Evaluation of Pomegranate Seed Extract and TGF-β3 on Chondrogenic Differentiation of Human Adipose-Derived Stem Cells

Document Type : Original Article


1 Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Iran

2 Clinical Research Development Centre, Najafabad Branch, Islamic Azad University, Najafabad, Iran

3 Department of Genetics, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran


Introduction: The repair of the cartilage continues to be a big challenge. Autologous cartilage is the gold standard, but enough sources are not available. The development of stem cells, biomaterials, and bioactive factors has led to cartilage tissue engineering becoming a promising strategy for the regeneration of cartilage defects. In this study, the effect of Transforming Growth Factor Beta 3 (TGF-β3) and Pomegranate Seed Extract (PSE), as chondrogenesis inducers, were evaluated.
Materials and Methods: Human Adipose-Derived Stem Cells (ADSCs) were seeded in alginate scaffolds and cultivated for two weeks in chondrogenic media. Finally, the MTT assay was used to test the effect of TGF-β3 and PSE on the survival of differentiated cells. The mRNA levels of the cartilage-specific markers such as SRY-box9 (SOX9), Collagen type II (COLII), Collagen type X (COLX), and Aggrecan (ACAN) were determined by RT-PCR. Also, CD markers were evaluated by flow cytometry.
Results: Using both natural PSE and chemical TGFβ3 inducers simultaneously, had the best results in chondrogenesis by increasing the expression of the SOX9, COLII, and ACAN genes. Furthermore, the flow cytometry analysis indicated that the expression of CD14 marker of differentiated cells significantly increased, although the expression of CD44 marker decreased two weeks post differentiation.
Conclusions: PSE is a suitable inducer and its combined use with TGF-β3 can improve the efficiency of the chondrogenic potential of ADSCs. Our results suggest that PSE may have the potential to be used in tissue engineering as a replacement for TGF-β3.