Title Proteomska analiza epitelnoga i vezivnoga tkiva parodonta kod pacijenata s agresivnim parodontitisom Title (english) Proteomic analysis of the epithelial and connective periodontal tissue in patients with aggressive periodontitis Author Ana Badovinac Mentor Darko Božić (mentor)Mentor Lovorka Grgurević (komentor)Committee member Darije Plančak (predsjednik povjerenstva)Committee member Andrej Aurer (član povjerenstva)Committee member Marinka Mravak-Stipetić (član povjerenstva)Committee member Marija Ivić (član povjerenstva)Committee member Fran Borovečki (član povjerenstva)Granter University of Zagreb Defense date and country 2013-10-23, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 616.31 - Stomatology Abstract Proteomske analize omogućuju iscrpan prikaz ekspresije proteina u različitim biološkim uzorcima te predstavljaju napredan pristup u istraživanjima proteoma. Svrha ovog istraživanja bila je dokazati da epitelno i vezivno tkivo parodonta zdravih ispitanika (kontrolna skupina) i zdravog mjesta, mjesta s gingivitisom, parodontitisom i mjesta s parodontitisom nakon terapije kod pacijenata s generaliziranim agresivnim parodontitisom (ispitivane skupine) pokazuju različitu ekspresiju proteina.
U istraživanju je sudjelovalo osam pacijenata. Ispitivanu skupinu činila su četiri pacijenta s dijagnozom generaliziranog agresivnog parodontitisa (dva muškarca i dvije žene), dok su kontrolnu skupinu činila četiri ispitanika bez znakova parodontne bolesti kojima je razlog dolaska bio kirurško produljenje krune zuba (dva muškarca i dvije žene). Uzorci epitelnog i
vezivnog parodontnog tkiva uzeti su pri provođenju inicijalne parodontološke terapije u ispitivanoj skupini, a u kontrolnoj skupini tijekom indiciranog kirurškog zahvata. Uzorci su pohranjeni u RNAlater tekućinu prema uputama proizvođača do analiza. Proteini tkivnog lizata razdvojeni su 1D gel elektroforezom nakon koje je uslijedila digestija u gelu tripsinom te mjerenje proteina pomoću nano-skale HPLC sistema preko nano-elektrospreja ionizacijskog izvora i spektrometra mase. Dobiveni podaci obrađeni su u MaxQuant programu. Ukupan broj identificiranih proteina u istraživanju iznosio je 3420. Nakon statističke obrade podataka izdvojena su 193 proteina koji su identificirani i kvantificirani u svim uzorcima, te su u svim ispitivanim skupinama pokazali statistički značajnu promjenu ekspresije u odnosu na kontrolnu skupinu. Proteini koji bi mogli biti obilježje zdravog parodonta jesu: Multimerin-1, aneksini, 14-3-3 protein gamma; gingivitisa: Dedicator of cytokinesis protein 2, Band 3 anion transport protein; parodontitisa: Azurocidin, Myeloid cell nuclear differentiation antigen, histoni, imunoglobulini, Prolargin; stanja poslije terapije:
Oligosaccharyltransferase complex subunit i Mimecan. Najzanimljiviji protein istraživanja je Azurocidin koji pokazuje jaku ekspresiju u gingivitisu i parodontitisu. Rezultati istraživanja pokazali su različitu ekspresiju proteina u ispitivanim skupinama te potvrdili da bi proteomske analize parodontnog tkiva mogle pridonijeti boljem razumijevanju patogeneze parodontnih bolesti, možda čak i napretku u dijagnostici i prognozi ovih bolesti.
Abstract (english) Proteomic analyses enable large scale descriptions of protein expression in a variety of biological samples, and provide an advanced approach in proteome research. The aim of this study was to establish the changes in protein expression between the epithelial and connective periodontal tissues samples in healthy subjects (control group) and healthy sites, sites with gingivitis, periodontitis, and post treatment sites in patients suffering from generalized aggressive periodontitis (test groups). The study was conducted on eight patients divided into two groups. The test group consisted of four patients with generalized aggressive periodontitis (two male, two female), and the control group consisted of four subjects (two male, two female) with no signs of periodontal disease, referred to the Department of Periodontology for crown lengthening procedure. The epithelial and connective periodontal tissue samples in the test group were harvested during initial periodontal treatment, and in the control group during the surgical procedure. Tissue samples were stored in the 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. The obtained raw data was processed by MaxQuant software.
A total number of proteins identified in the study was 3420. After statistical processing of data, a total of 193 proteins were identified and quantified in all samples, and showed statistically significant changes in expression level between the control group and test groups. Proteins that could possibly be a mark of periodontal health are: Multimerin-1, annexins, 14-
3-3 protein gamma; of gingivitis: Dedicator of cytokinesis protein 2, Band 3 anion transport protein; of parodontitis: Azurocidin, Myeloid cell nuclear differentiation antigen, histones, immunoglobulins, Prolargin; and of post treatment condition: Oligosaccharyltransferase complex subunit and Mimecan. The most interesting protein in the study was Azurocidin, showing the highest fold change in periodontitis and gingivitis. The 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 even contribute to the improvements in diagnosis and prognosis of these diseases.
Keywords
agresivni parodontitis
gingivitis
proteomika
spektrometrija mase
biopsija
identifikacija proteina
proteini
Keywords (english)
aggressive periodontitis
gingivitis
proteomics
mass spectrometry
biopsy
protein identification
proteins
Language croatian URN:NBN urn:nbn:hr:127:469233 Study programme Title: dental medicine Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2017-03-10 11:27:59
