Dataset Information

Proteomic Analysis of Whole Saliva in Chronic Periodontitis

ABSTRACT: Periodontitis is a chronic inflammatory disease resulting from a dysbiosis of the dental biofilm and a dysregulated host response in susceptible individuals. It is characterized by periodontal attachment destruction, bone resorption, and eventual tooth loss. Salivary biomarkers have been sought to predict and prevent periodontitis. This comparative study analyzed the salivary proteome of 30 individuals with chronic periodontitis (CP) and 10 with periodontal health (PH), and correlated specific proteins with clinical parameters of disease by using mass spectrometry. Stimulated whole saliva was obtained and pooled for 5 healthy controls and 15 CP patients, precipitated with TCA, digested enzymatically with trypsin and analyzed by an LTQ Orbitrap Velos equipped with a nanoelectrospray ion source. A wide range of salivary proteins of various functions was significantly reduced in CP individuals, whereas salivary acidic proline-rich phosphoprotein, submaxillary gland androgen-regulated protein, histatin, fatty acid-binding protein, thioredoxin, and cystatin were predominant in diseased patients and correlated significantly with signs of periodontal attachment loss and inflammation. Specific salivary proteins were associated with PH and CP. These differences in salivary proteome profiles may contribute to the identification of disease indicators or signatures and the improvement of periodontal diagnosis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Saliva

DISEASE(S): Periodontitis

SUBMITTER: Proteomics Unit

LAB HEAD: Fabio Cesar Sousa Nogueira

PROVIDER: PXD015024 | Pride | 2020-05-20

REPOSITORIES: Pride

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Publications

Proteomic analysis of whole saliva in chronic periodontitis.

Hartenbach Fátima Aparecida Rocha Resende FARR Velasquez Érika É Nogueira Fábio C S FCS Domont Gilberto B GB Ferreira Eliane E Colombo Ana Paula Vieira APV

Journal of proteomics 20191204

Periodontitis is a chronic inflammatory disease resulting from a dysbiosis of the dental biofilm and a dysregulated host response in susceptible individuals. It is characterized by periodontal attachment destruction, bone resorption and eventual tooth loss. Salivary biomarkers have been sought to predict and prevent periodontitis. This comparative study analyzed the salivary proteome of individuals with chronic periodontitis (CP) and periodontal health (PH) and correlated specific proteins with ...[more]

PMID: 31809901

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Project description:Phase 1: The whole saliva samples were obtained from 66 systemically healthy subjects consisting of patients with chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP), gingivitis and individuals with periodontal health. The use of humans for study satisfied the requirements of the Ege University Institutional Review Board (Ethics number 16-12.1/16) and and was conducted in accordance with the guidelines of the World Medical Association Declaration of Helsinki. It is confirmed that this cross-sectional study conforms to STROBE guidelines for observational studies. Complete medical and dental histories were taken from all participants. Systemic exclusion criteria were the presence of cardiovascular and respiratory diseases, diabetes mellitus, HIV infection, systemic inflammatory conditions or non-plaque-induced oral inflammatory conditions, immunosuppressive chemotherapy, and current pregnancy or lactation. Smoking status was also recorded. None of the patients had taken medication such as antibiotics or anti-inflammatory drugs that could affect their periodontal status for at least 6 months before the study. Patients eligible for the study returned to the clinic for clinical measurement screening one-week after being pre-screened. Before being enrolled in the study, participants provided written and informed consent to use their saliva samples and individual data for scientific purposes. The clinical periodontal indices including probing depth (PPD), clinical attachment level (CAL), plaque index (PI) and bleeding on probing (BOP) were recorded by a manual periodontal probe by a trained and calibrated examiner (V.O.O). The extent and severity of alveolar bone support were evaluated radiographically in each patient. The participants were classified into four groups based on their periodontal conditions according to the criteria proposed by the 1999 International Workshop for a Classification of Periodontal Diseases and Conditions (REF). Generalized Aggressive Periodontitis (G-AgP): The G-AgP group included 17 patients with ≥16 teeth. The patients had a non- contributory medical history and demonstrated with an early age of clinical manifestations with a generalized pattern of rapid attachment loss and bone destruction. These individuals had minimum of CAL ≥ 5 mm and PD ≥ 6 mm on eight or more teeth; at least three of these were other than central incisors or first molars. Radiographic bone loss was above ≥ 30 % of root length affecting ≥3 permanent teeth other than first molars and incisors. Generalized Chronic Periodontitis (CP): The CP group included 17 patients with severe generalized chronic periodontitis. These individuals had a minimum four non-adjacent teeth with sites with CAL ≥ 5 mm and PPD ≥ 6 mm, and ≥50 % alveolar bone loss in at least two quadrants which was commensurate with the amount of plaque accumulation. Phase 2: We aimed to develop SRM-based workflow for detecting and quantitating the relative abundance of the candidate proteins in an independent saliva cohort . 82 subjects (48 females and 34 males; age range 24-59 years) were recruited for the study at the Department of Periodontology, School of Dentistry, Adnan Menderes University, Aydın, Turkey. Ethical clearance was obtained from the Ethics Committee of the School of Medicine, Ege University with the protocol number ((Ethics number 70198063-050.06.04). Each participant gave written and verbal informed consent after the purpose and procedures of the study were explained. A dental and medical history was compiled for all subjects as detailed in phase 1 and participants were clustered into four groups according to the criteria proposed by the 1999 International Workshop for a Classification of Periodontal Diseases and Conditions as detailed at Phase 1

Project description:The effect of oral microbiota on the intestinal microbiota has garnered growing attention as a mechanism linking periodontal diseases to systemic diseases. However, the salivary microbiota is diverse and comprises numerous bacteria with a largely similar composition in healthy individuals and periodontitis patients. Thus, the systemic effects of small differences in the oral microbiota are unclear. In this study, we explored how health-associated and periodontitis-associated salivary microbiota differently colonized the intestine and their subsequent systemic effects by analyzing the hepatic gene expression and serum metabolomic profiles. The salivary microbiota was collected from a healthy individual and a periodontitis patient and gavaged into C57BL/6NJcl[GF] mice. Samples were collected five weeks after administration. Gut microbial communities were analyzed by 16S ribosomal RNA gene sequencing. Hepatic gene expression profiles were analyzed using a DNA microarray and quantitative polymerase chain reaction. Serum metabolites were analyzed by capillary electrophoresis time-of-flight mass spectrometry. The gut microbial composition at the genus level was significantly different between periodontitis-associated microbiota-administered (PAO) and health-associated oral microbiota-administered (HAO) mice. The hepatic gene expression profile demonstrated a distinct pattern between the two groups, with higher expression of Neat1, Mt1, Mt2, and Spindlin1, which are involved in lipid and glucose metabolism. Disease-associated metabolites such as 2-hydroxyisobutyric acid and hydroxybenzoic acid were elevated in PAO mice. These metabolites were significantly correlated with Bifidobacterium, Atomobium, Campylobacter, and Haemophilus, which are characteristic taxa in PAO mice. Conversely, health-associated oral microbiota were associated with higher levels of beneficial serum metabolites in HAO mice. The multi-omics approach used in this study revealed that periodontitis-associated oral microbiota is associated with the induction of disease phenotype when they colonized the gut of germ-free mice.

Dataset Information

Proteomic Analysis of Whole Saliva in Chronic Periodontitis

Publications

Proteomic analysis of whole saliva in chronic periodontitis.

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