Optimizacija uvjeta za trodimenzionalni rast hrskavice in vitro

master's thesis

• Title (english): Optimization of conditions for in vitro three-dimensional cartilage growth
• Author: Mihaela Jagrić
• Mentor: Inga Marijanović (mentor)
• Committee member: Inga Marijanović (predsjednik povjerenstva)
• Committee member: Jasna Hrenović (član povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2016-02-29, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Artikularna hrskavica je tkivo koje nema sposobnost spontane regeneracije. Tkivno inženjerstvo, kao novo područje regenerativne medicine, nudi potencijalna učinkovita rješenja u liječenju ozljeda i oštećenja artikularne hrskavice. Cilj ovoga istraživanja bio je optimizirati uvjete za trodimenzionalni uzgoj hrskavice in vitro. U istraživanju su korištena tri različita tipa staničnih kultura: hondrociti, mezenhimske matične stanice iz koštane srži (hBMSCs) te kombinacija hondrocita i hBMSCs u omjeru 2:1. Stanice su uzgajane u diferencijacijskom mediju, inkorporirane u RADA peptidni hidrogel, u normoksičnim (20% O2) ili hipoksičnim (5% O2) uvjetima. Dexametazon, jedna od komponenti diferencijacijskoga medija, bila je dodana u medij ili inkorporirana sa stanicama u peptidnom hidrogelu. Nakon 19 dana metodom kvantitativnog PCR-a ispitana je razina ekspresije gena SOX9 i ACAN, karakterističnih za hrskavicu. Među korištenim tipovima stanica hondrociti su pokazali najviši, a hBMSCs najniži hondrogeni potencijal. Hipoksija je značajno poboljšala hondrogenezu jedino u slučaju hondrocita, dok izvor dexametazona nije bitno utjecao na razinu ekspresije gena SOX9 i ACAN.

Abstract (english)

Articular cartilage is a tissue that shows no capacity for effective spontaneous regeneration. Tissue engineering, as a new important field of regenerative medicine, emerges as potential effective solution for treatment of articular cartilage injuries and defects. The aim of this study was to optimize conditions for 3D in vitro chondrogenesis. In the experiment, three different cell culture types were used: chondrocytes, human bone marrow mesenchymal stem cells (hBMSCs) and combination of chondrocytes and hBMSCs in 2:1 ratio. Cells were grown in a chondrogenic medium, incorporated in a peptide hydrogel RADA, in either normoxic (20% O2) or hypoxic (5% O2) conditions. Dexamethasone, one of the chondrogenic medium components, was added in a differentiation medium or it was incorporated with cells within a peptide hydrogel. After 19 days, expression levels of two cartilage marker genes, SOX9 and ACAN, were evaluated by quantitative PCR. Chondrocytes showed the highest and hBMSCs the lowest chondrogenic potential among used cell types. Hypoxia enhanced chondrogenesis significantly only in the case of chondrocytes, while dexamethasone source did not considerably influence on expression levels of SOX and ACAN genes.

• Keywords: tkivno inženjerstvo; peptidni hidrogel; hondrogeneza; hondrociti; mezenhimske matične stanice; hipoksija
• Keywords (english): tissue engineering; peptide hydrogel; chondrogenesis; chondrocytes; mesenchymal stem cells; hypoxia
• Language: croatian
• URN:NBN: urn:nbn:hr:217:793257
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-05-05 11:49:44