Elif Ece AKGÜN, Eda DEMİRTAŞ, Özlem ÖZDEN AKKAYA, Metin ERDOĞAN, Korhan ALTUNBAŞ

BMP-9 ve TGF-ß3 Eklenmiş Kondrojenik Farklılaşma Medyumunun Transwell Ko-kültürde Hipertrofi Üzerine Etkisi

Mezenkimal kök hücreler, kıkırdağa farklılaşma yetenekleri nedeniyle osteoartrit dahil birçok hastalığın tedavisinde yaygın olarak kullanılmaktadır. Sinoviyal sıvı kökenli mezenkimal kök hücrelerinin kondrojenik farklılaşma potansiyellerinin fazla olması bu hücrelerin osteoartrit tedavilerindeki önemini artırmaktadır. Kondrojenik farklılaşma medyumuna ilave edilen BMP-9 ve TGF-ß3 büyüme faktörleri kondrojenik farklılaşmayı artırır ve aynı zamanda kondrositlerde hipertofik etkilere sebep olur. Çalışmamızda in vitro kondrojenik farklılaşma esnasında kondrojenik bazal medyuma ilave edilen BMP-9 ve TGF-ß3’ün hücre hipertrofisi üzerine etkilerinin gösterilmesi amaçlandı. Çalışmada transwell ko-kültür sisteminde 5. pasajdaki kök hücreler ve pasaj 1'deki kondrositler birlikte kültüre edildi ve altı deney grubu oluşturuldu. Hücre hipertrofisi, transwell ko-kültüründe kondrogenezin indüklendiği kök hücrelerde MMP-13 ve RUNX-2 gen ekspresyonları incelenerek gösterildi. BMP-9 ve TGF-ß3'ün kondrojenik ortama eklenmesi, kontrol grubuna göre deney gruplarında hipertrofik gen ekspresyonlarını artırmasına rağmen bu artış istatistiksel olarak anlamlı değildi. Kök hücrelerin kondrojenik farklılaşması esnasında BMP-9 ve TGF-ß3’ün ayrı ayrı veya birlikte kullanılması önemli derecede kıkırdak hücresi hipertrofisine neden olmamaktadır.

Anahtar Kelimeler:

Sinoviyal Sıvı, Kıkırdak, Hipertrofi, Transwell Kokültür, Kondrogenezis

Effect of Chondrogenic Differentiation Medium Supplemented with BMP-9 and TGF-ß3 on Hypertrophy in Transwell Co-Culture

Mesenchymal stem cells are widely used in the treatment of many diseases, including osteoarthritis, due to their ability to differentiate into cartilage. The high chondrogenic differentiation potential of synovial fluid-derived mesenchymal stem cells increases the importance of these cells in osteoarthritis treatments. Addition of BMP-9 and TGF-ß3 into chondrogenic differentiation medium, increases chondrogenic differentiation and they also cause hypertrophic effects on chondrocytes. In our study, it was aimed to demonstrate the effects of BMP-9 and TGF-ß3 on cell hypertrophy by adding them into the chondrogenic basal medium during in vitro chondrogenic differentiation. In the study, stem cells in passage 5 and chondrocytes in passage 1 were cultured in a transwell co-culture system and six experimental groups were formed. Cell hypertrophy was demonstrated by examining MMP-13 and RUNX-2 gene expressions, in stem cells where chondrogenesis were induced in transwell co-culture. Although the addition of BMP-9 and TGF-ß3 to the chondrogenic medium increased hypertrophic gene expressions in experimental groups compared to control, the results were not statistically significant. The addition of BMP-9 and TGF-ß3, separately or in combination, during the chondrogenic differentiation of stem cells does not cause significant chondrocyte hypertrophy.

Keywords:

Synovial Fluid, Cartilage, Hypertrophy, Transwell Coculture, Chondrogenesis,

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