Utjecaj BMP-2, BMP-6 i fluoksetina na ekspresiju diferencijacijskih biljega osteoblasta u kulturi stanica MC3T3-E1

master's thesis

• Title (english): Impact of BMP-2, BMP-6, and fluoxetine on expression of osteoblast differentiation markers in MC3T3-E1 cell culture
• Author: Nikolina Martinčić
• Mentor: Donatella Verbanac (mentor)
• Committee member: Donatella Verbanac (predsjednik povjerenstva)
• Committee member: Marija Grdić Rajković (član povjerenstva)
• Committee member: Slobodan Vukičević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2021-08-30, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Uvod: Kronična upotreba fluoksetina u svrhu liječenja depresije povezana je s povećanim rizikom od loma kostiju. Osteoblasti su stanice koje izgrađuju koštani matriks, tj. zaslužne su za rast, razvoj i regeneraciju koštanog tkiva. Koštani morfogenetski proteini potiču proliferaciju i diferencijaciju osteoblasta. Ovaj rad ispituje učinak fluoksetina na osteoblaste, te zajednički učinak fluoksetina i koštanih morfogenetskih proteina na osteoblaste .
Metode: Stanice preosteoblasta MC3T3-E1 su nasađene na tri ploče i tretirane koštanim morfogenetskim proteinima 2 i 6 u dvije koncentracijske razine, fluoksetinom, te kombinacijom koštanog morfogenetskog proteina i fluoksetina. Iz stanica na prvoj ploči izolirana je RNA i provedena analiza ekspresije odabranih diferencijacijskih markera osteoblasta lančanom reakcijom polimeraze u stvarnom vremenu. Druga ploča je obojana na alkalnu fosfatazu, a treća po von Kossi. Rezultati bojanja su fotografirani, a fotografije su računalno obrađene u svrhu određivanja ekspresije alkalne fosfataze i mineralizacije.
Rezultati: Koštani morfogenetski proteini 2 i 6 su u obje koncentracijske razine pokazali izrazito povećanu ekspresiju svih ispitivanih parametara. Fluoksetin nije pokazao značajan utjecaj na ispitivane parametre. Kombinacija fluoksetina i koštanog morfogenetskog proteina pokazuje pozitivan učinak na diferencijaciju osteoblasta, ali značajno slabiji od samih koštanih morfogenetskih proteina.
Zaključak: Fluoksetin vjerojatno ima inhibitorni učinak na signalizacijske puteve koštanih morfogenetskih proteina čime posredno uzrokuje sporiju regeneraciju fraktura i mikrofraktura kostiju.

Abstract (english)

Introduction: Chronic use of fluoxetine for treatment of major depresive disorder is in correlation with increased risk of bone fractures. Osteoblasts are cells that build up bone matrix, ie. they promote growth, development, and regeneration of bone tissue. Bone morphogenetic proteins promote proliferation and differentiation of osteoblasts. This research explores the effect of fluoxetine on osteoblasts, and cummulative effect of fluoxetine and bone morphogenetic protein on osteoblasts.
Methods: Preosteoblast cells MC3T3-E1 were seeded on three cell culture plates and treated with bone morphogenetic proteins in two concentration levels, fluoxetine, and combination of bone morphogenetic protein and fluoxetine. From the first plate, RNA was isolated to conduct analysis of expression of selected diferentiation markers of osteoblasts via real-time polymerase chain reaction. Second plate was stained for alkaline phosphatase, and the third one was stained by von Kossa method. Results of staining were photographed, and expression of alkaline phosphatase and mineralization was determined via computer analysis of photographs.
Results: Bone morphogenetic proteins 2 and 6 in both concentration levels caused significant increase of expression of all inquired parameters. Fluoxetine did not show significant impact on inquired parameters. Combination of fluoxetine and bone morphogenetic protein showed positive effect on differentiation of osteoblasts, but this effect was significantly weaker than the effect of bone morphogenetic proteins alone.
Conclusion: Fluoxetine likely has inhibitory impact on signalization pathways of bone morphogenetic proteins, whereby indirectly inhibits regeneration of bone fractures and microfractures.

• Keywords: BMP-2; BMP-6; fluoksetin; SSRI; osteoblasti
• Keywords (english): BMP-2; BMP-6; fluoxetine; SSRI; osteoblasts
• Language: croatian
• URN:NBN: urn:nbn:hr:163:784637
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije i laboratorijske medicine (magistar/magistra medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-02-02 09:58:17