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) MBZ: 230266
Granter University of Zagreb School of Dental Medicine (Department of Periodontology) Zagreb
Defense date and country 2013-10-23, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Dental Medicine Periodontology
Universal decimal classification (UDC ) 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 Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo, polje dentalna medicina (doktor/doktorica znanosti, područje biomedicine i zdravstvo, polje dentalna medicina)
Type of resource Text
File origin Born digital
Access conditions Open access
Terms of use
Created on 2017-03-10 11:27:59