Methodology Involved in the Osteogenic Differentiation of Mesenchymal Stem Cells on Chitosan-Collagen Nanofibers Incorporated with Titanium Dioxide Nanoparticles.

Tissue regeneration especially in case of bones is a complex process as a repair involved is often inadequate. The electrospun chitosan nanofibers incorporated with titanium dioxide and collagen due to their ability to enhance biomineralization have been widely explored for bone tissue regeneration. Moreover, the mesenchymal stem cells (MSCs) possessing the properties of both self-renewal and multipotency offer a suitable recourse for cell-based regeneration strategies. This chapter summarizes the fabrication steps involved in the synthesis of titanium dioxide nanoparticles using sol-gel technique and their subsequent loading into chitosan/collagen nanofibers using the electrospinning process. Further on, the protocol involved in isolation of MSCs from bone marrow, seeding on fabricated nanofibers, and differentiation into osteoblasts is reported. The methods and techniques involved such as MTT assay, qRT-PCR, ALP activity, and immunofluorescence staining are also highlighted to investigate the potential of multifunctional nanofibers for the development of bony tissues.