Vpliv termoreverzibilnega hidrogela na sposobnost hondrogene diferenciacije primarnih mezenhimskih matičnih/stromalnih celic

Kalan, Lanna (author), Zupan, Janja (mentor)

Mezenhimske matične/stromalne celice (MSC) veljajo za terapevtsko izredno privlačno orodje, saj je njihova uporaba etično manj vprašljiva od uporabe embrionalnih matičnih celic, poleg tega pa jih je mogoče pridobiti iz mnogih tkiv. Glavni izziv v tkivnem inženirstvu hrustanca je vzpostavitev tridimenzionalnega (3D) okolja, ki bi omogočilo ustrezno diferenciacijo in tvorbo stabilnega zunajceličnega matriksa. Namen magistrske naloge je bil raziskati vpliv termoreverzibilnega hidrogela na sposobnost hondrogene diferenciacije primarnih humanih MSC, pridobljenih iz kostnega mozga. Sposobnost hondrogeneze smo določali na dva načina, in sicer v obliki mikromas in v obliki 3D pelet. Sposobnost hondrogeneze celic v obliki pelet smo ocenili tako, da smo izmerili njihov premer, jih ocenili z metodo po Bern-u in določili stopnjo izražanja kolagena tipa II. Rezultati so pokazali, da je hondrogena diferenciacija izrazitejša pri vzorcih, tretiranih s kombinacijo hondrogenega medija in termoreverzibilnega hidrogela. Sklepamo lahko, da hidrogel predstavlja 3D nosilec s potencialom za spodbujanje hondrogene diferenciacije. Rezultati podpirajo nadaljnjo uporabo termoreverzibilnega hidrogela v tkivnem inženirstvu hrustanca, hkrati pa odpirajo vprašanja o optimizaciji pogojev diferenciacije in dodatnih kvantitativnih metodah.

• Language: Slovenian
• Keywords: mezenhimske matične celice, hondrogeneza, termoreverzibilni hidrogel, 3D pelete
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2025
• PID: 20.500.12556/RUL-172156
• Publication date in RUL: 06.09.2025

Secondary language

• Language: English
• Title: Effect of thermoreversible hydrogel on chondrogenic differentiation of primary mesenchymal stem/stromal cells

Abstract

Mesenchymal stem/stromal cells (MSC) are considered a highly promising therapeutic tool due to their less controversial ethical profile compared to embryonic stem cells and their availability from various tissue sources. A major challenge in cartilage tissue engineering is the establishment of a three-dimensional (3D) environment that enables proper differentiation and the formation of a stable extracellular matrix. The aim of this master’s thesis was to investigate the effect of a thermoreversible hydrogel on the chondrogenic differentiation potential of primary human MSCs isolated from bone marrow. Chondrogenic capacity was assessed in two models: in the form of micromasses and 3D pellet cultures. The chondrogenic potential of the pellets was evaluated by measuring their diameter, performing histological scoring based on the Bern scale, and assessing the level of type II collagen expression. The results demonstrated that chondrogenic differentiation was more pronounced in samples treated with a combination of chondrogenic medium and thermoreversible hydrogel. It can be concluded that hydrogel has a potential as three-dimensional scaffold for stimulating chondrogenesis. The results support further use of this gel in cartilage tissue engineering, while also raising important questions regarding the optimization of differentiation conditions and the application of additional quantitative analytical methods.

• Keywords: mesenchymal stem cells, chondrogenesis, thermoreversible hydrogel, 3D pellets