Document Type : Original Article(s)
Authors
1
Assistant Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
2
Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
3
Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
4
Assistant Professor, Department of Physics and Medical Engineering, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
5
Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
6
Assistant Professor, Department of Embryology, Cell Sciences Resaerch Center (ACECR), Royan Institute (Isfahan Campus), Isfahan, Iran.
7
Assistant Professor, Department of Pathology and Hematology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
8
General Practitioner, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Abstract
Background: Cartilage damages and diseases such as osteoarthritis are rather worldwide problems of many people. In addition, articular cartilage has a limited ability to repair. Current treatment methods for cartilage tissue injuries lead to formation of fibrous tissue, apoptosis, and further cartilage degeneration. Therefore, engineering of cartilage tissue with adult stem cells is considered necessary. This study aimed to investigate the effect of TGF-β3 and BMP-6 growth factors on chondrogenic differentiation of adipose-derived stem cells in alginate scaffold. Methods: Stem cells were isolated from human subcutaneous adipose tissue with collagenase enzyme digestion. The isolated cells were treated with chondrogenic medium supplemented with TGF-β3 and BMP-6 on alginate scaffold then harvested after 3 weeks. Histological, immunohistochemical and RT-PCR methods were performed for assessment of cartilage–specific markers. Findings: Our results revealed that chondrogenesis of ADSCs on alginate scaffold induced by TGF-β3 and BMP-6 growth factors. Immunohistochemical method was shown deposition of typical cartilage extracellular matrix components such as type II collagen and aggrecan in constructs. RT-PCR analysis of cartilage-specific genes, also confirmed the induction of the chondrocytic phenotype upon stimulation with TGF-β3 and BMP-6. Also, TGF-β3 and BMP-6 together increased the expression of aggrecan gene in compare with one growth factor alone. Conclusion: It can be concluded that TGF-β3 and BMP-6 promote chondrogenesis of ADSCs in 3-D culture and the use of both growth factors together increase the expression of aggrecan gene in compare with one growth factor alone. Key words: Adipose-derived stem cells, Alginate, Tissue engineering of cartilage, BMP-6, TGF-β3.
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