Osteogenic differentiation of bone marrow derived stem cells in gelatin-hydroxyapatite nanocomposite

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel- HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel- HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel- HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores (20-40 mu m). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel- HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel- HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.