Title ANALIZA DENTALNOG BIOFILMA NA
POLITETRAFLUORETILENSKIM
MEMBRANAMA VISOKE GUSTOĆE
Title (english) DENTAL BIOFILM ANALYSIS OF DENSE
POLYTETRAFLUOROETHYLENE
MEMBRANES
Author Gabrijela Begić
Mentor Ivana Gobin (mentor)
Mentor Davor Kuiš (komentor)
Committee member Marina Bubonja Šonje (predsjednik povjerenstva)
Committee member Branka Bedenić (član povjerenstva)
Committee member Jelena Prpić (član povjerenstva)
Granter University of Rijeka Faculty of Medicine (Department of Microbiology and Parasitology) Rijeka
Defense date and country 2023-09-06, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Clinical Medical Sciences Medical Microbiology
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Dental Medicine
Universal decimal classification (UDC ) 61 - Medical sciences
Thesaurus (MESH - Medical Subject Headings )
Biofilms
Dentin
Hydroxyapatites
Abstract Ciljevi: Analizirati sposobnost adherencije i stvaranja biofilma određenih vrsta oralnih streptokoka i parodontopatogenih bakterija na guste politetrafluoroetilenske membrane (d PTFE) koje se koriste u postupku prezervacije alveole. Ispitati utjecaj antimikrobnih sredstava, koamoksiklava i klorheksidina na adheziju i stvaranje biofilma oralnih bakterija te na bakterije u biofilmu. Metode: U istraživanju su korištene dvije d-PTFE membrane, Permamem i Cytoplast. Mikrostruktura d-PTFE membrane je ispitivana rendgenskom difrakcijom (XRD), diferencijalnom skenirajućom kalorimetrijom (DSC) i infracrvenom Fourierovom transformacijom (FTIR). Svojstva površine analizirana su mikroskopijom atomske sile (AFM), skenirajućom elektronskom mikroskopijom (SEM) i mjerenjem slobodne energije površine (SFE). Stvaranje biofilma u mikrotitar pločici je model korišten u istraživanju. Kvantificirane su adherirane i bakterije u pojedinačnom i miješanom biofilmu. Kvalitativna analiza biofilma provedena je pomoću SEM i konfokalne laserske skenirajuće mikroskopije (CLSM). Učinak antimikrobnih sredstava određen je metodom mikrodilucije u bujonu. Rezultati: Utvrđeno je da ispitivane bakterije snažnije adheriraju i stvaraju biofilm na Cytoplast u odnosu na Permamem membrane. Otkrivena je razlika u mikrostrukturi polimera što je rezultiralo različitim antiadhezivnim učincima kemijski identičnih membrana. Analiza miješanog biofilma pokazala je da S. salivarius K12 u interakciji s drugim vrstama ima antagonistički učinak na rast i dominaciju oportunističkih patogena u dentalnom biofilmu. Utvrđene su značajno veće inhibitorne i baktericidne koncentracije koamoksiklava i klorheksidina prema bakterijama u biofilmu u odnosu na planktonske oblike. Subinhibitorne koncentracije koamoksiklava i klorheksidina mogu utjecati na povećanje broja bakterija u biofilmu. Zaključak: Dvije vrste komercijalnih d-PTFE membrana imale su isti kemijski sastav, ali su im se površinske mikrostrukture uvelike razlikovale, naročito u stupnju kristalnosti i hrapavosti u nanometarskoj skali, što je rezultiralo različitom bakterijskom adhezijom. Ovi rezultati daju doprinos razumijevanju adhezije ranih oralnih mikrobnih kolonizatora koji mogu pomoći kliničarima u odabiru membrane u regenerativnoj dentalnoj medicini. S. Salivarius K 12 u miješanom biofilmu može inhibirati rast parodontopatogena i pomoći u održavanju fiziološke ravnoteže u oralnom mikrobiomu i unaprjeđenju oralnog zdravlja.
Abstract (english) Objectives: To analyze the ability of adherence and biofilm formation of certain types of oral streptococci and periodontopathogenic bacteria on dense polytetrafluoroethylene membranes (d-PTFE) used in the socket preservation process. Our goal was to examine the influence of antimicrobial agents, amoxicillin/clavulanic acid, and chlorhexidine digluconate on oral bacteria's adhesion and biofilm formation and on the bacteria in the biofilm. Materials and methods: Two d-PTFE membranes, Permamem and Cytoplast, were used in the research. The microstructure of the d-PTFE membrane was investigated by X-ray diffraction (XRD), differential scanning calorimetry (DSC), and infrared Fourier transform (FTIR). Surface properties were analyzed by atomic force microscopy (AFM), scanning electron microscopy (SEM) and surface free energy (SFE) measurements. Biofilm formation of selected oral bacteria in a microtiter plate model was used in the research. Adherents were quantified in individual and mixed biofilms. Qualitative analysis of the biofilm was performed using SEM and confocal laser scanning microscopy (CLSM). The effect of antimicrobial agents was determined by the broth microdilution method. Results The research demonstrated that the examined more potent bacteria adhere and form a biofilm on Cytoplast compared to Permamem membranes. A difference in the polymer's microstructure was discovered, resulting in different anti-adhesive effects of chemically identical membranes. Analysis of the mixed biofilm showed that S. salivarius K12, in interaction with other species, has an antagonistic effect on the growth and dominance of opportunistic pathogens in the dental biofilm. Significantly higher inhibitory and bactericidal concentrations of coamoxiclav and clorhexidine against bacteria in the biofilm compared to planktonic forms were found. Subinhibitory concentrations of coamoxiclav and clorhexidine can lead to an increase in the number of bacteria in the biofilm. Conclusion: Two commercial d-PTFE membranes had the same chemical composition, but their surface microstructures differed significantly, especially in the degree of crystallinity and roughness on the nanometer scale, which resulted in different bacterial adhesion. These results contribute to understanding of the adhesion of early oral microbial colonizers that may assist clinicians in membrane selection in socket preservation procedures. S. salivarius K 12 in a mixed biofilm can inhibit the growth of periodontal pathogens, help maintain the physiological balance in the oral microbiome, and improve oral health.
Keywords
dentalni biofilm
d-PTFE membrana
mikrostruktura polimera
oralne bakterije
parodontopatogene bakterije
vođena regeneracija kosti
Keywords (english)
dental biofilm
d-PTFE membrane
guided bone regeneration
microstructure of polymers
oral bacteria
Language croatian
URN:NBN urn:nbn:hr:184:372711
Promotion 2023
Project Number: IP-2020-02-7875 Title: Usporedba reparacijskog odgovora koštanog tkiva upotrebom dentina, ksenogenog biomaterijala i autologne kosti Acronym: MgXBiomBone Leader: Olga Peloza Jurisdiction: Croatia Funder: HRZZ Funding stream: IP
Study programme Title: Biomedicine Postgraduate (doctoral) study programme Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo (doktor/doktorica znanosti, područje biomedicine i zdravstvo)
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Created on 2023-10-11 07:22:48