Tissue engineering is a new technique to help damaged cartilage treatment using cells and scaffolds. In this study we tried to evaluate electrospun scaffolds composed of gelatin/ glycosaminoglycan (G/GAG) blend nanofibers in chondrogenesis of bone marrow-derived mesenchymal stem cells (BMMSCs). Scaffolds were fabricated by electrospinning technique with different concentration of glycosaminoglycan (0%, 5%, 10%, and 15%) in gelatin matrix. BMMSCs were cultured on the scaffolds for chondrogenesis process. MTT assay was done for scaffold's biocompatibility and cells viability evaluation. Alcian blue staining was carried out to determine the release of GAG and reverse transcription polymerase chain reaction (RT-PCR) was done for expression of COL2A1 and also immunocytochemistry assay were used to confirm expression of type II collagen. Scaffold with 15% GAG showed better result for biocompatibility (p =0.02). Scanning electron microscopy (SEM) micrographs showed that MSCs have good attachment to the scaffolds. Alcian blue staining result confirmed that cells produce GAG during differentiation time different from GAG in the scaffolds. Also the results for RT-PCR showed the expression of COL2A1 marker. Immunocytochemistry assay for type II collagen confirm that this protein expressed. Scaffold comprising 15% GAG is better results for chondrogenesis and it can be a good applicant for cartilage tissue engineering.

Tissue engineering is a technique that can be used for repairing damaged cartilage.The aim of this study was chondro-differentiation of mesenchymal stem cells (MSCs) on polyvinyl alcohol/gelatin/chondroitin sulfate (PVA/GT/CS) blend nanofibers. Nanofibers were fabricated by electrospinning method with different concentrations of CS (10%, 15%, and 20%). MSCs were seeded on nanofibers for biocompatibility and cell viability test by MTT assay. Alcian blue staining assay was done to show that MSCs were differentiated to chondrocyte by staining the glycosaminoglycans (GAGs) that was produced by cells. Reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (ICC) assay were done to confirm type II collagen expression. Scaffolds with different concentrations of CS were tested for biocompatibility, and the nanofiber with 15% of GAG showed better results (P = 0.02) compared with other nanofibers; therefore, it was chosen as the main sample for chondrogenesis tests. On the basis of scanning electron microscopy (SEM), hMSCs had a proper attachment to the nanofiber with 15% CS. Alcian blue staining showed the different rates of blue coloration, which means cells produced different types of GAGs from the GAG we used in the nanofiber. RT-PCR and ICC showed that COL2A1 was expressed in mRNA and protein levels. This study indicates that the PVA/GT/CS nanofiber containing 15% CS can be a good candidate for cartilage tissue engineering. K E Y W O R D S cartilage, chondroitin sulfate, electrospun, mesenchymal stem cells, tissue engineering