Effect Of TGF-β3on The Differentiation Of Adipose-derived Stromal Cell Into Fibrocartilage Cells

Objective:The repair of temporomandibular joint (TMJ) disc defect remains challenges despite a large number of efforts to engineer functional fibrocartilage. Tissue engineering may provide a promising solution for clinical repair and reconstruction of TMJ cartilage. However, cell source of fibrous cartilage is still a problem to be solved. Pluripotent adipose-derived stromal cells (ADSCs) has shown its advantages as the cell source of fibrous cartilage and brought potential promising for repair of fibrocartilage-like tissue. The aim of this study is to investigate the effect of transforming growth factor-β3(TGF-β3) on the cell growth morphology, proliferation, and fibrocartilage differentiation potential of ADSCs.Methods:In this study, TGF-β3of10μg/ml was used to treat ADSCs from SD rats. Cell morphology was observed by microscope. Types Ⅰ and Ⅱ collagen and glycosaminoglycan (GAG) were visualized by alican blue staining, immunohistochemistry and histology. RT-PCR was further used to characterize the expression of cartilagenic-related genes.Results:The alican staining result of GAG extracellular matrix indicates distinct deep blue in the treated ADSCs group with the presence of TGF-β3, which suggests that ADSCs treated by TGF-β3produced a large amount of GAG. Immunohistochemistry results show that ADSCs in TGF-β3treatment group produced type Ⅰ and Ⅱ collagen, and RT-PCR result further confirm higher gene expression of type Ⅰ and Ⅱ collagen in TGF-β3treatment group, compared to non-treatment group. After TGF-(33treatment for2weeks, ADSCs show stellar-like or polygonal-like cell morphology. After3weeks, ADSCs in treated group show significant trend to hyaline cartilage differentiation through the expression of type Ⅰ and Ⅱ collagen, compare to the non-treated control group. These results suggested that TGF-β3could promote the differentiation of ADSCs into fibrocartilage-like cells.Conclusion:In this study, ADSCs could differentiate into fibrocartilage-like cells with the treatment of TGF-β3and possess potential as a promising cell source for fibrocartilage repair and regeneration.