Primerjava hondrogenega potenciala človeških mezenhimskih matičnih/ stromalnih celic iz kostnega mozga in sinovije v pogojih in vitro

Strašek, Larisa (author), Zupan, Janja (mentor), Stražar, Klemen (comentor)

Regeneracija hrustanca je ključna pri zdravljenju degenerativnih bolezni kot je osteoartroza, in pri obvladovanju poškodb sklepov, zato je iskanje optimalnih virov celic za tovrstne terapevtske aplikacije izjemnega pomena. V magistrski nalogi smo podrobno raziskali potencial mezenhimskih matičnih celic (MSC) iz dveh različnih virov, kostnega mozga (KM) in sinovijske ovojnice (S), za hondrogeno diferenciacijo. Primerjali smo tudi vpliv gojenja celic, tako da smo uporabili MSC iz kostnega mozga, ki so bile gojene v kvantnem bioreaktorju ali na standardni način. Postavili smo štiri hipoteze: 1. MSC iz KM, gojene v bioreaktorju, imajo višjo sposobnost hondrogeneze v testu z mikromasami kot enake celice, gojene na standardni način. Absorbanci vzorca MSC iz KM gojenih v bioreaktorju sta znašali 0,452 in 0,445; absorbance vzorca MSC iz KM gojenih na standardni način pa 1,315; 0,445 in 0,516. Naši rezultati nakazujejo, da gojenje MSC v bioreaktorju ne izboljša sposobnosti hondrogeneze v primerjavi z gojenjem teh celic na standardni način. 2. MSC iz KM imajo višjo oceno po Bernu kot MSC iz S. Ocene po Bern-u MSC iz KM so znašale 5, 4, 5, 5, 4; ocena po Bern-u MSC iz S pa 5. MSC iz KM so v našem primeru dosegle nižjo oceno po Bernu kot MSC iz S. 3. MSC iz KM tvorijo večje hondrogene pelete kot MSC iz S. MSC iz KM so tvorile 822,311 μm; 1060,192 μm; 893,317 μm; 802,279 μm in 475,987 μm velike hondrogene pelete. MSC iz S so tvorile hondrogeno peleto veliko 842,116 μm. Naši rezultati nakazujejo, da MSC iz KM tvorijo večje hondrogene pelete kot MSC iz S. 4. MSC iz KM imajo boljše izražanje kolagena tipa II kot MSC iz S. Naši rezultati niso pokazali razlik v prisotnosti kolagena tipa II med vzorci, vendar zaradi pozitivnih kontrolnih vzorcev ne moremo trditi o zanesljivosti naših rezultatov. Na ta način smo dokazali hondrogeni potencial preiskovanih MSC iz KM. Da pa bi dokazali razlike med vzorci MSC iz KM in S, bi jih morali vključiti več ter izvesti ustrezno statistično analizo. Naša raziskava je pomemben korak naprej pri pripravi MSC za uporabo v celičnih terapijah za regeneracijo hrustanca. Kljub temu pa ostaja še veliko neodgovorjenih vprašanj, ki bodo zahtevala nadaljnje preučevanje, da bi lahko v celoti izkoristili potencial MSC v klinični praksi in zagotovili boljšo kakovost življenja bolnikov s poškodbami hrustanca.

• Language: Slovenian
• Keywords: mezenhimske matične celice, kostni mozeg, sinovijska ovojnica, hondrogena diferenciacija, regeneracija hrustanca
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2024
• PID: 20.500.12556/RUL-162889
• Publication date in RUL: 28.09.2024

Secondary language

• Language: English
• Title: In vitro chondrogenic potential comparisonl of bone-marrow and synovial membrane-derived human mesenchymal stem/ stromal cells

Abstract

Cartilage regeneration is important in the treatment of degenerative diseases such as osteoarthritis and in managing joint injuries, making the search for optimal cell sources for those therapeutic applications of high importance. In this master's thesis, we investigated the potential of mesenchymal stem cells (MSCs) from two different sources: bone marrow (BM) and the synovial membrane (S) for chondrogenic differentiation. We also compared the impact of cell cultivation by using BM-derived MSCs that were cultured in a quantum bioreactor or using standard cultivation methods. We formulated four hypotheses: 1. BM-derived MSCs cultured in the bioreactor have a higher capacity for chondrogenesis in the micromass assay compared to the same cells cultured using standard approach. The absorbances of the BM-derived MSCs cultured in the bioreactor were 0.452 and 0.445, while the absorbances of those cultured using standard approach were 1.315, 0.445 and 0.516. Our results suggest that culturing of MSCs in a bioreactor does not improve their chondrogenic potential compared to the cells cultured in a standard way. 2. BM-derived MSCs have a higher Bern score than S-derived MSCs. The Bern scores of BM-derived MSCs were 5, 4, 5, 5 and 4, while the Bern score of S-derived MSCs was 5. In our case, BM-derived MSCs achieved a lower Bern score than S-derived MSCs. 3. BM-derived MSCs form larger chondrogenic pellets than S-derived MSCs. BM-derived MSCs formed chondrogenic pellets measuring 822.311 μm, 1060.192 μm, 893.317 μm, 802.279 μm and 475.987 μm. S-derived MSCs formed a pellet measuring 842.116 μm. Our results suggest that BM-derived MSCs form larger pellets than S-derived MSCs. 4. BM-derived MSCs show higher type II collagen expression than S-derived MSCs. BM-derived MSCs show better expression of type II collagen compared to S-derived MSCs. Our results did not show differences in the presence of type II collagen between samples, but due to positive control samples, we cannot claim the reliability of our results. Here, we have demonstrated the chondrogenic potential of the tested cells, i.e. BM-derived MSCs. To evaluate the differences between BM and S-derived MSCs, more samples should be included and statistical analysis performed. Despite remaining unanswered questions, this research is a significant step toward understanding MSC use in cartilage regeneration, with further studies required to fully realize their clinical potential and improve patient outcomes.

• Keywords: mesenchymal stem cells, bone marrow, synovial membrane, chondrogenic differentiation, cartilage regeneration