Human Bone Marrow MSCs form Cartilage and Mineralized Tissue on Chitosan/Polycaprolactone (CS/PCL) Combined Nanofibrous Scaffolds

Human bone marrow mesenchymal stem cells (MSCs) are well established for their in vitro differentiation, however a protocol for transplanting the cells at the site of injury is needed. Here we present our recent investigation on chondrocyte and osteoblast differentiation of the hMSCs which are grown on Chitosan/Polycaprolactone (CS/PCL) combined Nanofibrous Scaffold. We used hMSC-TERT-CL1 cells that were grown on Chitosan (CS)/Poly-(epsilon-caprolactone) (PCL) Nanofibrous Scaffold, 50000 cells were cultured on each piece of Nanofibrous Scaffold. Cell viability and proliferation was assets by Alarmar Blue staining. Cell attachment and differentiation was assets by scanning electron microscopy (SEM) and specific stains for osteoblasts and chondrocytes. We demonstrated that CS/PCL nanoscaffold supports the attachment of hMSCs. We discovered that hMSCs proliferate on CS/PCL scaffold with or without differentiation; however the proliferation was lower on scaffold compared to cells on plastic when hMSCs were differentiated to osteoblasts. Contrary when hMSC were differentiated to chondrocytes we did not see significant difference in proliferation as 3D or cells on scaffold. Furthermore, we showed that hMSCs differentiate at a higher rate to chondrocyte when grown on CS/PCL scaffold with high expression of differentiation specific gene including SOX9, COMP and collagen II, further the cells stained better with Alcian Blue stain. Our results indicate that CS/PCL nanoscaffold is a useful scaffold for transplantation in patients with critical size bone defect.