In vitro assessment of the differentiation potential of bone marrow-derived mesenchymal stem cells on genipin-chitosan conjugation scaffold with surface hydroxyapatite nanostructure for bone tissue engineering.

摘要

Increasing evidence has revealed that the surface characteristics of biomaterials, such as chemical composition, stiffness, and topography, especially nanotopography, significantly influence cell growth and differentiation. In this study, we examined the effect of surface biomimetic apatite nanostructure of a new hydroxyapatite-coated genipin-chitosan conjugation scaffold (HGCCS) on cell shape, cytoskeleton organization, and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells in vitro. Cell shape and cytoskeleton organization showed significant differences between cells cultured on genipin-cross-linked chitosan framework and those cultured on HGCCS with surface apatite network-like nanostructure after 7 days of incubation in the osteogenic medium. The result of specific alkaline phosphatase activity as an indicator of osteogenic differentiation showed that the alkaline phosphatase activity of rat bone marrow-derived mesenchymal stem cells was higher on HGCCS. Based on quantitative real-time polymerase chain reaction, HGCCS induced highest mRNA expression of osteogenic differentiation makers, runt-related transcription factor 2 by 7 days, osteopontin by 7 days, and osteocalcin by 14 days, respectively. The enhanced ability of cells on HGCCS to produce mineralized extracellular matrix and nodules was also assessed on day 14 with Alizarin red staining. The results of this study suggest that the surface biomimetic apatite nanostructure of HGCCS is a critical signal cue to promoting osteogenic differentiation in vitro. These findings open a new research avenue to controlling stem cell lineage commitment and provide a promising scaffold for bone tissue engineering.