Survey differentiation of mesenchymal stem cell to chondrocyte using synovial fluid and cold atmospheric plasma treated nanofibrous pcl substrate

Abstract

Tissue engineering has developed strategy for the repair and regeneration of a variety of tissues. The demand for effective treatment strategies of cartilage cause to autologous human Mesenchymal stem cells (hMSC) used for the repair of cartilage defects. In this study effect of some factors on human MSCs for differentiation to chondrocyte were surveyed such as Present of Poly Ɛ-caprolactone (PCL) nanofibrous scaffold and cold atmospheric plasma (CAP), also effect and amount of synovial fluid were evaluated. In this experimental study cell culture for differentiation assay performed in 6 well pellets by DMEM high glucose medium with a supplement of 10% FBS and synovial fluid 5% and 10% of medium add to pellets for 21 days. Total cellular RNA extracted was used for analysis by RT-PCR. Alcian blue staining confirmed the chondrogenic differentiation. MTT assay results in this study showed that the cells proliferation gradually increased from 1st day to 21st days on the electrospun nanofibrous scaffold. Cellular attachment and proliferation of hMSCs showed higher rate on PCL substrate, modified with Helium cold atmospheric plasma, compared to the unmodified and control group at the first day to 21st days. Chondrogenic differentiation confirmed by alcian blue staining, and show high degree of conversion to the chondrocyte. MTT assay and the expression of chondrogenic specific genes Collagen type П and Has-2 in comparison with control groups demonstrated cell survival ability, proliferation and cell differentiation by synovial fluid.