Osteogenic differentiation of human bone marrow mesenchymal stem cells seeded on melt based chitosan scaffolds for bone tissue engineering applications

摘要

The purpose of this study was to evaluate the growth patterns and osteogenic differentiation of human bonemarrow mesenchymal stem cells (hBMSCs) when seeded onto new biodegradable chitosan/polyester scaffolds.Scaffolds were obtained by melt blending chitosan with poly(butylene succinate) in a proportion of 50% (wt)each and further used to produce a fiber mesh scaffold. hBMSCs were seeded on those structures and cultured for3 weeks under osteogenic conditions. Cells were able to reduce MTS and demonstrated increasing metabolicrates over time. SEM observations showed cell colonization at the surface as well as within the scaffolds. Thepresence of mineralized extracellular matrix (ECM) was successfully demonstrated by peaks corresponding tocalcium and phosphorus elements detected in the EDS analysis. A further confirmation was obtained when carbonateand phosphate group peaks were identified in Fourier Transformed Infrared (FTIR) spectra. Moreover, by reversetranscriptase (RT)-PCR analysis, it was observed the expression of osteogenic gene markers, namely, Runt relatedtranscription factor 2 (Runx2), type 1 collagen, bone sialoprotein (BSP), and osteocalcin. Chitosan-PBS (Ch-PBS) biodegradable scaffolds support the proliferation and osteogenic differentiation of hBMSCs cultured attheir surface in vitro, enabling future in vivo testing for the development of bone tissue engineering therapies.