engleski

Proteomic Analyses of Gingival Tissue in Patients with Aggressive Periodontitis

Objectives: The aim of this study was to establish changes in protein expression between gingival tissues samples in healthy subjects (HH, control group) and healthy sites, sites with gingivitis, periodontitis, and post treatment sites in patients with generalized aggressive periodontitis (GAgP, test groups). Methods: The study enrolled 8 patients, 4 with GAgP, and 4 with no signs of periodontal disease referred for crown lengthening. Gingival tissue samples in GAgP were harvested during initial periodontal treatment and sampled as healthy (PD≤3mm, BOP-), gingivitis (PD≤3mm, BOP+), periodontitis (PD≥4mm), 3 months post treatment, and in HH during surgical procedures. Tissue samples were stored in RNAlater reagent according to the manufacturer’s instructions until further analyses. Proteins from tissue lysate were separated by 1D gel electrophoresis, in-gel digested with trypsin, and measured by nano-scale HPLC system coupled to a mass spectrometer through a nano-electrospray ionization source. Shotgun label-free quantitative proteomics was used to identify proteins. False discovery rate was set to 5%. The obtained raw data was processed by MaxQuant software. Results: The total number of identified proteins was 3420. After statistical data processing, 193 proteins showed statistically significant changes in expression levels between the control groups and test groups. Proteins that could possibly be a mark of periodontal health are: Multimerin-1, Annexins, Prolargin ; of gingivitis: Dedicator of cytokinesis protein 2, Band 3 anion transport protein ; of periodontitis: Azurocidin, Myeloid cell nuclear differentiation antigen, Histones, Immunoglobulins ; and of post treatment condition: Oligosaccharyltransferase complex subunit and Mimecan. The most interesting protein was Azurocidin, showing the highest fold change in periodontitis and gingivitis. Conclusion: Results showed different protein expressions in tested groups and confirmed that proteomic analyses of periodontal tissue could contribute to a better understanding of the pathogenesis of periodontal diseases, and might contribute to the improvements in diagnosis and prognosis of GAgP.

aggressive periodontitis ; gingivitis ; proteomics ; mass spectrometry ; biopsy ; protein identification ; proteins

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