzed from 17 subjects with periodontal disease.
Project description:Aggressive periodontitis (AP; Grade C, Stage 3-4) is rapid destructive condition of periodontium triggered by an imbalanced immune reaction with dysbiosis of periodontal pathogens. AP is characterized by extensive tooth loss in early ages due to its severe nature and rapid progression, significantly impacting the affected individual's quality of life. Despite the severity and refractoriness of the disease, the precise pathophysiology of this hyper-reactive phenotype remains unclear. In this study, we dissected the distinct immunological features in AP by establishing the comprehensive human gingival cell atlas on individuals with AP, chronic periodontitis (CP), and healthy controls (HC). Iterative clustering analysis based on the single-cell transcriptome revealed the distinct populations in B and plasma cells upon the periodontal conditions. AP group exhibited a significant rise in B cells including a unique cell cluster dominantly expressing ID3, a key suppressor of plasma cell development. This observation is coordinated with the impaired phenotypes of antibody class switching in AP patients. Remarkably, the secreted IgM gingival crevicular fluid from AP patients exhibited the autoreactivity to gingival tissues, suggesting a potential autoimmune mechanism that involved in AP pathogenesis. These findings highlight distinct immune cell and antibody profiles in AP, providing the insights into the groundwork for personalized monitoring and proactive intervention approaches to mitigate irreversible tissue damage.